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Acta Agron Sin ›› 2017, Vol. 43 ›› Issue (11): 1622-1631.doi: 10.3724/SP.J.1006.2017.01622


Pyramiding Pi-ta, Pi-b and Wx-mq Genes by Marker-assisted Selection in Rice (Oryza sativa L.)

YAO Shu, CHEN Tao,ZHANG Ya-Dong,ZHU Zhen,ZHAO Qing-Yong,ZHOU Li-Hui,ZHAO Ling,ZHAO Chun-Fang,WANG Cai-Lin*   

  1. Institute of Food Crops, Jiangsu Academy of Agricultural Sciences/Jiangsu High Quality Rice R&D Center,/Nanjing Branch of China National Center for Rice Improvement, Nanjing 210014, China
  • Received:2017-03-03 Revised:2017-07-23 Online:2017-11-12 Published:2017-08-02
  • Contact: 王才林, E-mail: clwang@jaas.ac.cn, Tel: 025-84390307
  • Supported by:

    This study was supported by the Agricultural Science and Technology Independent Innovation Fund of Jiangsu (CX [12]1003), the Key Research and Development Projects of Jiangsu Province (BE2016370), and the Special Funds for the Construction of Modern Agricultural Industrial Technology System (CARS-01-47).


In recent years, japonica rice variety Nanjing 46, Nanjing 5055 and Nanjing 9108 were planted in a large area in Jiangsu province because of their good eating quality. However, these varieties are not resistant to rice blast, and there is no japonica rice variety with good eating quality and suitable for planting in Huaibei area, Jiangsu province. The goal of modern rice breeding is to develop new varieties with good quality, high yield and multi-resistance. Marker-assisted selection combined with traditional breeding methods can greatly improve breeding efficiency. In this study, the high yield rice variety “Wujing 15” contained two blast resistance genes of Pi-ta and Pi-b was used as gene recipient to be crossed with the rice variety “Nanjing 5055”, which carries the low-amylose content gene Wx-mq. Self-designed multiple PCR system to detect rice blast resistance genes of Pi-ta and Pi-b and Tetra-primer Amplification Refractory Mutation PCR System for Wx-mq were used to detect the target loci in separation generation. Combining with many generations of breeding in the field, resistance identification, and classification of the grain endosperm appearance, a new japonica rice line “Nanjing 0051” with excellent agronomic traits and suitable for planting in Huaibei area, Jiangsu province was bred by multi-generational selection on resistance and endosperm appearance. This study successfully used the three sets of PCR detection system in the marker-assisted selection, providing a rapid selection method in rice breeding also important intermediate materials for quality and disease-resistance.

Key words: Rice, Blast, Low-amylose content, Pi-ta, Pi-b, Wx-mq, Marker-assisted selection

[1] 王世维, 郑文静, 赵家铭, 魏松红, 王妍, 赵宝海, 刘志恒. 辽宁省稻瘟病菌无毒基因型鉴定及分析. 中国农业科学, 2014, 47: 462–472 Wang S W, Zheng W J, Zhao J M, Wei S H, Wang Y, Zhao B H, Liu Z H, Identification and analysis of Magnaporthe oryzae avirulence genes in Liaoning province. Sci Agric Sin, 2014, 47: 462–472 (in Chinese with English abstract) [2] 李祥晓, 王倩, 罗生香, 何云霞, 朱苓华, 周永力, 黎志康. 黑龙江省稻瘟病菌无毒基因分析及抗病种质资源筛选. 作物学报, 2012, 38: 2192–2197 Li X X, Wang Q, Luo S X, He Y X, Zhu L H, Zhou Y L, Li Z K. Analyzing avirulence genes of Magnaporthe oryzae from Heilongjiang province and screening rice germ plasm with resistance to blast fungus. Acta Agron Sin, 2012, 38: 2192–2197 (in Chinese with English abstract) [3] Shen M G, Lin J Y. The economic impact of rice blast disease in China. In: Zeigler R S, Leong S A, Teng P S. Rice Blast Disease. England, UK: CAB Internationa1, 1994. pp 321–331 [4] Hittalmani S, Parco A, Mew T V', Zeigler R S, Huang N. Fine mapping and DNA marker assisted pyramiding of the three major genes for blast resistance in rice. Theor Appl Genet, 2000, 100: 1121–1128 [5] 时克, 雷财林, 程治军, 许兴涛, 王久林, 万建民. 稻瘟病抗性基因Pi-ta和Pi-b在我国水稻主栽品种中的分布. 植物遗传资源学报, 2009, 10: 21–26 Shi K, Lei C L, Cheng Z J, Xu X T, Wang J L, Wan J M. Distribution of two blast resistance genes Pi-ta and Pi-b in major rice cultivars in China. J Plant Genet Resour, 2009, 10: 21–26 (in Chinese with English abstract) [6] Bryan G T, Wu K S, Farrall L, Jia Y I, Hershey H P, McAdams S A, Fault K N, Donaldson G K, Renato T, Valent B. A single amino acid difference distinguishes resistant and susceptible alleles of the rice blast resistance gene Pi-ta. Plant Cell, 2000, 12: 2033–2045 [7] 陈涛, 骆名瑞, 张亚东, 朱镇, 赵凌, 赵庆勇, 周丽慧, 姚姝, 王才林. 利用四引物扩增受阻突变体系PCR技术检测水稻低直链淀粉含量基因Wx-mq. 中国水稻科学, 2013, 27: 529–534 Chen T, Luo M R, Zhang Y D, Zhu Z, Zhao L, Zhao Q Y, Zhou L H, Yao S, Wang C L. Detection of Wx-mq gene for low-amylose content by tetra-primer amplification refractory mutation system PCR in rice. Chin J Rice Sci, 2013, 27: 529–534 (in Chinese with English abstract) [8] 朱昌兰, 沈文彪, 翟虎渠, 万建民. 水稻低直链淀粉含量基因育种利用的研究进展. 中国农业科学, 2004, 37: 81–88 Zhu C L, Shen W B, Zhai H Q, Wan J M. Progresses in researches of the application of low-amylose content rice gene for breeding. Sci Agric Sin, 2004, 37: 81–88 (in Chinese with English abstract) [9] 姚姝, 陈涛, 张亚东, 朱镇, 赵庆勇, 赵凌, 周丽慧, 王才林. 利用分子标记辅助选择聚合水稻暗胚乳突变基因Wx-mq和抗条纹叶枯病基因Stv-bi. 中国水稻科学, 2010, 24: 341–347 Yao S, Chen T, Zhang Y D, Zhu Z, Zhao Q Y, Zhao L, Zhou L H, Wang C L. Pyramiding of translucent endosperm mutant Wx-mq gene and Stv-bi for resistance to rice stripe virus by Marker-assisted selection in rice (Oryza sativa L.). Chin J Rice Sci, 2010, 24: 341–347 (in Chinese with English abstract) [10] 王才林, 张亚东, 朱镇, 陈涛, 赵凌, 林静. 通过分子标记辅助选择培育优良食味水稻新品种. 中国水稻科学, 2009, 23: 25–30 Wang C L, Zhang Y D, Zhu Z, Chen T, Zhao L, Lin J. Breeding of a new rice variety with good eating quality by marker assisted selection. Chin J Rice Sci, 2009, 23: 25–30 (in Chinese with English abstract) [11] 王才林, 张亚东, 朱镇, 陈涛, 赵庆勇, 赵凌, 周丽慧, 姚姝. 优良食味粳稻新品种南粳5055的选育及利用. 农业科技通讯, 2012, (2): 84–87 Wang C L, Zhang Y D, Zhu Z, Chen T, Zhao Q Y, Zhao L, Zhou L H, Yao S. Breeding and application of a new rice variety Nanjing 5055 with good eating quality. Bull Agric Sci & Technol, 2012, (2): 84–87 (in Chinese) [12] 王才林, 张亚东, 朱镇, 姚姝, 陈涛, 赵庆勇, 周丽慧, 赵凌. 优良食味粳稻新品种南粳9108的选育与利用. 江苏农业科学, 2013, 41(9): 86–88 Wang C L, Zhang Y D, Zhu Z, Yao S, Chen T, Zhao Q Y, Zhou L H, Zhao L. Breeding and application of a new rice variety Nanjing 9108 with good eating quality. Jiangsu Agric Sci, 2013, 41(9): 86–88 (in Chinese) [13] 姚姝, 刘燕清, 张亚东, 朱镇, 陈涛, 赵庆勇,周丽慧, 赵春芳, 于新, 王才林. 水稻抗稻瘟病基因Pi-ta和Pi-b多重PCR体系的构建与应用. 作物学报, 2014, 40: 1565–1571 Yao S, Liu Y Q, Zhang Y D, Zhu Z, Chen T, Zhao Q Y, Zhao L, Zhou L H, Zhao C F, Yu X, Wang C L. Establishment and application of multiplex PCR system for two blast resistance genes Pi-ta and Pi-b in rice. Acta Agron Sin, 2014, 40: 1565–1571 (in Chinese with English abstract) [14] Li W, Lei C L, Cheng Z J, Jia Y L, Huang D Y, Wang J L, Wang J K, Zhang X, Su N, Guo X P, Zhai H Q, Wan J M. Identification of SSR markers for a broad-spectrum blast resistance gene Pi20(t) for marker-assisted breeding. Mol Breed, 2008, 22: 141–149 [15] 王忠华, 贾育林, 吴殿星, 夏英武. 水稻抗稻瘟病基因Pi-ta的分子标记辅助选择. 作物学报, 2004, 30: 1259–1265 Wang Z H, Jia Y L, Wu D X, Xia Y W. Molecular marker-assisted selection of the rice blast resistance gene Pi-ta. Acta Agron Sin, 2004, 30: 1259–1265 (in Chinese with English abstract) [16] Fjellstrom R, Conaway-Bormans C A, McClung A M, Marchetti M A, Shank A R, Park W D. Development of DNA markers suitable for marker assisted selection of three Pi genes conferring resistance to multiple Pyricularia grlsea pathotypes. Crop Sci, 2004, 44: 1790–1798 [17] 刘洋, 徐培洲, 张红宇, 徐建第, 吴发强, 吴先军. 水稻抗稻瘟病Pi-b基因的分子标记辅助选择与应用. 中国农业科学, 2008, 41: 9–14 Liu Y, Xu P Z, Zhang H Y, Xu J D, Wu F Q, Wu X J. Marker-assisted selection and application of blast resistant gene Pi-b in rice. Sci Agric Sin, 2008, 41: 9–14 (in Chinese with English abstract) [18] 罗楚平, 倪磊, 陈志谊, 刘永锋, 刘邮洲, 聂亚锋. 水稻稻瘟病接种技术及2009年江苏省区试品种抗性鉴定. 江苏农业科学, 2009, (6): 178–179 Luo C P, Ni L, Chen Z Y, Liu Y F, Liu Y Z, Ne Y F. Inoculation technology and identification of resistance to rice blast in regional tests in 2009 of Jiangsu Province. Jiangsu Agric Sci, 2009, (6): 178–179 (in Chinese) [19] Huang N, Angeles E R, Domingo J, Magpantay G, Singh S, Zhang G, Kunaravdiel N, Bennett J, Khush G S. Pyramiding of bacterial blight resistance genes in rice: marker-assisted selection using RFLP and PCR. Theor Appl Genet, 1997, 95: 313–320 [20] Sugiura N, Tsuji T, Kato T, Saka N, Touyama T, Hayano S Y, Izawa T. Molecular marker assisted selection in a recurrent backcross breeding for the incorporation of resistance to rice stripe virus and panicle blast in rice (Oryza sativa L.). Breed Res, 2004, 6: 143–148 [21] Narayanan N N, Baisakh N, Cruz C M V, Gnanamanickam S S, Datta K, Datta S K. Molecular breeding for the development of blast and bacterial blight resistance in rice cv.IR50. Crop Sci, 2002, 42: 2072–2079 [22] Jiang G H, Xu C G, Tu J M, Li X H, He Y Q, Zhang Q F. Pyramiding of insect-and disease-resistance genes into an elite indica. cytoplasm male sterile restorer line of rice ‘Minghui 63’. Plant Breed, 2004, 123: 112–116 [23] 倪大虎, 易成新, 李莉, 汪秀峰, 张毅, 赵开军, 王春连, 章琦, 王文相, 杨剑波. 分子标记辅助培育水稻抗白叶枯病和稻瘟病三基因聚合系. 作物学报, 2008, 34: 100–105 Ni D H, Yi X C, Li L, Wang X F, Zhang Y, Zhao K J, Wang C L, Zhang Q, Wang W X, Yang J B. Developing rice lines resistant to bacterial blight and blast with Molecular marker-assisted selection. Acta Agron Sin, 2008, 34: 100–105 (in Chinese with English abstract) [24] 柳武革, 王丰, 金素娟, 朱小源, 李金华, 刘振荣, 廖亦龙, 朱满山, 黄慧君, 符福鸿, 刘宜柏. 利用分子标记辅助选择聚合Pi-1和Pi-2基因改良两系不育瘟病. 作物学报, 2008, 34: 1128–1136 Liu W G, Wang F, Jin S J, Zhu X Y, Li J H, Liu Z R, Liao Y L, Zhu M S, Huang H J, Fu F H, Liu Y B. Improvement of rice blast resistance in TGM S Line by pyramiding of Pi-1and Pi-2 through molecular marker-assisted selection. Acta Agron Sin, 2008, 34: 1128–1136 (in Chinese with English abstract)

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