KERALA STATE COUNCIL FOR SCIENCE, TECHNOLOGY AND ENVIRONMENT

“BACK TO LAB PROGRAMME”

(WOMEN SCIENTISTS DIVISION)

Final Technical Report 2016-18

EMPOWERMENT OF RURAL WOMEN FOR CULTIVATION OF

ECONOMICALLY VIABLE MEDICINAL PLANTS THROUGH

AGRO-BIOTECHNOLOGICAL INTERVENTIONS

Project Reference No. : File No. 068/WSD-BLS/2015/CSTE

Post Doctoral Fellow: Dr. Resmi J

Scientist Mentor: Dr.M.R.Bindu (Professor)

Onattukara Regional Agricultural Research Station (ORARS),

Kayamkulam P.O., Alappuzha - 690 502

1. Authorization

We hereby declare that this project report entitled “Empowerment of rural women for cultivation of economically viable medicinal plants through agro-

biotechnological interventions” is a bonafide record of research work done by us during

the course of research and that the project report has not previously formed the basis

for the award of any degree, diploma, associateship, fellowship or other similar title, of

any other university or society.

Kaymkulam,

15 -03-2018.

RESMI. J. Dr. M. R. BINDU

Post Doctoral Fellow Scientist Mentor

2. Acknowledgement

“By the grace of God I am What I am”

I am deeply thankful to God for the numberless blessings he showered upon my

life, I bow my head before the mighty power for his eternal love and grace for

rendering help through various hands in completing the work successfully.

I would like to place on record our deep appreciation and thanks to Dr.

K.R.Lekha, Head, Women Scientists Division, Kerala State Council of Science,

Technology and Environment for her keen interest in promoting and co-ordinting

support activities for women scientists in Kerala.

It is my proud privilege to have worked under the guidance ofDr. M. R. Bindu,

Professor, Department of Plant Breeding and Genetics, Onattukara Regional

Agricultural Research Station, Kayamkulam. I feel immense pleasure in extending my

heartfelt gratitude and sincere thanks for her kind treatment, ever-willing help,

valuable guidance, expertise, patient and critical scrutiny of the manuscript,

constructive suggestion and motivation throughout the course of this research work and

in the preparation of the project report. I owe much to her for the moral support and

affection which helped me to complete this endeavor successfully.

From deep within my heart, I owe my indebtedness to Dr. Suja, G., Professor and

Head, Onattukara Regional Agricultural Research Station, Kayamkulam, for her

genuine interest, periodic suggestions, constructive perusal of manuscript and whole

hearted help.

My heartfelt gratitude to Dr. Mini. V., Assistant Professor, Department of Soil

Science and Agricultural Chemistry for her valuable comments, timely help and

encouragement throughout the study.

I convey my deep sense of gratitude to Dr. Sushamakumary, Dr. Susamma P.

Georgeand Dr. Indira for their valuable suggestions and wholehearted approach during

theresearch.

I owe my immense thanks to Sri. Muraleedharan, Farm Superintendent, Onattukara

Regional Agricultural Research Station, Kayamkulam for the sincere help rendered

during the course of investigation.

I sincerely acknowledge each and every non-teaching staff and labourer of the

ORARS, for their whole hearted co-operation and sincere efforts for the successful

completion of my research work.

I am grateful to NBPGR, Thrissur, IISR, Calicut, JNTBGRI, Palode for their help

for providing planting materials of turmeric and kacholammngenotypes for conducting

the study.

I feel special pleasure in expressing my whole hearted thanks to my dearest

colleaguesSunil, Remya, Seeja, Shemi, Fathima, Dhanya, Sreeparvathy, Jishnu, Abhijith, Divya,

Athira and Janeesh for their affection, moral support, selfless help and pleasant company.

I acknowledge KSCSTE for providing Post Doctoral Fellowship and Kerala

Agricultural University for providing the facility to conduct project work.

My loving mother Mrs. N. Ratnaveni (now late) initiated me into the scientif ic

lore. My in-laws (now late) were also to be remembered in this occasion. My hearty

prayers for the departed soul.

Lastly I thank my husband, Dr. Jijo. S., for his constant moral support,

encouragement and co-operation; our son Harikiran and Daughter Parvathy, for never

being demanding; and other family members for their commendable sacrifice, mental

support, prayers and for bearing the inconveniences caused during the study.

May God’s chicest blessing be showered on all of them.

RESMI J.

3. ABSTRACT

The research project “Empowerment of rural women for cultivation of economically viable medicinal plants through agro-biotechnological interventions” was carried out at Onattukara Regional Agricultural Research Station, Kayamkulamduring 2016-

2018. The objectives of the study were to collect and catalogue the genotypes based on the

IBPGR descriptor for turmeric and kacholam, to estimate the genetic parameters for different

traits in the germplasm for identifying superior lines based on yield and quality, to standardize

and optimize cost effective micropropagation protocol for the identified superior genotype of

selected medicinal plants and to initiate homestead cultivation of the target plant species and

assist the beneficiaries in forming their own Self Help Groups (SHGs) to sustain and promote

the medicinal plant cultivation programme.

Evaluation and characterisation of thirty genotypes of turmeric and twenty one

genotypes of kacholam collected from various sources pointed out wide variation for several

morphological characters. Analysis of variance revealed significant differences among the

genotypes for all the characters studied namely, plant height (cm), number of tillers per plant,

number of mother rhizomes per plant, number of primary rhizomes per plant, number of

secondary rhizomes per plant in turmeric, length of mother rhizome (cm), core diameter of

mother rhizome (cm), length of primary rhizome (cm), core diameter of primary rhizome

(cm), weight of mother rhizome per plant (g), weight of primary rhizome per plant (g),

weight of secondary rhizome per plant (g), days to maturity, wet rhizome yield per plant

(g/plant) and dry rhizome yield per plant in turmeric.

Among the turmeric genotypes, Rasmi recorded the maximum rhizome and

curcumin yieldwhich will be attractive forthe Onattukara farmers for its adoption and

cultivation.The variety is medium in duration (matures 8 months after planting). The

accession produces bold, medium sized rhizomes with closer internodes which have high

market preference. The rhizome powder is yellowish brown in colour, slightly grainy in

texture and have pleasing aroma. Fresh rhizome yield is 55.03 t/ha with dry recovery

percentage is 22% and curcumin content is 6.1 %.

In the case of kacholam, IC 373593 recordedthe highest yield (13.95t/ha) among

the twentyone genotypes evaluated and can be recommended for general cultivation in

Onattukara tract. The high yield in IC 373593 may be attributed to the higher number of

rhizomes and weight of mother as well as primary rhizomes. The genotype is collected from

NBPGR Regional Station, Thrissur. It is medium in duration and produces orange coloured

thick, round, plumpy mother rhizomes.

In the experiment on organic nutrient management in turmeric and kacholam,

quantity of vermicompost25 t/ha and 28 t/ha respectively (100 % replacement of nitrogen by

vermicompost) can be recommended. This recommendation will ensure the highest fresh

turmeric and kacholam yield.

In the experiment conducted to standardise cost effective micropropagation

protocol for identified superior genotype of turmeric and kacholamrevealed that the cost of

media can be reduced by substituting analytical grade sucrose with table sugar,Cytokinin –BAP and Kinetin with tender coconut water anddouble distilled waterwithtap water. The cost

effective protocol developed in this study by usingcheap media constituents, equipments and

culture containers can be utilized in any other crop species.

For starting homestead cultivation of superior lines of turmeric and kacholam, 15

women beneficiaries of Muthukulam and Chunakarapanchayats were selected and imparted

training in the scientific cultivation and post harvest handling. Various inputs like tissue

culture plantlets (Curcuma longa and Kaempferia galanga), organic manure, plant protection

chemicals etc were supplied. Appropriate guidelines were given for starting the field

demonstration in each panchayath.

4. INTRODUCTION AND REVIEW OF LITERATURE

Nationally, Kerala state is identified as a major consumer of medicinal plants. Apart from

the age old practices of Ayurvedic systems in its purest form, the state promotes herbal tourism,

which is a dwelling activity at present and generates employment potential. It is needless to note

that, there is tremendous scope for medicinal plant cultivation in Kerala for the development of

herbal industry and enhancement of economic background of rural unemployed women.

Production of ideal type and quality planting material for large-scale cultivation is also an

important task. The planting material therefore should be of good quality, rich in active

ingredients, pest- and disease-resistant, suitable for the agro-climatic condition of the proposed

land and pharmaceutical demand. It is a well determined fact that there are technical reasons

hindering the development of medicinal plant cultivation such as non –availability of quality

planting material, lack of agro-technology know-how and sustainable market

linkage.Considering the importance for promotion of cultivation of important medicinal plants

and income generation for unemployed rural women in Kerala state, this project is proposed with

the objectives of homestead cultivation of selected high value and industrially demanded

medicinal plants [Kaempferiagalanga (Kacholam), Curcuma longa (Manjal/Turmeric)] through

appropriate agro-technologies and increase the income of rural women. An improved, cost

effective micropropagation protocol will be standardized. The major outcome of the project will

be livelihood enhancement and empowerment of rural women and supply of genuine raw

materials for drug preparation.

Turmeric is a very important spice in India, which is obtained from rhizomes of plant

Curcuma longa, a member of the Zingiberaceae (ginger) family. Turmeric forms a part of most

Indian curry powder. It is a natural antiseptic. The spice is sometimes also called the ‘Indian saffron’ thanks to its brilliant colour. Kacholam (Kaempferia galangal L.) belongs to the family

Zingiberaceae commonly known as resurrection lily, is a short-stemmed herb which has flat,

green and round leaves. It is regarded as a cash plant as its rhizomes are used for the essential oil

extraction as well as for direct uses in the preparation of Ayurvedic drugs, perfumery and

cosmetics, spices. Further, bulky leaves of the plant are used for flavouring foodstuffs, preparing

mouthwashes and ahair tonic, locally; leaves are antinociceptive and antiulcerative.

Conventional breeding and crop improvement programmes are difficult in turmeric and

kacholamdue to low seed set, slow multiplication rate, limited availability of high yielding

genotypes and expensive field maintenance of planting material; and hence their genetic

improvement is limited to germplasm selection and chance variability arising during vegetative

propagation. Indiahas a high degree of variability of turmericandkacholam.No research work has

been conducted to evaluate Kacholam and turmeric genotyoes suited to Onattukara region.

Cultivation can find an alternative to solve problems of conservation, species

misidentification, genetic and phenotypic variability, variability and instability of extracts, toxic

components and contaminants. Information on the propagation of medicinal plants is available

for less than 10% and agro-technology is developed for 1% of total known plants globally. The

main factors behind the slow pace of domestication of medicinal plants sourced from the wild

are the absence of knowledge on cultivation practices and lack of sustainable marketing

systems.Indian turmeric is preferred due to its high Curcumin content as compared to other

countries.Organic farming assumes significant globally towards sustainable production and

quality up gradation of turmeric and kacholam. The adverse effects of continuous use of high

dose of chemical fertilizers on soil health and environment were realized; hence, the farmers are

also showing considerable inclination towards traditional farming with least usage of fertilizers.

The role of organic manures in improving soil structure and fertility is well understood. Organic

manures have positive influence on soil texture and structure, improve water holding capacity

and drainage which in turn help for better growth and development of rhizomatous crop like

turmeric and kacholam.

Conventional propagation of turmeric and kacholam by the splitting of rhizome is not

sufficiently rapid to meet the need of planting materials, and it takes years to build up for the

commercial quantities. In the recent years micropropagation techniques are being profitably used

to overcome the present demand of medicinal plants. Considering the present demand (both for

economic and medicinal values) and propagation problem of the plant, development of suitable

cost effective protocols for rapid multiplication of existing elite cultivars has become crucial for

meeting the market demand and to replenish highly impoverished populations.

Women mainstreaming through quality planting material production of medicinal plants

will be a step towards prosperity of farming community. Empowering farm women should focus

on socio-economic empowerment, technical empowerment, access to inputs and empowerment

through institutional mechanism. Also cultivation of medicinal plants is a sustainable alternative

to the arduous collection of medicinal plants from the wild and an income generating option for

unemployed rural women.

Taking into consideration of all these aspects, the present study was undertaken with the

following objectives:

a. Collection and germplasm establishment of the selected medicinal plants (Kaempferia

galanga and Curcuma longa) at Onattukara Regional Agricultural Research Station (ORARS).

b. To refine a cost effective micropropagation protocol for the identified superior

genotype of selected medicinal plants (Kaempferia galanga and Curcuma longa).

c. To select unemployed women beneficiaries from the selected village in Alappuzha

district and train them on cultivation of commercially valuable selected medicinal plants

(Kaempferia galanga and Curcuma longa) by adopting appropriate agro-technologies.

d. To initiate homestead cultivation of the target plant species and assist the beneficiaries

in forming their own Self Help Groups (SHGs) to sustain and promote the medicinal plant

cultivation programme.

e. To establish market linkage with local raw drug shops and Ayurvedic firms to generate

sustainable and steady income.

REVIEW OF LITERATURE

Study I: Turmeric (Curcuma longa L.)

Turmeric is the common name used for dried rhizome of Curcuma longa L., a

monocotyledonous plant belonging to the family Zingiberaceae. Turmeric powder, curcumin and

its derivatives and many other extracts from the rhizome were found to be bioactive. Studies on

the extent of variation in morphological and quality characters of turmeric are important for

selection of genotype with higher yield and better quality. This chapter gives the comprehensive

description about the work done in the field of evaluation, selection and characterization of

turmeric genotypes and curcumin standardization.

4.1.1 Evaluation and selection of turmeric genotypes

Philip (1983) assessed thirty-two types of Curcuma longa for plant height, tillers/plant,

leaves/plant, leaves/tiller, leaf length, width and leaf area, yields, finger mother rhizome ratio,

curing percentage, curcumin content, and the yield of green turmeric, cured produce and

curcumin. Mannuthy Local gave the highest yield of curcumin (391.5 kg/ha).

Datta and Chatterjee (2001) studied on 11 turmeric germplasms under rainfed condition

in the New Alluvial Zone of West Bengal, India revealed that among the different germplasms,

Kasturi produced the highest fresh yield (49.63 t/ha) and dry yield (10.67 t/ha), but curcumin

content was very low (2.1% i.e., lower than appreciable level as spice). Roma produced the

second highest dry yield (9.77 t/ha) and gave the highest level of dry recovery (29.89%) and

curcumin content (9.12%). Investigation also revealed that germplasms Roma, Armoor, PTS-8

and PTS-62 can be made popular for their higher yield and curcumin content instead of local

cultivars.

Kanjilal et al. (2002) examined and characterized the morphological and chemical

properties of cultivars of turmeric (Lakadong, Nongstein, Dalu, Baghmara, Rongjeng and

Shillong type), collected from different growing regions in Meghalaya, India. Among the

turmeric cultivars, Lakadong recorded the longest fingers (15.3 cm) and highest curcumin

content (6.8%). Shillong type recorded the highest number of fingers (7.5) and weight of

rhizome per clump (345 g).

Singh et al. (2015) evaluated seven genotypes of turmeric (Curcuma longa L.) under

irrigated condition for twoconsecutive years at Fruit Research Station, Banswara (Rajasthan).

The genotypes varied in their productionpotential, growth characters and curcumin content.

Pooled data revealed that variety Suroma produced maximum plant height (134.17 cm), number

of tillers per plant (5.39), single plant yield (0.431 kg) and yield perhectare of fresh rhizome

(231.11 q/ha) and it was at par with the production of Roma (216.40 q/ha). These twovarieties

viz., Suroma and Roma were significantly superior among all the varieties during both the

seasonsand are suitable for general cultivation in the southern parts of Rajasthan, whereas,

Pratibha variety exhibited highest curcumin content (3.12 g/100g) followed by Roma (2.69

g/100g).

Dodamani et al. (2017) evaluated six turmeric cultivars and observed that yield and

contributing characters, i.e. weight of mother rhizomes per plant, per plot and per hectare, weight

of fresh fingers per plant, per plot and per hectare were found maximum in Krishna.

4.1.2 Curcumin content in Curcuma spp.

Curcumin is responsible for the biological actions of turmeric and comprises of

curcumin, demethoxycurcumin and bisdemethoxycurcumin. Generally, the commercially

produced curcumin is a mixture of the above with curcumin as the main constituent.Several

methods have been standardized by researchers for determination of curcumin content of

turmeric rhizomes (Franco Cavaleri, 2018; Pawar et al., 2014).

Muthuswamy and Shah (1982) evaluated the comparative quality of Salem and Erode

turmeric types for curcumin content of mother and finger rhizomes of turmeric. Curcumin

content was 4.75% compared with 3.9% in Erode.

Philip (1982) evaluated the sixteen-turmeric cultivars for the variation of yield and

quality. The highest yield of curcumin was 560.6 kg/ha and oleoresin was 1470.3 kg/ha.

Jayaprakasha et al. (2002) developed an improved HPLC method for the determination of

curcumin, demethoxycurcumin, and bisdemethoxycurcumin. Curcuminoids were isolated by

column chromatography and identified by spectroscopic studies. The purity of the curcuminoids

was analyzed by an improved HPLC method. HPLC separation was performed on a C (18)

column using three solvents, methanol, 2% AcOH, and acetonitrile, with detection at 425 nm.

Four different commercially available varieties of turmeric, namely, Salem, Erode, Balasore, and

local market samples, were analyzed to detect the percentage of these three curcuminoids. The

percentages of curcumin, demethoxycurcumin, and bisdemethoxycurcumin as estimated using

their calibration curves were found to be 1.06 +/- 0.061 to 5.65 +/- 0.040, 0.83 +/- 0.047 to 3.36

+/- 0.040, and 0.42 +/- 0.036 to 2.16 +/- 0.06, respectively, in four different samples.

Kumar et al. (2015) reported thatout of the twelve varieties of turmeric evaluated, the

highest curcumincontent (6.3 %) was recorded in Roma, followed byRajendra Sonia (6.1 %).

They also reported that curcumin content ofturmeric varies from place to place due to genetic

andinfluence of environment and agro-climatic conditions.

4.1.3 Effect of organic manures on growth and rhizome yield of turmeric

Previous studies reported that organic fertilizer improved soil productivity and fertility,

which improved yield and quality of long duration crop like turmeric (Hossain and Ishimine,

2007; Velmurugan et al., 2007; Mohapatra and Das 2009; Roy et al., 2010; Dinesh et al., 2010).

Moreover, Manhas and Gill (2010) found that application of FYM increased the growth, dry

matter accumulation, yield and quality of turmeric.

Application of vermicompost alone or in combination with inorganic fertilizers was

found to increase the yield of turmeric (Vadiraj et al., 1996).

In a study on bio-organic inputs for production of organic turmeric grown as intercrop in

arecanut plantation, the highest projected yield 28.94 t/ha was recorded with vermicompost +

Azospirillum + Arbuscularmycorrhiza, followed by compost + Azospirillum +

Arbuscularmycorrhiza (26.93 t/ha) as compared to 24.11 t/ha under inorganic management (Roy

and Hore, 2009).

Kamal and Yousuf (2012) evaluated the effect of different organic manures on turmeric

withreference to vegetative growth, biomass production, rhizome yield and its attributes of

turmeric and reported that turmeric showed better response to the application of organic

manures. Among the organic manures, neem cake followed by poultry manure was superior to

cowdung with regard to yield and quality parameters.

Sahoo et al. (2015) reported that the scientific organic cultivation of turmeric recorded

significantly higher fresh (18.093 t ha-1) and dry rhizome yields (4.682 t ha-1) than the

respective yields from farmer’s cultivation practice (11.337 and 3.071 t ha-1). Such significant

and positive result in the former cultivation practice could possibly be due to the double rate of

application of FYM along with adequate organic plant protection measures compared to the

latter cultivation practice without any plant protection.

4.1.4 In-vitro studies on turmeric

Tissue culture, an important area of biotechnology can be used to improve the

productivity of planting material through enhanced availability of identified planting stock with

desired traits.

Pandey (2017) reported that MS media with 2.0 mg/l BAP+ 2.0 mg/l NAA was found to

be good for shoot initiation and multiplication in turmeric.

Sit and Tiwari (1997) standardized micropropagation of turmeric using buds of newly

developed turmeric rhizome and were cultured on MS medium supplemented with BA (0.0, 0.5

or 1.0 mg/l), Kinetin (0, 1, 2, 3, or 4 mg/l) and IBA (0.2 mg/l). Shoot proliferation after 15 and

21 days of subculture was highest on MS medium supplemented with Kinetin 3 mg/l and BA at

1.0 mg/l. However, shoot length after 15 and 21 days of subculturing was greatest on MS

medium supplemented with BA at 1.0 mg/l and Kinetin at 1 and 2 mg/l, respectively. Rooting

did not occur in absence of IBA and the number of roots per shootlet was proportional to the IBA

concentration.

Balachandran et al. (1990) reported in-vitro clonal multiplication of turmeric by using

rhizome buds as explant, inoculated on MS medium with 3% sucrose and 0.8% agar,

supplemented with BAP or the combination of BAP and kinetin for shoot multiplication. 2.5

mg/l BAP was best. In vitro plants were successfully established in the field and were

morphologically uniform.

Nadgauda et al. (1978) reported rapid multiplication of turmeric in-vitro using young

sprouting rhizome buds as explants. Rhizome buds were inoculated on MS medium with varying

levels of BAP and kinetin. The combination of 0.1 mg/l kinetin and 0.2 mg/l BAP was best

resulting in formation of 7-8 healthy green shoots. The survival rate of plants transferred to the

field was 80%.

Study II: Kacholam (Kaempferiagalanga L.)

Kacholam is an endangered medicinal plant with potent medicinal activities. The leaves,

and rhizome of the plant possess a number of medicinal applications such as stimulant,

expectorant, carminative, diuretic and are commercially used for the production of about 300

common ayurvedic preparations. K. galanga is one among 100 Red listed medicinal plants to be

conserved in Southern India [Revikumar and Ved, 2000)]. It is currently recognized as an ideal

medicinal plant for intercropping in the coconut gardens of Kerala. The present review highlights

agronomy and in vitro conservation methods of K. galanga.

4.2.1 Evaluation and selection of kacholam genotypes

According to Rahmanet al. (2004), a well-managed plantation of K. galanga yielded

about 4-6 tonnes of fresh rhizomes per hectare. Generally insects and pests are not reported in

this crop.

‘Kasthuri’ and ‘Rajani’, two high yielding varieties with rich flavour have been developed through clonal selection in kacholamby Kerala Agricultural University. The rhizomes

of ‘Kasthuri’ are large and light brown with a yield of 2.52 tonnes of dry rhizomes per hectare with a driage of 32.78%. It has high volatile oil content and total extractive of 3.4%. The

rhizomes of ‘Rajani’ are medium bold, creamy white and yields 2.55 tonnes of dry rhizomes per

hectare with a volatile content of 1% with high total extractives of 7.68%. Both varieties differ

morphologically and biochemically(Indrayan et al., 2007).

4.2.2 Effect of organic manures on growth and rhizome yield of kacholam

The plants which are used for medicinal purposes should not have any residues of

chemical fertilizer or pesticides; otherwise, they may become more of poison than a medicine

(Smitha et al., 2010). Moreover, the medicinal plants have several active ingredients which may

be affected by excessive use of chemical fertilizer, pesticide and low quality irrigation water.

Thus, as far as possible, they have to be grown in near-to-nature condition, so that both the

quality of medicinal plants and the socio-ecological balance can be maintained for which organic

production of the medicinal plants is the only option.Higher population of microbes under

organic treatments act as an index of soil fertility because it serves as temporary sink of nutrients

flux.

Maheswarappa et al. (1999) reported that farm yard manure and major nutrient

combinations recorded significantly higher yield compared to farm yard manure, vermicompost,

composted coir pith and major nutrients applied alone in kacholamgrown as intercrop in coconut.

Smitha et al. (2015) tried varied levels of farm yard manure (FYM - 30 and 40 t ha-1),

vermicompost (VC - 1 and 2 t ha-1), neem cake (NC - 1 and 2 t ha-1) and bio-fertilizers

(Azospirilum, phosphate solubilizing bacteria and vesicular arbuscularmycorrhizae each at 10 kg

ha-1) in sixteen different combinations in long pepper. The results after 3 years revealed that the

application of organic manures has a significant impact on plant and soil nutrient status after

three years of cropping. The highest tissue nutrient content and nutrient uptake were recorded in

the treatment FYM 40 t + VC 2.0 t + NC 2.0 t + BF 10 kg ha-1 where the organic manures and

bio-fertilizers were applied at highest level. The same treatment has also increased the organic

carbon, available NPK and microbial population in the soil after three years of cropping. All

these parameters were lowest in treatment FYM 30 t + VC 1.0 t + NC 1.0 t ha-1 where manures

were applied in lowest quantity without bio-fertilizers.

4.2.3 In-vitro studies on kacholam

In vitro techniques have been successfully applied to solve some of theproblems of

conventional clonal propagation in kacholam. These methods have the advantage of requiring

very little plant tissueto initiate propagation and very high multiplication rates are

typicallyobtained. Propagation can be carried out at any time of the year anduniform, disease-

free plantlets suitable for planting are produced on a largescale.

An efficient in vitro propagation protocol was standardized in K. galanga, wherein shoot

cultures were raised from rhizome with axillary bud explants in MS medium supplemented with

different hormonal regimes. Maximum 10.6±0.83 multiple shoots per explants were obtained in

MS medium supplemented with 4.0 mgl-1 BA along with 1.0 mgl-1 each of NAA and kinetin

(Preetha et al., 2014).

An efficient and direct single medium based protocol for multiple shoot and

rootinduction has been achieved from the rhizome explants ofkacholam when cultured on full

strength MS media with Kn(2.0 mg/l) + NAA (1.0 mg/l) with a high level ofsucrose of 6%.

Maximum number of 7±0.258 shoots and maximum number 6.7±0.291 of roots were induced in

the mediumwith cent percent regeneration. The regenerated plantlets were acclimatized

andestablished on the soil with 95% of success(Ibemhal et al., 2012).

Shirin et al. (2000) showed that BA (2.7 mgl-1) and NAA (0.5 mgl-1) was effective for

the multiplication of K. galanga.

4.3.1 Optimization of low cost micropropagation techniques in medicinal plants

Micropropagation is a capital-intensive technology involving energy and labour. Cost of

chemical inputs, media, energy, labour and capital counts on production cost. The high cost of

tissue culture raised plants continues to challenge their wider acceptance by the farmers.

Therefore, low cost alternatives are needed to reduce cost production of tissue cultured plants.

Low cost technology means an advanced generation technology in which cost reduction is

achieved by improving process efficiency and better utilization of resources (Savangikar, 2002).

Low cost options should lower the cost of production without compromising the quality of the

micropropagules.Few attempts have been made to standardize the different factors for lowering

the cost of production of in-vitro regeneration of medicinal plants.

Proper choice of media and containers can reduce the cost of micropropagation (Prakash

et al., 2002)

Raghu et al. (2007) reported that cost of media ingredients was reduced by using

household sugar instead of laboratory grade sucrose and tap water instead of double distilled

water and liquid media instead of agar for micropropagation of Centellaasiatica.

4.4.1 Women empowerment in medicinal plant cultivation

Recently, Women empowerment has been recognized as a central issue in determining

the status of women and has acquired a very important place in government development

programmes. The Word Bank has suggested that empowerment of women should be a key aspect

of social development programs (World Bank, 2001). In Kerala, the catalyst behind this venture

is the Kudumbashree, women based participatory poverty eradication programme launched on 1st

April 1998 by the Kerala State Government with the financial support of NABARD (National

Bank for Agriculture and Rural Development) and the Central Government (Dhanya and

Sivakumar, 2010). About 2.5 lakh women in the State constituted in 30,000 groups are engaged

in collective farming in the State (Rajagopal, 2011). Medicinal plant cultivation can be treated as

an alternative income generation source for the rural unemployed women without hampering

their ongoing income generating activities. The Department of Science and Technology (DST) in

collaboration with National Botanical Research Institute (NBRI) has initiated an All India

Coordinated Project on Plant Based Health System involving Women. Under this, the women are

trained on different improved techniques related to the cultivation of medicinal plants,

preparation of herbal products and their marketing. Several NGOs have also been involved in

this project and they have been instrumental in implementing it in the different states of India. In

addition, projects were sponsored in diverse technology areas such as nursery techniques for

medicinal plants cultivation, tissue culture etc. that have made visible impact for the

empowerment of women and sustainable livelihood options for rural women by Central

Government funding agencies like Department of Biotechnology (DBT) and Kerala State

Government agencies like WGC (Western Ghats Cell), SC/ST Department etc. A novel

participatory programme for self income generation of rural women in Alapuzha district of

Kerala through cultivation of high value medicinal and aromatic plants under DBT programme

was a model project that created a general awareness on medicinal plant cultivation and

promoted homestead cultivation in the state (Sudha, 2010). Motivating the woman groups in

interiors of Mandi district in Himachal Pradesh to cultivate medicinal plants by the officials of

Himalayan Research Group (HRG) - an NGO instead of Cannabis crop has yielded positive

results. The hardwork and dedication of women groups has led to selling of a huge consignment

of 2500 Kgs of Chirayita – a medicinal plant found in Himalayas, sold to Dabur India Ltd for

manufacturing herbal medicines at their Ghaziabad factory (Anonymous 1). Thus cultivation of

medicinal plants is a sustainable alternative to the arduous collection of medicinal plants from

the wild and an income generating option for unemployed rural women. Cultivation of medicinal

plants was done effectively through a five year project in Idukki District of Kerala (Anonymous

2).

5. OBJECTIVES

a. Collection and germplasm establishment of the selected medicinal plants (Kaempferiagalanga

and Curcuma longa) at Onattukara Regional Agricultural Research Station (ORARS).

b. To refine a cost effective micropropagation protocol for the identified superior genotype of

selected medicinal plants (Kaempferiagalanga and Curcuma longa).

c. To select unemployed women beneficiaries from the selected village in Alappuzha district and

train them on cultivation of commercially valuable selected medicinal plants

(Kaempferiaalanga and Curcuma longa) by adopting appropriate agro-technologies.

d. To initiate homestead cultivation of the target plant species and assist the beneficiaries in

forming their own Self Help Groups (SHGs) to sustain and promote the medicinal plant

cultivation programme.

e. To establish market linkage with local raw drug shops and Ayurvedic firms to generate

sustainable and steady income.

6. MATERIALS AND METHODS

6.1.1 Collection/purchase, establishment and characterization of germplasm :

Planting material (Rhizomes) of 30 genotypes of Curcuma longa L. were collected from

Indian Institute of Spices Research (IISR), Kerala Agricultural University (KAU) and Jawaharlal

Nehru Tropical Botanical Garden (JNTBGRI) and 21 genotypes of Kaempferia galanga L. from

National Bureau of Plant Genetic Resources (NBPGR), KAU, JNTBGRI and Farmer’s fields

(Table 1 and 2). Characterisation was done based on standard descriptors for turmeric and

kacholam during 2016-17.

6.1.2 Evaluation of germplasm for two consecutive years at ORARS, Kayamkulam:

Planting material (Rhizomes) of 30 genotypes of Curcuma longa L. were collected from

IISR, KAU and JNTBGRI and 21 genotypes of Kaempferia galanga L. from NBPGR, KAU,

JNTBGRI and Farmer’s fields. The experiment was laid out in a randomized block design with

two replications to evaluate the genotypes of turmeric and kacholam during the periods 2016-17

and 2017-18. Planting material used was healthy rhizome bits having 2-3 buds pretreated with

Pseudomonas fluorescence and management practices adopted were according to package of

practices recommendations of Kerala Agricultural University (POP, 2016) (Plate 1). Biometrical

observations of fifteen characters namely plant height (cm), number of tillers per plant, number

of mother rhizomes per plant, number of primary rhizomes per plant, number of secondary

rhizomes per plant, length of mother rhizome (cm), core diameter of mother rhizome (cm),

length of primary rhizome (cm), core diameter of primary rhizome (cm), weight of mother

rhizome per plant (g), weight of primary rhizome per plant (g), weight of secondary rhizome per

plant (g), days to maturity, wet rhizome yield per plant (g/plant) and dry rhizome yield per plant

were recorded and analysed statistically (Panse and Sukatme, 1985).

6.1.3 Estimation of rhizome curcumin content using HPLC:

A simple, precise, rapid and accurate, high performance liquid chromatographic method

has been developed for the determination of curcumin in 30 samples of turmeric with short run

time. One (1.0) g of ground turmeric sample of each accession was extracted with 200 mL of N-

hexane using a Soxhlet extractor for 30 min. The hexane extract was discarded and the sample

was further re-extracted with 200 mL of methanol for two hours. Duplicate extractions were

done for each sample accession. A 1-mL aliquot of the methanolic extract transferred to a 10-mL

volumetric flask and diluted to the mark with methanol was used for HPLC injection. For the

analytical standard, a known amount of curcumin (98% purity) was weighed in a 10-mL

volumetric flask, dissolved and diluted to the mark with methanol to arrive at a 1000 ppm stock

solution. Aliquots were taken from the stock solution to prepare the working standard solutions

needed to establish the standard curve. Twenty microliters (20 μL) of each of the samples and the working standard solutions (containing active ingredient ranging from 0.0078 μg to 1.9600 μg) was injected into Shimadzu Liquid Chromatograph 10AT. The chromatographic column used

was a Novapak C18, 3.9 x 150 mm, 4 μm column and detection was done using Shimadzu SPD-

10 AV - UV detector at 254 nm wavelength. Elution was carried out with a solvent system

Table 1. Particulars of varieties / accessions of Curcuma longa used in the study and their sources

Sl. No. Variety/ Accession Source

1 Kanthi Kerala Agricultural University

2 Shobha Kerala Agricultural University

3 Sona Kerala Agricultural University

4 Varna Kerala Agricultural University

5 IISR Prabha Indian Institute of Spices Research, Calicut

6 IISR Prathibha Indian Institute of Spices Research, Calicut

7 IISR Alleppy Supreme Indian Institute of Spices Research, Calicut

8 IISR Suvarna Indian Institute of Spices Research, Calicut

9 IISR Suguna Indian Institute of Spices Research, Calicut

10 IISR Sudarsana Indian Institute of Spices Research, Calicut

11 IISR Kedaram Indian Institute of Spices Research, Calicut

12 Roma (High Altitude Research Station, Odhisa) Indian Institute of Spices Research, Calicut

13 Suroma (High Altitude Research Station, Odhisa) Indian Institute of Spices Research, Calicut

14 Renga (High Altitude Research Station, Odhisa) Indian Institute of Spices Research, Calicut

15 Resmi (High Altitude Research Station, Odhisa) Indian Institute of Spices Research, Calicut

16 BSR-I (Tamil Nadu Agricultural University) Indian Institute of Spices Research, Calicut

17 BSR-II (Tamil Nadu Agricultural University) Indian Institute of Spices Research, Calicut

18 Rajendra Sonia (Tirhut College of Agriculture,

Bihar)

Indian Institute of Spices Research, Calicut

19 Mega Turmeric (ICAR Research Complex for NEH

Region, Shillong)

Indian Institute of Spices Research, Calicut

20 Punjab Haldi I (Punjab Agricultural University) Indian Institute of Spices Research, Calicut

21 Punjab Haldi II (Punjab Agricultural University) Indian Institute of Spices Research, Calicut

22 Pant Peetab (GB Pant Univ. of Agri. & Tech.,

Uttarakhand)

Indian Institute of Spices Research, Calicut

23 Duggirala Red (YSR Andhra Pradesh Hort. Univ.,

Jagtial)

Indian Institute of Spices Research, Calicut

24 Suranjana (Uttar Bengal Krishi Viswa Vidyalaya) Indian Institute of Spices Research, Calicut

25 Narendra Haldi (Narendra Deva University of Agri.

& Tech, Faizabad)

Indian Institute of Spices Research, Calicut

26 TBGRI Local Palode

27 Kasaragod Local Kasaragod

28 Kanjikuzhy Local Kanjikuzhy

29 Kuttipuram Local Kuttipuram

30 Aluva Local Aluva

Table 2. Particulars of accessions of Kaempferia galanga used in the study and their sources

Sl. No. Accession Source

1 Haripad Local Haripad

2 IC 582810 NBPGR Regional station, Thrissur

3 IC 373589 NBPGR Regional station, Thrissur

4 IC 373592 NBPGR Regional station, Thrissur

5 IC 087830 NBPGR Regional station, Thrissur

6 IC 210748 NBPGR Regional station, Thrissur

7 IC 572784 NBPGR Regional station, Thrissur

8 IC 373588 NBPGR Regional station, Thrissur

9 IC 373590 NBPGR Regional station, Thrissur

10 IC 582794 NBPGR Regional station, Thrissur

11 IC 373594 NBPGR Regional station, Thrissur

12 IC 582811 NBPGR Regional station, Thrissur

13 IC 550136 NBPGR Regional station, Thrissur

14 IC 087831 NBPGR Regional station, Thrissur

15 IC 582798 NBPGR Regional station, Thrissur

16 IC 373593 NBPGR Regional station, Thrissur

17 IC 087826 NBPGR Regional station, Thrissur

18 IC 550152 NBPGR Regional station, Thrissur

19 TBGRI Local Palode

20 Wayanad Local Wayanad

21 Kannur Local Kannur

consisting of methanol:acetonitrile (80:20 v/v mixture) at a flow rate of 0.8 mL min-1. A

calibration curve was plotted between peak area and concentration. The curcumin content was

estimated by calibration curve and the results were expressed in percentage.

6.2.1 Experiment on organic nutrient management in turmeric and kacholam:

Field experiment was conducted at ORARS, Kayamkulam using high yielding variety of

turmeric Rasmi, using different amounts of vermicompost to replace recommended dose of

nitrogen. The experiment was laid out in a randomized block design (RBD) with four

replications having five treatments: T1- 25% replacement of Nitrogen with vermicompost,

T2- 50% replacement of Nitrogen with vermicompost, T3 - 75% replacement of Nitrogen with

vermicompost, T4 - 100% replacement of Nitrogen with vermicompost, T5 – Package of

Practices recommendation (Plate 2). Vermicompost application was done as basal dose. Urea,

rock phosphate and Muriate of potash were used as inorganic source of N, P and K respectively.

1/3rd N andfull P were applied as basal, rest nitrogen and potassium were applied in 2 equal split

at 45 and 90 days after planting (DAP). Mulching and other intercultural operations were

practiced uniformly to all treatments. The crop was harvested after complete maturity, as

indicated by the drying of leaf and plant as a whole. To evaluate the effect of vermicompost,

biometric and yield characters viz. plant height (cm), number of tillers per plant, number of

mother rhizomes per plant, number of primary rhizomes per plant, number of secondary

rhizomes per plant, length of mother rhizome (cm), core diameter of mother rhizome (cm),

length of primary rhizome (cm), core diameter of primary rhizome (cm), weight of mother

rhizome per plant (g), weight of primary rhizome per plant (g), weight of secondary rhizome per

plant (g), days to maturity, wet rhizome yield per plant (g/plant) and dry rhizome yield per plant

of five randomly selected plants in each of the treatment were recorded and mean was

computed.

6.3.1 Standardisation of cost effective micropropagation protocol of the selected medicinal

plants:

Explants used in the present study were young fresh rhizome bits with emerged and not

emerged buds, leaf segments and root segments. The surface sterilization procedure followed

was washing in tap water containing Tween 20, followed by drenching in 0.2% Bavistin for 10

minutes. Then washed thoroughly and treated with 0.1% mercuric chloride solution for 15 min in

the laminar air flow. The materials were then washed three times with sterile distilled water to

remove all the traces of mercuric chloride. Sterilized explants were cut into 2-3 mm sized pieces

using sterile scalpel under laminar air flow chamber and inoculated into MS media with different

growth regulators singly or in combinations to standardize bud initiation and multiplication

(Table 3, 4 and 5). The regenerated microshoots were placed on half-strength MS medium

supplemented singly with various concentrations of NAA or IBA for rooting (Table 6). Standard

procedures (Murashige and Skoog, 1962) were followed for the preparation of the plant tissue

culture media. The details of the MS media composition are given in Appendix I. Specific

quantities of the stock solutions were pipetted out into a 1000 ml beaker. Sucrose and inositol

Table 3. Treatments tried to study the effect of plant growth regulators on shoot

initiation in turmeric

(Medium –MS + inositol 100 mg l-1

+ sucrose 30.00 g l-1

+ agar 7.50 g l-1

)

Treatment No. Plant growth regulators (mg l-1

)

BAP + IAA

1 0.5 + 0.0

2 1.0 + 0.0

3 1.5 + 0.0

4 2.0 + 0.0

5 2.5 + 0.0

6 3.0 + 0.0

7 2.0 + 0.1

8 2.0 + 0.2

9 2.0 + 0.3

10 2.0 + 0.4

11 2.0 + 0.5

12 2.0 + 0.6

13 2.0 + 0.7

BAP + NAA

14 2.0 + 0.1

15 2.0 + 0.2

16 2.0 + 0.3

17 2.0 + 0.4

18 2.0 + 0.5

19 2.0 + 0.6

20 2.0 + 0.7

21 3.0 + 0.1

22 3.0 + 0.2

23 3.0 + 0.3

24 3.0 + 0.4

25 3.0 + 0.5

BAP+IBA

26 2.0 + 0.1

27 2.0 + 0.2

28 2.0 + 0.3

29 2.0 + 0.4

30 2.0 + 0.5

Table 4. Treatments tried to study the effect of plant growth regulators on shoot Proliferation in

turmeric

(Medium –MS + inositol 100 mg l-1

+ sucrose 30.00 g l-1

+ agar 7.50 g l-1

)

Table 5. Treatments tried to study the effect of plant growth regulators on shoot initiation and

multiplication in Kacholam

(Medium –MS + inositol 100 mg l-1

+ sucrose 30.00 g l-1

+ agar 7.50 g l-1

)

Treatment No. Plant growth regulators (mg l-1

)

BAP + IAA + ADS 2.0 + 0.5 + 0

BAP + IAA + ADS 2.0 + 0.5 + 25

BAP + IAA + ADS 2.0 + 0.5 + 50

BAP + IAA + ADS 2.0 + 0.5 + 100

Treatment No. Plant growth regulators (mg l-1

)

BAP

1 0.5

2 1.0

3 1.5

4 2.0

5 2.5

6 3.0

BAP + Kin

7 2.0 + 1.0

8 2.0 + 2.0

Table 6. Treatments tried to study the effect of plant growth regulators on rooting in turmeric

and kacholam

(Medium –Half MS + inositol 100 mg l-1 + sucrose 30.00 g l-1 + agar 7.50 g l-1)

Treatment No. Plant growth regulators (mg l-1

)

NAA

1 0.2

2 0.5

3 1.0

4 2.0

IBA

6 0.2

7 0.5

8 1.0

9 2.0

were added fresh. After making up the volume to 1000 ml using distilled water, the pH of the

medium was adjusted to 5.7 using 0.1 N NaOH/HCl. The basal medium used for all the

experiments were Murashige and Skoog (1962) mineral formulation containing standard salts

and vitamins, 30 g/l sucrose and 7.5 g/l agar. The medium was autoclaved at 15 Ibs/sq. inch for

20 min at 121ºC. Cultures were incubated at a temperature of 20±2°C and 16 h photoperiod.

Observations were recorded after a month and analysed statistically (Panse and Sukhatme, 1985).

6.3.2Optimization of Cost Effective Micropropagation Protocol for identified Superior

Genotype of Selected Medicinal Plants:

Cost-reduction and simplfication of conventional tissue culture procedures were

standardised for the micropropagation of Curcuma longa and Kaempferiagalanga.

LOW-COST OPTIONS USED IN THIS STUDY

Equipment:

Sterilization of glassware and culture media was carried out using a large pressure cooker

(Make: Prestige. 22 Litres capacity) and an LPG instead of autoclave. pH paper strips were used

instead of pH meter for pH measurement. Double distilled water was replaced with tap water for

preparation of culture media.

Chemicals:

Laboratory Reagent (LR) grade chemicals were used wherever possible as an alternative

to Analytical Reagent (AR) Grade chemicals that is used conventionally for micropropagation.

Plant growth regulators and organic additives including vitamins were available only in AR

grade. Tender coconut water was used as low cost alternative to plant growth regulators. The

Grocery sugar/ table sugar were used instead of sucrose as a carbon source in the all

combination.

Culture containers:

The following alternative to the test tube and conical flasks were used as culture

containers in this study. Culture bottles with Polypropylene (PP) caps and wide mouths were

used.

The experiment was carried out, each with 10 replications. The days to initiation, number

and length of shoots and number and length of roots were calculated. Cultures were observed

daily and the data were recorded at an interval of 10 days and finally after 4 weeks.

6.3.3 Hardening of microplantlets :

In vitro grown plants having well developed shoots and roots were washed gently under

running water to remove agar from roots and planted in small polybags containing soil,

vermicompost and sand mixture in the 1:1:1 proportion. The plantlets were kept in shade net

house under 50% sunlight for acclimatization to the outside environmental conditions for 30

days with regular watering and the percentage of survival was noted. Morphological parameters

for hardened plants like number and length of shoots; number and length of roots; number of

leaves as well as leaf length and width were recorded.

6.4.1 Initiation of Project Activities in the Selected GramaPanchayaths and Selection of

Women Beneficiaries:

Detailed discussions were carried out with the Panchayath Committee members and

officials of the selected two panchayaths viz. Muthukulam and Chunakkara to stream line the

programme successfully with their co operation. Local awareness programmes were conducted

in the selected villages on scientific mode of medicinal plant cultivation and sustainable income

generation. Selection of 15 beneficiaries of two panchayats were carried out through

Krishibhavan and with assistance of people’s representatives in the selected Panchayats.

Selection forms (Appendix II) based on the criteria like age, education, size of the family, socio-

economic status, social activities, entrepreneurship, land and irrigation facilities possessed and

their interest in the programme from the target site were distributed for the purpose.

Subsequently segregate them to groups and a leader beneficiary were selected from each group.

6.4.2 Initiation of Homestead Cultivation for Macropropagation of Superior Line:

To initiate cultivation, sufficient number of rhizomes as well as healthy and established

tissue culture microplantlets (Curcuma longa and Kaempferia galanga), organic manure, plant

protection chemicals etc were distributed to selected women beneficiaries of each panchayath

viz. Muthukulam (Sandy loam) and Chunakara (Laterite). The experiment on performance of

rhizome generated plants as well as microplantlets were laid out in farmer’s field in each panchayat and supervised by leader farmers successfully.

7. RESULTS AND DISCUSSIONS

7.1.1 Collection/purchase, establishment and characterization of germplasm:

Characterisation of Curcuma longa showed distinct variations among the accessions with

respect to vegetative, floral and rhizome characters (Table 7, 8, 9) (Plate 3). Twenty one

genotypes were categorized as moderate to high in plant growth habit. Leaf size observed were

small (3 accessions), medium (12 accessions), large (10 accessions) and very large (5

accessions). Leaf shapes are lanceolate, elliptic oblong or slender elongated. Twenty three

accessions had acuminate - caudate leaf tip while seven accessions had tapering leaf tip. All the

genotypes had erect leaf orientation, obtuse-cuneate leaf base shape and glabrous type of

pubescence. Twenty two accessions had green leaf colour while dark green and light green leaf

color was observed for 4 accessions in each case. Petiole size also varied from small to large.

Turmeric is shy in flowering and only six genotypes flowered. Both pink (2 accessions namely

Roma and Kasaragod Local) and pale yellow flower colour (4 accessions namely IISR Alleppy

Supreme, Suroma, BSR-I and Suranjana) was noticed among the accessions that flowered during

the study. Variability was more pronounced for rhizome characters and wide variation was

noticed in inner core colour of rhizomes ranging from orange to reddish yellow. Twenty four

accessions had brownish orange rhizome peel color, while 4 accessions possessed dull brown

and 2 accessions had golden yellow colour. Sensory characters of rhizomes like colour, flavour

and texture also showed variability among the accessions. The high variability among turmeric

accessions denotes the scope for its genetic improvement.

Twenty one accessions of kacholam were genetically catalogued for vegetative, leaf and

rhizome characters as per NBPGR descriptor (Table 10, 11) (Plate 4). All the accessions were

having very short stem with large simple leaves spreading flat on the ground. Three accessions

were flowered and the flower was white in colour with lilac spots. Variability was more

pronounced for leaf and rhizome characters. Leaf shape was either suborbicular, obtuse, ovate,

lanceolate, cuneate or elliptic. Leaf colour ranged from light green (12 accessions) to dark green

(6 accessions). Sixteen accessions were categorized as medium to large leaf size. Leaf margin

was entire in 19 accessions of kacholam. Eighteen accessions had cylindrical rhizome shape but

tubular shape was also found in three accessions. Wide variation was noticed in rhizome colour

ranging from brown to brownish yellow of which eight accessions were having reddish brown

coloured rhizomes. The rich variability for rhizome nature among kacholam accessions have

many value-added characters and will bring fruits in its improvement. If they are considered

while breeding, no doubt kacholam will be a promising medicinal plant.

7.1.2 Evaluation of germplasm for two consecutive years at ORARS, Kayamkulam:

The mean values of 30 accessions of turmeric and 21 accessions of kacholam for

different characters are presented in Table 12 and Table 13 respectively.

In the evaluation of germplasm for two consecutive years, analysis of variance revealed

considerable genetic diversity among elite accessions of Curcuma longa for most the characters

7. Vegetative and Leaf characters in Curcuma longa L. varieties/accessions

Accession No. Growth habit

Leaf Shape Leaf tip Leaf size Leaf colour Petiole

size

T 1 High Lanceolate Other (Tapering) Large Dark green Medium

T 2 High Elliptic oblong Acuminate - caudate Medium Green Long

T 3 High Elliptic oblong Acuminate - caudate Large Green Short

T 4 High Lanceolate Acuminate - caudate Large Green Short

T 5 Low Other (Slender Elongated) Acuminate - caudate Large Green Medium

T 6 Moderate Lanceolate Acuminate - caudate Large Green Short

T 7 Moderate Other (Slender Elongated) Other (Tapering) Very large Green Long

T 8 Moderate Lanceolate Acuminate - caudate Large Green Short

T 9 Low Elliptic oblong Acuminate - caudate Medium Light green Medium

T 10 Moderate Lanceolate Other (Tapering) Medium Green Medium

T 11 Moderate Elliptic oblong Acuminate - caudate Small Light green Medium

T 12 High Elliptic oblong Other (Tapering) Medium Green Medium

T 13 High Lanceolate Acuminate - caudate Large Green Short

T 14 Moderate

Elliptic oblong Acuminate - caudate Medium Green Very

long

T 15 Low Lanceolate Acuminate - caudate Very large Green Medium

T 16 High Lanceolate Acuminate - caudate Large Dark green Long

T 17 High Other (Slender Elongated) Other (Tapering) Medium Green Short

T 18 Low Lanceolate Other (Tapering) Medium Green Long

T 19 High Elliptic oblong Acuminate - caudate Medium Green Medium

T 20 High Other (Slender Elongated) Acuminate - caudate Very large Green Medium

T 21 Low Elliptic oblong Other (Tapering) Medium Dark green Long

T 22 High Lanceolate Acuminate - caudate Small Light green Medium

T 23 Moderate Other (Slender Elongated) Acuminate - caudate Very large Green Long

T 24 Low Lanceolate Acuminate - caudate Very large Dark green Medium

T 25 High Other (Slender Elongated) Acuminate - caudate Medium Green Long

Leaf orientation was erect, Leaf base shape was obtuse-cuneate and glabrous type of pubescence for all

the 30 turmeric accessions.

T 26 High Elliptic oblong Acuminate - caudate Medium Green Long

T 27 Low

Elliptic oblong Acuminate - caudate Small Green Very

long

T 28 Moderate Lanceolate Acuminate - caudate Large Green Medium

T 29 Low Lanceolate Acuminate - caudate Large Green Short

T 30 Low Lanceolate

Acuminate - caudate Medium Light green Very

long

Table 8 . Rhizome characters in Curcuma longa L . var ieties/accessions

Acc.

No.

Branching

of Rhizome Rhizome nature

Rhizome Peel

color

Inner core colour

of rhizome

T 1 Horizontal Other (Medium sized rhizomes and closer internodes) Brownish orange Orange

T 2 Horizontal Other (Medium sized rhizomes and closer internodes) Brownish orange Yellow

T 3 Horizontal Other (Bigger mother rhizomes and medium sized fingers) Brownish orange Orange

T 4 Horizontal Other (Medium sized rhizomes) Brownish orange Yellow

T 5 Curved Plumpy Brownish orange Orange

T 6 Horizontal Plumpy Brownish orange Orange

T 7 Horizontal Other (Medium sized mother rhizome and slender fingers) Brownish orange Yellow

T 8 Horizontal Other (Bold, medium sized rhizomes) Brownish orange Other (Reddish

Orange)

T 9 Horizontal Other (Bold mother rhizomes and slender fingers) Brownish orange Orange

T 10 Horizontal Other (Bold mother rhizomes and slender fingers) Brownish orange ReddishYellow

T 11 Horizontal Plumpy Dull brown Orange

T 12 Curved Plumpy Brownish orange Orange

T 13 Horizontal Other (Very big, conical mother rhizomes and slender

fingers)

Brownish orange Orange

T 14 Horizontal Other (slender spindle shaped fingers) Brownish orange Light Yellow

T 15 Curved Other (Bold, medium sized rhizomes with closer

internodes)

Brownish orange Orange

T 16 Horizontal Other (Thick, round, plumpy mother rhizomes) Brownish orange Orange

T 17 Horizontal Other (Very slender spindle shaped fingers) Brownish orange Orange

T 18 Horizontal Plumpy Brownish orange Orange

T 19 Horizontal Other (Bold rhizomes with closer internodes) Golden yellow Orange

T 20 Horizontal Other (Medium sized rhizomes) Brownish orange Yellow

T 21 Horizontal Other (Medium sized mother rhizomes and plumpy

fingers)

Brownish orange Orange

T 22 Horizontal Other (Slender mother rhizomes with plumpy fingers) Brownish orange Other (Reddish

Orange)

T 23 Horizontal Other (Plumpy mother rhizomes and medium sized Brownish orange Orange

fingers)

T 24 Curved Plumpy Brownish orange Orange

T 25 Curved Other (Very bold rhizomes with closer internodes) Golden yellow Yellow

T 26 Horizontal Other (Slender rhizomes with closer internodes) Brownish orange Yellow

T 27 Horizontal Plumpy Dull brown ReddishYellow

T 28 Horizontal Plumpy Dull brown Orange

T 29 Horizontal Other (Bigger and plumpy mother rhizomes with slender

fingers)

Dull brown Orange

T 30 Horizontal Slender Brownish orange Yellow

Table 9 . Sensory Rhizome characters in Curcuma longa L . var ieties/accessions

Acc.

No.

Sensory properties of rhizomes

Acceptability of Colour Flavour Texture

T 1 Attractive Yellow Good aroma Fine

T 2 Attractive Yellow Pleasing aroma Slightly grainy

T 3 Yellowish brown Pleasing aroma Slightly grainy

T 4 Unattractive Pale brown Pleasing aroma Coarse

T 5 Attractive Yellow Good aroma Very Fine

T 6 Yellowish brown Pleasing aroma Fine

T 7 Yellowish brown Pleasing aroma Coarse

T 8 Attractive Yellow Pleasing aroma Very Fine

T 9 Attractive Yellow Pleasing aroma Fine

T 10 Unattractive Brown Unpleasant aroma Coarse

T 11 Attractive Yellow Pleasing aroma Very Fine

T 12 Unattractive Pale brown Pleasing aroma Coarse

T 13 Yellowish brown Pleasing aroma Slightly grainy

T 14 Attractive Yellow Pleasing aroma Fine

T 15 Yellowish brown Pleasing aroma Slightly grainy

T 16 Unattractive Pale brown Pleasing aroma Coarse

T 17 Yellowish brown Pleasing aroma Slightly grainy

T 18 Unattractive Brown Pleasing aroma Coarse

T 19 Yellowish brown Pleasing aroma Slightly grainy

T 20 Unattractive Brown Unpleasant aroma Coarse

T 21 Unattractive Brown Pleasing aroma Slightly grainy

T 22 Yellowish brown Pleasing aroma Slightly grainy

T 23 Unattractive Pale brown Unpleasant aroma Coarse

T 24 Yellowish brown Pleasing aroma Coarse

T 25 Attractive Yellow Pleasing aroma Fine

T 26 Attractive Yellow Good aroma Very Fine

T 27 Attractive Yellow Good aroma Fine

T 28 Yellowish brown Pleasing aroma Fine

T 29 Yellowish brown Good aroma Fine

T 30 Unattractive Pale brown Unpleasant aroma Coarse

Table 10. Leaf characters in Kaempferia galanga L. accessions

Accession No. Leaf Shape Leaf colour Leaf margin Leaf size

K1 Obtuse Light green Entire

K2 Suborbicular Dark green Slightly wavy

K3 Obtuse Light green Entire

K4 Suborbicular Light green Entire

K5 Obtuse Green Slightly wavy

K6 Suborbicular Light green Entire Small

K7 Ovate Light green Entire

K8 Suborbicular Light green Entire

K9 Suborbicular Light green Entire

K10 Lanceolate Light green Entire

K11 Elliptic Light green Entire Small

K12 Suborbicular Dark green Entire

K13 Suborbicular Light green Entire

K14 Cuneate Light green Entire

K15 Elliptic Dark green Entire Small

K16 Elliptic Light green Entire

K17 Lanceolate Dark green Entire

K18 Suborbicular Dark green Entire

K19 Cuneate Dark green Entire

K20 Elliptic Green Entire Small

K21 Obtuse Green Entire Small

Table 11 . Rhizome characters in Kaempferia galanga L. accessions

Acc. No. Rhizome

shape Rhizome nature Rhizome colour

K1 Other (Medium sized rhizomes and closer internodes) Orange

K2 Other (Medium sized rhizomes and closer internodes) Yellow

K3

Other (Bigger mother rhizomes and medium sized

fingers) Orange

K4 Other (Medium sized rhizomes) Yellow

K5 Plumpy Orange

K6 Plumpy Orange

K7

Other (Medium sized mother rhizome and slender

fingers) Yellow

K8 Other (Bold, medium sized rhizomes) Other (Reddish Orange)

K9 Other (Bold mother rhizomes and slender fingers) Orange

K10 Other (Bold mother rhizomes and slender fingers) ReddishYellow

K11 Plumpy Orange

K12 Plumpy Orange

K13

Other (Very big, conical mother rhizomes and slender

fingers) Orange

K14 Other (slender spindle shaped fingers) Light Yellow

K15

Other (Bold, medium sized rhizomes with closer

internodes) Orange

K16 Other (Thick, round, plumpy mother rhizomes) Orange

K17 Other (Very slender spindle shaped fingers) Orange

K18 Plumpy Orange

K19 Other (Bold rhizomes with closer internodes) Orange

K20 Other (Medium sized rhizomes) Yellow

K21

Other (Medium sized mother rhizomes and plumpy

fingers) Orange

Table 12. Mean value of biometric characters in Curcuma longa L.

Variety /

Accession

Plant Height (cm) No. of Tillers No. of Mother Rhizomes

per plant

No. of Primary Rhizomes

per plant

No. of Secondary

Rhizomes per plant

2016 2017 Mean 2016 2017 Mean 2016 2017 Mean 2016 2017 Mean 2016 2017 Mean

Kanthi

Shoba

Sona

Varna

IISR Prabha

IISR Prathibha

IISR All.Supreme

IISR Suvarna

IISR Suguna

IISR Sudarsana

IISR Kedaram

Roma

Suroma

Renga

Resmi

BSR I

BSR II

Rajendra Sonia

Mega Turmeric

Punjab Haldi I

Punjab Haldi II

Pant Peetab

DuggiralaRed

Suranjana

Narendra Haldi

TBGRI Local

Kasaragod Local

Kanjikuzy Local

Kuttipuram Local

Aluva Local

101.20

102.50

75.20

58.40

71.10

51.70

125.80

59.50

86.10

82.90

56.80

74.80

84.50

93.90

76.70

74.80

71.70

53.80

54.90

73.00

96.60

60.10

86.90

51.70

86.60

87.50

93.00

63.90

46.10

105.80

104.67

104.32

83.34

64.67

73.33

53.67

128.5

41.11

92.00

84.00

60.50

79.15

88.50

95.34

82.84

78.17

69.5

46.00

53.17

70.17

96.17

62.84

86.67

50.83

85.84

85.84

92.00

64.67

41.00

109.34

102.93

103.41

79.27

61.54

72.20

52.69

127.15

50.31

89.05

83.45

58.65

79.98

86.50

94.62

79.77

76.49

70.6

49.9

54.04

71.59

96.39

61.47

86.79

51.27

86.22

86.67

92.50

64.29

43.55

107.57

4.90

5.20

4.30

4.10

1.50

2.50

2.10

2.30

1.30

1.50

2.30

3.20

2.50

2.20

1.60

2.80

3.10

1.90

4.00

3.70

1.90

3.80

1.90

1.50

4.00

3.10

1.30

1.90

1.30

1.10

4.84

5.33

4.84

3.84

1.83

2.33

2.34

2.17

1.49

1.84

2.17

3.34

2.84

2.17

1.67

3.17

2.84

1.84

3.17

3.50

1.84

3.84

1.83

1.17

4.00

2.84

1.17

2.00

1.00

1.17

4.87

5.27

4.57

3.97

1.67

2.40

2.22

2.24

1.39

1.67

2.17

3.34

2.67

2.19

1.64

2.99

2.97

1.87

3.59

3.60

1.87

3.82

1.87

1.34

4.00

2.97

1.24

1.95

1.05

1.24

1.30

1.30

2.60

2.90

3.10

2.00

1.10

1.00

1.50

1.30

3.00

2.50

1.00

1.70

1.10

2.40

1.10

3.60

1.40

3.50

2.10

2.10

2.40

2.50

3.70

2.00

1.90

1.30

1.50

2.00

1.84

1.50

2.67

3.34

3.50

2.17

1.17

1.00

1.50

1.17

3.17

2.50

1.00

2.17

1.17

2.17

1.17

3.00

1.17

2.84

2.17

2.19

1.50

2.50

3.17

1.17

2.00

1.00

1.34

1.99

1.57

1.40

2.64

3.12

3.30

2.09

1.14

1.00

1.50

1.24

3.09

2.85

1.00

1.94

1.14

2.49

1.14

3.30

1.29

3.17

2.14

2.14

1.95

2.50

3.44

1.59

1.95

1.15

1.67

1.25

5.90

7.40

6.50

8.50

5.50

6.50

6.10

8.30

7.00

7.20

6.70

4.80

5.40

7.50

8.20

6.70

5.20

5.80

7.10

4.50

4.40

5.00

4.70

2.90

7.40

2.40

5.90

6.10

9.20

4.90

7.00

7.84

6.84

9.84

6.34

7.34

8.49

7.83

6.67

7.17

5.50

6.50

4.67

6.83

8.00

6.67

4.84

4.84

7.00

5.00

4.17

4.67

4.83

2.84

7.5

2.00

7.00

5.17

8.00

3.82

6.45

7.62

6.67

9.17

5.92

6.92

7.29

8.32

6.84

7.19

6.10

5.65

5.04

7.17

8.10

6.69

5.02

5.32

7.05

4.75

4.29

4.84

4.77

2.87

7.45

2.20

6.45

5.64

8.60

4.36

25.50

17.10

15.50

15.50

8.20

9.70

9.50

22.00

9.60

15.40

12.80

9.60

11.80

15.60

26.60

8.20

17.70

9.80

33.30

8.10

8.50

15.3

10.00

11.70

22.90

14.10

9.90

7.90

14.10

8.70

22.34

17.67

19.67

19.84

9.34

12.67

9.00

18.34

7.34

14.34

12.67

7.67

10.00

14.67

23.34

6.83

15.34

8.83

33.17

10.5

7.34

14.00

9.83

12.00

23.50

14.34

8.50

7.00

13.5

9.84

23.92

17.39

17.59

17.67

8.77

11.19

9.25

20.17

8.47

14.87

12.74

8.64

10.9

15.14

24.97

7.52

16.52

9.32

33.24

9.00

7.92

14.65

9.92

11.85

23.20

14.22

9.20

7.45

13.80

9.27

Table 12 Continued. Mean value of biometric characters in Curcuma longa L.

Variety /

Accession

Length of Mother

Rhizome (cm)

Length of Primary

Rhizome (cm)

Core diameter of Mother

Rhizome (cm)

Core diameter of Primary

Rhizome (cm)

Weight of Mother Rhizome /

Plant (g)

2016 2017 Mean 2016 2017 Mean 2016 2017 Mean 2016 2017 Mean 2016 2017 Mean

Kanthi

Shoba

Sona

Varna

IISR Prabha

IISR Prathibha

IISR All.Supreme

IISR Suvarna

IISR Suguna

IISR Sudarsana

IISR Kedaram

Roma

Suroma

Renga

Rasmi

BSR I

BSR II

Rajendra Sonia

Mega Turmeric

Punjab Haldi I

Punjab Haldi II

Pant Peetab

Duggirala Red

Suranjana

Narendra Haldi

TBGRI Local

Kasaragod Local

Kanjikuzy Local

Kuttipuram Local

Aluva Local

6.92

5.82

5.94

5.99

8.68

5.98

6.63

5.80

5.88

4.90

12.45

4.73

13.20

5.95

7.28

9.26

6.05

7.34

7.94

5.79

8.30

6.03

7.16

9.69

4.05

7.19

8.26

7.23

12.17

4.58

7.00

6.14

5.92

6.28

8.75

6.87

6.62

6.09

6.14

5.09

12.51

4.67

13.82

6.25

7.97

9.30

6.37

7.39

8.22

5.20

7.60

5.55

6.64

9.20

3.84

6.70

8.08

6.65

12.1

4.40

6.96

5.98

5.93

6.14

8.72

6.43

6.63

5.95

6.01

4.99

12.48

4.70

13.50

6.10

7.63

9.28

6.21

7.37

8.08

5.49

7.95

5.79

6.90

9.45

3.95

6.95

8.08

6.94

12.14

4.49

8.54

8.11

8.92

7.83

7.56

6.34

7.72

7.92

9.14

10.32

5.93

7.81

8.56

8.49

8.66

9.66

6.23

6.97

10.21

8.81

10.56

8.25

6.94

7.61

8.04

5.45

4.30

7.72

5.65

3.87

8.47

7.92

9.07

7.95

7.69

7.67

8.15

8.10

8.27

10.67

6.35

7.47

8.50

8.80

8.92

10.04

6.18

6.34

9.50

8.50

10.15

8.24

6.33

7.75

7.80

5.40

4.12

6.80

5.04

3.30

8.51

8.02

8.99

7.89

7.63

6.99

7.94

8.01

8.23

10.49

6.14

7.89

8.53

8.65

8.79

9.85

6.21

6.64

9.86

8.66

10.34

8.25

6.64

7.68

7.92

5.43

4.21

7.26

5.35

3.59

8.74

11.60

11.85

11.28

13.49

11.13

9.68

5.58

8.39

7.31

11.38

8.65

14.29

9.33

11.48

14.41

10.65

8.39

13.43

8.73

9.78

8.91

13.00

12.27

10.32

10.28

12.47

10.93

13.29

7.85

8.87

12.25

12.17

11.47

14.43

10.05

10.44

5.40

8.52

7.13

12.38

9.00

13.63

9.60

12.39

14.28

11.02

8.14

12.45

8.75

9.07

8.57

12.49

11.9

10.02

8.43

11.07

10.05

12.12

7.74

8.81

11.93

12.01

11.38

13.96

10.59

10.06

5.49

8.46

7.22

11.88

8.83

13.96

9.47

11.94

14.35

10.84

8.27

12.94

8.74

9.43

8.74

12.75

12.09

10.17

9.36

11.77

10.49

12.71

7.79

5.66

6.35

5.25

4.97

5.98

6.00

5.07

5.80

6.98

6.22

6.40

6.90

7.73

5.14

4.66

6.45

6.02

5.47

7.05

7.30

7.43

6.23

4.72

6.15

7.32

5.16

5.98

7.12

4.78

4.06

5.48

6.20

5.45

4.47

6.45

4.75

5.20

5.75

6.57

6.62

6.70

6.94

7.32

5.03

5.77

6.38

5.97

5.20

7.20

7.18

7.05

6.44

4.77

6.07

6.65

4.50

5.47

6.37

4.27

4.27

5.57

6.28

5.35

4.65

6.22

5.38

5.23

5.78

6.78

6.42

6.55

6.92

7.53

5.09

5.22

6.42

5.99

5.34

7.13

7.24

7.24

6.34

4.75

6.11

6.99

4.83

5.73

6.75

4.53

4.17

29.15

40.65

23.87

29.29

72.12

33.05

27.22

31.90

19.30

12.24

46.05

22.77

118.02

16.77

36.24

60.72

35.16

20.75

60.04

21.40

26.55

20.21

40.35

57.15

28.20

38.45

48.10

43.40

26.37

12.94

23.97

37.39

22.15

26.75

67.10

30.35

25.95

31.30

18.30

10.95

44.95

22.45

117.35

14.95

37.90

58.30

33.75

20.05

58.80

20.35

24.20

19.40

39.85

54.95

25.15

26.90

45.45

42.35

25.40

10.95

26.56

39.02

23.01

28.02

69.56

31.70

26.59

31.60

18.80

11.59

45.50

22.61

117.69

15.86

37.07

59.91

34.46

20.04

59.42

20.88

25.38

19.80

40.10

56.05

26.68

32.68

46.78

42.88

25.89

11.95

Table 12 Continued. Mean value of biometric characters in Curcuma longa L.

Variety /

Accession

Weight of Primary

Rhizome /Plant (g)

Weight of Secondary

Rhizome / Plant (g)

Days to Maturity Wet Rhizome Yield per plant

(g/plant)

Dry Rhizome yield (%)

2016 2017 Mean 2016 2017 Mean 2016 2017 Mean 2016 2017 Mean 2016 2017 Mean

Kanthi

Shoba

Sona

Varna

IISR Prabha

IISR Prathibha

IISR All.Supreme

IISR Suvarna

IISR Suguna

IISR Sudarsana

IISR Kedaram

Roma

Suroma

Renga

Rasmi

BSR I

BSR II

Rajendra Sonia

Mega Turmeric

Punjab Haldi I

Punjab Haldi II

Pant Peetab

DuggiralaRed

Suranjana

Narendra Haldi

TBGRI Local

Kasaragod Local

Kanjikuzy Local

Kuttipuram Local

Aluva Local

12.18

19.19

10.83

11.87

17.08

12.44

13.17

16.92

19.11

15.42

16.44

16.23

23.39

13.38

17.65

28.41

9.40

16.75

25.53

20.09

30.35

20.50

11.90

18.40

26.20

26.41

20.27

23.40

6.16

5.69

10.26

18.76

9.35

9.97

15.45

11.20

12.30

16.80

18.20

15.05

15.40

15.30

21.80

16.25

19.05

26.35

8.55

15.90

24.05

18.05

27.30

20.05

11.05

15.15

25.45

24.25

19.75

22.50

5.40

4.45

11.22

18.98

10.09

10.92

16.27

11.82

12.74

16.80

18.66

15.24

15.92

15.77

22.59

14.82

18.35

27.38

8.18

16.33

24.79

19.07

28.83

20.28

11.48

16.78

25.83

25.33

20.01

22.95

5.78

5.07

11.28

6.48

2.36

4.41

2.69

1.67

5.16

14.70

6.47

5.35

5.08

7.69

5.26

3.95

16.68

4.01

7.39

3.19

5.57

17.05

7.77

9.73

1.84

3.12

6.17

16.16

9.12

5.71

9.57

8.63

10.59

8.01

1.84

3.85

1.75

1.33.3

3.45

13.85

6.35

3.89

4.44

7.00

5.25

2.85

18.25

3.15

6.95

2.50

4.90

16.60

6.00

9.05

1.90

3.05

4.95

16.20

8.75

4.70

9.25

6.40

10.94

7.25

2.10

4.13

2.22

1.51

4.31

14.28

6.41

4.62

4.76

7.35

5.26

3.40

17.47

3.58

7.17

2.85

5.24

16.83

6.89

9.39

1.87

3.09

5.56

16.18

8.94

5.21

9.41

7.32

248.20

255.40

249.50

264.00

220.00

231.50

223.50

234.50

230.50

231.00

229.00

259.00

263.00

254.00

243.50

276.50

237.00

229.00

284.50

249.50

256.00

235.50

218.00

238.00

248.50

249.50

252.00

231.50

233.00

249.00

238.50

240.50

240.17

249.50

215.00

222.00

216.00

227.75

224.00

220.85

216.15

230.50

239.50

230.50

230.00

243.00

228.50

218.85

256.34

233.15

234.35

223.75

214.20

223.85

225.30

237.80

238.30

219.50

218.50

226.00

243.35

247.95

244.84

256.75

217.00

226.75

219.75

231.13

227.25

225.93

222.58

244.75

251.25

242.25

236.75

259.75

232.75

223.93

270.42

241.33

245.18

229.63

216.10

230.93

236.90

243.65

245.15

225.50

225.75

237.50

404.10

309.00

166.00

253.00

340.00

163.50

160.50

494.50

224.00

202.00

315.00

205.50

300.00

217.00

669.50

356.90

217.50

298.50

437.50

328.00

252.50

278.50

177.00

232.50

435.00

644.00

302.00

244.00

233.00

131.00

285.58

297.38

145.88

238.41

334.09

144.07

140.89

398.09

192.74

187.91

281.82

182.51

269.58

181.39

619.09

320.43

186.02

256.99

397.84

318.68

208.51

266.64

126.12

214.20

385.58

310.29

279.88

199.96

219.49

111.80

294.84

303.19

155.94

245.71

337.05

153.79

150.69

396.29

208.37

194.96

298.41

194.01

284.79

199.19

644.29

338.67

201.76

277.75

417.67

323.34

230.51

272.57

151.56

223.35

410.29

327.15

290.94

221.98

226.25

121.40

20.15

19.38

18.88

19.05

19.10

16.60

14.80

21.30

18.50

13.70

17.30

17.40

18.80

22.50

22.10

22.60

19.20

21.70

19.40

24.60

18.40

22.50

12.60

15.90

28.50

21.90

20.60

15.10

19.30

11.20

19.75

19.19

18.86

18.54

18.52

16.28

13.86

20.77

18.35

12.82

16.28

17.19

18.35

21.65

21.50

22.15

18.55

20.70

18.25

23.45

16.10

21.00

10.30

15.30

27.30

20.60

18.40

14.45

17.95

10.15

19.95

19.29

18.87

18.79

18.81

16.44

14.33

21.04

18.43

13.26

16.79

17.29

18.58

22.08

21.80

22.38

18.88

21.20

18.83

24.03

17.25

21.75

11.45

15.60

27.90

21.25

19.50

14.78

18.63

10.68

studied namely, plant height (cm), number of tillers per plant, number of mother rhizomes per

plant, number of primary rhizomes per plant, number of secondary rhizomes per plant, length of

mother rhizome (cm), core diameter of mother rhizome (cm), length of primary rhizome (cm),

core diameter of primary rhizome (cm), weight of mother rhizome per plant (g), weight of

primary rhizome per plant (g), weight of secondary rhizome per plant (g), days to maturity, wet

rhizome yield per plant (g/plant) and dry rhizome yield per plant. Hence the accessions with high

mean value can directly be used for cultivation or as parents in hybridization in future research.

Among the turmeric accessions, Kanthi, Shoba, Punjab Haldi I and NarendraHaldi recorded

better growth in respect of plant height and number of tillers. Plant height varied from 46.10 cm

(Kuttipuram Local) to 125.80 cm (IISR Alleppy Supreme). The highest tillers per clump were

recorded in Shoba (5.20) which was significantly superior with all other cultivars.

Crop duration is an important factor to determine the cropping sequence of the specific

region. In the present investigation, the accessions IISR Prabha, IISR Kedaram, Rajendra Sonia

and Duggirala Red were found to be early maturing types (200 to 230 days), local accessions

from Aluva, Kuttipuram and Kanjikuzhy as well as Suranjana, NarendraHaldi were found to be

medium in maturation (230 to 250 days) whereas Roma, Suroma, Renga and Punjab Haldi II

were fallen in late maturation group (250 to 270 days). These findings are in confirmations with

Reddy et al. (1988), Sasikumar et al. (1994) and Shanmugasundaram and Thangaraj (2001) who

have recorded considerable variation in the duration of different accessions of turmeric.

Significant variation in wet rhizome yield per plant was observed. Among the turmeric

accessions, Rasmi is the highest yielder followed by Mega Turmeric and NarendraHaldi(Plate 5).

The accession NarendraHaldi got highest number of mother rhizomes per plant while

Kuttippuram Local got highest number of primary rhizomes per plant. The accession IISR

Suroma was characterized by the longest and heaviest mother rhizome.

A perusal of ANOVA data presented in Table13 showed highly significant variations

among the different kacholam accessions for all the 12 characters studied namely plant height

(cm), number of mother rhizomes per plant, number of primary rhizomes per plant, length of

mother rhizome (cm), diameter of mother rhizome (cm), length of primary rhizome (cm),

diameter of primary rhizome (cm), weight of mother rhizome per plant (g), weight of primary

rhizome per plant (g), days to maturity and wet rhizome yield per plant (g/plant).

Among the twentyonekacholam accessions, number of mother rhizomes as well as

heaviest mother rhizomes was observed in IC 582811. A wide range of variation was observed

for rhizome yield per plant and IC 373593 had the highest yield (155.03 g/plant) followed by IC

582811(151.22 g/plant) (Plate 6). The high yield in IC 373593 may be attributed to the higher

number of rhizomes and weight of mother as well as primary rhizomes. This confirms the fact

that rhizome yield is a complex trait and is the ultimate expression of many component

characters.

Scoring for pests and diseases

In the present study, shoot borer and leaf roller attack was observed in turmeric at early

stages and was controlled by spraying 0.05 per cent dimethoate. Among diseases, leaf blotch of

turmeric was observed in lower leaves during October and November and was controlled by 0.2

per cent mancozeb spray. In case of Kacholam, leaf rot disease incidence was noticed and was

controlled by drenching the soil with 0.2% Pseudomonas followed by spraying three times at

weekly intervals during first year experimentation(Plate 7).

7.1.3 Estimation of rhizome curcumin content using HPLC:

Curcumin, the yellow pigment can be considered as ideal “spice of life” which is the most important fraction of turmeric that is responsible for its biological activity. Table 14 shows

the result of the analysis for curcumin content of the 30 turmeric accessions. Results revealed

variation in the curcumin content among the accessions. Rasmi yielded the highest curcumin

content (3675 kg/ha) while BSR II gave the lowest (293.63 kg/ha). Expressed in terms of percent

curcumin, the range would be from 1.5 to 7.8 %.

Variation was exhibited not only among the different accessions evaluated but also with

the range of curcumin content reported. The study of Garg et al. (1999) resulted to a 0.61 to

1.45% range in curcumin content among the 27 accessions evaluated while Pandey and Katiyar

(2010) reported curcumin content to vary from 0.15 to 1.87% in the 22 genotypes analyzed.

Jayaprashka et al. (2002) obtained a range of 1.06 to 5.65% curcumin in four commercially

available varieties of turmeric. On the other hand, a range of 0.3 to 3.24% curcumin was

obtained by Pathak et al. (2010) in turmeric samples from different zones in India, while

Kulkarni et al. (2012) reported a curcumin yield of 12.39% from turmeric samples collected from

a Satara district in India. The significant differences in curcumin content among the 30 turmeric

accessions observed in our study may be an expression of genetic diversity among the turmeric

accessions. Since all the accessions were planted in the same environment and subjected to the

same growth conditions, any variation in the curcumin content could be attributed either to a

possible difference in the genotypes of these accessions, particularly the genes coding for the

enzymes in the biosynthetic pathway leading to the synthesis of curcumin as well as the varied

expression of these enzymes.

7.2.1 Experiment on organic nutrient management in turmeric and kacholam:

India shares around 90 per cent of the global turmeric production. But the share of

organic turmeric is only 11 per cent compared to the conventional turmeric. There is a great

demand for organic turmeric in the world market and also the growing demand for natural

colours in industry, fast food chains, pharmaceuticals offer a potential scope for organic

production of turmeric. Considering the economic importance of turmeric and environmental

problems caused by chemicals application, it is important to cultivate turmeric using organic

fertilizer. Different organic manures influence differently in terms of yield and quality of

turmeric. In view of this background, this study was aimed to evaluate the effect of

organic manure namely vermicompost on turmeric and kacholam yield.

Table 14. Curcumin yield of the 30 turmeric accessions

Sl No Variety / Accession Curcumin Percentage Average curcumin yield

(Kg/ha)

1 Kanthi 3.9 1067.39

2 Shoba 2.9 806.49

3 Sona 3.8 567.72

4 Varna 3.8 865.25

5 IISR Prabha 3.8 1162.80

6 IISR Prathibha 3.2 462.24

7 IISR All.Supreme 4.9 707.81

8 IISR Suvarna 3.2 645.12

9 IISR Suguna 4.7 947.52

10 IISR Sudarsana 4.7 854.46

11 IISR Kedaram 5.2 1474.20

12 Roma 5.8 1072.71

13 Suroma 6.3 1701.00

14 Renga 2.7 527.31

15 Resmi 6.1 3675.55

16 BSR I 1.6 513.93

17 BSR II 1.5 293.63

18 Rajendra Sonia 7.1 1907.42

19 Mega Turmeric 7.8 3071.25

20 Punjab Haldi I 4.4 1298.88

21 Punjab Haldi II 7.0 1590.75

22 Pant Peetab 6.2 1554.03

23 DuggiralaRed 2.8 446.04

24 Suranjana 4.2 878.85

25 Narendra Haldi 7.0 2740.50

26 TBGRI Local 7.3 2260.08

27 Kasaragod Local 7.5 2038.50

28 Kanjikuzy Local 5.3 1163.88

29 Kuttipuram Local 2.4 503.28

30 Aluva Local 3.0 353.70

The plants grown in the treatment namely 100 % replacement of Nitrogen with

vermicompost had the maximum plant height, number of tillers/plant, fresh weight and dry

weight of rhizome per plant compared to that of control in turmeric (Table 15). These plants

remained green longer which ultimately provided longer and higher photosynthesis process and

resulted in a higher rhizome yield of turmeric. The application of 100 % replacement of nitrogen

with vermicompost produced superior yield attributes like more number of mother rhizomes per

plant (1.75), more number of primary rhizomes per plant (5.19) and secondary rhizomes per

plant (18.03).Similarlythe same treatment expressed the best in terms of size of mother rhizome

(7.69 cm), primary rhizome (21.86 cm) and secondary rhizomes (7.05 cm). The lowest yield with

10 t/ha in the treatment with 25 % replacement of nitrogen with vermicompost may be due to

shortage of required nitrogen at the initial active growth stage which might have delayed plant

growth at early stages.

In kacholam (Table 16), POP recommendation recorded significantly higher yield

compared to vermicompost. The application of 100 % of recommended dose of nitrogen through

vermicompost (basal) resulted in significantly higher plant height, maximum number of mother

(5.6) and primary rhizomes per plant (25.92) and the highest rhizome yield(156.23 g/plant). It

was followed by the crop grown in 75 % replacement with vermicompost. The lowest content

was recorded in the crop grown with 25 % replacement with vermicompost. The vermicomposts

contain plant growth regulating substances including plant growth hormones and humic acids

which are probably responsible for increase in germination, growth and yield of plants.

Similar trends were noticed with respect to fresh rhizome yield per plot, which recorded

no significant difference among the various treatments (Table 4.5). As in the case of fresh

rhizome yield per plant, a higher mean value of 2.98 kg of fresh rhizome yield per plot was noted

in T2 (FYM + biofertilizers) followed by T4 (FYM + biofertilizers + Kalanchoepinnata) with

2.62 kg per plot. Lowest yield was recorded in T10 (2.16 kg) followed by the control plot,

recording 2.22 kg fresh rhizomes per plot.

The positive effect of biofertilizers on the various growth and yield parameters observed

in the present study might be due to enhanced uptake of nutrients by the plants. A possible

explanation for the beneficial effect of vermicompost may be due to the accumulation of mobile

substances in earthworm casts. Earth worms are reported to excrete plant growth promoting

substances into castes (Nielson, 1965). Kale et al.(1992) observed that vermicompost application

enhanced the activity of beneficial microbes like N2 fixers and colonization by mycorrhizal fungi

and hence play a significant role in N2 fixation and phosphate mobilization leading to better

uptake by the plant. Thus the increased availability of nutrients and uptake by the plants would

have resulted in better growth and yield in plots treated with vermicompost. The enriched

vemicompost might have provided nutrients to the plants andmayimprove edaphic factors, which

resulted in higher vegetative growth parameters. In conclusion, consideringall the above aspects

together 100 % replacement of nitrogen by vermicompost can be recommended. This

recommendation will ensure the highest fresh turmeric and kacholam yield.

7.3.1 Standardisation of micropropagation protocol of the selected medicinal plants

Establishment of aseptic cultures was found to be very difficult in both target species viz.,

turmeric and kacholam, because the explants were taken from underground rhizomes. In case of

turmeric, nearly 52% of the cultures were found to be contaminated and culture establishment

was only 48%. Culture establishment was only 44% in the case of Kacholam. In the present

study, different explants viz. leaf, root and rhizome (having emerged and not emerged buds) was

used. Growth was observed only in rhizomes.

In case of turmeric, the rhizome segments with emerged buds were found to be the best

explants for direct regeneration. Bud growth was noticed 11-13 days after inoculation. The

initiation of shoot growth occurred in almost all the media tried. But the rate of initiation was

found to be relatively higher in the media containing BA and IAA combinations. Highest

percentage of shoot initiation i.e., 95 % was recorded in MS supplemented with BAP (2 mg/l)

and IAA (0.5 mg/l) media (Plate 19). Average number of shootlets is two and mean shoot length

of 4.7 cm. The second best medium is MS + BAP (2 mg/l) and NAA (0.5 mg/l) in which 85% of

shoot initiation with an average of 1.2 shootlets and a mean shoot length of 4.1 cm (Table 17)

was recorded. However, increase in BAP concentration in the medium had adverse effects on

plant growth. Higher concentration of BAP (3.0 mg/l) with NAA (0.5 mg/l) showed callusing of

explants with fewer numbers of shoots. The present investigation has well demonstrated the

ability of BAP to induce axillary branching in turmeric. In consistence with these results, the

present study also revealed the role of BAP (singly or in combination with NAA) in shoot

initiation. In this study IBA has no effect in shoot initiation.

In order to further standardise the multiple shoot development, BAP and IAA

combinations along with Adenine sulphate treatments were tested. Combined use of BAP and

IAA did not help to maximize shoot multiplication in turmeric. However, the best production

response (4.8 shoots) under joint action BAP, IAA and Adenine sulphate was recorded on BAP

(2 mg/l) and IAA (0.5 mg/l) combination along with ADS (100 mg/l), showing improvement

over their individual use (Table 18) (Plate 8). Whereas, synergistic effect of ADS with other

cytokinins has been reported to increase shoot regeneration in Korarima (Tefera and

Wannakrairoj, 2006), Ochlandrawightii (Bejoy et al., 2012), A. hypoleucum (Bejoy et al., 2010)

etc. Maximum shoot elongation of average 21.8 cm growth was obtained from explants

inoculated onto MS medium with this combination in 35 days.

In the case of Kacholam, multiple shoots were found to develop from the sprouted

rhizome buds in MS medium supplemented with BA and also in BA + Kin, four weeks after

inoculation. The best medium identified for initiation of shoot buds was MS supplemented with

2 mg/l BA and 1 mg/l Kin (Table 19) (Plate 9). In this treatment, 90% of the cultured explants

showed initiation and 95% multiple shooting.

Rooting of the proliferated shoots in Turmeric and Kacholam

Root development was induced in the in vitro proliferated shoots by culturing them on

half strength of MS medium with different concentrations of NAA and IBA. Among different

Table 17. Effect of different concentrations of BAP, IAA and NAA on the shoot initiation of turmeric in

MS media

Hormonal

concentrations

(mg/l)

No. of

explants

cultured

Mean number

of explants

regenerated

Percentage of

regeneration

Days to

shoot

initiation

Number of

shoots per

explant

Shoot

length

(cm)

BAP + IAA

0.5 + 0.0 20 12 60 25-27 1 3.2

1.0 + 0.0 20 14 70 20-25 1 3.5

1.5 + 0.0 20 15 75 15-20 1 3.6

2.0 + 0.0 20 19 95 13-14 1 4.0

2.5 + 0.0 20 18 90 15-18 1 3.5

3.0 + 0.0 20 17 85 20-25 1 3.2

2.0 + 0.1 20 11 55 18-20 1 3.4

2.0 + 0.2 20 12 60 15-18 1 3.9

2.0 + 0.3 20 14 70 14-18 1.6 3.9

2.0 + 0.4 20 16 80 12-15 1.8 4.0

2.0 + 0.5 20 19 95 11-13 2 4.7

2.0 + 0.6 20 13 65 15-18 1.1 4.5

2.0 + 0.7 20 1 5 20-25 1 3

BAP + NAA

2.0 + 0.1 20 12 60 25-29 1 3.1

2.0 + 0.2 20 10 50 25-27 1 3.5

2.0 + 0.3 20 14 70 20-25 1 3.7

2.0 + 0.4 20 16 80 13-15 1 4.0

2.0 + 0.5 20 17 85 12-14 1.2 4.2

2.0 + 0.6 20 15 75 15-20 1 3.8

2.0 + 0.7 20 14 70 18-21 1 3.6

3.0 + 0.1 20 15 75 20-22 1 3.5

3.0 + 0.2 20 13 65 21-23 1 3.3

3.0 + 0.3 20 12 60 23-25 1 2.5

3.0 + 0.4 20 16 80 25-28 1 1.9

3.0 + 0.5 20 14 70 26-30 Callusing -

Table 18: Effect of different concentrations of BAP, IAA and NAA on the shoot proliferation of turmeric

in MS media.

Hormonal

concentrations

(mg/l)

No. of

explants

cultured

Mean number

of explants

regenerated

Percentage of

regeneration

Days to

shoot

proliferation

Number of

shoots per

explant

Shoot

length

(cm)

BAP + IAA + ADS

2.0 + 0.5 + 0 20 11 55 48-50 1 9.4

2.0 + 0.5 + 25 20 12 60 35-38 1 14.9

2.0 + 0.5 + 50 20 14 70 33-38 1.6 15.7

2.0 + 0.5 + 100 20 18 90 30-35 4.8 21.8

Table 19: Effect of different concentrations of BAP and Kin on the initiation and shoot proliferation of

Kacholam in MS media.

Hormonal

concentrations

(mg/l)

No. of

explants

cultured

Mean

number of

explants

regenerated

Percentage

of explants

showing

proliferation

Days to

shoot

initiation

Number of shoots per

explant

Average

length of

shoot per

culture Initiation Proliferation

BAP

0.5 20 13 65 35-37 1 1.20+ 0.19 1.7

1.0 20 14 70 32-35 1 3.40+ 0.12 2.3

1.5 20 15 75 30-32 1 4.00+ 0.35 2.5

2.0 20 18 90 28-30 3 5.20+ 0.45 2.7

2.5 20 16 80 30-31 1 3.80+ 0.31 2.0

3.0 20 14 70 33-35 1 2.60+ 0.35 1.8

BAP + Kin

2.0 + 1.0 20 19 95 28-30 3 6.40+ 0.17 3.4

2.0 + 2.0 20 14 70 30-35 1 3.80+ 0.22 2.6

Table 20. Effect of different concentrations of auxins on adventitious root formation from in vitro

regenerated shoots

Concenration

of auxin

(mg/l)

Percentage of shoots

rooted

Number of roots per

rooted shoots

Average length of Roots

Turmeric Kacholam Turmeric Kacholam Turmeric Kacholam

NAA

0.2 65 75 5.40+ 0.16 2.17+ 0.02 3.74 +0.24 3.18+ 0.14

0.5 75 80 6.00+ 0.23 2.25+ 0.07 4.10 +0.24 3.59+ 0.05

1.0 80 90 7.60+ 0.42 3.54+ 0.06 5.22 +0.46 3.84+ 0.07

2.0 60 70 4.40+ 0.52 3.10+ 0.04 3.34 +0.30 3.61+ 0.04

IBA

0.2 74 88 6.40+ 0.35 2.63+ 0.07 4.32 +0.27 3.22+ 0.01

0.5 88 93 8.20+ 0.23 3.38+ 0.11 5.64 +0.23 4.23+ 0.08

1.0 96 98 9.40+ 0.74 4.71+ 0.12 6.30 +0.34 6.21+ 0.02

2.0 83 80 5.20+ 0.28 3.88+ 0.09 5.28 +0.56 5.90+ 0.05

types of auxin used in the experiment, IBA was found to be the most effective at different

concentrations tested for producing roots on bases of micro-shoots (Table 20) (Plate 10). Among

different concentrations of IBA, 1 mg/l was found to be the best concentration of auxin for

proper rooting of turmeric and kacholam in which 96% and 98% shoots rooted within 4 weeks of

culture respectively.

7.3.2 Optimization of Cost Effective Micropropagation Protocol for identified Superior

Genotype of Selected Medicinal Plants:

The conventional micropropagation laboratory is typically a capital intensive and

sophisticated facility. These laboratories have rigorous procedures for maintenance of sterility.

The use of high purity chemicals, expensive equipments and specialized labware is the rule.

Therefore, low cost alternatives are needed to reduce cost of production of tissue-cultured plants

without compromising the quality of the micropropagules and plants. Successful

micropropagation of turmeric and kacholam were demonstrated using a combination of the

simple and low-cost alternatives in equipment and techniques adopted.

LOW-COST OPTIONS USED IN THIS STUDY

EQUIPMENT

When sterilization of glassware and culture media was carried out using a large pressure

cooker and an LPG burner contamination was high when compared to autoclave sterilization.

While the initial investment and running costs are compared, using of pressure cooker and LPgas

was found to be more economical than electrically operated autoclave. pH paper strips instead of

the pH meter for pH measurement.

Water is the main compound of all tissue culture media. Distilled water made through

electrical distillation is very expensive and use of alternative water sources like tap water was

experimented in this study to lower the cost of medium. Shoot multiplication rates and normal

growth was maintained in Kacholam when glass distilled water was replaced with tap water in

preparation of media. Since operating a glass distillation still is highly energy intensive,

significant savings in cost of production will result with the use of tap water. However it has to

be anticipated that the quality of tap water will vary with the location and the efficiency of the

water treatment system. Dissolved impurities and suspended matter and varying levels of

chlorine are to be expected in the tap water. Hence boiling and filtering of the tap water is

suggested if required or single distilled or deionized water.

CHEMICALS:

Tender coconut water (TCW) is natural and cheaper souce of plant hormones and

minerals. It contains mainly water (94%) and growth promoting substances that can influence in

vitro cultures including inorganic ions, amino acids, organic acids, vitamins, sugars, sugar

alcohols, lipids, nitrogenous compounds and phytohormones (Yong et al., 2009). The ability of

coconut water to support plant growth in vitro is due to its ability to stimulate cell division and

morphogenesis. Moreover, the pH of coconut water is within the range of 4.6 - 5.9 which falls

within the pH, 5.8 of MS medium. Not only does coconut water support plant growth in vitro but

George (1993) has attributed the robustness and high survival rate of plants cultured on coconut

water to the high carbohydrate content which could be used to meet the respiratory demands

while surviving the physiological shocks of ex-vitro procedures.

In the experiment on turmeric, addition of 100 ml/L CW along with IAA produced a

maximum of 5.38 shoots with 85% shoot regeneration ability (Table 21) (Plate 11). Increase in

concentration to 150 ml of CW promotes elongated shoots (23.72 cm) compared to BAP + IAA

+ ADS combination. It can be concluded that tender coconut water is a complex combination of

compounds, containing a number of amino acids, organic acids, nucleic acids, several vitamins,

sugars and sugar alcohols, plant hormones (auxins, cytokinins), minerals, and other unidentified

substances and none of which alone is totally responsible for growth promoting qualities. These

results suggest that complex natural supplements can efficiently support shoot induction,

multiplication and multiple root formation. Also, Direct rooting of in vitro shoots was obtained

when subcultured on same media combination (MS + 100 ml/l coconut water + IAA (0.5 mg/L).

In the present study on kacholam, from Table 22, it was observed that 50 per cent

explants differentiated into shoots when the coconut water was not supplemented to the medium

and produced 5.20 shoots per explant and in this condition shoot length was 2.7 cm on an

average. When the coconut water was added in the medium, a significant increase in the

frequency of explants showing shoot regeneration was observed and 90 per cent of explants

showed shoot regeneration in the presence of 150 ml L-1 coconut water followed by 80 per cent

explants in 100 ml L-1 coconut water in the medium. Use of coconut water significantly affected

multiple shoot regeneration from the explants. Whereas 5.84 shoots per explant were observed in

the medium containing 150 ml L-1 coconut water followed by 5.56 shoots per explant in medium

containing 100 ml L-1 coconut water. Developed shoots showed significant increase in length as

the concentration of coconut water was increased up to 150 ml L -1. Highest shoot length i.e.

4.52 cm was observed in the medium containing 150 ml L-1 coconut water. Increasing the

concentration of coconut water in the medium above 150 ml L-1 did not show any significant

effect on shoot regeneration (Plate 12). Hence in this study, tender coconut water can be

considered itself as a nutrient medium because it provides undefined nutrients to growing plants

and is a source of mainly growth hormones and vitamins viz., zeatin, inositol and reduced

nitrogen compounds. The results obtained support the earlier reports of Kalpona et al. (2000)

and Aktar et al. (2008). They also reported that the addition of coconut water as organic growth

supplement in the media could improve plant’s growth in tissue culture.

Similarily, the idea of replacing purified sucrose with market sugar holds analogous

promise. Comparative cost assessment reveals sucrose price varies from 220.00 to 450.00 / 500g

(grade dependent) while the cost of market sugar ranging from 30-40/ 500g.

Other chemical sterilants commonly used are calcium or sodium hypochlorite. Since

freshly made calcium or sodium hypochlorite is not readily available, mercuric chloride was

preferred although it is less safe to use and dispose.

Table 21. Effect of coconut water on shoot proliferation in turmeric

(Medium –MS + inositol 100 mg l-1 + Table sugar 30.00 g l-1 + agar 7.50 g l-1 + 1000 ml Tap water)

MS + IAA (mg/L) +

CW (ml/L)

Frequency (%) of

explants showing

shoot regeneration

Number of shoots

regenerated per

explants

Shoot length

(cm)

5 + 0 50 2.45 12.32

5 + 50

70 3.28 15.48

5 + 100 85 5.38 20.52

5 + 150 74 3.84 23.72

5 + 200 73.2 3.80 18.85

Table 22. Effect of coconut water on shoot proliferation in Kacholam

(Medium –MS + inositol 100 mg l-1 + Table sugar 30.00 g l-1 + agar 7.50 g l-1 + 1000 ml Tap water)

MS + CW (ml/L) Frequency (%) of

explants showing

shoot regeneration

Number of shoots

regenerated per

explants

Shoot length

(cm)

0 50 5.20 2.70

50

70 4.17 2.78

100 80 5.56 3.84

150 90 5.84 4.52

200 73.2 3.90 2.85

In case of kacholam, the difference in plant growth between plantlets that were cultured on BAP

along with kinetin and those on coconut water and table sugar was not wide. Plants on coconut

water (150 ml/l) with table sugar performed satisfactorily compared with those on the BAP (2

mg/l) + Kin (1 mg/l) along with sucrose. This confirms that coconut water contain plant growth

hormones which can equally support the growth of plants in vitro. Since coconut water contains

endogenous cytokinins, it is expected that adding it to the medium should have the same or

similar effect on growth or morphogenesis.

CULTURE CONTAINERS

The type of culture vessel influences the efficiency of transfer during subculture and

production of propagules per unit area. The disadvantage in the use of test tubes is that in the

later stages of cultures transfers will be impeded by the small diameter of the vessel. Conical

flasks, also made of borosilicate, is the commonly used container when larger volumes are

required. The narrow neck of such flasks also makes it extremely difficult to carry out transfers.

Being expensive makes it unsuited for large scale micropropagation. Cotton plugs used as

closures for test tubes and flasks are reusable for a few times if they have not been in contact

with media. In long term cultures cotton plugs accumulate dust and spores which will trickle

down into the vessel and pose a risk of contamination. In the present study, the cost savings was

achieved by use of jam bottles having wide mouths with Polypropylene (PP) caps and thereby

avoiding borosilicate glassware as culture containers (Plate 13).

7.3.3Hardening of microplantlets : The in vitro regenerated plants were transferred in small

polybags containing soil, vermicompost and sand mixture in the 1:1:1 proportion (Plate 14). The

plantlets were kept in shade net house under 50% sunlight for acclimatization to the outside

environmental conditions for 30 days with regular watering and the percentage of survival was

noted. It was observed that 86% of turmeric plantlets and 92% of kacholam plantlets could be

established under ex vitro conditions. The use of sufficiently porous potting mix that allows

adequate drainage and aeration has been recommended for fast acclimatization of in vitro

regenerated plants. Rest of the transplants could not survive either due to wilting caused of hot

and dry atmosphere or microbial overgrowth that caused damping off and necrosis of transplants.

These results demonstrate that turmeric and kacholam can be micropropagated readily with

above said methods and medium.

7.4.1 Initiation of Project Activities in the Selected GramaPanchayaths and Selection of

Women Beneficiaries:

As per the suggestion of 3rd group Monitoring Workshop of Back to lab projects

regarding site selection, two locations were selected for initiation of project activities in

Alappuzha district. The selected gramapanchayaths were Muthukulam (Sandy loam soil) and

Chunakara (Lateritic soil). Detailed discussion with the selected Panchayath officials and

People’s representatives for the successful implementation of the project were conducted. The

discussions were fruitful as it had conveyed the objective of this project and the successful

implementation of the present project will have dual benefits: upliftment of livelihood of

unemployed rural women and cultivation of economically viable medicinal plants. Besides, it

can create self reliance and empowerment for rural women.

Basic data of women farmers entered in Farmer Database of these selected krishibhavans

were collected from the concerned agricultural officers. Local awareness campaigns were

conducted in both the panchayaths to provide a platform for dissemination of the latest

information on scientific mode of medicinal plant cultivation and sustainable income generation

(Plate 15). Two leaflets on scientific cultivation of Turmeric and Kacholam were prepared in

Malayalam and were distributed to the participants during the campaigns. Selection forms based

on the criteria like age, education, size of the family, socio-economic status, social activities,

entrepreneurship, land and irrigation facilities possessed and their interest in the programme

from the target site were distributed for the purpose of selection of women beneficiaries. After

verification of the selection forms, 8 women beneficiaries from Muthukulampanchayath and 7

women beneficiaries from Chunakarapanchayath were selected. Subsequently leader beneficiary

was selected for each group for streamlining the project activities. Separate registers were kept

for each group for documenting the minutes of meetings, day today activities of beneficiaries in

the experiment field, observations etc.

7.4.2 Initiation of Homestead Cultivation for Macropropagation of Superior Line:

Hands-on trials will be given to the leader beneficiaries to initiate homestead cultivation

and was given to co ordinate the activities. To initiate cultivation, sufficient number of rhizomes

as well as healthy and established tissue culture microplantlets (Curcuma longa and

Kaempferiagalanga), organic manure, plant protection chemicals etc was distributed for each

pancahayath. Appropriate guidelines were given for starting the field experimentation in each

panchayath. Separate fields were laid out for evaluation of rhizomes as well as microplantlets of

turmeric and kacholam (Plate 16).

8.

SUMMARY

In the experiment on germplasm evaluation of turmeric and kacholam conducted at

ORARS, Kayamkulam revealed that the turmeric variety Rasmirecorded thehighestrhizome yield

andcurcumincontent.This variety is having a pleasing sensory characters andtolerance to pest and

diseases. Rasmi is a clonal selection from Rajapore local released by the High Altitude Research

Station, Odisha and is medium in duration (matures 8 months after planting). The accession

produces bold, medium sized rhizomes with closer internodes which have market preference.

The rhizome powder is yellowish brown in colour, slightly grainy in texture and have pleasing

aroma. Fresh rhizome yield is 55.03 t/ha. The dry recovery percentage is 22%. Curcumin content

is 6.1 %. On the basis of best performance, Rasmi is adjudged as the most promising and suitable

turmeric variety for general cultivation in Onattukara tract. The high yields in terms of rhizome

and commercially valuable curcumin along with other desirable field characteristics will make

the variety “Rasmi” acceptable to the Onattukara farmers for its adoption for cultivation.

In the case of kacholam, IC 373593 recorded the highest yield (13.95 t/ha) among the

twentyone genotypes evaluated and can be recommended for general cultivation in Onattukara

tract. The high yield in IC 373593 may be attributed to the higher number of rhizomes and

weight of mother as well as primary rhizomes. The genotype is collected from NBPGR Regional

Station, Thrissur. It is medium in duration and produces orange coloured thick, round, plumpy

mother rhizomes.

In the experiment on organic nutrient management in turmeric and kacholam, quantity of

vermicompost 25 t/ha and 28 t/ha respectively (100 % replacement of nitrogen by

vermicompost) can be recommended. This recommendation will ensure the highest fresh

turmeric and kacholam yield.

In the experiment on standardisation of cost effective micropropagation protocol for

identified Superior Genotype of turmeric and kacholam observed that low cost substitutes such

as market sugar, tap water and tender coconut water did not adversely affect the proper growth of

the micro plants. The shoot multiplication rate obtained in the low cost options was not less

when compared to the conventional in vitro multiplication trials. The cost on media ingredients

was reduced by using household sugar instead of analytical grade sucrose, tender coconut water

instead of Cytokinin –BAP and Kinetin as well as tap water instead of double distilled water.

The cost effectiveness by use of alternate media constituents, equipments and culture containers

established during this study using Kacholam and turmeric shoot culture as a model system can

be utilized in any other crop species.

In the case of initiation of homestead cultivation for macropropagation of superior lines

of turmeric and kacholam, 15 women beneficiaries of muthukulam and chunakarapanchayats

were provided with sufficient number of rhizomes as well as healthy and established tissue

culture microplantlets (Curcuma longa and Kaempferiagalanga), organic manure, plant

protection chemicals etc. Appropriate guidelines were given for staring the field experimentation

in each panchayath. Separate fields were laid out for evaluation of rhizomes as well as

microplantlets of turmeric and kacholam.

9. Outcomes of the project (Brief summary)

i) Salient findings (in bullet points) including technical details and innovations

• On characterization and evaluation of germplasm of turmeric and kacholam, superior

lines namely RASMI (turmeric variety) and IC 373593 (Kacholam accession) was

identified as economically viable medicinal plants and are suitable for agro-climatic

conditions for Onattukara tract.

• On evaluation of the effect of organic manure on the yield of turmeric and kacholam

,vermicompost (………)can replace N (as chemical fertilizer) without any reduction

in yield

• The standardization of a cost effective tissue culture methodology for Kaempferia

galangal and Curcuma longa had developed a protocol for the mass production of

these medicinal plants to satisfy the demand in the market in Kerala state.

• Women farmers became well trained on scientific mode of medicinal plant cultivation

which include appropriate selection and identification of planting material,

propagation methods and cultivation techniques.

• Formation of Self Help Groups has raised the programme to a sustainable level which

will be effective after the completion of market linkage activities in the project.

ii) Publications

a. Journals (a. International b. National) :

National

a. Genetic variability of turmeric (Curcuma longaL.) genotypes in India. Sent for

publication in Indian Journal of Horticulture (NAAS rating : 6.2)

b. Genetic variabilityanalysis in Kacholam genotypes. Sent for publication in Journal

of Horticultural Sciences (NAAS rating : 4.4)

c. In vitro multiplication of kaempferiagalanga l.- an important medicinal plant.

Journal of applied horticulture (NAAS rating : 4.6)

b. Papers presented in Conferences

1. “Genetic cataloguing of Turmeric (Curcuma longa L.)” published in National seminar on “Modern trends in conservation, utilization and improvement of plant geneic

resources during November 23 and 24, 2017 at University of Kerala, Kariavattom and

GregorMendalfoundation.Proceedings. p.18

“Genetic cataloguing of Kacholam (Kaempferiagalanga L.) published in 30th

Kerala

Science Congress at Government Brunnen College, Thalassery on January 28-30,

2018.Abstracts. pp.40-41

c. Other publications

1. “Rasmi: a gifted turmeric variety for Onattukara tract.” e-Kerala

KarshakanFebruary 2018 pp. 4-9

2.“All about Turmeric” Kisan World November 2017, pp. 46-47

3.“Production Technology of Kacholam (Kaempferiagalanga)” Sent for publication

in Indian Farmer’s Digest.

4.“Kacholam- Keralathinteparambaragathasasyam”- Sent for publication in Kerala

Karshakan

5.“KacholamKrishi” – Malayalam leaflet prepared

6.“Manjalkrishi” – Malayalam leaflet prepared

10. SCOPE OF FUTURE WORK

Medicinal plants occupies an important position in the socio-cultural, spiritual and

medicinal arena of rural people of India. Their sustainable management and harvesting can

conserve biodiversity, sustain human and environmental health, generate employment and

enhance export earnings.

Systematic cultivation of many medicinal plants needs specific cultural practices and

agronomical requirements. These are species-specific and are dependent on soil, water and

climatic conditions. Hence research and development work has to be done to formulate good

agricultural practices which will include appropriate selection and identification, propagation

methods, cultivation techniques, harvesting, stepwise quality control of raw material

uptoprocessing stage, post-harvest treatment, storage and safety. These aspects have to be

incorporated into protocols for the cultivation of economically viable medicinal plants.

Organic farming is another practice that is gaining wide acceptance as world demand

particularly in developed countries for organically grown crops is rapidly on the increase.

Farmers have to be trained in all aspects of organic farming of medicinal plants including

obtaining certification from associations that do the monitoring starting from cultivation to final

harvesting. Organic farming which is labour-intensive gives the developing countries the

comparative advantage to be competitive.

Voucher specimens in pots of each species collected during the project is maintained in

ORARS germplasm which will help in future cross-references. Vouchers provide a verifiable

means to identify and distinguish those plants from one another, especially if they share the same

common name.Previously identified vouchers can also be valuable tools to assist researchers in

confirming the identity of an unknown plant after dichotomous keys have been used to narrow

down the possibilities to several plausible options.

Low cost tissue culture techniques standardized can be utilized for large scale production

of healthy plant material which is essential for maintaining product quality.The data obtained in

this work could serve as an important resource for further studies on these medicinal plants.This

attempt will certainly encourage young researchers to work on various areas of research for the

development and promotion of economically viable medicinal plants of Kerala.

The project is envisaged for technological as well as economic empowerment of women

through training, education and participatory approach involving self help women groups.

Emphasis was also laid on increasing decision making capabilities, improving skill through

transfer of technologies of agro-biotechnological aspects and marketing. The project will make

democratic approach to aware rural women about income generation through homestead

cultivation of commercially valuable medicinal plants and exploring its potential in livelihood

improvement. The sector is expected to generateadditional employment to women farmers in the

process of cultivation/regeneration,collection, drying, grading and processing of these medicinal

plants. To fulfill their true potential as a potent women empowerment tool SHGs should broaden

their horizon and view empowerment as a multi-dimensional social process and therefore it is

proposed that greater in depth research needs to be undertaken into SHGs so that more evidence

can be obtained in order to ascertain the consistency and long-term accomplishments of SHGs as

a potent tool for holistic women empowerment.

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