European project « 5th PCRDT » GARLIC AND HEALTH Third year progress report Half year meeting...
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Transcript of European project « 5th PCRDT » GARLIC AND HEALTH Third year progress report Half year meeting...
![Page 1: European project « 5th PCRDT » GARLIC AND HEALTH Third year progress report Half year meeting Montpellier 1-2 october 2002.](https://reader030.fdocuments.us/reader030/viewer/2022032723/56649f525503460f94c76281/html5/thumbnails/1.jpg)
European project « 5th PCRDT »
GARLIC AND HEALTH
Third year progress report
Half year meeting Montpellier 1-2 october 2002
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ADVANCEMENT OF THE Works
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Scientific and administrative staff
• L. FEREOL scientist– Duties: somatic embryogenesis,
histology and flow cytometry
• S. CAUSSE technical assistant– Duties: somatic embryogenesis,
histology and flow cytometry
• M. Roux-Cuvelier– Duties: field trials in Reunion
• R. Kahane– Duties: management
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Objectives
• Setting a somatic embryogenesis procedure
• Comparative characterisation of plantlets from somatic embryos and from classic multiplication by meristem culture (as control)
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Work plan
• Establishment of somatic embryogenesis procedure for four varieties: Rouge Reunion, Messidrome, Morasol, Printanor
• Production and provision of somatic embryos via callus and via cell suspensions cultures of these varieties
• Genetic evaluation of somatic embryos derived plants by : flow cytometry (by P8), biochemistry (by P5), molecular markers (by P1), morphological and physiological analyses (by P8, P9)
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Milestones of the first year
Production and germination of somatic embryos « Rouge Reunion » from callus and cell suspension cultures
√
Provision of 200 plantlets from embryogenic calli, and from cell suspension cultures « Rouge Reunion »
√
Production of embryogenic callus, and initiation of embryogenic cell suspension cultures « Messidrome, Morasol »
√
Production and germination of somatic embryos from callus and from cell suspension culture « Messidrome, Morasol »
√
√ = achieved; partly achieved; to be done)�
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Milestones of the second year
Genetic characterisation of these plants by flow cytometry « Messidrome », « Morasol »
Provision of 200 plantlets from embryogenic calli, and from embryogenic cells suspension cultures of « Messidrome », « Morasol »
√Genetic characterisation of somatic embryos derived plantlets « Rouge Reunion »:
By flow ctytometry(P8), finger printing(P1), dry matter and sulphur content(P5), morpho-physiology(P9)
√
√
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Milestones of the second year (continue)
Production and germination of somatic embryos from callus and cell suspension cultures « Printanor »
√
√
Production of embryogenic callus, and Initiation of embryogenic cell suspension cultures « Printanor »
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Milestones of the third year
Promotion of the embryogenesis procedure
Genetic characterisation of these plants by flow cytometry « Printanor »
√Provision of 200 plantlets from embryogenic calli, and from embryogenic cells suspension cultures of « Printanor »
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Embryogenesis from callus
Friableembryogeniccallus
Globular somatic embryos
h
Matureembryo
MORPHOLOGY HISTOLOGY
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Embryogenesis from cell suspension culture
Embryogeniccell suspensionculture established
Conversion into plantlets and then bulbification
Regeneration30 days after plating
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Regeneration of cell suspension after plating
Globular embryos
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Varietal effect on somaticembryogenesis from callus
induction of embryogenic tissues
0%
10%20%
30%
40%
50%60%
70%
80%
1
varieties
% e
mbr
yoge
nic
callu
s RR
MES
MOL
PRI
Induction of embryogenic tissues
Embryo induction
0102030405060708090
1
varieties
Mean
nbr
of s
.e./1
50 m
g of
ca
llus
RR
MES
MOL
PRI
Somatic embryo induction
Conversion of embryos into plantlets
0
1020
30
40
5060
70
80
1
varieties
% ge
rmin
atio
n
RR
MES
MOL
PRI
Conversion into plantlets
Significant effect on the different parameters of somaticembryogenesis from callus
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Varietal effect on somaticembryogenesis from cell
suspension culture
Growth of cell suspension culture
1
2
3
1
Varieties
Grow
th ra
te
RR
MES
MOL
PRI
Growth of the cell suspension
Regeneration after plating
0
2040
60
80
100120
140
160
1
varieties
nbre
of s
.e. /
ml P
CV
RR
MES
MOL
PRI
Regeneration after plating
Conversion of embryos into plantlets
0102030405060708090
1
varieties
% o
f ger
min
atio
n
RR
MES
MOL
PRI
Conversion into plantlets
- No varietal effect on the growth rate- But a significative effecton regeneration and conversion into plantlets
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2 X4 X
2 X+4 X
Flow cytometric histogramOf diploid and tetraploid
Somatic embryos
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Genetic characterisation by flow cytometry
Nbr. of plants tested
Nbr. of diploid
Nbr.of tetraploid
MES control
12 12 0
MOL control
16 16 0
MES cal 102 93 9
MOL cal 100 97 3
MOL ecs 100 96 4
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DeliverablesDP 5: paper on: Evidence of a somatic embryogenesis process and plant regeneration and acclimatisation in garlic (Allium sativum L.)L. FEREOL, V. CHOVELON, S. CAUSSE, N. MICHAUX FERRIERE and R. KAHANEPlant Cell Report, september 2002
DP13: paper on: Establishment of garlic (Allium sativum L.) cell suspensions and plant regenerationL. FEREOL, V. CHOVELON, S. CAUSSE, D. TRIAIRE, N. MICHAUX FERRIERE and R. KAHANE
in progressDP21: paper on the use of somatic embryogenesis for rapid multiplication of garlic in vitro.In progress
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What will be done
• Promotion of the embryogenesis procedure– Histological studies– Looking for more convenient
conditions for somatic embryogenesis– Maintenance of the material
• MES ecs, PRI ecal and PRI ecs: – Genetic characterisation of plants from
these material, by flow cytometry
• Rouge Reunion ecal and ecs:Field evaluationFlow cytometry of the material after field
culture