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Identification of Molecular Markers for a Aphid Resistance Gene in Sorghum and Selective Efficiency Using These Markers |
CHANG Jin-hua, CUI Jiang-hui, XUE Wei, ZHANG Qing-wen |
1.College of Agriculture and Biotechnology, China Agricultural University, Beijing 100096, P.R.China
2.College of Agriculture, Agricultural University of Hebei, Baoding 071001, P.R.China
3.Student Employment Guidance Center, Baoding Vocational and Technical College, Baoding 071001, P.R.China |
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摘要 In this study, an F2 segregated population obtained by hybridization between the aphid-sensitive sorghum strain Qiansan and aphid-resistant cultivar Henong 16 was used to establish an aphid-resistant pool and an aphid-sensitive pool. 192 pairs of AFLP (amplified fragment length polymorphism) marker primers were screened in these pools using BSA (bulked segregant analysis). Three pairs of EcoR I-CTG/Mse I-CCT, EcoR I-CTG/Mse I-CAT, and EcoR I-AGT/Mse I-CCC showed linkage with aphis resistance. EcoR I-CTG/Mse I-CCT-475, EcoR I-CTG/Mse I-CAT-390, and EcoR I-AGT/Mse I-CCC- 350 (E42/M52-350) were mapped within 6, 10, and 13 cM distances with the aphid-resistant gene by using Mapmaker 3.0 software. The bands amplified by EcoR I-CTG/Mse I-CCT-475 and EcoR I-CTG/Mse I-CAT-390 were extracted, cloned, and sequenced. Specific primers of SCAR (sequence characterized amplified regions) were then designed from these bands. A specific band of 300 bp was amplified by a pair of SCAR primers designed based on the sequence obtained from the EcoR I-CTG/Mse I-CAT-390 marker. The SCAR marker was named SCA50. The marker was used to detect the F2, BC1, and F2:3 populations. The selective efficiency was 86.8, 91.1, and 86.3% in the BC1, F2, and F2:3 populations, respectively. The average selective efficiency was 88.2%.
Abstract In this study, an F2 segregated population obtained by hybridization between the aphid-sensitive sorghum strain Qiansan and aphid-resistant cultivar Henong 16 was used to establish an aphid-resistant pool and an aphid-sensitive pool. 192 pairs of AFLP (amplified fragment length polymorphism) marker primers were screened in these pools using BSA (bulked segregant analysis). Three pairs of EcoR I-CTG/Mse I-CCT, EcoR I-CTG/Mse I-CAT, and EcoR I-AGT/Mse I-CCC showed linkage with aphis resistance. EcoR I-CTG/Mse I-CCT-475, EcoR I-CTG/Mse I-CAT-390, and EcoR I-AGT/Mse I-CCC- 350 (E42/M52-350) were mapped within 6, 10, and 13 cM distances with the aphid-resistant gene by using Mapmaker 3.0 software. The bands amplified by EcoR I-CTG/Mse I-CCT-475 and EcoR I-CTG/Mse I-CAT-390 were extracted, cloned, and sequenced. Specific primers of SCAR (sequence characterized amplified regions) were then designed from these bands. A specific band of 300 bp was amplified by a pair of SCAR primers designed based on the sequence obtained from the EcoR I-CTG/Mse I-CAT-390 marker. The SCAR marker was named SCA50. The marker was used to detect the F2, BC1, and F2:3 populations. The selective efficiency was 86.8, 91.1, and 86.3% in the BC1, F2, and F2:3 populations, respectively. The average selective efficiency was 88.2%.
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Received: 18 March 2011
Accepted:
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Fund: This work was supported by the Natural Science Foundation, Hebei Province, China (C2010000758) and the Science and Technology Department of Hebei Province, China (06547004D-2). |
Corresponding Authors:
ZHANG Qing-wen, Tel/Fax: +86-10-62733016, E-mail: zhangqingwen@263.net
E-mail: zhangqingwen@263.net
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Cite this article:
CHANG Jin-hua, CUI Jiang-hui, XUE Wei, ZHANG Qing-wen.
2012.
Identification of Molecular Markers for a Aphid Resistance Gene in Sorghum and Selective Efficiency Using These Markers. Journal of Integrative Agriculture, 12(7): 1086-1092.
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[1]Chang J H, Xia X Y, Zhang L, Li R G, Liu G Q, Luo Y W. 2006. Analysis of the resistance gene to the sorghum aphid, Melanaphis sacchari, with SSR marker in Sorghum bicolor. Acta Prataculturae Sinica, 115, 113-118. (in Chinese) [2]Concibido V C, Denny R L, Lange D A. 1996. RFLP mapping and marker-assisted selection of soybean cyst nematode resistance in PI 209332. Crop Science, 36, 1643-1650. [3]Doggett H. 1988. Sorghum. 2nd ed. Longman Scientific and Technical, John Wiley and Sons, New York, USA. pp. 4-5. [4]Gao A L, He H G, Chen Q Z. 2005. Pyramiding wheat powdery mildew resistance genes Pm2, Pm4a and Pm21 by molecular marker-assisted delection. Acta Agronomica Sinica, 131, 1400-1405. [5]He F G, Xin W M, Yan F Y, Wang Y Q, Li X P. 1991. Sorghum aphid resistance identification method of sorghum. Liaoning Agricultural Science, 3, 6-9. (in Chinese) [6]Jena K K, Mackill D J. 2008. Molecular markers and their use in marker-assisted selection in rice. Crop Science, 48, 1266-1276. [7]Kosambi D. 1994. The estimation of map distances from recombination values. Annals of Eugenics, 12, 172-175. [8]Lande R, Thompson R. 1990. Efficiency of marker-assisted selection in the improvement of quantitative traits. Genetics, 124, 743-756. [9]Lander E S, Green P, Abrahamson J. 1987. Mapmaker: an interactive cumputer for constructing primary genetics maps of experimental and natural populations. Genomics, 1, 174-182. [10]Li Y Y, Zhao M H, Yang L G. 2003. Study on the molecular markers linked to aphid resistance gene of sorghum. Acta Agronomica Sinica, 29, 534-540. (in Chinese) [11]Pei Q L, Wang C L, Liu P Q, Wang J, Zhao K J. 2011. Markerassisted selection for pyramiding disease and insect resistance genes in rice. Chinese Journal of Rice Science, 25, 119-129. [12]Radehenko E E. 1999. Identification of genes for resistance to greenbug in sorghum. Journal of Genetics, 36, 408-417. [13]Ramasamy P, Menz M A, Mehta P J, Katilé S, Gutierrez-Rojas L A, Klein R R, Klein P E, Prom L K, Schlueter J A, Rooney W L, et al. 2009. Molecular mapping of Cg1, a gene for resistance to anthracnose (Colletotrichum sublineolum) in sorghum. Euphytica, 165, 597-606. [14]Sabina A, Bhat M A, Wani S A, Bhat K A, Chalkoo S, Mir M R, Wani S A. 2010. Marker assited selection in rice. Journal of Phytology, 2, 66-81. [15]Saghai-Maroof M A, Jeongs S C, Gundnz I, Tucker D M, Buss G R, Tolin S A. 2008. Pyramiding of soybean mosaic virus resistance genes by marker assisted selection. Crop Science, 48, 517-526. [16]Saghai-Maroof M A, Soliman K M, Jorgesen R A, Allard R W. 1984. Ribosomal DNA spacer-length polymorphisms in barly mendelian inheritance, chromosomal location and population dynamics. Proceedings of the National Academy of Sciences of the United States of America, 81, 8014-8018. [17]Sambrook J, Russell D W. 2002. The Condensed Protocols from Molecular Cloning: A Laboratory Manual. 3rd. ed. Science Press, Beijing. p. 512. (in Chinese) Staub J E. 1997. Comparation of isozyme and random amplified polymorphic DNA data for determining in transpacific variation in cucumis. Genetic Resources and Crop Evaluation, 44, 257-269. [18]Stromberg L D, Dudley J W, Rufener G K. 1994. Comparing conventional early generation selection with molecular marker assisted selection in maize. Crop Science, 34, 1221-1225. [19]Vos P, Hogers R, Bleeker M, Reijans M, van de Lee T, Homes M, Freijters A, Pot J, Peleman J, Kuiper M, et al. 1995. AFLP: a new technique for DNA fingerprinting. Nucleic Acids Research, 23, 4407-4414. [20]Wang J, Yang J, Chen Z D, Fan F J, Zhu J Y, Yang J H, Zhong W G. 2011. Pyramiding resistance gene Pi-ta, Pi-b, and Stv-bi by marker-assisted selection in rice (Oryza sativa L.). Acta Agronomica Sinica, 37, 975-981. [21]Wang Z, Jia Y L, Xia Y. 2004. Molecular markers-assisted selection of the rice blast resistance gene Pi-ta. Acta Agronomica Sinica, 30, 1259-1265. (in Chinese) [22]Wu Y Q, Huang Y H. 2008. Molecular mapping of QTLs for resistance to the greenbug Schizaphis graminum (Rondani) in Sorghum bicolor (Moench). Theoretical and Applied Genetics, 117, 117-124. |
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