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L. saligna L. aculeata
Collecting and study genetic diversity in natural
populations of wild Lactuca spp.
Alex Beharav Institute of Evolution, University of Haifa
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● Collecting, conservation, studying ● Morphological traits, taxonomic validation ● Identification natural putative hybrids ● Bolting control (L. georgica) ● Genetic diversity ● Out-crossing rate ● species definition ● Diseases resistance - Bremia Lactucae ● Chemical composition ● "Maror” מרור
Objectives
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Categorization of Lactuca species into genes pools following McGuire et al. (1993) concept.
L. sativa is part of a reproductively isolated group that includes the wild species L. serriola, L. saligna, and L. virosa (in order of decreasing sexual compatibility with L. sativa).
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● Only L. serriola representing the primary gene pool ● Only low samples (generally 1-2) representing each species in most studies
Main bottlenecks
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Plant geographical territories and
district of Israel
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Lactuca serriola חסת המצפן
(2n=2x=18), the progenitor of cultivated lettuce (Zohary
1991)
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Unique germplasm of 562 accessions that
morphologically seemed L. saligna was
collected from 41 natural populations
(locations) throughout the Israel.
Beharav et al. Isr. J. Pl. Sci. 2008
Metulla, Israel
Kafer Kela, Lebanon
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Six other wild species are taxonomically close to the cultivated lettuce:
● L. aculeata
● L. scarioloides
● L. azerbaijanica
● L. Georgica
● L. dregeana
● L. altaica
However, they are less widespread, and the status of some of them as a distinct species is unclear (Doležalová et al. 2003; Lebeda et al. 2004a,b; Zohary 1991).
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Seeds from 151 plants that looked like L. aculeata were collected from 11 localities
(nine across the Golan Heights and two from the
Hula Plain)
Beharav et al. Genet. Resour. Crop Evol. 2010
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(i) to collect and validate the taxonomy of germplasm of five wild species collected throughout their distribution regions:
● L. scarioloides
● L. azerbaijanica
● L. Georgica
● L. dregeana
● L. altaica
(ii) to study the status and diversity of above wild species on a wider scale among individuals from different natural populations.
Objectives
Learning morphology of rare wild Lactuca spp.
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3 samples with respective labels of L.
dregeana (seeds supplied by various
gene banks)
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3 samples with respective labels of L. altaica
(2n=2x=18; seeds supplied by various gene banks)
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Sample with respective labels of L. georgica
(2n=2x=18; seeds supplied by DEU gene bank)
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Seeds from 51 plants (4 of them were marked as
natural putative hybrids with L. serriola) that looked like L. georgica (2n=2x=18)
were collected from 7 localities during collecting missions throughout four
regions of Armenia.
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Unique germplasm of 109 L. tuberosa
פרחים-כחולתחסה
(2n=2x=18) accessions, collected from 39 natural populations
(locations) throughout the Israel.
Afula-‘Illit
24.04.2010 Rosh-Pinna
21.04.2011
Shemurat Hermon II (Har Kahal)
28.05-24.06.2011
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L. Orientalis חסה מזרחית
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L. Viminea
חסת השבטים
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L. undulata
חסה גלונית
South Sinai
02.04.1971 (Danin)
Wadi Musa, Edom
17.04.1929 (Naftolsky)
Wadi Bed, Edom
11.05.2009 (Saifan)
Sabah
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Cichorium pumilum Cichorium calvum
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Leafy vegetables Number of accessions (old accessions)
Genus Crop name IGB (ISR) IOE (ISR)
Asparagus Asparagus 10 (6)
Atriplex French spinach 32 (7)
Chenopodium Good King Henry, Mercury 13 (3)
Chrysanthemum Garland chrysanthemum 28 (5)
Cichorium Chicory, Endive 20 (59) 40
Cynara Artichoke 18 (10)
Diplotaxis Rocket salad 26 (11)
Eruca Rocket salad 42 (28)
Lactuca Lettuce 73 (157) 1249
Lepidium Garden cress 9 (32)
Portulaca Purslane 10 (2)
Rheum Rhubarb 3
Rumex Garden sorrel, Sour grass 41
Spinacia Spinach 0 (1)
Taraxacum Dandelion 2
Tetragonia New Zealand spinach 0 (4)
Valerianella Lambs lettuce, Corn salad 9
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Multiplication and taxonomic validation of L. saligna accessions
according to morphological characters following Doležalová et al. 2002 (Hort. Sci. (Prague) 29: 56-83).
Beharav et al. Isr. J. Pl. Sci. 2008
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Multiplication and taxonomic validation of L. aculeata accessions according to morphological characters following Doležalová et al.
2002 (Hort. Sci. (Prague) 29: 56-83).
Beharav et al. Genet. Resour. Crop Evol. 2010
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Lebeda et al. submitted
Genetic polymorphism in L. aculeata populations and occurrence of natural putative hybrids
between L. aculeata and L. serriola
L. aculeata (376-15)
putative hybrid L. aculeata × L. serriola (343-8A)
L. serriola f. serriola (377-8)
Lebeda et al. Biochem. Syst. Ecol. 2012
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Bolting control of L. georgica by low temperatures
Beharav, Hellier, Harutyunyan, Hovhannisyan, running
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Kitner et al. Isr. J. Pl. Sci. 2008
Analysis of 490 AFLP loci (Jaccard´s
similarity) from 67 L. saligna accessions
collected from four European and three
Middle Eastern countries
Genetic diversity of wild Lactuca spp.
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Lebeda et al. submitted
36 accessions 21 allozyme loci Dice similarity
Genetic polymorphism in L. aculeata populations and occurrence of natural putative hybrids
between L. aculeata and L. serriola
Lebeda et al. Biochem. Syst. Ecol. 2012
Species definition ?
Open to discussion
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Genetic and phenotypic structure in Israeli
natural populations of three predominantly self-pollinating wild Lactuca species
Beharav et al. In preparation
Fig... Correlation between geographical distance (Meter) and genetic distance (Simple match; 235 AFLP
markers), in six wild Lactuca spp. populations (Mantel test; P = probability values (9999 permutations))
y = 0.0002x + 0.0169r² = 0.0188; P = 0.169
0.000
0.020
0.040
0.060
0.080
0 15 30 45 60
Sim
ple
mat
ch
Meter
L. serriola - population 365
y = 0.0012x + 0.0117r² = 0.2425; P = 0.000
0.000
0.050
0.100
0.150
0.200
0 30 60 90 120
Sim
ple
mat
ch
Meter
L. serriola - population 377
y = 1E-05x + 0.0043r² = 0.0027; P = 0.351
0.000
0.003
0.005
0.008
0.010
0 15 30 45 60
Sim
ple
mat
ch
Meter
L. aculeata - population 367
y = 0.0002x + 0.0186r² = 0.2996; P = 0.001
0.000
0.015
0.030
0.045
0.060
0 40 80 120 160
Sim
ple
mat
ch
Meter
L. aculeata - population 380
y = 0.0005x + 0.0631r² = 0.1995; P = 0.012
0.000
0.040
0.080
0.120
0.160
0 35 70 105 140
Sim
ple
mat
ch
Meter
L. saligna - population 369
y = 0.0007x + 0.0255r² = 0.5444; P = 0.000
0.000
0.040
0.080
0.120
0.160
0 30 60 90 120
Sim
ple
mat
ch
Meter
L. saligna - population 379
Fig... Correlation between geographical distance (Meter) and dissimilarity coefficient (Cavalli-Sforza and
Edwards´chord distance Dc; 11 EST-SSR loci), in six wild Lactuca spp. populations (Mantel test; P = probability
values (9999 permutations).
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Genetic and phenotypic structure in Israeli
natural populations of three predominantly self-pollinating wild Lactuca species
Beharav et al. In preparation
0.426
0.045 0.048 0.062
0.110
0.153
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
L. serriola Avi'el
L. serriola Kaukab
L. aculeata Hamappalim
L. aculeata Nov
L. saligna Metulla
L. saligna Binyamina
t (out-crossing rate)
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Genetic polymorphism in L. Georgica populations
using TRAP technique
Beharav, Hellier, Kisha. Running
- L. Georgica (Armenia)
- L. serriola (Armenia; Israel)
- L. aculeata (Israel)
- L. saligna (Armenia; Israel)
- L. virosa. (Various)
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Frequency of different degrees of infection (i.e. percentages of a pathogen sporulation intensity) among all L. saligna-B. lactucae
interactions (N=1460).
Bremia lactucae
Petrželová et al. Ann. App. Biol. 2011
0
10
20
30
40
50
0 5 10 15 20 25 30 35 40 45 50 55 60 65 70
Degree of infection (%)
Fre
qu
en
cy (
%)
Conclusion: to produce a mapping population within L. saligna?
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Reaction pattern to Bremia lactucae California Pathotype VIII 879 (Avr 17, 36, 37, 38) in seedlings from natural
populations of three wild Lactuca spp. collected in Israel
Beharav, Michelmore, Ochoa, unpublished data
0% 20% 40% 60% 80% 100%
L. aculeata
L. serriola
L. saligna
36
0
83
33
60
0
Resistance
Susceptible
Species definition ?
Open to discussion
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Beharav, Michelmore, Ochoa, unpublished data
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Genetic variability of L. aculeata germplasm expressed by resistance variation against five Bl races of B. lactucae
Jemelková et al. WPP conference (Poster), Olomouc, 2013
● Susceptible reactions were prevailing, however surprisingly there was recorder also full resistance what is showing that this species could be considered as interesting source of new race-specific resistance for lettuce breeding
Conclusion: to produce a mapping population within L. aculeata?
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Variation of sesquiterpene lactones in L. aculeata natural populations from Israel, Jordan and Turkey
Beharav et al. Biochem. Syst. Ecol. 2010
Total sesquiterpene lactone contents (mg/g of dry plant material)
1
3
5
7
Num
ber
of accessio
ns
2 4 6 8 10 12 14
L. sativa lines JOR224-22 ISR343-3 a
1
2
3
4
5
Num
ber
of accessio
ns
0 5 10 15
b L. sativa lines,
ISR343-3
JOR224-22
Total sesquiterpene lactone contents (mg/g of dry plant material)
Analysis of seven sesquiterpene lactones from roots (a) and leaves (b) of 23
accessions collected from 11 localities
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Since Israel is not a signatory to the SMTA Treaty, Israeli collections are not defined as European collections. So, topic of signature is currently not under the Israeli scientists.
Finally…
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ACKNOWLEDGEMENTS
Members from: ● Institute of Evolution, University of Haifa, Israel. ● Flora of Israel Online. ● National Agrarian University of Armenia, yerevan, Armenia. ● National Center for Agricultural Research and Extension (NCARE), Baqa‘, Jordan. ● Department of Botany, Palacký University, Olomouc-Holice, Czech Republic. ● Western Regional Plant Introduction Station, Washington State University, Pullman, WA. ● The Genome Center, University of California, Davis, CA. ● Department of Phytochemistry, Institute of Pharmacology, Polish Academy of Sciences, Krakow, Poland. ● Rijk Zwaan Breeding B.V., De Lier, The Netherlands.
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