基于RIL群体鉴定棉花抗黄萎病相关QTLs
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摘要
【目的】鉴定出能够稳定表达的棉花抗黄萎病相关数量性状位点(Quantitative trait loci,QTLs)。【方法】以抗落叶型黄萎病棉花品种常抗棉和感黄萎病品种TM-1为亲本配制的111个重组自交系家系为作图群体,筛选出多态性简单序列重复(Simple sequence repeat,SSR)标记,并用于构建遗传图谱。用完备复合区间作图法对该群体在安阳大田、新疆重病地及病圃等多个环境下的黄萎病病情指数进行QTLs检测。【结果】构建了1张含有12个连锁群、40个标记、总长212.5 cM(厘摩)的遗传图谱。获得了6个与抗黄萎病基因相关的QTLs,对数优势比(Logarithm of the odd score,LOD)分布在2.51～5.55,贡献率最大为20.34%,最小为6.93%。其中,qVR-D05-1能够在安阳大田2015年7月15日和新疆南疆重病地2016年7月9日2个环境中检测到,贡献率分别为12.96%和20.34%。【结论】本研究得到的qVR-D05-1能够为定位出稳定的棉花抗黄萎病相关QTLs提供参考。
[Objective] The objective of this study was to identify the stable quantitative trait loci(QTLs) related to Verticillium wilt resistance in cotton. [Method] In this study, a population of 111 recombinant inbred lines(RILs) strains was developed by crossing a highly resistant parental line "Changkangmian" to Verticillium wilt and the susceptible parent TM-1. The complete composite interval mapping method was adopted to detect QTLs by Verticillium wilt disease index in multiple environmental conditions and periods in Anyang and Verticillium wilt affected areas of Xinjiang. Simple sequence repeat(SSR) markers of polymorphism were screened for genetic mapping. [Result] The genetic map was constructed by 40 simple sequence repeat(SSR) markers, consisted of 12 linkage groups with total length of 212.5 centimorgan(cM). A total of six QTLs related to the resistance to Verticillium wilt were obtained. The likelihood of odd(LOD) values ranged from 2.51 to 5.55. The maximum phenotypic variation explained(PVE) 20.34%, and the minimum PVE 6.93% were recorded. Among detected QTLs,q VR-D05-1 was detected in both Verticillium wilt affected fields in Anyang and Xinjiang in July 2015 and July 2016 with PVE of 12.96% and 20.34%, respectively. [Conclusion] This study can provide a potential reference for mapping stable QTLs related to resistance to Verticillium wilt.
[Objective] The objective of this study was to identify the stable quantitative trait loci(QTLs) related to Verticillium wilt resistance in cotton. [Method] In this study, a population of 111 recombinant inbred lines(RILs) strains was developed by crossing a highly resistant parental line "Changkangmian" to Verticillium wilt and the susceptible parent TM-1. The complete composite interval mapping method was adopted to detect QTLs by Verticillium wilt disease index in multiple environmental conditions and periods in Anyang and Verticillium wilt affected areas of Xinjiang. Simple sequence repeat(SSR) markers of polymorphism were screened for genetic mapping. [Result] The genetic map was constructed by 40 simple sequence repeat(SSR) markers, consisted of 12 linkage groups with total length of 212.5 centimorgan(cM). A total of six QTLs related to the resistance to Verticillium wilt were obtained. The likelihood of odd(LOD) values ranged from 2.51 to 5.55. The maximum phenotypic variation explained(PVE) 20.34%, and the minimum PVE 6.93% were recorded. Among detected QTLs,q VR-D05-1 was detected in both Verticillium wilt affected fields in Anyang and Xinjiang in July 2015 and July 2016 with PVE of 12.96% and 20.34%, respectively. [Conclusion] This study can provide a potential reference for mapping stable QTLs related to resistance to Verticillium wilt.
引文
[1]汪若海,承泓良,宋晓轩.中国棉史概述[M].北京:中国农业科学技术出版社, 2017.Wang Ruohai, Cheng Hongliang, Song Xiaoxuan. History of China's cotton overview[M]. Beijing:China Agricultural Science and Technology Press, 2017.
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[24]宋国立,崔荣霞,王坤波,等.改良CTAB法快速提取棉花DNA[J].棉花学报, 1998, 10(5):273-275.Song Guoli, Cui Rongxia, Wang Kunbo, et al. A rapid improved CTAB method for extraction of cotton genomic DNA[J]. Cotton Science, 1998, 10(5):273-275.
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[2]简桂良,邹亚飞,马存.棉花黄萎病连年流行的原因及对策[J].中国棉花, 2003, 30(3):13-14.Jian Guiliang, Zou Yafei, Ma Cun. The outbreak reason and integral control of cotton Verticillium wilt[J]. China Cotton, 2003,30(3):13-14.
[3]汪敏,焦睿,邢小萍,等.棉花黄萎病菌致病的分子机理[J].棉花学报, 2011, 23(3):272-278.Wang Min, Jiao Rui, Xing Xiaoping, et al. The molecular mechanism of pathogenicity for Verticillium dahlia Kleb.[J]. Cotton Science, 2011, 23(3):272-278.
[4]李淑英,吴德祥,路曦结,等.棉花黄萎病致病机理及抗病育种研究进展[J].安徽农业科学, 2001, 29(4):477-479.Li Shuying, Wu Dexiang, Lu Xijie, et al. Advanced researches on the mechanism of virulence of Verticillium dahliae and resistance breeding in cotton[J]. Journal of Anhui Agricultural Sciences, 2001, 29(4):477-479.
[5]潘家驹,张天真,蒯本科,等.棉花黄萎病抗性遗传研究[J].南京农业大学学报, 1994, 12(3):8-18.Pan Jiaju, Zhang Tianzhen, Kuai Benke, et al. Studies on the inheritance of resistance to Verticillium dahliae in cotton[J]. Journal of Nanjing Agricultural University, 1994, 12(3):8-18.
[6]徐理,朱龙付,张献龙.棉花抗黄萎病机制研究进展[J].作物学报, 2012, 38(9):1553-1560.Xu Li, Zhu Longfu, Zhang Xianlong. Research on resistance mechanism of cotton to Verticillium wilt[J]. The Crop Journal,2012, 38(9):1553-1560.
[7]房卫平.棉花黄萎病抗性机制、遗传及分子标记的研究[D].杭州:浙江大学, 2001.Fang Weiping. Studies on the mechanism, inheritance and molecular markers of cotton resistance to Verticillium dahliae[D]. Hangzhou:Zhejiang University, 2001.
[8]高玉千,聂以春,张献龙.棉花抗黄萎病基因的QTL定位[J].棉花学报, 2003, 15(2):73-78.Gao Yuqian, Nie Yichun, Zhang Xianlong. QTL mapping of genes resistant to Verticillium wilt in cotton[J]. Cotton Science,2003, 15(2):73-78.
[9] Said J I, Lin Z X, Zhang X L, et al. A comprehensive meta QTL analysis for fiber quality, yield, yield related and morphological traits, drought tolerance, and disease resistance in tetraploid cotton[J]. BMC Genomics, 2013, 14(1):776. http://dx.doi.org/10.1186/1471-2164-14-776
[10] Munis M F, Tu L L, Deng F L, et al. A thaumatin-like protein gene involved in cotton fiber secondary cell wall development enhances resistance against Verticillium dahliae and other stresses in transgenic tobacco[J]. Biochemical&Biophysical Research Communications, 2010, 393(1):38-44.
[11]肖松华,赵君,刘剑光,等.转天麻抗真菌蛋白基因提高棉花对黄萎病抗性[J].作物学报, 2016, 42(2):212-221.Xiao Songhua, Zhao Jun, Liu Jianguang, et al. Transgenic upland cotton lines of Gastrodia antifungal protein gene and their performance of resistance to Verticillium wilt[J]. The Crop Journal, 2016, 42(2):212-221.
[12]邵冰欣.基于转录组测序筛选及克隆棉花抗黄萎病相关基因[D].北京:中国农业科学院, 2015.Shao Bingxin. Screening and cloning resistance genes to cotton Verticillium wilt based on the transcriptome sequencing[D].Beijing:Chinese Academy of Agricultural Sciences, 2015.
[13] Bidochka M J, Burke S, Ng L. Extracellular hydrolytic enzymes in the fungal genus Verticillium:adaptations for pathogenesis[J]. Canadian Journal of Microbiology, 1999, 5(10):856-864.
[14]肖红利.棉花组织诱导体系中大丽轮枝菌分泌蛋白分析及其致病性研究[D].北京:中国农业科学院, 2014.Xiao Hongli. Secretome analysis and pathogenicity study of Verticillium dahliae in cotton-extract induction system[D]. Beijing:Chinese Academy of Agricultural Sciences, 2014.
[15]王红梅.棉花抗黄萎病遗传及分子标记研究[D].武汉:华中农业大学, 2005.Wang Hongmei. Study on the inheritance and QTLs mapping of Verticillium wilt in cotton[D]. Wuhan:Huazhong Agricultural University, 2015.
[16] Wang H M, Lin Z X, Zhang X L, et al. Mapping and quantitative trait loci analysis of Verticillium wilt resistance genes in cotton[J]. Journal of Integrative Plant Biology, 2008, 50(2):174-182.
[17]杜威世,杜雄明,马峙英.棉花黄萎病抗性基因SSR标记研究[J].西北农林科技大学学报(自然科学版), 2004, 32(3):20-24.Du Weishi, Du Xiongming, Ma Zhiying. Studies on SSR markers of resistance gene of Verticillium wilt in cotton[J]. Journal of Northwest Sci-Tech University of Agriculture and Forestry(Natural Science Edition), 2004, 32(3):20-24.
[18]葛海燕,汪业春,郭旺珍,等.陆地棉抗黄萎病性状的遗传及分子标记研究[J].棉花学报, 2008, 20(1):19-22.Ge Haiyan, Wang Yechun, Guo Wangzhen, et al. Inheritance and molecular tagging of resistance against Verticillium wilt in upland cotton[J]. Cotton Science, 2008, 20(1):19-22.
[19] Zhang J F, Yu J W, Pei W F, et al. Genetic analysis of Verticillium wilt resistance in a backcross inbred line population and a meta-analysis of quantitative trait loci for disease resistance in cotton[J]. BMC Genomics, 2015, 16(1):577. https://doi.org/10.1186/s12864-015-1682-2
[20]蒋锋.陆地棉黄萎病抗性的遗传分析与QTLs定位[D].南京:南京农业大学, 2008.Jiang Feng. Inheritance and QTLs analysis for resistance to Verticillium wilt in upland cotton[D]. Nanjing:Nanjing Agricultural University, 2008.
[21]孔祥瑞.棉花黄萎病抗性的分子标记研究[D].北京:中国农业科学院, 2011.Kong Xiangrui. Study on cotton Verticillium wilt resistance by molecular marker technology[D]. Beijing:Chinese Academy of Agricultural Sciences, 2011.
[22]王芙蓉,刘任重,王留明,等.陆地棉品种抗黄萎病性状的分子标记及其辅助选择效果[J].棉花学报, 2007, 19(6):424-430.Wang Furong, Liu Renzhong, Wang Liuming, et al. Molecular markers of Verticillium wilt resistance in upland cotton(Gossypium hirsutum L.)cultivar and their effects on assisted phenotypic selection[J]. Cotton Science, 2007, 19(6):424-430.
[23]杨昶,郭旺珍,张天真.陆地棉抗黄萎病、纤维品质和产量等农艺性状的QTL定位[J].分子植物育种, 2007, 5(6):797-805.Yang Chang, Guo Wangzhen, Zhang Tianzhen. QTL mapping for resistance to Verticillium wilt, fiber quality and yield traits in upland cotton(Gossypium hirsutum L.)[J]. Molecular Plant Breeding, 2007, 5(6):797-805.
[24]宋国立,崔荣霞,王坤波,等.改良CTAB法快速提取棉花DNA[J].棉花学报, 1998, 10(5):273-275.Song Guoli, Cui Rongxia, Wang Kunbo, et al. A rapid improved CTAB method for extraction of cotton genomic DNA[J]. Cotton Science, 1998, 10(5):273-275.
[25]中国农业部.棉花抗病虫性评价技术规范第5部分:黄萎病:GB/T 22101.5―2009[S].北京:中国标准出版社, 2009.Ministry of Agriculture, China. Technical specification for evaluating resistance of cotton to diseases and insect pests Part 5:Verticillium wilt[S]. Beijing:Standard Publishing House of China, 2009.
[26] Zhu T, Liang C Z, Meng Z G, et al. CottonFGD:an integrated functional genomics database for cotton[J]. BMC Plant Biology,2017, 17(1):101. http://dx.doi.org/10.1186/s12870-017-1039-x
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