1

UNIVERSITY OF AGRICULTURAL SCIENCES AND VETERINARY MEDICINE

CLUJ-NAPOCA

FACULTY OF AGRICULTURE

Doctoral School

Ing. CHEŢAN SIMONA CLAUDIA (TSOULOS)

ABSTRACT OF PhD THESIS

RESEARCH UPON THE POSSIBILITIES OF INTRODUCING TRUFFLES IN CULTURE IN XIROMERO REGION IN GREECE

SCIENTIFICAL COORDINATOR,

Prof. univ. dr. ing. GAVRIL Ă MORAR

CLUJ-NAPOCA

2011

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CONTENT

Page thessis

Page abstract

INTRODUCTION………………………………………………………… 7 4 Cap. 1. HISTORY OF TRUFFLES……………………………………………… 11 5 Cap. 2. IMPORTANCE AND USAGE OF TRUFFLES…………………………. 16 6

2.1. Ecological, economical and social importance…………………………. 16 - 2.1.1. News and perspectives in truffles cultivation………………………….. 11 -

2.2. Chemical composition and alimentary importance of truffles………… 19 - Cap.3. TRUFFLES SPREADING WORLDWIDE……………………………… 23 6

3.1. Forestry associations with truffles………………………………………. 25 - 3.2. Description of truffles species with economical importance……………. 27 -

3.2.1. Black truffle (Tuber melanosporum)…………………………………….. 27 - 3.2.2. Truffle of the summer or summer truffle (Tuber aestivum)……………… 29 - 3.2.3. Tuber uncinatum or Tuber aestivum variety uncinatum…………….. 29 - 3.2.4. White truffle (Tuber magnatum pico)………………………………….. 30 - 3.2.5. Truffle of tautumn or autumn truffle (Tuber brumale)……………….. 31 - 3.2.6. Mesenteric truffle (Tuber mesentericum)……………………………….. 31 - 3.2.7. Chinesse truffle (Tuber indicum)……………………………………….. 32 - 3.2.8. Whity truffle – bianchetto (Tuber borchii sau Tuber albinum pico)…… 33 -

Cap.4. BOTANICAL CLASSIFICATION AND TRUFFLES MORPHOLOGY 34 7 4.1. Botanical classification at truffles………………………………………. 34 - 4.2. Morphology of truffles………………………………………………….. 34 -

4.2.1. Morphology of the specie -Tuber melanosporum-Vitt. – black truffle… 36 - 4.2.2. Morphology of the specie - Tuber aestivum-Vitt. – summer truffle….. 38 - 4.2.3. Morphology of the specie - Tuber uncinatum-chatin (vurgundia)………. 39 - 4.2.4. Morphology of the specie - Tuber magnatum pico – white truffle………. 39 -

Cap.5. BIOLOGY OF TRUFFLE……………………………………………….. 41 8 5.1. Stages of development …………………………………………………… 41 -

5.1.1. Germinative period ………………………………………………………. 41 - 5.1.2. Reproduction period……………………………………………………… 43 -

Cap.6. TARTUFIGEN PLANTS PRODUCTION………………………………. 46 8 6.1. Tartufigen plants innoculation – types of innoculation………………… 50 8

6.1.1. Micelium innoculation……………………………………………………. 50 - 6.1.2. Sporal innoculation…………………………………………………….. 51 - 6.1.3. Shyntesis grafting……………………………………………………….. 53 -

Cap.7. OBJECTIVES OF THE DOCTORAL THESIS, MATERIAL AND METHODS OF RESEARCH…………………………………………….

57

9

7.1. Research goals…………………………………………………………… 57 - 7.2. Research objectives……………………………………………………… 58 9 7.3. Methods of research…………………………………………………….. 59 9

Cap.8. STUDY OF TE NATURAL CONDITIONS OF THE AREA IN WHICH THE RESEARCH TOOK PLACE………………………………………

60

10

8.1. Truffles requests towards the vegetation conditions…………………… 60 10 8.1.1. Main ecological factors involved in cultivating and development of

truffles…………………………………………………………………….

61

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8.1.2. Xiromero area characterization – geopedological and ecological aspects 67 11 Cap.9. TRUFFLES REQUEST TOWARDS SOIL AND CLIME AND THE

WAY THEY ARE FULLFILLED IN XIROMERO AREA……………….

73

11

9.1. THE REQUESTS OF DIFFERENT SPECIES TOWARDS SOIL AND THE WAY THEY ARE FULLFILLED IN THE XIROMERO AREA….

73

11

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9.1.1. Characterization of the fields in the Xiromero area – Katunei area…….. 76 11 9.2. Requests of truffles towards clime and the way they are fulfilled in

Xiromero area………………………………………………………………

79

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9.2.1. Characterization of the climatic conditions in Greece………………….. 80 12 9.2.2. Spreading of the truffle cultivation in Greece………………………….. 84 12

9.3. Pedo climatic characterization of the experimental area………………… 86 13 9.3.1. Suitability of soil in Xiromero area………………………………………. 86 13 9.3.2. Suitability of clime conditions in Xiromero area……………………… 89 13

9.4. Suitability study of several fields pretable for truffles cultivation in Etoloacarnania county…………………………………………………….

95

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9.4.1. Research of several pretable fields for truffles culture………………… 96 14 9.4.2. Pedological and agrochemical study of pretabile soils for truffles cultures.. 98 15

Cap.10. PREPARING AND STARTING THE EXPERIMENTAL FIELD……… 108 17 10.1. Planting the innoculating sampling………………………………………. 111 18

10.1.1. Planting consistency……………………………………………………….. 113 - 10.2. Tillage for to maintain and care the experimental research field……… 116 18 10.3. Growing and development stages for the inoculated saplings…………. 117 19

Cap.11. AGRICOL PRACTICES FOR TRUFFLES CULTIVATION…………… 131 22 11.1. Cultivation field establishment………………………………………….. 131 22

11.1.1. Field tillage………………………………………………………………… 131 - 11.1.2. Planting phase……………………………………………………………. 135 - 11.1.3. Planting method………………………………………………………….. 133 -

11.2. Tillage…………………………………………………………………….. 134 22 11.2.1. Soil tillage………………………………………………………………… 134 - 11.2.2. Fertilisers appliance……………………………………………………… 135 - 11.2.3. Weeds control…………………………………………………………….. 136 - 11.2.4. Cutting of the trees………………………………………………………. 136 - 11.2.5. Irrigation…………………………………………………………………… 137 - 11.2.6. Gathering leaves from te forestry association……………………………. 139 - 11.2.7. Correction of soil pH…………………………………………………….. 139 - 11.2.8. Plant protection……………………………………………………………. 139 - 11.2.9. Pests control………………………………………………………………... 140 -

Cap.12. TRUFFLES HARVEST……………………………………………………. 141 23 12.1. BURN AREA INDICATOR FOR TRUFFLES FRUCTIFICATION…… 141 23 12.2. PERIODS FOR HARVESTING TRUFFLES……………………………. 142 23 12.3. HARVESTING – HARVESTING METHODS…………………………. 143 24 12.4. CONDITIONING AND PRICE OF TE HARVESTED TRUFFLES……. 146 24

Cap.13. ECONOMIC STUDY FOR THE POSSIBILITIES TO INTRODUCE TRUFFLES IN GREECE………………………………………………….

148

25

Cap.14. ECONOMIC ANALYSIS FOR TRUFFLES CULTURE………………. 150 26 14.1. ECONOMIC EFFICIENCY FOR TRUFFLES CULTURE IN THE

STUDIED AREA…………………………………………………………

156

26

14.1.1. Investment, amortization and profit of the Xiromero area……………… 157 27 CONCLUSIONS…………………………………………………………. 162 28 BIBLIOGRAPHY………………………………………………………… 164 30 ANNEXES……………………………………………………………….. 168 32 ABSTRACT RO………………………………………………………….. 1 - ABSTRACT EN………………………………………………………….. 1 -

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INTRODUCTION

What are truffles ?

Fig 1. Black truffle (source: www. tartufaipicentini.it)

Truffles are edible mushrooms with exceptional taste and flavor. The fame of

these gifts of nature is due primarily to their values great food, accompanied by a

delicious taste and a pleasant fragrance.

If the products of animal origin, caviar is placed first, then certainly of plant

products for food, truffles can occupy the same place with honor.

Truffles, some of the rarest and most demanded of the species of fungi are

living underground in symbiosis developed only with roots of certain trees or shrubs,

and requiring specific climatic conditions.

These mushrooms are good at all, enriching with their delicate inimitable

flavor. But it is best to prepared them so as not to lose the flavor by heating

treatments, either in salads or grated over pasta drowned in butter, foie-gras etc. For

such delicacies there were paid far enough wealth. Thus a white truffle (Tuber

magnatum) of 1.3 kg was found in Croatia in 1999 sold with $ 212,000 in November

2007, another giant white truffle 1.5 kilograms found near Pisa was auctioned and

awarded price of $ 300,000. And the race continues, the more fierce as pollution and

deforestation to make truffles are harder to find.

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In reality, we can not talk about "truffle" in general, because there are a variety

of species of truffles, and not all are high quality and not all have the same

commercial value.

There are types of truffles that have very low selling price, such as Chinese

truffle (Tuber indicum) and species such as black truffle (Tuber melanosporum), the

sale price is forty times higher.

In general, the truffle is a black fungus that is wild in nature, but is very

difficult to find because it is not visible. It has gastronomic qualities (flavor, taste), but

mostly because of high market demand, research conducted in recent decades have

been taking large scale, and after many trials and studies concerning their

development and reproduction in the wild in recent years has developed a systematic

culture of innoculated truffles by getting in the laboratory and the establishment of

mycorrhizal truffles (Gatsios K. - 2007).

In Greece, in recent years people have begun to create small areas of truffle

culture, however, there isn`t a deep study on the factors that influence the cultivation

of this species (S. Diamantis 2003).

These factors such as climatic conditions suitable for cultivation, soil types that

may evolve suitable and types of truffle trees that can grow in those regions.

CHAPTER 1.

HISTORY OF TRUFFLE

In chapter one it is presented teh history of truffle form ancient times until

nowadays and also its evolution on phases up to the obtaining in laboratory conditions

of the first symbiosis between mycorrhized sterile plants and truffle, leading that way

to the possibility of establishing truffle culture by using myccorrhized sampling in

specialized plants nurseries.

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CHAPTER 2. IMPORTANCE AND USAGE OF TRUFFLE

In this chapter is given the importance of culture truffles (made either in natural

forests or in the cultivated species) that is primarily important economic income in

mountain and submountain, grown or cultivated. The economic importance of these

fungi is due on the one hand to high nitrogen content, minerals, lipids, carbohydrates,

organic acids, cellulose, coloring, and on the other hand to their taste and pleasant

flavor thus inimitable taste all the culinary that comes in contact (R. Coli, AM coli, B.

Granett and P. Damiani, 1990).

CHAPTER 3. SPREADING OF TRUFFLES IN THE WORLD

There are over 50 species of truffles but few have commercial gastonomic

importance: white truffle or truffle of Piedmont (Tuber magnatum) black truffle

(Tuber Melanosporum), summer truffle (Tuber aestivum), winter truffle (Tuber frost)

Chinese truffle (Tuber indicum) white truffle (Tuber borchi). (Gatsios K. - 2007).

From statistical data analysis and bibliographical data, that production is obtained in 3

major European countries, namely Italy, France and Spain (Hall IR, Brown GT,

Alessandra Zambonelli, 2007). In Greece so far, according to experts who have

studied some parts of the country was estimated to be very suitable for proper

establishment of truffle cultivation in terms of climate and soil conditions. Growing

real truffle is only possible by creating a home for them, tartufigene plants. These are

plants whose roots are specially designed to associate with truffles that will then live

in symbiosis. (Chatin A., 1869) namely: various species of oak (Quercus pubesccens,

pedunculate Quercus, Quercus Ilex, Quercus coccifera,), hazel (Corylus avellana),

hornbeam (Carpinus betulus), beech (Fagus sylvatica), poplar (Populus alba), linden

(Tilia sylvestris), etc.

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CHAPTER 4. BOTANICAL CLASSIFICATION AND MORPHOLOGY OF TRUFFLE

Truffle belongs to the genus Tuber, which comprises several species, some

with high food value (of those mentioned above) and some low value or non-edible

(with odor, foul-smelling Tuber rufum or radish or mustard - Tuber rapaeodorum) .

(trufflesitaly.com). Like all fungi, truffles have a mycelium represented by structural

vegetative and reproductive organs that it bears. The mycelium consists of filamentous

cells, thin, simple and branched hyphae called mycelia. Hyphae are small, micron, due

to which the mycelium is never visible to the naked eye. It is only visible to optical

microscopy and appears as a set of extended thin hyphae. Fruit body is a "carpoforo"

hypogeum which normally grows in the ground at a depth ranging from a few cm. up

to 40-50 cm. near the plants living in symbiosis. Truffles when mature, evolve and

penetrating aroma typical for the different species (Gatsios Kassandra, 2007).

The truffle belongs to the Ascomycete class, fungi which form their

reproductive organs that spores, in some small bags, which are called ASCAE.

Ascosporus is thus the means of dissemination of truffle, and are contained in

ASCAE in a variable number from 1-8 depending on the species.

Fig. 2. Black truffle (source:www.manitariazagora.blogspot.com)

Fig. 3. Summer truffle (source:www.ayme-truffe.com )

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Fig. 4 White truffle (source: manitariazagora.blogspot.com)

CHAPTER 5. BIOLOGY OF TRUFFLE

In chapter 5 it is described the biological cycle for truffle,with the two periods

of growing and development (the germinative and reproduction period) and the factors

that influnce the formation of the fungus.

CHAPTER 6. PRODUCTION OF TARTUFIGENE

For the production of the mycorrhyzed plants, nowadays there are laboratory

methodsin safe conditions,that allow a production of sampling infected with truffle

sporus on a large scale. These results were obtained due to the progress registered in

the domain of truffle (germinative rate of sporus, functional aspects of the mycorrizal

symbiosis), plant biology (cultivation of in vitro condition) and technological

(polifunctional greenhouses etc.).

6.1 INOCULATION OF MYCORRIZED PLANTS (TARTUFIGENE) – TYPES

OF INOCULATION

Inoculation is the most delicate operation and THE most important of all

operations are carried out to reach mycoryzal phase.

There are three methods of inoculation to achieve synthesis with truffles:

- micelian-inoculation (starting from pure culture of the fungus)

- Inoculation increased (starting from spores probably germ)

- Inoculation by grafting (from parts already mycorrhizal roots)

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CHAPTER 7. OBJECTIVES OF THE DOCTORAL THESIS, MATERIAL AND

MOTHODS OF RESEARCH

7.2. OBJECTIVES OF THE RESEARCH

Objectives of the research are:

- study of the clime and characterization of the area.

- pedological study and pretability of the truffle species to the soils in the area.

- adaptability of the truffle species to the conditions in Aitoloakarnan area.

- development of the tree species used in the culture field, species on which

truffles develop in the conditions of the research area.

- establishing the aproximate production potential for the truffle in this area.

- calculation for the economical efficacy and the profit from the establishing of

the culture in a systematic way until they becomme productive and during the

evolution of culture.

- establisging the growing potential and development for plants from two

truffle culture in different conditions.

- identification of several myccorhizal areas in Etoloakarnania county alike

under the aspect of full filling the pedoclimatic requests.

- possibility to introduce truffles in Xiromero area

7.3. METHODS OF REASEARCH USED

In the studies and research we have used several methods such as:

1) Studies on the climatic characteristics of soil and climate of the study compared with traditional areas of the truffle countries - Italy and France.

2) studies and determination of annual increases of three fruit species used for mycorrhizal culture.

3) Suitability of species for growing truffles in these areas.

4) Observations on the influence of irrigation on the crop.

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5) there were located two experiences of culture with different distances

depending on plant species and culture system

CHAPTER 8. STUDY OF THE NATURAL CONDITION OF THE AREA IN WHICH THE

RESEARCH TOOK PLACE

8.1.TRUFFLE NEED TOWARDS VEGETATION CONDITIONS

Ecological research conducted on natural truffles, made possible the

description of the natural environments where truffles grow spontaneously, also it

made possible the understanding, the most important ecological factors responsible for

their cultivation.

As the main important issues to be considered are those concerning the nature

of soil, climate, exposition, vegetation and woody species that live in symbiosis with

truffles. Investigations made should show which of them have distinctive ecological

character, which will help promote the further development of the biological cycle of

the fungus and may be leading to fructification (www.agri.marche.it)

Geopedological factors

The study envisages geopedologic geomorphological aspects of planning and

physical-chemical soil characteristics of land generally classifies as mycorrizal plants

(tartufigen), by emphasizing the physical-structural, which influence enough the

production capacity of truffles.

Microbiological factors

Symbiotic interaction does not only refer to plants and fungi, and bacteria but

also to the existing symbiotic bacteria involved in playing an active role in the

formation of mycorrhiza (tartufaie.it)

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Botanical factors –Ecosystem

Truffles need a general outline appropriate vegetation as forest and grass

species have a key role in the synthesis of mycorrhyza.

8.1.2.Characterization for Xiromero area– geopedological and ecological aspects

This chapter describes Xiromero area where it was located field experience,

geopedological characterization and ecological aspects of the region (Vlami V.

Zogkaris St., Dimopoulos DP, 2003).

CHAPTER 9. TRUFFLES NEEDS TOWARDS SOIL AND CLIME AND THE WAY THEY

ARE FULFILLED IN XIROMERO REGION

9.1 NEEDS OF DIFFERENT SPECIES OF TRUFFLES TOWARDS SOIL AND THE WAY THEY ARE FULFILLED IN XIROMERO REGION

We can not talk in general about a suitable soil for truffles, because physical

and chemical soil characteristics are not the same for all species of truffle. This

chapter presents the data in terms of physicochemical properties of soils considered as

good for growth and development of truffles (black truffle Cultivo JABonet, D. & C.

Oliach Colinas).

The land that will choose to have such guidance to avoid frost damage to

winter truffle, but especially the first big frost in November or January and February

frosts.

9.1.1 Characterization of land in the region Xiromero – Katuna region

Characterization of land in the area of experimental field

(news.blogspot.com), plays that they correspond in terms of soil types, their structure,

fertility and cultural landscape for the summer truffles especially Tuber aestivum

Tuber melanosporum and black truffle. (www.xiromeronews blogspot.com).

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9.2. TRUFFLES NEEDS TOWARDS CLIME AND THE WAY THEY ARE

FULFILLED IN XIROMERO REGION

9.2.1Characterization of climatic conditions in Greece

Greece is between 340 and 420 parallel of the northern hemisphere and bathed

in the Mediterranean Sea from the East. Greece is typical Mediterranean climate: mild

and wet winters, relatively dry summers and generally sunny and long periods in most

of the year (agro-tour.net)

Geographical elements required for a succesful truffle crop

Altitude, field exposion and choosing carrying truffles, (IPLA S.p.A. 2001) are

geographical recommended elements necessary to establish a crop with truffles. These

elements are met in Xiromero geographical area, namely:

- Truffle crop is established at an altitude of 350 m.

- An exhibition of land south-west

- Wood species used are oak, Quercus pubescen establishment culture, t-

Coryllus avellana hazel and lime-Tilia sylvestris.

9.2.2. Spreading of truffle crop in Greece

Given the climatic conditions existing in Greece, according to French experts'

views and knowledge, there are mountain and submontane regions that meet the

conditions of climate and soil necessary for growth and development of the truffle.

Although so far no major attempts have been made on this culture, Forest Research

Institute promotes truffle culture, organizing information courses in recent years. So

far there have been established truffle crops in different regions of the country namely

Agrinio (County Etoloacarnania) Sidironero (N-drama), Kozani, Grevena, Larisa,

Florina, Anilio-Pilio, Kilkis, Chalkidiki, etc. (S. Diamandis, 2003)

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9.3. PEDOCLIMATIC CHARACTERIZATION OF THE EXPERIMEN TAL

FIELD

9.3.1.Characterization of soil in the crop area

Fig. 5 Soils in the research area (original)

Soils that cover the region comprising on the one hand are characterized by

age and recent alluvial soils on the other hand argiloiluviarea soil erosion and older.

All soils are highly susceptible to structural degradation due to low humus content and

acidity. The main soils in the study area are: preluvosoil, luvosoil, cambisoil,

podzoluvisoil, regosoil and fluvisoil.

9.3.2. Characterization of the clime conditions in Xiromero region

The general climate is characteristic exclusively Mediterranean area, with mild

winters and wet and dry prolonged summers.

The climate of the last years, with data taken from weather stations Agrinio, are

presented in table 1.

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Table1.

Meteorological results – Agrinio – for the years 2003 - 2008

YEAR AVERAGE AIR TEMPERATURE

PRECIPITATION

(mm)

PERIOD OF SUN SHINING

Annual sum 6438 865,5 2548 h 2003

Annual sum 17,8

Annual sum 6075 730,5 2504 h 2004

Annual sum 16,8

Annual sum 6207 1516 2355 h 2005

Annual sum 17,3

Annual sum 6180 823,2 2693 h 2006

Annual sum 17,1

Annual sum 6765 1046 2796 h 2007

Annual sum 18,7

Annual sum 6483 588,9 2370 h 2008

Annual sum 18

Source:meteorological station, Agrinio airport

9.4. SUITABILITY STUDY OF SEVERAL FIELDS PRETABLE F OR

TRUFFLES CULTIVATION IN ETOLOACARNANIA COUNTY

9.4.1. Research towards several pretable fields for truffle crop

With regards to soil characteristics, truffles fail on any ground, they prefer deep

soil texture Light-Middleweight, well ventilated, with a pH between 7.5 to 8.5,

containing sufficient nutrients, C / Ν in about 10 land rich in limestone.

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9.4.2. Pedological and agro chemical study for soils pretable for truffle crop

Table 2.

Physical-chemical soil feature from the experimental field compared to physical- chemical recommended

A/A PARAMETERS PHYSICAL- CHEMICAL

GEATURES RECOMMENDED

RESULTS OBTAINED

1 Ph 7- 8,5 % 7,77%

2 Organic substance 1,5 – 8 % 1,30 %

3 Total Calcium % 4150 mg/l

4 Phosphorus % P Olsen (mg/Kg ) 5

5 Potasium (Potasium oxide) % K+( mg/Kg) 154

6 Asimilable magnesium % Mg + (mg/kg ) 96

7 C/N rate 5 -20 % ; ideal 10 -11 % 6 %

8 Calcium carbonate % 0.5 -80% ; ideal 35% Urme

9 Sand % 20 – 80 % 19 %

10 Clay 5 -50 % (depending on teh specie of truffle )

60 %

11 Clay % Mud %

5 -80 % 21 %

Soil analysis was made with those of field experience to set up four more trufle

crop about the same period and all within a radius of 50 km. Xiromero area.

The five land used in experience have surfaces between 30 and 80 acres,

minimum area proposed in this new culture.

Comparing the results of the analysis of field experience and soil physical and

chemical characteristics of soil suitable for the recommended establishment of a

truffle crop, one can see that from the results obtained from field experience soil type

is clay with an appropriate pH,, but an important issue is a very low percentage of

calcium carbonate, important and necessary for the establishment of a truffle crop

(Table 2).

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Following the results of chemical analysis performed Xiromero soils in the

region corresponding to truffle culture (S. Cheţan-2011).

Table 3.

pH evaluated in the field analyzed

Experimental field

Field 1 Field 2 Field 3 Field 4

Strongly acid Sufficient acid Acid Light acid Neutre Light alkaline Sufficient alkalin Strongly alkaline

Table 4.

Appreciation of the calcium carbonate (CaCO3 total %) in the analyzed fields

Experimental field

Field 1 Field 2 Field 3 Field 4

Lack CaCO3

Poor Moderate Sufficient Rich Very rich Marnae Limestone

Table 5.

Organic substance % determined in the analyzed fields

Experimental field

Field 1 Field 2 Field 3 Field 4

Very poor Poor Moderate Sufficient Rich Very rich

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RESULTS AND CONCLUSIONS – regarding the climatic and pedological

conditions, but also regarding the trees species for the natural region,are as follow:

1. The average annual temperature of the region Xiromero (of 16 to 18.70 C) is

closer to the average annual temperatures of the regions in other areas (Italy,

France 15-16 0C), where truffles are grown successfully.

2. The amount of rainfall in the area of research (700-900 mm) falls within the

growing truffles (500-1400 mm), being close to the area trufocultivatorilor of

Italy (600-800 mm) and France (700-800mm).

3. Altitude of 350 m from the experimental field is located is within the limits

of cultivating truffles (150-400 m).

4. Forest species enter into symbiosis with truffles ithaburensis Quercus,

Quercus coccifera, Quercus pubescens increase spontaneously their normal

area and vegetation creates optimal conditions for association with truffles.

5. The soil is suitable in terms of acidity (pH = 7.1 to 7.9) and in terms of

structure, texture and content in some of the major nutrients.

CHAPTER 10. PREPARING AND ESTABLISHING THE EXPERIMENTAL FIELD

After soil tests, there were conducted as the next operation was the choice of

tree species with truffle mycorrhiza chosen, depending on climate, soil, etc..

In the Xiromero - Katun village was located two truffle crops -field research

area and land area of 2. 25 areas, located at a distance of 6 km. Planting was done in

both cultures fall in October with fruit species seedlings (-recommended by the

company Raggi-viva) of oak (Quercus pubescens), hazel (Coryllus avellana) and lime

(Tilia sylvestris) inoculated species with spores of black truffles (Tuber

melanosporum), white truffle (Tuber magnatum) and summer truffles (Tuber

aestivum).

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10.1. PLANTING THE INOCULATED SAPLING TREES

Planting density depends on soil composition and depth of work provided

(irrigation, tillage), economic factors, etc.

S

S S S S S S S S S

A

A A A A A A A A A

S

S S S S S S S S S

T

T T T T T T T T T

A

A A A A S S S S S

Fig. 6. Planting the experimental field 0,1 ha. with oak, hazelnut and lime at distances of 6x3m., (50 trees /0.1ha) (Che�an S. 2011)

Experimental field planting was carried out at distances of 6m x 3m, with a

total of 250 plants from 0.5 ha (fig.6) and on the ground witnessed 5m x 5m distance

to peanuts and lime and oak a distance of 6m x 5m planting, with a number of plants

from 90 to 0.25 hectares.

10.2. TILLAGE AND MAINTAINCE FOR THE RESEARCH FIELD

Experimental field planting was carried out at distances of 6m x 3m, with a

total of 250 plants from 0.5 ha (fig.6) and on the ground witnessed 5m x 5m distance

to peanuts and lime and oak a distance of 6m x 5m planting, with a number of plants

from 90 to 0.25 hectares.

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10.3.GROWING AND DEVELOPMENT OF INOCULATED SAMPLING

A compared development of the two trufoculturi trees is different, because the

conditions of maintenance and care enough especially that influenced their growth in

height.

The most important role so far is the influence on growth and development of

water plants inoculated. It appears that two species, hazel and especially lime, can not

succeed in our area only under optimum irrigation.

Fig. 7. Quercus pubescent /oak (original 2010)

Evolution of the vegetation at the trees species used (oak, hazelnut and lime) in

the experimental field - Regarding the evolution of growth in height and perimeter of

the trunk, repeated measurements were made in 2010 and 2011 to determine the

growth rate of each fruit species.

y = 31,667x2 - 745x + 4583,3R2 = 0,5662, r=0,39

150170190210230250270

10.5 11 11.5 12 12.5 13

Heig

ht(

cm

)

Hazelnut 5 years

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Fig. 8. Correlation between the perimeter of the trunk and the hight at hazelnut (Corylus avellana) of 5 years (2010)

Stejar 5 ani

y = -1,25x2 + 36,831x - 18,081R2 = 0,932, r=0,95

150

170

190

210

230

250

270

6 7 8 9 10 11 12 13 14

Perimetru (cm)

Înălţ

imea

(cm

)

Fig. 9. Correlation between the primeter of the trunk and hight at oak

(Quercus pubescens) of 5 years (2010)

Measurements taken in the sixth year of culture - since 2011 - on trees

inoculated, showed that growth and fruit species was significant compared to the

measurements made in the fifth year of cultivation.

y = -19,839x2 + 573,39x - 3868,4R2 = 0,9148

200210220230240250260270280290

12 13 14 15 16

Hei

ght (

cm)

Hazelnut 6 years

Fig.10 Correlation between the perimeter of the trunk and the hight at

hazelnut (Corylus avellana) of 6 years (2011)

Oak 5 years

Perimeter

H

e

i

g

h

t

21

y = -8,5714x2 + 301,43x - 2360R2 = 0,5635, r=0,53

200

220

240

260

280

300

320

14 15 16 17 18 19

Hei

ght(

cm)

Oak 6 years

Fig. 11 Correlation between the perimeter of the trunk and hight at oak

(Quercus pubescens) of 6 years (2011)

The correlation coefficient between average heights / species and the average

perimeter of the trunk / species in the two years is significant, which shows that the

two variables are directly related, with increasing height of the plant grows and trunk

area. (Cheţan Simona, 2011).

Fig. 12 Aspects from the experimental field (year 2011)

22

CHAPTER 11. AGRICOL PRCATICE IN CULTIVATING TRUFFLE

11.1. CROP ESTABLISHMENT

In this chapter there are presented agricol practices necesary to prepare the field

for planting, period (autumn or spring) and the method of planting for truffle crop

(monocrop or policrop).

11. 2. TILLAGE

In order to assure the growing and development of truffle crop there are necessary

the following tillage:

- Soil tillage

- fertilizer appliance

- weeds control (there are te following methods):

a ) Mulching the field with gravel

b ) Mulching the soil with izolant material

c ) With the help of herbicide (applied only in extreme cases)

d ) By digging the soil (by turning it)

- Cutting trees (there is no documentation as there would be a real link between felling

and production of truffles.

- Irrigation must be regular in the first year after planting, because of the risk of

destroying the mycorrhyza seedlings due to drought.

- Assembly of the Association leaf forest

- Correcting the soil pH, if necessary, the pH is corrected

- Plant protection

- Pest control

23

CHPATER 12 TRUFFLE HARVEST

12. 1. BURNT AREA INDICATOR FOR TRUFFLE FRUCTIFICATION

Before the advent the fructification in trees inoculated with some truffles species,

there is the phenomenon of "burnt area" (Figure 13) under the tree trunk and usually

spread on the same surface as the roots do.

Fig. 13.Burn area a. (Source: trufflesitaly.com)

12. 2. PERIODS FOR TRUFFLE HARVEST

The periods in which the different species of truffles rippen and are harvested

by the farmers are presented in table 6.

Table 6.

Periods of the fructification at the main species of truffles (Sources: www.jardineries-jeanrey.fr )

Perioadele fructific ării principalelor specii de trufa Mai Iun. Iul. Aug. Sept. Oct. Noem Dec Ian Feb Mar Apr

T. aestivum T. uncinatum

T. mesentericum T. magnatum

T. melanosporum T. brumale T. indicum

The periods for fructification of the main truffles species

24

12.3. HARVEST – METHODS OF HARVEST

Truffles are usually found at a depth of 10-25 cm, but up to 40-50 cm from soil

surface.

When truffles are found in very shallow, shallow, land is a slightly raised

mound or form some cracks in its surface, or other obvious signs above the ground,

identifying mature nose. (www.trovatartufi.com).

To harvest truffles one need to use an animal smell. More than with the pig,

today for harvesting truffles there are often used trained dogs.

12.4.CONDITIONING AND PRICE OF THE TRUFFLE HARVESTE D

Truffles can be stored in the refrigerator for a week, six months in the freezer at

a temperature of minus 18 degrees while others keep them in olive oil, goose or duck

fat. But the safest and THE best way is TO keep them frozen in vacuum packs and

preservE the form of products, generally only with flavors of truffles.

The fact is that any method of maintenance truffles used, some of their flavor

and overall image of truffles is destroyed.

Table 7.

Retail price for selling truffles up to March 2009 for different species of

fresh truffles depending on their size gr/kg.

(source: Coltivazione –Tartufo.com)

Specia trufei 0-15 gr./kg 15-50gr./kg >50 gr./kg

Tuber magnatum Pico 2.000 euro 2500 euro 3500-4500 euro

Tuber melanosporum 350 euro 400 euro 450-500 euro

Tuber borchii-vitt 180 euro 210 euro 250 euro

Tuber aestivum-vitt 300 euro 350 euro 350-450 euro

25

CHAPTER 13. SWOT ANALYSIS FOR THE POSSIBILITIES OF INTRODUCING

TRUFFLES IN GREECE

In general, making a rational analysis (SWOT) of the truffle as an economic

product, one can find the following:

a) The strengths of the truffle:

� requires less surface for efficient culture

� requires less manual labor

� has no problems distributing the product on the market (high demand

and high prices)

� In some cases there is double production (peanuts, lime flowers)

� it can help to develop local industry and tourism

� there is high demand on foreign markets.

b ) Weaknes of truffle:

• it isn`t sufficient known and appreciated in the Greek cousine

• it is a complicated crop,for long term

• the crop depends on the climate conditions which are

unpredictible and nor balanced

• there is always a risk for the truffle to be stolen

c ) Oportunities

� it is a biological crop, favorable for the environment

� it is a good stimulent in order for the farmers to stay in the agricol

areas

d) Threats

� there aren`t any disadvantages because truffle crop doesn`t affect

the environment.

26

CHAPTER 14.

ECONOMIC ANALYSIS FOR A TRUFFLE CROP

Before establishing a truffle crop, it is important that people interested to

cultivate this kind of crop to have knowledge first of all about (beside the pedological

ones mentioned above), the financial part necessary to establish the crop and also the

annual cost of maintaince for such a crop.

Cost of establishing the experimental field (for 0,5 ha)

The cost of establishing a truffle crop on a certain surface, contains the cost of

the main tillage for establishing a tree crop (soil analysis, field cleaning, fertilizers

appliance, planting), and furthermore the expenses for irigation system, fencing and

the price for buying the mycorrhized sampling. The economical analysis of the

experimental field in order to establish a truffle crop of 0,5 ha. Was about 7.600,00

euro, so for a ha it can be estimated that it will reach 14.000 - 17.000 euro, depending

on the machines, materials methods used (Che�an Simona, 2011).

Annual cost to maintain a truffle crop (of 0,5 ha.)

- for the first three years was 285.00 euro/ year

- After the forth year and continue approximately euro 335.00/year

As seen from the results of calculations made after the establishment of a

truffle crop, maintenance costs / year are reduced, and efforts by farmers for their

maintenance are minimal.

DURATION OF CULTURE - the duration of production depends on species

and tree size, planting distance, soil quality and cultivation methods.

14.1 ECONOMIC EFICACY FOR TRUFFLE CROP IN THE

EXPERIMENTAL FIELD

From existing data, known and experienced entry bearing plant will be gradual,

according to the species of trees, namely starting with the 5-6th year of culture- hazel,

27

lime starting with the 6-7th year, oak, starting with the 8-10th year of culture

(fig.14.)

Fig. 14 Graphic representation for the production of truffle yearly in kg depending on the carrying species of plants

Also the quantity of truffles obtained per hectare or per tree basis is

progressive (fig.14), respectively in the first year being found in trace amounts

(several grams / tree) and later followed in the coming years, namely 10 years -25 to

reach peak production (leading to even over one kg / tree). The probability of non

productive treeas presence in a culture can not be anticipated from year to year

14.1.1 Investment, amortization and profit for the experimental crop

These calculations are approximate, representing data from field experience,

our expectations about the trajectory and evolution that the culture will take over the

years.

oak

lime

hazelnut

Year 1 - 35

28

0 €

5,000 €

10,000 €

15,000 €

20,000 €

25,000 €

30,000 €

35,000 €

anii 1-

6

anii 6-

8

anul 9 anul

11

anul

15

anul

20

anul

30

anul

35

Investiții

Amortizare lineară a

investiției

Profit

Fig.15. Graphic representation from the economical- financial

point of view

Data presented in chart above (fig.15.) represent a case study, in which up to

now known exact values of financial investments, while accounting for depreciation

and profit values are approximate.

CONCLUSIONS

• In the agricultural production, in the context of reorganization of the

Xiromero area where there are several crops that are aliminated as a result of

European agricultural policies (tobacco, sugar beet, cotton), introducing a viable

cultivation for truffles can be a solution for the development of agricultural activity

in the area.

• Existing plant associations in ecosystem in the region and the presence

of dense different species of oak, beech, chestnut, lime, are an important premise

to establish truffle cultures, having �n vegetation woody species that live right �n

simbiosis with truffles.

Investment

Linear

amortization of

investment

Years 1-Years 6 - Year Year Year Year Year Year 6 8 9 11 15 20 30 35

29

• Pedoclimatic resemble of Xiromero climatic zone with growing areas of

Italy and France, and the possibility of increasing hazel, lime, and especially olive,

represents a strong argument for introducing truffles �n the area mentioned above.

• The most spread oak species are Q. pubescens and Q. cocifera, are

spontaneous taken into study in the region and are used more frequently and

elsewhere in the world because they are good indicators for truffles presence, shall

fall faster �n symbiosis with fungus. Only after 4-5 years they produce truffles

flavored, highly valued by subscribers consumers.

• Geomorphological studies of the territory, carried out on orographical

slope, structure and nature of subsoil, associated with physicochemical soil

features, as soil texture, pH, C / N, humus, content in NPK, Ca, Mg, are favorable

the recommended limits for truffles culture.

• Limestone brown soils and brown soils of the region presents Xiromero

with favorable physical and chemical features for truffle development, such as

glomerural structure, good water retention capacity �n the ground, a balance of

elements that determine the soil texture (clay, clay-sand), a good drainage

capacity, a favorable pH (7.0 to 8.5), a ratio C / N according to (10-11).

• Katun zone with colluvial soils and lowland area located �n the

mountain and Trifu area with alluvial soils are located in plain and view different

structural, morphologic and fertility features, but truffles are positive culture,

especially Tuber aestivum summer truffles and black truffles, Tuber

melanosporum. The climate of the area studied is characteristic of the

Mediterranean.

• Average annual temperatures in the region are included �ntre 17-180C,

with an average of 8.0 to 8.30 C in January, and 26-270 C as average in July.

Absolute minimum rarely go under 0C in January, and peaks are high, 33-350C

�n July-August. Rainfall in winter period (November-December-January) are

more abundant (100-150 mm per month) and lower in summer (30-50 mm), their

annual sum in some years exceeding 1000 mm.

• Frequently, the annual amount of precipitations is between 600 and

1100 mm, the value for liquid and truffles respond to the needs during the whole

30

year. These climatic conditions correspond to fully formation and and growth for

truffles and climatic conditions in regions located �n truficole traditional areas of

Italy and France. Region Xiromero stands at an average altitude of 350 m, the

altitude value is alike the one met in cultures of truffle in Spain, France or Italy

(150-400 mm).

• Inoculated seedlings growth and development depend on climatic

conditions and on the maintaining conditions in the first 4-5 years of vegetation.

The factors of vegetation, water seems to have predominant role for the increase of

the species inoculated and lime peanut especially the annual increases through the

size but also the thickness determined by the trunk perimeter and the normal

functional vegetation symbiosis with fungus. Peanuts, but especially lime are

species dependent on irrigation in Xiromero region.

• Among the known appreciated species of truffles, the most appropriate

for the region of Xiromero, Greece would be black truffle (Tuber melanosporum)

and summer truffle (Tuber aestivum), which were identified in natural conditions.

• Economic analysis of an investment of truffle culture suppose

significant investment until the plantation starts fructification phase, hazel �n 5-6

years, 6-7 to 8-10 lime and oak, but there can be obtained significant profits in 10-

25 years, estimated at 60 000 -70 000 euros / year / ha.

• Orographic conditions, geopedological, climatic, vegetation species are

suitable economically favorable for the evaluation of establishing and exploiting a

culture of truffles in Xiromero, Greece.

SELECTIVE BIBLIOGRAPHY

1. CHETAN, SIMONA CLAUDIA, MORARU G., 2011 Economically and alimentary important truffle species, establishment and development of the truffle culture in Xiromero region University of Agricultural Sciences and Veterinary Medicine of Cluj Napoca

2. CHEŢAN, SIMONA CLAUDIA, MORAR G., 2011, Introduction in Trufficulture in Xiromero Area Greece, University of Agricultural Science and Veterinary Medicine, Cluj Napoca.

3. COLI R., A. M. COLI, B. GRANETT AND P. DAMIANI . 1990. Composizione chimica e valore nutritivo de tartufo nero (Tuber melanosporum Vitt) e del tartufo bianco (Tuber magnatum Pico) raccolti in Umbria. P 511 – 516. In: M. Becivenga and

31

B. Granetto (eds.) Atti del secondo congresso internazionale sul tartufo. Spoleto.Italy. 1988. Com. Mont. dei Martini. Italy.

4. DIAMANTIS S., 2003, Trufa şi cultura ei, Ed.Pa-Thiras 5. GATSIOS K., 2007, Trufa şi cultura ei, Ed. Agro Tipos, Atena. 6. HALL I.R., BROWN G.T ., Alessandra Zambonelli 2007, Taming the

Truffle: The History , Lore , and Science of the Ultimate Mushroom , Timber Press 7. KHANAQA DR ., 1996, La culture de plantes mycorhizees 8. LETIZI H. C ., 1977, Ecologia trufei 9. MANNAZZIO – TORIN, L., 1984, Il Tartufo e la sua Coltivazione,

Edagricole, Bologna. 10. PLANFOR . Truffes-Densite de planttion des plants truffiers. 11. The Guardian 2007-article history. Italian mongrel leads owner to record-

breaking truffle 12. VLAMI V., S. ZOGKARIS, P.D. DIMOPOULOS , 2003, Pădurea de stejar

Xiromero-Etoloakarnania (Facultatea din Ioanina – secţia de management a resurselor naturale şi de mediu).

13. ZAMBONELLI A., C.GOVI , 1983, Micorrizazione en semenzaio di Quercus pubesceus Wild.con specie del genere Tuber, Micol Ital. 12:77-22.

14. *** http:// www.xiromeronews blogspot.com-Căutând trufa sălbatecă în pădurea de stejar Xiromero-articol-2009)

15. ***http://www. euro-trufa.net. -Asociaţia Trufa Net 16. ***http://www.agri.marche.it -Centro Sperimentale di Tartoficoltura 17. ***http://www.agro-tour.net 18. ***http://www.trovatartufi.com -Il Portale sul tartufo fatto dai ricercator di

tartufi italiani 19. ***http://www.trufamania.com/truffles-stories.htm 20. ***http://www.truffe-haute –provence.fr, La truffe de Haut – Provence (Cea

de-a 22 sărbătoare a trufelor din Haut-Provence/5-6/02/2011) 21. *** http://www.trufflesitaly.com/it/Tartufo /micologia.htm 22. ***INRA, 1999, Les truffes, Station de recherché sur les champignons,

Bordeaux 23. ***IPLA: Manuale di tartuficoltura. Le posibilita di coltivazione del tartufo in

Piemonte

32

ANEXE

Fig. 1. Truffle consevation

(www.deliziatartufi.com /il-tartufo.htm)

1. Truffle salat

We can use three kinds of lettuce. We must prepare the salad with olive oil in the

morning, to serve it in the evening. We brush and cut truffles in thin slices, then add

salt, pepper, etc., but not vinegar. We cover the truffles and mix them with salad just

before serving.

Fig. 2. Truffle salat (Source: www.beaugraviere.com/fr)