Development of Chromosome Segment Substitution Lines in two...

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Development of Chromosome Segment Substitution Lines in two interspecific Oryza sativa x Oryza glaberrima populations Gutiérrez Andrés Gonzalo 1 , Alvarez Maria Fernanda 1 , Martínez César Pompilio 1 , Giraldo Olga Ximena 1 , Tohme Joe 1 and Lorieux Mathias 1,2 1 Centro Internacional de Agricultura Tropical (CIAT) AA 6713 Cali, Colombia. 2 Institut de Recherche pour le Développement (IRD), France e-mail: [email protected], [email protected] 2. IR64 (indica) x TOG5681 cross CONCLUSIONS AND PERSPECTIVES RESULTS AND DISCUSSION METHODOLOGY INTRODUCTION Genotyping of 312 lines with 200 SSRs (125 SSRs automatically selected by CSSL Finder program) 64 lines covering the whole Oryza glaberrima genome Strong segregation distortion on Chr.6 1. Caiapó (japonica) x MG12 cross BC 3 F 3 and BC 2 F 4 population. 363 lines genotyped with 143 SSRs selected from the Universal Rice Core Map (115 SSRs automatically selected by CSSL Finder program) 61 lines covering 95% O. glaberrima genome. This work allowed us to advance significantly in the construction of CSSLs between the two cultivated species of rice, in both indica and japonica genetic backgrounds. Fifteen QTLs in each interspecific population were located Furthermore, CIAT also has available CSSLs populations with Wild species to explore allelic diversity, using the same SSRs from the Universal Rice Core Map and the same genetic background (acc. Curinga, tropical japonica elite line): Curinga x O. meridionalis acc. W2112/OR44 (CIAT, Colombia) Curinga x O. barthii acc. IRGC101937 (WARDA, Africa) Curinga x O. rufipogon acc. IRGC105491 (Fedearroz and CIAT, Colombia) Curinga x O. glumaepatula acc. GEN1233 (CNPAF, Brazil) BAC libraries for each of the Wild species have been constructed (Arizona University) Gene mapping of underlying quantitative traits have progressed considerably however the development of new population structures has been poorly documented (Eshed and Zamir, 1995). New mapping populations called Chromosome Segment Substitution Lines (CSSLs) have been developed in rice (Ebitani et al., 2005). These lines are characterized to contain one or a few contiguous chromosomal segments of the donor genotype in the genetic background of a recurrent genotype. Two populations of CSSLs between the two cultivated species of rice, Oryza sativa L. and O. glaberrima Steud were developed. We present a QTLs detection analysis for yield, yield components and resistance to Rice Stripe Necrosis Virus (RSNV) in order to illustrate the advantages of using this kind of materials in genetic analysis and breeding of rice. Two interspecific crosses: Oryza sativa x Oryza glaberrima 1.Caiapó (japonica) x MG12 (acc. IRGC103544) 2.IR64 (indica) x TOG5681 (2 populations) 4200 BC 4 F 2 lines in field and molecular identification of target lines. REFERENCES Eshed Y. and Zamir D. (1995) An Introgression Line Population of Lycopersicon pennellii in the Cultivated Tomato Enables the Identification and Fine Mapping of Yield-Associated QTL. Genetics 141: 1147-1162 Ebitani T., Takeuchi Y., Nonoue Y., Yamamoto T., Takeuchi K. and Yano M. (2005) Construction and Evaluation of Chromosome Segment Substitution Lines Carrying Overlapping Chromosome Segments of indica Rice Cultivar “Kasalath” in a Genetic Background of japonica Elite Cultivar “Koshihikari”. Breeding Science 55: 65-73 http://mapdisto.free.fr/ QTLs Mapping DNA ISOLATION AND QUANTIFICATION PCR AGAROSE AND PAGE ELECTROPHORESIS STATISTICAL ANALYSIS True CSSLs Yield, yield components and RSNV Rice Stripe Necrosis Virus (RSNV) QTL on Chr. 11 Characteristic symptoms of the disease CSSLs with extreme phenotype For example: Plant height Height Yield Tillering 1000 Grain weight % Health plants

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Development of Chromosome Segment Substitution Lines in twointerspecific Oryza sativa x Oryza glaberrima populations

Gutiérrez Andrés Gonzalo1, Alvarez Maria Fernanda1, Martínez César Pompilio1, Giraldo Olga Ximena1, Tohme Joe1 and Lorieux Mathias1,21 Centro Internacional de Agricultura Tropical (CIAT) AA 6713 Cali, Colombia.

2 Institut de Recherche pour le Développement (IRD), Francee-mail: [email protected], [email protected]

2. IR64 (indica) x TOG5681 cross

CONCLUSIONS AND PERSPECTIVES

RESULTS AND DISCUSSION

METHODOLOGY

INTRODUCTION

Genotyping of 312 lines with 200 SSRs (125 SSRs automaticallyselected by CSSL Finder program)64 lines covering the whole Oryza glaberrima genomeStrong segregation distortion on Chr.6

1. Caiapó (japonica) x MG12 cross

BC3F3 and BC2F4 population. 363 lines genotyped with 143SSRs selected from the Universal Rice Core Map(115 SSRs automatically selected by CSSL Finder program)61 lines covering 95% O. glaberrima genome.

This work allowed us to advance significantly in the constructionof CSSLs between the two cultivated species of rice, in bothindica and japonica genetic backgrounds.

Fifteen QTLs in each interspecific population were located

Furthermore, CIAT also has available CSSLs populations with Wild species toexplore allelic diversity, using the same SSRs from the Universal Rice Core Mapand the same genetic background (acc. Curinga, tropical japonica elite line):Curinga x O. meridionalis acc. W2112/OR44 (CIAT, Colombia)Curinga x O. barthii acc. IRGC101937 (WARDA, Africa)Curinga x O. rufipogon acc. IRGC105491 (Fedearroz and CIAT, Colombia)Curinga x O. glumaepatula acc. GEN1233 (CNPAF, Brazil)

BAC libraries for each of the Wild species have been constructed (ArizonaUniversity)

Gene mapping of underlying quantitative traits have progressedconsiderably however the development of new populationstructures has been poorly documented (Eshed and Zamir,1995).

New mapping populations called Chromosome SegmentSubstitution Lines (CSSLs) have been developed in rice (Ebitaniet al., 2005). These lines are characterized to contain one ora few contiguous chromosomal segments of the donor genotypein the genetic background of a recurrent genotype.

Two populations of CSSLs between the two cultivated speciesof rice, Oryza sativa L. and O.  glaberrima Steud weredeveloped. We present a QTLs detection analysis for yield,yield components and resistance to Rice Stripe Necrosis Virus(RSNV) in order to illustrate the advantages of using this kindof materials in genetic analysis and breeding of rice.

Two interspecific crosses: Oryza sativa x Oryza glaberrima 1.Caiapó (japonica) x MG12 (acc. IRGC103544) 2.IR64 (indica) x TOG5681 (2 populations)

4200 BC4F2 lines in fieldand molecular identificationof target lines.

REFERENCESEshed Y. and Zamir D. (1995) An Introgression Line Population of Lycopersicon pennellii in the CultivatedTomato Enables the Identification and Fine Mapping of Yield-Associated QTL. Genetics 141: 1147-1162

Ebitani T., Takeuchi Y., Nonoue Y., Yamamoto T., Takeuchi K. and Yano M. (2005) Construction and Evaluationof Chromosome Segment Substitution Lines Carrying Overlapping Chromosome Segments of indica Rice Cultivar“Kasalath” in a Genetic Background of japonica Elite Cultivar “Koshihikari”. Breeding Science 55: 65-73

http://mapdisto.free.fr/

QTLs Mapping

DNA ISOLATION ANDQUANTIFICATION

PCR

AGAROSE AND PAGEELECTROPHORESIS

STATISTICAL ANALYSIS

True CSSLs

Yield, yield components and RSNV

Rice Stripe Necrosis Virus (RSNV) QTL on Chr. 11

Characteristic symptoms of the disease

CSSLs with extreme phenotypeFor example: Plant height

HeightYield

TilleringSterility1000 Grain weight% Health plants