Construction and Genetic Background Recovery of Blast-resistant Near-isogenic Lines from Japonica Variety, Yunyin
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摘要: 通过对稻瘟病抗性近等基因系的构建和评价，为抗稻瘟病基因的抗性分析和遗传学研究提供良好载体。分别以广谱抗稻瘟病品种云引和普感稻瘟病品种丽江新团黑谷(LTH)为抗病基因的供体亲本和轮回亲本，结合分子标记辅助选择和人工接种抗性鉴定，经连续4代回交和8代自交，获得7个以LTH为遗传背景的稻瘟病抗性近等基因系。选取均匀分布于水稻12条染色体上的184对SSR标记，对所获得的近等基因系进行遗传背景分析、评价。结果表明：在亲本云引和LTH之间具多态性的SSR引物有72对，标记多态性频率为39.1%。以在亲本间具有多态性的标记对7个近等基因系与LTH间的遗传多态性进行分析，发现其中W15、W17的遗传背景恢复率为97.61%，W18的遗传背景恢复率为97.22%，其他4个株系的遗传背景恢复率均大于98%。成功构建并获得了以LTH为背景的含有云引稻瘟病抗性基因的近等基因系，且近等位基因系的遗传背景与表型值均恢复良好。Abstract: Construction of near-isogenic lines(NILs) is an effective mean for analyzing blast-resistance genes as well as genetic research in general. This study applied the broad-spectrum blast-resistant japonica variety, Yunyin, as the donor parent and the blast-susceptible variety, LTH, as the recurrent parent. Combining the molecular markers assisted selection and artificial inoculation, 7 blast-resistant NILs were bred with LTH for the genetic background analysis after 4 generations of backcrossing and 8 generations of self-crossing.184 pairs of SSR markers evenly distributed on 12 chromosomes of the rice were selected to evaluate the genetic background of the NILs. The results showed that 72 pairs of SSR primers to be polymorphic with a frequency of 39.1%. The polymorphisms between 7 NILs and LTH were analyzed by markers among the parents. The background recovery rates of W15 and W17 were 97.61%, that of W18, 97.22%, and those of other 4 lines, greater than 98%. It appeared that the construction of NILs carrying Yunyin blast-resistance gene with LTH as background were successful, and that all phenotypic indices of the NILs recovered well in this study.
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Key words:
- rice /
- rice blast-resistance /
- near-isogenic lines /
- genetic background
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图 1 近等基因系中目标基因分子标记检测
注：M为20bp DNA marker；1为供体亲本云引；2为轮回亲本LTH；3~9为田间接种抗病近等基因系，对应编号为：W13、W14、W15、W17、W18、W19、W22；10为感病近等基因系S1；11为阴性对照(水)。
Figure 1. Detection of molecular markers of target genes in NILs
图 2 供体亲本云引、受体亲本LTH及其近等基因系W22的株型表现
Figure 2. Plant types of donor parent Yunyin, recipient parent LTH and their NIL W22
图 3 用SSR标记对近等基因系进行遗传背景分析
注：M为20 bp marker，A1~A12、B1、B2均为水稻品种鉴定技术规程(NY/T 1433-2014)中的引物。样品顺序为LTH、云引、分子标记为阳性株、分子标记为阴性株。
Figure 3. Genetic background analysis of NILs by SSR markers
表 1 近等基因系和亲本的人工注射接菌及分子标记检测
Table 1. Artificial inoculation and molecular marker detection of NILs and parents
品种(系) 注射接菌结果(抗性) 分子检测结果 云引 高抗(HR) + LTH 高感(HS) - W13 抗病(R) + W14 高抗(R) + W15 抗病(R) + W17 抗病(R) + W18 中抗(MR) + W19 中抗(MR) + W22 抗病(R) + S1 高感(HS) - 注：①人工注射接菌时对接种叶片造成了伤口，病情略微偏重，本研究将调查结果中抗及以上的均记为抗病表型；②+表示阳性，-表示阴性。 
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表 2 供体亲本、受体亲本及近等基因系材料的主要农艺性状
Table 2. Agronomic traits of donor parents, receptor parents and NILs
品种(系) 株高/cm 分蘖数/个 穗长/cm 株总粒数/粒 株实粒数/粒 穗总粒数/粒 穗实粒数/粒 结实率/% 千粒重/g 云引 128.6 12.7 22.4 1967.3 1678.0 154.6 131.9 85.3 29.3 LTH 134.7 18.3 21.1 2584.6 2149.7 141.0 117.2 83.1 26.6 W13 133.2 17.6 22.2 2306.8 1923.4 130.5 108.8 83.3 27.8 W14 134.5 19.3 23.0 2798.6 2302.2 144.7 119.1 82.2 27.2 W15 131.2 18.6 20.5 2524.6 2083.8 135.2 111.6 82.5 26.7 W17 135.4 17.8 20.8 2396.3 2009.0 133.9 112.3 83.8 27.1 W18 131.9 18.3 21.4 2448.6 2035.7 133.5 111.0 83.1 26.9 W19 135.8 17.9 22.5 2502.9 2062.5 139.5 115.0 82.4 27.0 W22 130.9 18.6 21.1 2675.0 2209.9 143.2 118.3 82.6 27.1
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表 3 近等基因系与轮回亲本LTH之间的多态性标记及其遗传背景恢复率
Table 3. Genetic background recovery rates and polymorphic markers between NILs and LTH
品系 多态性标记数 恢复率/% W13 2 98.61 W14 2 98.61 W15 3 97.61 W17 3 97.61 W18 4 97.22 W19 1 99.31 W22 1 99.31
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[1] 薛恒.米业产业化经营:米业产品的营销创新[J].中国稻米, 2003, 9(5):28-29. doi: 10.3969/j.issn.1006-8082.2003.05.016 [2] 刘萌.培育水稻空育131BL多系品种供体亲本抗稻瘟病性鉴定及目的基因分子标记分析[D].哈尔滨: 黑龙江大学, 2012. [3] 马玉银, 左示敏, 张再金, 等.水稻近等基因系构建及其应用[J].安徽农业科学, 2008, 36(17):7167-7168. doi: 10.3969/j.issn.0517-6611.2008.17.049 [4] 章琦, 杨文才, 施爱农, 等. 3个粳稻抗白叶枯病近等基因系的构建[J].作物学报, 1998, 24(6):799-804. doi: 10.3321/j.issn:0496-3490.1998.06.026 [5] 褚晋, 何鹏飞, 赵正龙, 等. 3套水稻近等基因系对两个病圃稻瘟病菌的抗性评价[J].江西农业学报, 2014, 26(7):50-54. doi: 10.3969/j.issn.1001-8581.2014.07.014 [6] 朱小源, 杨祁云, 杨健源, 等.抗稻瘟病单基因系对籼稻稻瘟病菌小种鉴别力分析[J].植物病理学报, 2004, 34(4):361-368. doi: 10.3321/j.issn:0412-0914.2004.04.013 [7] 赵霏, 任三娟, 郭泽建, 等.利用近等基因系研究豌豆铁蛋白基因对水稻重要生物学特性的影响[J].农业生物技术学报, 2011, 19(1):63-68. doi: 10.3969/j.issn.1674-7968.2011.01.009 [8] 凌忠专, 蒋琬如, 王久林, 等.水稻品种丽江新团黑谷普感特性的研究和利用[J].中国农业科学, 2001, 34(1):116-116. doi: 10.3321/j.issn:0578-1752.2001.01.025 [9] TEMNYKH S, DeCLERK G, LUKASHOVA A, et al. Computational and experimental analysis of microsatellites in rice (Oryza sativa L.):frequency, length variation, transposon associations, and genetic marker potential[J]. Genome Research, 2001, 11(8):1441-1452. doi: 10.1101/gr.184001 [10] 中华人民共和国农业部.水稻品种鉴定技术规程SSR标记法: NY/T 1433-2014[S].北京: 中国标准出版社, 2014: 9-13. [11] Cold Spring Harbor Laboratory, Oregon State University, EMBL-EBI. SSR Markers Resource-Microsatellite Markers[R/OL].(2016-08-26)[2018-06-13].http://www.gramene.org/microsat/ssr.html [12] 陈锦文, 朱永生, 张建福, 等.四个杂交水稻骨干恢复系的多态性分析[J].福建农业学报, 2014, 29(4):319-323. doi: 10.3969/j.issn.1008-0384.2014.04.005 [13] 阳海宁, 韦绍丽, 李孝琼, 等.标记辅助培育水稻抗稻褐飞虱和稻白叶枯病基因聚合系[J].分子植物育种, 2010, 8(1):11-19. http://d.old.wanfangdata.com.cn/Periodical/fzzwyz201001004 [14] 沈新莲, 张天真.作物分子标记辅助选择育种研究的进展与展望[J].高技术通讯, 2003, 13(2):105-110. doi: 10.3321/j.issn:1002-0470.2003.02.023 [15] XU Y, CROUCH J H. Marker-assisted selection in plant breeding from publication to practice[J]. Crop science, 2008, 48(2):391-407. doi: 10.2135/cropsci2007.04.0191 [16] LUO Y C, SANGHA J S, WANG S H, et al. Marker-assisted breeding of Xa4, Xa21 and Xa27 in the restorer lines of hybrid rice for broad-spectrum and enhanced disease resistance to bacterial blight[J]. Molecular Breeding, 2012, 30(4):1601-1610. doi: 10.1007/s11032-012-9742-7 [17] 郑康乐, 黄宁.标记辅助选择在水稻改良中的应用前景[J].遗传, 1997, 19(2):40-44. http://www.wanfangdata.com.cn/details/detail.do?_type=perio&id=QK199700879646 -
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