Apport de la Cytogénétique Moléculaire. àl analyse du Génome de la Canne à sucre

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Apport de la Cytogénétique Moléculaire àl analyse du Génome de la Canne à sucre Maguy Rodier, Lolita Triaire, Angélique D Hont in collaboration with BSES, Australia : Nathalie & George Piperidis USP, Brazil : Douglas Silva & Marie Anne Van Sluis

FISHing with sugarcane Maguy Rodier, Lolita Triaire, Angélique D Hont in collaboration with BSES, Australia : Nathalie & George Piperidis USP, Brazil : Douglas Silva & Marie Anne Van Sluis

Sugarcane = polyploid suspected large-scale duplication event Inferred polyploidy events during the evolution of angiosperms. Branch lengths are not to scale. Numbers indicate roughly estimated dates (in millions of years) since the duplication event.

70% of worldwide raw sugar production High biofuel production potential

S. officinarum 2n = 80 X =?? S. barberi S. spontaneum 2n = 40-128? X =?? modern cultivars 2n ~ 120??

Origin of S. barberi S. officinarum S. spontaneum rdna Paunra (2n=82) Interspecific hybrids between S. officinarum and S. spontaneum Two genomes are involved in cultivated clones, directly or through S. barberi

S. officinarum 2n = 80 X =? S. barberi S. spontaneum 2n = 40-128? X =?? modern cultivars 2n ~ 120??

Modern cultivar chromosome complement Cultivar R570 Cultivar NCo376 rdna 80% S. officinarum 10% S. spontaneum 10% recombinants 70% S. officinarum 20% S. spontaneum 10% recombinants

S. officinarum 2n = 80 X =? S. barberi S. spontaneum 2n = 40-128? X =? modern cultivars 2n ~ 85-95 + 17-30

Basic chromosome numbers in S. officinarum S. officinarum (2n=80) x = 10 rdna 45S rdna 5S 2n = 80 8 X = basic chromosome number = number of chromosomes number of sites per locus

Basic chromosome numbers in S. spontaneum S. spontaneum with 2n=80 x = 8 rdna 45S rdna 5S 2n = 80 10 X = basic chromosome number = number of chromosomes number of sites per locus

Sugarcane cultivars = complex polyploids S. spontaneum S. officinarum recombinants Homology group Chromosome pairing : - mainly bivalent - combination of polysomy and preferential pairing Monoploid genome size =950 Mb Total genome size (=2C)=10 Gb

S. spontaneum S. officinarum recombinants Canneàsucre 2n = c.a.115 2C = 10 000 Mbp Sorgho 2n = 2x = 20 2C = 1 600 Mbp Riz 2n = 2x = 24 2C = 800 Mbp Maïs 2n = 2x (4x) = 20 2C = 5 500 Mbp

Sorgho Canne Sorgho Canne From Grivet et al, 1996; Dufour et al, 1997; D Hont et Paulet, pers comm

Structural variation between parental species S. spontaneum S. officinarum (x=8) (x=10) recombinants Homology group VIII

Genetic map :Homology group VIII S. spontaneum (x=8) S. officinarum (x=10) VIII1 VIII2 VIII3 VIII4 VIII5 VIII6 VIII7 VIII8 (3) C105 B304 C107(4) C148 D57 U15 rdna Adh1 R63 C62 R35 C92 (4) B304 D57 C105 U15 C148 Adh1 rdna S54 C62 R35 C92 B1209 U6 U44 R125 C105 D57 VIII9 R35 C92 C105 U15 C107(4) VIII10 R35 C92 U42 U49 Adh1 C148 B1209 R125 U44 C86 2.7 U49 Adh1 C92 B1209 R125 C86 U42 C148 C92 B1209 B1209 B1206 U42 Adh1 VIII11 VIII12 VIII13 VIII14 (3) C86 U44

Homology group VIII S. spontaneum (x=8) S. officinarum (x=10) VIII1 VIII2 VIII3 VIII4 VIII5 VIII6 VIII7 VIII8 BAC 52F03 BAC 63D09 (3) C105 B304 C107(4) C148 D57 U15 rdna Adh1 R63 C62 R35 C92 (4) B304 D57 C105 U15 C148 Adh1 rdna S54 C62 R35 C92 B1209 U6 U44 R125 C105 D57 VIII9 R35 C92 C105 U15 C107(4) VIII10 R35 C92 U42 U49 Adh1 C148 B1209 R125 U44 C86 2.7 U49 Adh1 C92 B1209 R125 C86 U42 C148 C92 B1209 B1209 B1206 U42 Adh1 VIII11 VIII12 VIII13 VIII14 (3) C86 U44 BAC sorgho

rdna Cytogenetic data? S. spontaneum S. officinarum rdna 45S /BAC 63D09 VIII1 VIII2 VIII3VIII4 VIII5 VIII6 VIII7 VIII8 rdna rdna VIII9 VIII10VIII11 VIII12 VIII13VIII14 Genetic mapping data

rdna Cytogenetic data? S. spontaneum S. officinarum rdna 45S /BAC 63D09 VIII1 VIII2 VIII3VIII4 VIII5 VIII6 VIII7 VIII8 rdna rdna VIII9 VIII10VIII11 VIII12 VIII13VIII14 Genetic mapping data

S. spontaneum S. officinarum U15/BAC 52F03 /BAC 63D09 VIII1 VIII2 VIII3VIII4 VIII5 VIII6 VIII7 VIII8 U15 U15 U15 VIII9 VIII10VIII11 VIII12 VIII13VIII14 Genetic mapping data

Monitoring introgression with related genus S. spontaneum (2n=40 to 128) S. robustum (2n=60, 80 + up to 200) S. edule (2n=40 to 128) S. sinense (2n=116 to 120) S. barberi (2n=81 to 124) S. officinarum (2n=80) Modern cultivars (2n=100 to 120) Saccharum Miscanthus (2n=38, 40, 76) Erianthus (2n=20, 30, 40 and 60) Cultivated Wild

Chromosome transmission in intergenric hybrids S.officinarum (Badila) 2n=80 x n+n F1 Expected if n + n : 40 + 30 Observed: 40 + 30 E.arundinaceus (HN92-77) 2n=60 x 2n+n BC1 Expected if n + n : 76 + 15 Observed : 92 + 30 Expected if 2n + n : 96 + 30 Cultivar ~ 112 Saccharum (estimation) x n+n Cultivar ~ 112 Saccharum (estimation) BC2 QuickTime et un décompresseur TIFF (LZW) sont requis pour visionner cette image. Expected if n + n : 104 + 15 Observed: 94-96 + 10-12 No recombination

Distribution of repeated sequences Erianthus tandem specific sequence EaCIR1 EaCIR8 Subtelomeric

Distribution of repeated sequences Saccharum transposable element (retro copia like, TE197) Saccharum spp = cultivar LTR PBS gag pr int RT-RNaseH PPT TE 197 LTR 1763bp San Paolo University

Saccharum-Miscanthus hybrid TE197 LTR PBS gag pr int RT-RNaseH PPT TE 197 LTR 1763bp San Paolo University

Let s go FISHing