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ELI KHAYAT A Breed apart In-vitro mutagenesis, a tool for germplasm improvement
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Sources of variability Somaclonal variation: A genetic variation that is observed in plants regenerated by tissue culture of somatic cells.
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3 % of population Kg per bunch 25 35 45 55 65 Genetic drift Unselected Selected
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4 Clonal genetic diversity
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5 micropropagationmutagenesis Hyper amplification of BR-1 In-vitro mutagenesis
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Genetic mechanisms
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5’LTR Gag prot. integ. RT RN H 3’LTR DR IR IR IR IR DR PBS PPT 5’LTR Gag prot. integ. RT RN H env 3’LTR DR IR IR IR IR DR PBS PPT Copia-like element Retrovirus P O L A comparison between the structures of Copia-like Retro- element and a retrovirus
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RETROTRANPOSON ACTIVATION RNA molecules Reintegration to Chromosomal DNA Episomal DNA molecules GENE I GENE I Insertion Retro transposon Transcripts LNL WT DNA METHYLATION Activation of retro- transposing element
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TE Pro ORF Pro TE Translation RT enzyme Integrase nuclease RNA Amplification of Retro elements in mutated genomic DNA. Southern hybridization blot of banana DNA cut with EcoR1 enzyme (lanes 2,4,6,8,10) or uncut (lanes 1,3,5,7,9). The blotted DNA was hybridized to BR-1 probe encoding 344 ntds of the RT enzyme from Copya -1 retro-transposable element, Isolated by PCR according to Khayat et al. 2004. Lanes: 1,2 DNA from plants after exposure to high BAP and TDZ for 12 cycles In TC; Lanes 3,4 DNA from plants after exposure to high BAP and TDZ for 9 cycles In TC; Lanes 5,6 DNA from plants after exposure to high BAP and TDZ For 6 cycles In TC; 7,8 DNA from a dwarf mutant grown in the field and 9.10 a normal Plant grown in the field, 1 2 3 4 5 6 7 8 9 10 Episomal DNA Linear DNA LTR Southern Hybridization blot
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Retro transposons activation in rice
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Southern Analysis of TOS 17
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13 RA – Grand Nain
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Selection of a Fusarium (TR4) resistant banana clone
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Masada Mutation Irregular variegation Normal n = 33 Chromosomes Masada n = 34 Chromosomes
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Effect of GA3 in the growth medium on internode length of dwarf vs. normal plants **) length in cm
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GAI (GA insensitive) gene Landing pad GAI gene 250-300 Kbp Genomic DNA Bi-Bac
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18 “Adi” - yield and Size a revolution in “Cavendish breeding” Grande Naine Zelig ADI
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Martinique July 17 2013 (Post Hurrican) GAL ADI
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Early flowering in Jaffa(2014) Banana “King “ Ecuador 2013 Guadeloupe – French Caribbean Islands
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21 Goal : lower stature Santa Marta Colombia
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Effect of Ancymidol and GA 3 on elongation of TC plants “GN”, “ADI” and “JAFFA” '84' - 250 p.p.m of Ancy. '3342' control no added Anc. or GA 3 '3343' - 1 000 p.p.m of GA 3. '3344' -250 p.p.m of Ancymidol and 1000 p.p.m of GA 3. ADI JAFFA ADI- Defective GA synthesis JAFFA- Insensitive to ANC
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