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Selection of new eggplant (Solanum

melongena, L.) lines

Doktor rerum horticulturarum

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Zusammenfassung

in vitro

in vitro

in vitro

,

in vitro

in vitro

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Abstract

in-vitro

in-vitro

in-vitro

in-vitro

in-vitro

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Table of contents

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4.3 in vitro

in vitro Invitro

In vitro

in vitro

in vivo

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Acknowledgements

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Dedicated To

my family, who offered me unconditional love and support throughout the course of my studies,

and

special thanks to MS for his love and patience.

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Abbreviations

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1 Introduction

Solanum melongena

Solanaceae Solanum

et al

et al et

al

et al

et al

et al

et al

et al

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et al

et al

et al

et al

et al

et al

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2 Literature review

2.1 Economical importance, world production and commercialisation

of eggplant

Solanum melongena

Solanaceae

2.1.1 Field cultivation of eggplant

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et al

2.1.2 Greenhouse and tunnel production system

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6,4

7,5

10,5

5,0

4,4

9,6

14,9

5,5

6,2

9,8

18,8

4,6

0 5 10 15 20 25 30 35 40 45 50

Import (Mt)

Import (1000 $)

Export (Mt)

Export (1000 $)

2001

2002

2003

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et al

et al

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2.2 Biological characterisation of eggplant

2.2.1 Origin

Solanum melongena

et al

S.

melongena

S scabrum S macrocarpom S aethiopicum Solanum

2.2.2 Systematic

Solanum Species

Plantarum

S. incanum S. melongena

et al

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Solanum et al

Incaniformia

S. incanum S.

campylacanthumsensu ampliore S. bojerisensu ampliore

S. incanumsensu ampliore

S. incanum et al

Incaniformia

et al

Plantarum

S incanum S

melongena

Solanum

Solanum

Solanum

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Solanum

et al

Solanum

et al Solanum

Solanum melongena

Solanum

S.

melongena

S incanumsensulato Smelongena

S. campylacanthum

S. melongena(S. insanum)

S. panduriforme S. melongena(S. cumingii)

S. incanum

S. melongena(S. ovigerum)

S. lichtensteinii S. melongena(S. melongena)

Solanum

scabrumSolanum

macrocarpon

Solanum

aethiopicum

Solanum

melongena

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Solanum melongena orientale pekinensis occidentalis

falcatumet al

et al et al

Magnoliopsida

Solanaceae Solanum

Solanoideae Leptostemonum

Solaneae melongena

Solanum melngena

orientale

meridionale

suspontaneum

agrestis

depressum

pekinense

serpentinum

palestinicum

esculentum

americnum

arabicum

leuconum

occidentali

zhukowskii

falcatum

izmir

bulgaricum

azerbaijanicum

europaeum

kashgaricum

ramosissimum

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2.2.3 Description of the botanical characteristics

melongena

S melongenaserpentium

S melongena

S melongena esculentum

S. melongena

esculentum

et al

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etal

etal

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2.2.4 Botanical description of eggplant

tomentum

etal

et

al

Solanummelongena esculentum

etal

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etal

2.3 Nutritional value and marketable quality of eggplants fruits

et al

etal

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Solanum

Solanum

etal

et al

et al

et al

et al

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et al et al

et al et al

Solanacea

et al et al

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et al et al

et al

2.4 Review about breeding methods applied to eggplant

et al

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2.4.1 Amplification fragment length polymorphism (AFLP)

et al

et al

et al

et al

et al

et al

Solanum et al

et al

et al

et al

et al

et al

2.4.2 Microsatellites or single sequence repeats (SSR)

et al

et al et al

et al

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et al

Arabidopsis et al

in situ

et al

2.4.3 Random amplified polymorphic DNA (RAPD)

et

al et al

et al

et al et al

et al

et al et al et al

et al

et al

et al et al et al

et al et al

et al

et al

et al

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et al

et al et al et al et al

et al et al

et al et al

et al et al

et al et al

et al

et al et al

et al et al

et al

et al

2.5 Conventional and unconventional breeding methods applied to

eggplant

et al

et al

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Solanum melongena

et al et al

et al

Fusarium oxysporum melongenae Ralstonia solanacearum

et al et al

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etal

etal

etal

etal

invitro

invitro

invitro

Agrobacterium

thumefaciens

Bt

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2.6 Growing conditions of eggplant in the geographical centres of

origin

Solanum melongena

et al

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2.7 Climatic requirement and fertilization in open cultivation system

2.7.1 Light and temperatures

et al

et al

et al

et al

et al

2.7.2 Soil and water consumption

et al

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et al et al

et al

et al et al et al

et al

et al

et al

etal

2.7.3 Nutrients and fertilization

et al

et al

et al et al

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etal et al

( et al, et al )

et al

et al et al

2.8 Growing conditions for eggplant in greenhouse cultivation system

et al AVRDC

et al

et al

etal et al

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2.8.1 Types of Greenhouses

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2.8.2 Cultivation Systems in Greenhouses: Soil and soilless

et al

etal

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substrate layers

hydroponic

Substrate layer

Hydroponic

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2.8.3 Greenhouse production systems

C. Eggplant cultivars for protected cultivation are usually trained in a V

2.9 Young plant production

2.9.1 Generative propagation

etal et al

et

al

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et al et al

et al

etal et al

etal et al

etal

et al

et al

2.9.2 Vegetative propagation

et al

et al

Solanaceae

Solanum

melongena S. integrifolium S. torvum

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et al

et al

In vitro

in vitro

et al

In vitro

et al

et al

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in vitro et al

et al

in vitro

et al et al

et al

in vitro

et al

in vitro

et al et al

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2.10 Cultivation systems for eggplant

2.10.1 Cultural practices common to eggplant production

2.10.2 Density and branching of plants

etal

etal

etal et al

etal

et al

etal

etal

etal et al

etal

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2.11 Open issues and problem statement

etal

et al

etal

etal

et al

etal etal etal

etal

et al et al

Solanum

melongena et

al Solanum

etal etal etal

et al et al et al

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etal etal

Colletotrichumgloeosporioides

S torvum S linneanum et al et al

et al

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3 Aim of the thesis

3.1 General objective

Smelongena

3.2 Specific objectives

in vitro

in vitro

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3.3 Hypothesis

In vitro

3.4 Research questions

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4 Material and Methods

4.1 Overview of the experimental programme

Experimental phase I

in vitro

in vitro

in vitro

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Experimental phase II

in vitro

Experimental phase III

in vitro

Experimental phase IV and V

in vivo

in vitro

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invitro

in

vitro

invitro

invitro

invitro

invitro

in

vitro

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4.2 Plant material

Solanum

melongena

in vitro

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4.3 Propagation of the plant material in vitro

4.3.1 Disinfection and explant selection

4.3.2 Seed inoculation and culture medium

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4.3.3 Callogenesis and indirect shoot regeneration

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4.3.4 Rooting, hardening and physical factors

in vitro

in vitro-

C and the 16

4.3.5 Evaluated parameters and statistical analysis

Micropropagation

Callogenesis and indirect shoot regeneration

Petri

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4.4 Molecular biology analysis (RAPD-PCR)

4.4.1 Plant material and DNA extraction

Total genomic DNA was extracted from 100 g of fresh leaves of the seedlings cloned in

vitro

4.4.2 DNA amplification

3 end

3 end

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4.4.3 DNA separation and RAPD-PCR analysis

4.5 Experimental design, materials and equipment in the greenhouse

4.5.1 Type of greenhouse and irrigation system

3]

S melongena

Thermus aquaticus E coli T aquaticus 3 synthesis of DNA, 3 exonuclease activity, and is reported to have

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4.5.2 Environmental conditions in the greenhouse

C) and

0

5

10

15

20

25

30

35

40

Apr

May

Jun

Jul

Aug

Sep

temperature(C)

0

10

20

30

40

50

60

70

80

90

100

relativehumidity

(%)

2002 2005 2002 2005(C) (C)(%) (%)

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4.5.3 Methodology and statistical methods

Global radiation

0

300

600

900

1200

1500

1800

2100

2400

2700

3000

3300

3600

Oct0

1

Dec

01

Feb

02

Apr0

2

Jun

02

Aug

02

Feb

03

Apr0

3

Jun

03

Aug

03

Feb

04

Apr0

4

Jun

04

Aug

04

Mar

05

May

05

Jul0

5

J/cm2

I Exp. phase

II Exp. phase

III Exp. phase

IV Exp. phase V Exp. phase

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4.5.4 Collection of the experimental data

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5 Results and Discussion

callogenesis organogenesis

hypocotyl cotyledon

5.1 Cultivation of eggplant in vitro

5.1.1 Invitrogermination

invitro

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Discussion

in vitro

in vitro

et al

5.1.2 In vitromultiplication

abcbcaab

c

a

b

c

bc

b

a

b

bb

a

0

1

2

3

4

5

6

Adon

a15

07

C

aoSan Do

k

VASI

5

Adon

a15

07

Cao

San Do

k

VASI

5

Adon

a15

07

C

aoSan Do

k

VASI

5

eggplant genotypes

mu

ltiplication

factor

subc.1 subc.2 subc.3

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in vitro

VASI-5

DokCao San1507

Adona

0

1

2

3

4

5

6

6 7 12 3 5 8 11 13 14 16 19 22 25 2 10 12 16 17 21 23 33 35 4 12 13 14 15 16 19 21 33 2 4 6 13 15 16 17 28 35 36

eggplant clones

mu

ltiplication

facto

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Discussion

et al in vitro

in vitro

in vitro

5.1.3 Contamination of the eggplant material propagated in vitro

in vitro

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Discussion

In vitro

et al et al

et al

et al

et al et al

et al et al

in vitro

in vitro

et al

in vitro

in vitro

in vitro

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5.1.4 Callogenesis and organogenesis of eggplant

callogenesis organogenesis

cotyledon hypocotyl

Experiment I Callogenesis organogenesis

hypocotyl cotyledon

callogenesis

cotyledons

hypocotyl

hypocotyl cotyledon

hypocotyl

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hypocotyl

hypocotyl

cotyledon

0,10,5

0,8

NAA

2,4 D0

20

40

60

80

100

NumberofCalli(%)that

regenerat

edintoshoots

Concentration in mg/L of PGR in callus

induction medium

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cotyledon

cotyledon

cotyledon

Experiment II Callogenesis organogenesis hypocotyl

0,10,5

0,8

NAA

2,4 D0

20

40

60

80

100

Num

bero

fCalli

(%)that

regenerate

dintos

hoots

Concentration in mg/L of PGR in callusinduction medium

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hypocotyl

callogenesis

hypocotyl

hypocotyl

Adona1507

Cao SanDok

VASI5

0,5 mg/L NAA

0,1 mg/L TDZ0

20

40

60

80

100

Percentage

ofcalli

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organogenesis

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hypocotyl

Discussion

et al

et al

et al

et al

et al

Solanum melongena et al S. torvum S. aculeatissimum S. aviculare S.

gilo et al S. torvum Khasianum et al

et al

in vitro

et al et

al et al

et al Paulownia tomentosa et al

invitro

et al et al et al

et al

et al

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0,2 m TDZ, but they were short and

et al

et al

et al

5.2 RAPD-PCR analysis

et al

et al

Taq

was increased to 3,5 l

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combined with 2.5 l 10X buffer without MgCl us 1.25 l of the primer. Preliminary

et al et al

et al

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3

S. Melongena

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S elongena

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Discussion

Solanum melongena

et al S. melongena

in vivo

Solanum et al et al., 2003; Skara et

al S. melongena

et al

et al

et al

et al

Sericea lespedeza et al H. rosa-

sinensis H. syriacus

et al

5.3 Cultivation of eggplant in vivo

5.3.1 Influence of the substrate on the vegetative development of

the genotypes

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aab

c

bcbc

a

bcab

c bc

0

10

20

30

40

50

Adona

1507

CaoSa

nDo

k

VASI

-5

Adona

1507

CaoSa

nDo

k

VASI

-5

Plantheight(cm)

organic sustrate perlite

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Discussion

et al

et al et al et al

et al

0

5

10

15

20

Adona 1507 Cao

San

Dok VASI-

5

Adona 1507 Cao

San

Dok VASI-

5

Leafnumb

er

organic sustrate perlite

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et al

et al

5.3.2 Morphological characteristics of the eggplant genotypes

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0

20

40

60

80

100

120

140

Adon

a15

07

Cao

San Do

k

VASI

-5

Adon

a15

07

Cao

San Do

k

VASI

-5

Plant

he

ight

(cm

)

0

10

20

30

40

Lea

fnum

ber

plant height (cm) leaf number

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ab

b

ab

aa

AB

B

AB

AA

0

20

40

60

80

100

120

140

Adona

1507

CaoSa

nDo

k

VASI

-5

Adona

1507

CaoSa

nDo

k

VASI

-5

Leafnum

ber

0

10

20

30

40

Plantheigh

t(cm)

plant height (cm) leaf number

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Discussion

et al

a

b

a a a

A

CBC

AB

BC

0

10

20

30

40

A

dona

1507

Ca

Tim

Dok

VAS

I-5

A

dona

1507

Ca

Tim

Dok

VAS

I-5

leaflength

(cm)

0

10

20

30

40

leafwidth(cm)

leaf length (cm) leaf width (cm)

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et al

etal

Coleus

et al et al

Solanum

melongena

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5.3.3 Generative development of the eggplant genotypes

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0%

20%

40%

60%

80%

100%

Adona 1507 Cao San Dok VASI-5

Floweredplan

ts

week 4 week 6 week 7

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0%

20%

40%

60%

80%

100%


Floweredplan

ts

week 4 week 5 week 6 week 7

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Discussion

etal

0%

20%

40%

60%

80%

100%

Adon

a/6

Adon

a/12

Cao

San/

12

Cao

San/

17

Cao

San/

21

Dok/

6

Dok/

8

Dok/

15

Floweredplants(%)

week 4 week 8 week 12

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et al

c

ab

b

a

ab

b b

aa

bbaa

0

1 0

2 0

3 0

4 0

Ad ona 1507 Cao San Dok VASI-5

Flo

wer

&Fru

itnum

ber

0%

20%

40%

60%

80%

100%

Fru

itset

flower number fr uit n umber fru it set (%)

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bb

b

a

b

cc

c

a

b

c

bc

b

a

ab

0

10

20

30

40

50

60

70


Flow

er&Fruitnumber

0%

20%

40%

60%

80%

100%

Fruitset

flower number fruit number fruit set (%)

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0

10

20

30

40

50

60

70

Ado

na/6

Adon

a/12

1507

/5

1507

/7

CaoSa

n/12

CaoSa

n/17

CaoSa

n/21

Dok/

6

Dok/

8

Dok/

15

VASI

-5/2

VAS

I-5/

16

Flower&Fruit

number

0%

20%

40%

60%

80%

100%

Fruitse

t


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Discussion

0

10

20

30

40

Adona

/6

Adona/

12

CaoSa

n/12

CaoSa

n/17

CaoSa

n/21

Dok

/6

Dok

/8

Dok/

15

Flower&Fruit

number

0%

20%

40%

60%

80%

100%

Fruitset


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et al

et al

et al et al

et al

et al et al

et al

5.3.4 Morphological characteristics of the fruits and yield of eggplant

in the greenhouse

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Discussion

etal

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b

b

a

a

bb

b

a

0

2

4

6

8

10

Adon

a15

07

Cao

San

Dok

VASI

-5

Adon

a15

07

Cao

San

Dok

VASI

-5

Fruitnumber/pla

nt

0

0,2

0,4

0,6

0,8

1

1,2

Fruitload(kg)/plant

fruit number fruit load (kg)

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cc

b

a

b

ccc

b

a

0

5

10

15

20

25

Adona

1507

CaoSa

nDo

k

VASI

-5

Adona

1507

CaoSa

nDo

k

VASI

-5

Fruitnumber/plant

0

0,5

1

1,5

2

2,5

3

3,5

Fruitload(kg)/plan

t


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0

5

10

15

20

Adona/6

Adona/12

Cao

San

/12

Cao

San

/17

Cao

San

/21

Dok/6

Dok/8

Dok/15

VASI

-5/2

VASI

-5/16

Adona/6

Adona/12

Cao

San

/12

Cao

San

/17

Cao

San

/21

Dok/6

Dok/8

Dok/15

VASI

-5/2

VASI

-5/16

Fruitnumbe

r/plant

0

0,5

1

1,5

2

2,5

3

3,5

Fruitload(k

g)/plant


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bccc

b c

b

ab aa

bbb

bbb

aa

0

5

10

15

Ado

na-6

Ad

ona-

12

CaoSa

n/12

CaoSa

n/17

CaoSan

/21

Dok/

6

Dok

/8

Dok/

15

Ado

na-6

Ad

ona-

12

CaoSa

n/12

CaoSa

n/17

CaoSa

n/21

Dok/

6

Dok/

8

Dok/

15

Fru

itnum

ber/p

lant

Fru

itloa

d(kg

)/p

lant

fru it n umbe r fruit lo ad (k g)

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a

b

b

c

b

c

bc

a

0

5

10

15

20

Ritmo Dok Cao

San

1507-

27

Ritmo Dok Cao

San

1507-

27

Fruitnumbe

r/plant

0

0,5

1

1,5

2

Fruitload(kg)/plant


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invitro

invitro

Discussion

etal

et al

et al

bbb

ab

a

0

5

10

15

20

25

Ritm

o

1507

/27(s)

1507

/27(i.v

.)

Dok/2

Dok/33

Ritm

o

1507

/27(s)

1507

(i.v.)

Dok/2

Dok/33

Fruitn

umber/plant

0

0,5

1

1,5

2

2,5

3

Fruitlo

ad(kg)/plant

fruit number fruit load (kg)/plant

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et al

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6 Conclusions and Recommendations

Background

et al et al., 2003; Skara et al

6.1 Summary of the main results

Chapter 3.

Aim of the research General and specific objectives

Research questions

Research Question 1:

in vitro

in vitro

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in vitro

Research Question 2:

et al

Research question 3:

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in vitro


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Research questions 6 and 7:

6.2 Conclusions

in vitro

in vitro

In vitro

in vitro

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in vitro

6.3 Recommendations

In vitro

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7 Literature list

Solanum melongena

in vitro

Solanumaethiopicum

Solanum melongena

Solanum melongena

Solanum melongena Ldecemlineata

Solanum melongena

Solanum

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ActaHorticulturae

ActaHorticulturae

ActaHorticulturae

Solanum melongena

ActaHorticulturae

Solanum melongena

ActaHorticulturae

Solanummelongena

ScientiaHorticulturae

ActaHorticulturae

ScentiaHorticulturae in vitro Lycopersicon esculentum

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ActaHorticulturae

Solanaceae Solanum Solanum melongena

Solanum melongena


Vaccinium vitisidaea in vitro

In Vitro Solanum melongena

Solanum melongena Solanaceae

Solanaceae Solanaceae

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Solanum melongena

Sechium edule Solanum melongena

Solanum melongena In Vitro

Solanum

melongena

esculentum

Picoidesborealis

Solanum

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Hibiscus sinensisHsyriacus

in vitro


Brassica oleracea

Arabidopsis



Solanum melongena Leucinodesorbonalis LepidopteraPyralidae

SchistosomamansoniBiomphalariaGlabrata

Solanum

Solanaceae Solanum melongena S incanum

ActaHorticulturae

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in vitro

Lactobacillus plantarum Corynebacterium Staphylococcus saprophyticus Pseudomonaspaucimobilis Hemerocallis ChoisyaDelphinium

plantarum

ActaHorticulturae

Solanum melongena

44 (2-3): 139-141


Agrobacterium

Solanummelongena

Solanummelongena

ActaHorticulturae

Solanum melongena

ScientiaHorticulture

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ActaHorticulturae

Coleus

ActaHorticulturae

ActaHorticulturae

ActaHorticulturae

in vitroPrunusglandulosaTiliacordata

Solanaceae

Solanumtuberosum

Solanum melongena

Solanum melongena

in vitro ActaHorticulturae

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bacillus

ActaHorticulturae

Solanum melongena Plantarum

Solanum melongena

Solanum melongena S incanum S marginatum

SolanumincanumsensulatoSolanaceae

Solanummelongena

Fragaria

chiloensisF virginianaF ananassaScientiaHorticulturae Skara A., S. Ce

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Solanum melongena

Solanum melongena ScientiaHorticulturae

ActaHorticulturae

SericeaLespedezaLespedezacuneata

Solanum melongena

in vitro Solanum melongena

solanum

Claosporiumfulvum

ActaHorticulturae

in vitro Musa ActaHorticulturae

ActaHorticulturae

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Index of figures

Solanum



Solanum melongena esculentum

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in vitro

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List of tables

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8 Attachment

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+ 500 l buffer AW

+ 400 l AP1+ 4 l R

+ 130 l AP2

in in a new tube+ 500 l buffer

+ 100 l previously warmed AE buffer at 65C

+ 100 l previously warmed AE buffer at 65C

650 l in white column

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Fruits

Nuts

Vegetables aubergines

Solanum melongena

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egg plant 2

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