Volume 4 • Issue 7 • 1000202J Bioremed Biodeg ISSN: 2155-6199 JBRBD, an open access journal

Open Access

Mohamed et al., J Bioremed Biodeg 2013, 4:7 DOI: 10.4172/2155-6199.1000202

Open Access

Research Article

Keywords: Jojoba; Micro-propagation

IntroductionJojoba (Simmondsia chinensis) is a perennial evergreen, dioecious

and an obligated cross pollinated shrub [1]. Because of its low water requirement, root system and economical potential, jojoba was introduced to cultivation in various semi-arid and arid regions of the world. The Jojoba seeds contain about 50-55% distinctive oil, which was used commercially in the cosmetic, pharmaceutical and lubricant industries [2]. Sexual propagation of Jojoba dose not reproduce true to type plants and asexual propagation of single jojoba plant is necessary to conserve the desirable traits. Tissue culture technique suited to the multiplication of elite crops by using explants of the plant leaf, root and steam. The genotypes/clones differ in their response when cultured in vitro [3,4]. The lower concentration of BA proved more effective for many shoot parameters [5]. MS media supplemented with BA and GA3 enhanced Shoot initiation of Jojoba, while MS+IBA supported root formation [6]. Bud initiation and shoot multiplication were found to be greatest on Murashige and Skoog's (MS) medium supplemented with BA and adenine [7]. The present study aims to investigate the responses of four jojoba genotypes to in vitro nodal segments and shoot tip culture under different growth regulators combinations and sucrose concentrations.

Materials and MethodsPlant materials

Four jojoba genotypes called ‘Hail A’, ‘Medina’, ‘Hail B’ and ‘Hada Al-sham’, and cultivated in the western regions of Saudi Arabia [8] were used. The 1-2 cm nodal segments were prepared from the one year established branches on the 5 years old jojoba plants (Figure 1). The actively growing shoot tips (0.5-1 cm), with both apical and auxiliary shoot buds were prepared from the newly performed branches on the jojoba plants (Figure 1).

Sterilization protocol

The explants were subjected to a sterilization protocol as following: a) thoroughly washed in running tap water for 30 min, b) dipped in98% ethanol for seconds, c) then 10% Clorox (v/v) solution+2 dropsof Tween-20 for 15 min, d) followed by four washings with sterile

distilled water, each for 5 min, and e) for nodal segments the explants were dipped in 0.1% mercuric chloride (w/v) solution for 15 min, and subsequently washed 4 times in sterile distilled water each for 5 min. The sterilization steps from b to e were conducted in a laminar air flow hood.

Culture medium and plant growth regulatorsFour experiments were conducted as followed: experiment (A)

was designed to study the effects of plant growth regulators on nodal segments, experiment (B) was conducted to study the effects of sucrose concentrations on nodal segments, experiment (C) was carried out to study the effects of plant growth regulators on shoot tips of jojoba genotypes, and experiment (D) was conducted to study the effects of sucrose on shoot tip culture. MS medium [9] containing 30 g/l sucrose, 8 g/l agar was used. The media was supplemented with BAP alone and BAP in combinations with NAA and IAA. Four sucrose concentrations of 20, 40, 50 and 60 were tested (Table 1). The pH of the media was adjusted to 5.8 prior to adding agar. The media were autoclaved at 121°C and 1.05 kg cm-2 for 15 min. The cultured tubes were incubated under 16 h light (white fluorescent light with intensity of 55 μmol m-2s-

1) and 8 h dark, and the temperature was 24 ± 1°C. A sub-culturing at 6 weeks interval was applied onto fresh medium of the same composition to preserve the cultures.

Assessment of genotype response to in vitro nodal segments and shoot tip culture

The following parameters were estimated: number of days to bud emerges, number of leafs per explants, number of nodes per shoot and length of primary shoot (cm).

*Corresponding author: Amr SM Mohamed, Department of Arid land Agriculture, Faculty of Meteorology, Environment and Arid land Agriculture, King AbdulazizUniversity, Saudi Arabia, E-mail: amrsalahw@yahoo.com

Received July 20, 2013; Accepted August 27, 2013; Published August 29, 2013

Citation: Mohamed ASM, Mousa MAA, Bakhashwain AAS (2013) Effects of Growth Regulators and Sucrose on in vitro Nodal Segments and Shoot Tip Culture of Jojoba (Simmondsia chinensis (Link) Genotypes. J Bioremed Biodeg 4: 202. doi:10.4172/2155-6199.1000202

Copyright: © 2013 Mohamed ASM, et al. This is an open-a ccess article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Effects of Growth Regulators and Sucrose on in vitro Nodal Segments and Shoot Tip Culture of Jojoba (Simmondsia chinensis (Link) GenotypesAmr SM Mohamed*, Magdi AA Mousa and Ahmed AS Bakhashwain

Department of Arid land Agriculture, Faculty of Meteorology, Environment and Arid land Agriculture, King Abdulaziz University, Saudi Arabia

AbstractThe responses of four jojoba genotypes to in vitro nodal segments and shoot tip culture under different growth

regulators combinations and sucrose concentrations were investigated. All experiments were laid out in Completely Randomized Blocks (CR) design using 5 replicates. The results revealed that the jojoba genotype ‘Al-Maddenah’ conveyed the highest response to nodal segments, with MS culture medium supplemented with BAP (3 mg) alone and BAP (3 mg), in combination with IAA (7 mg). Modifying sucrose concentration in the culture medium (20 g, 40 g, 50 g and 60 g) enhanced shoot formation of nodal segments of the genotype ‘Hada Al-Sham”. Concerning shoot tip culture, explants of the genotype ‘Hada Al-Sham’ were cultured on MS+BAP (5 mg)+IAA (5 mg) produced the highest percentages of bud sprouting and shoot formation. Increasing sucrose concentration from 20 g to 60 allowed the genotype ‘Hail-B’ to produced the highest percentage of bud sprouting and shoot formation.

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Citation: Mohamed ASM, Mousa MAA, Bakhashwain AAS (2013) Effects of Growth Regulators and Sucrose on in vitro Nodal Segments and Shoot Tip Culture of Jojoba (Simmondsia chinensis (Link) Genotypes. J Bioremed Biodeg 4: 202. doi:10.4172/2155-6199.1000202

Volume 4 • Issue 7 • 1000202J Bioremed BiodegISSN: 2155-6199 JBRBD, an open access journal

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a: Jojoba genotype ‘Al-Maddenah’ conveyed the highest response to nodal segments with MS culture medium supplemented with BAP (3mg) alone and BAP (3mg) in combination with IAA (7mg)

Figure 1b: Effect of Growth regulators combinations on in vitro nodal segment of jojoba genotypes: Bud sprouting

a: Jojoba genotype ‘Al-Maddenah’ conveyed the highest response to nodal segments with MS culture medium supplemented with BAP (3mg) alone and BAP (3mg) in combination with IAA (7mg)

b: Effect of Growth regulators combinations on in vitro nodal segment of jojoba genotypes: Bud sprouting

c: Effect of Growth regulators combinations on in vitro nodal segment of jojoba genotypes: shoot formation

Figure 1: a,b,c : Effects of plant growth regulators on nodal segments.

Conclusion and RecommendationsThe jojoba genotype ‘Al-Maddenah’ conveyed the highest response

to nodal segments, with MS culture medium supplemented with BAP (3 mg) alone, and BAP (3 mg) in combination with IAA (7 mg). Genotype ‘Hada Al-Sham’ conveyed the highest response to shoot tip culture with MS+BAP (5 mg)+IAA (5 mg). Bud sprouting and shoot formation of the genotypes ‘Hada Al-Sham’ and ‘Hail-B’ were enhanced by modifying sucrose concentrations in the culture medium. Shoot elongation and multiplication and rooting experiments are required to assess the efficiency of shoot tip and nodal segments tissue culture technique for micro-propagation of jojoba genotypes (Figure 1-4).

Acknowledgment

The author acknowledge with thanks the Deanship of High Graduate Studies (DGS), King Abd-ulaziz University, Jeddah. The author expresses his thanks to Dr. Ahmed AS Bakhashwain and Dr. Magdi AA Mousa for their valuable supervision and advices.

Treatments of effect of sucrose on nodal segment Treatments Code

Treatments of effect of growth regulators on nodal segmentMS0 MS/NS11

MS0+5 mg/L-1BAP MS/NS2 MS/NS2MS0+3 mg/ L-1BAP+5 mg/L-1IAA MS/NS3MS0+3 mg/ L-1BAP+7 mg/L-1IAA MS/NS4MS0+5 mg/ L-1AP+3 mg/L-1IAA MS/NS5

MS0+5 mg/ L-1BAP+7 mg/L-1IAA MS/NS6MS0+7 mg/ L-1BAP+7 mg/L-1IAA MS/NS7MS0+5 mg/ L-1BAP+7 mg/L-1NAA MS/NS8MS0+7 mg/ L-1BAP+7 mg/L-1NAA MS/NS9MS0+5 mg/ L-1BAP+7 mg/L-12,4-D MS/NS10

MS0+5 mg/L-1BAP+20 g/L-1Sucrose MS/NS1+S12

MS0+5 mg/L-1BAP+40 g/L-1 Sucrose MS/NS1+S2MS0+5 mg/L-1BAP+50 g/L-1 Sucrose MS/NS1+S3MS0+5 mg/L-1BAP+60 g/L-1 Sucrose MS/NS1+S4

MS0+5 mg/L-1BAP+20 g/L-1 Sucrose+0.5 g/L-1 AC MS/NS1+S5MS0+5 mg/L-1BAP+40 g/L-1 Sucrose+0.5 g/L-1 AC MS/NS1+S6MS0+5 mg/L-1BAP+50 g/L-1 Sucrose+0.5 g/L-1 AC MS/NS1+S7MS0+5 mg/L-1BAP+60 g/L-1 Sucrose+0.5 g/L-1 AC MS/NS1+S8MS0+3mg/L-1BAP+5 mg/L-1 IAA+20 g/L-1 Sucrose MS/NS5+S9MS0+3 mg/L-1BAP+5 mg/L-1 IAA+40 g/L-1 Sucrose MS/NS5+S10MS0+3mg/L-1BAP+5 mg/L-1 IAA+50 g/L-1 Sucrose MS/NS5+S11MS0+3 mg/L-1BAP+5 mg/L-1 IAA+60 g/L-1 Sucrose MS/NS5+S12

Treatments of effect of plant growth regulators on shoot tipMS0 MS/ST13

MS0+5 mg/L-1BAP MS/ST2MS0+5 mg/L-1 BAP+3 mg/L-1 IAA MS/ST3MS0+5 mg/L-1 BAP+5 mg/L-1 IAA MS/ST4

Treatments of effect of sucrose on shoot tip cultureMS0+5 mg/L-1 BAP+20 g/L-1 Sucrose MS/ST1+S5MS0+5 mg/L-1 BAP+40 g/L-1 Sucrose MS/ST1+S6MS0+5 mg/L-1 BAP+50 g/L-1 Sucrose MS/ST1+S7MS0+5 mg/L-1 BAP+60 g/L-1 Sucrose MS/ST1+S8

MS0+5 mg/L-1 BAP+3 mg/L-1 IAA+20 g/L-1 Sucrose MS/ST3+S1MS0+5 mg/L-1BAP+3 mg/L-1 IAA+40 g/L-1 Sucrose MS/ST3+S2MS0+5 mg/L-1 BAP+3 mg/L-1 IAA+50 g/L-1 Sucrose MS/ST3+S3MS0+5 mg/L-1 BAP+3 mg/L-1 IAA+60 g/L-1 Sucrose MS/ST3+S4

Table 1: Treatments applied to study the effects of plant growth regulators and sucrose on in vitro nodal segments and shoot tip culture of jojoba.

References

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2. Gentry HS (1958) The natural history of jojoba (Simmondsia chinensis) and its cultural aspects. Econ Bot 12: 261-295.

3. Llorente B, Apostolo NM (1998) Effect of different growth regulators and genotype on in vitro propagation of jojoba. N Z J Crop Hortic Sci 26: 55-62.

4. Prakash S, Agrawal V, Gupta SC (2003) Influence of some adjuvants on in vitro clonal propagation of male and female jojoba plants. In Vitro Cell Dev Biol Plant 39: 217-222.

5. Bashir MA, Rashid H, Anjum MA (2007) In vitro shoot multiplication of six promising strains of jojoba (Simmondsia chinensis). Biotechnology 6: 309-315.

6. Heba Allah AM, El-Bahr MK, Adam ZM, Moursy HA, Solliman ME (2009) In vitro clonal propagation of jojoba (Simmondsia chinensis (Link) Schn.). Aust J Basic Appl Sci 3: 3128-3136.

7. Singh A, Reddy MP, Patolia JS (2008) An improved protocol for micropropagation of elite genotype of Simmondsia chinensis (link) Schneider. Biol Plant 52: 538-542.

8. Osman HE, Abo Hassan A, Samarraie SM (1997) Jojoba (Simmondsia chinesis (link) Schneider): A potential shrub in the arabian desert. I. Overall performance of seven jojoba ecotypes. JKAU Met Env Arid Land Agri Sci 8: 85-96.

9. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473-497.

Citation: Mohamed ASM, Mousa MAA, Bakhashwain AAS (2013) Effects of Growth Regulators and Sucrose on in vitro Nodal Segments and Shoot Tip Culture of Jojoba (Simmondsia chinensis (Link) Genotypes. J Bioremed Biodeg 4: 202. doi:10.4172/2155-6199.1000202

Volume 4 • Issue 7 • 1000202J Bioremed BiodegISSN: 2155-6199 JBRBD, an open access journal

Page 3 of 5

Figure 2a:

a:

Figure 2 :

b: Modifying sucrose concentration in the culture medium enhanced the bud sprouting and shoot formation of jojoba genotype ‘Hada Al-Sham’.

Figure 2: a,b: Effects of sucrose on nodal segments.

Citation: Mohamed ASM, Mousa MAA, Bakhashwain AAS (2013) Effects of Growth Regulators and Sucrose on in vitro Nodal Segments and Shoot Tip Culture of Jojoba (Simmondsia chinensis (Link) Genotypes. J Bioremed Biodeg 4: 202. doi:10.4172/2155-6199.1000202

Volume 4 • Issue 7 • 1000202J Bioremed BiodegISSN: 2155-6199 JBRBD, an open access journal

Page 4 of 5

Figure 3a:

Figure 3b

a:

b:

c: Jojoba genotype ‘Hada Al-Sham’ conveyed the highest response to shoot tip culture with MS culture medium supplemented with BAP (5mg) in combination with IAA (5mg)

Figure 3: a,b,c: Effects of plant growth regulators on shoot tip culture.

Citation: Mohamed ASM, Mousa MAA, Bakhashwain AAS (2013) Effects of Growth Regulators and Sucrose on in vitro Nodal Segments and Shoot Tip Culture of Jojoba (Simmondsia chinensis (Link) Genotypes. J Bioremed Biodeg 4: 202. doi:10.4172/2155-6199.1000202

Volume 4 • Issue 7 • 1000202J Bioremed BiodegISSN: 2155-6199 JBRBD, an open access journal

Page 5 of 5

a:

Figure 4ab: Modifying sucrose concentration in the culture medium enhanced the bud sprouting and shoot formation of jojoba genotype ‘Hail A’ and ‘Hail B’.Figure 4a

Figure 4: a,b: Effects of sucrose on shoot tip culture.