基于BSA-seq法的水稻稻瘟病抗性基因定位
收稿日期: 2023-12-13
网络出版日期: 2024-11-19
基金资助
宁夏回族自治区自然科学基金(2022AAC03459);农业高质量发展和生态保护科技项目(NGSB-2021-13-3-3);宁夏重点研发计划项目“水稻种质资源耐直播抗逆精准鉴定与创新利用”
Mapping of Rice Blast Resistance Gene Based on BSA-seq
Received date: 2023-12-13
Online published: 2024-11-19
为挖掘水稻稻瘟病抗性相关性状基因位点，于2022—2023年以水稻2013ZJP-3×京宁11杂交得到的重组自交系群体为试验材料，采用人工接种和自然诱发相结合的方法鉴定其对叶瘟、穗颈瘟的抗性，并采用BSA-seq法进行QTL定位分析。表型鉴定结果表明，F11 群体的稻瘟病抗性呈连续性分布，且大部分材料表现为中抗，偏向于抗病亲本京宁11。筛选出19个抗性株系和17个感性株系材料，构建了抗感池。经过测序、变异检测和关联分析，共得到2个与稻瘟病抗性相关的区间，主要分布在5号和7号染色体上，区间总长度为1.45 Mb，共筛选出94个差异基因，其中，移码突变基因12个，非同义突变基因82个，移码和非同义突变基因9个。最终预测，基因LOC_Os05g10630、LOC_Os05g10650和LOC_Os07g08960与水稻稻瘟病抗性的相关性较大。
陈丽, 孙建昌, 王昕 . 基于BSA-seq法的水稻稻瘟病抗性基因定位[J]. 中国稻米, 2024 , 30(6) : 35 -41 . DOI: 10.3969/j.issn.1006-8082.2024.06.006
In order to explore the genetic loci of blast resistance-related traits in rice, the recombinant inbred lines of rice 2013ZJP-3×Jingning 11 hybridization were used as experimental materials. The resistance to leaf blast and neck blast was identified by artificial inoculation and natural induction during 2022—2023, and QTL was analyzed by BSA-seq method. Phenotypic identification showed that the resistance of F11 population to rice blast was distributed continuously, and most materials showed moderate resistance, preferring to the resistant parent jingning 11. 19 resistant lines and 17 sensitive lines were screened to construct the resistant pool. After sequencing, mutation detection and association analysis, a total of 2 regions to blast resistance were obtained, mainly distributed on chromosome 5 and 7, with a total length of 1.45 Mb. 94 different genes, including 12 frame shift mutations, 82 non-synonymous mutations, 9 frame shift and non-synonymous mutations, were screened out. LOC_Os05g10630, LOC_Os05g10650 and LOC_Os07g08960 genes were predicted to have the greatest correlation with blast resistance in rice.
Key words： rice; blast resistance; BSA-seq; gene mapping
| [1] | OU S H. Rice diseases, eds[M]. Kew, Surrey: Common Wealth Mycological Institute, UK, 1985: 109-201. |
| [2] | SKAMNIOTI P, GURR S J. Against the grain: safeguarding rice from rice blast disease[J]. Trends in Biotechnology, 2009, 27(3): 141-150. |
| [3] | MOYTRI R, JIA Y L, RICHARA A C. Structure, function, and blast resistance genes[J]. Acta Agronomica Sinica, 2012, 38(3): 381-393. |
| [4] | LIANG T M, GUO X R, CHEN Z Q, et al. Identification and gene mapping of a blast disease resistance gene in rice line IR65482[J]. Molecular Plant Breeding, 2018, 16(13): 4 308-4 313. |
| [5] | DENG Y W, ZHAI K R, XIE Z, et al. Epigenetic regulation of antagonistic receptors confers rice blast resistance with yield balance[J]. Science, 2017, 355(6328): 962-965. |
| [6] | 杨小林, 戚华雄, 殷得所, 等. 水稻稻瘟病抗性 QTL 的定位分析[J]. 植物病理学报, 2012, 42(6):600-607. |
| [7] | 刘文强, 李小湘, 王淑红, 等. 水稻稻瘟病抗性基因研究进展[J]. 杂交水稻, 2009, 24(3):1-7. |
| [8] | YANG H M, JIA M H, JIA Y L, et al. Molecular mappig of four blast resistance genes using recombinant inbred line of 93-11 and Nipponbare[J]. Plant Biology, 2013, 56(4): 91-97. |
| [9] | RAHIM H A, BHUIYAN M A, LIM L S, et al. Identification of quantitative trait loci for blast resistance in BC2F3 and BC2F5 advanced backcross families of rice[J]. Genetics and Molecular Research, 2012, 11(3): 3 277-3 289 |
| [10] | KARAMIAN F, NEZHAD A, ZAEIM A N, et al. Blast disease in rice: A review[J]. International Journal of Scientific Research in Science and Technology, 2015, 5(3): 228-232. |
| [11] | 刘佳, 李志, 高冠军, 等. 利用高世代回交群体定位水稻光谱抗稻瘟病 QTL[J]. 分子植物育种, 2015, 13(10):2155-2 162. |
| [12] | 辛威. 寒地粳稻种质资源稻瘟病抗性鉴定及基因定位[D]. 哈尔滨: 东北农业大学, 2017. |
| [13] | 杨德卫, 王莫, 韩利波, 等. 水稻稻瘟病抗性基因的克隆、育种利用及稻瘟菌无毒基因研究进展[J]. 植物学报, 2019, 54(2):265-276. |
| [14] | 李仕贵, 王玉平, 黎汉云, 等. 利用微卫星标记鉴定水稻的稻瘟病抗性[J]. 生物工程学报, 2000, 16(3):324-327. |
| [15] | 董巍, 李信, 晏斌, 等. 利用分子标记辅助选择改良培矮64S的稻瘟病抗性[J]. 分子植物育种, 2010, 8(5):853-860. |
| [16] | 文绍山, 高必军. 利用分子标记辅助选择将抗稻瘟病基因Pi-9( t)渗入水稻恢复系泸恢17[J]. 分子植物育种, 2012, 10 (1):42-47. |
| [17] | 潘素君. 广谱抗稻瘟病基因Pi9的应用和进化研究[D]. 长沙: 湖南农业大学, 2006. |
| [18] | DEAN R, VANKAN J A L, PRETORIUS Z A, et al. The top10 fungal pathogens in molecular plant pathology[J]. Molecular Plant Pathology, 2012, 13(4): 414-430. |
| [19] | 王云锋, 王长秘, 李春琴, 等. 稻瘟病菌侵染时水稻防御体系对外源茉莉酸的响应分析[J]. 南方农业学报, 2018, 49(7):1324-1 331. |
| [20] | 农业部种子管理局. NY/T 2646—2014,水稻品种试验稻瘟病抗性鉴定与评价技术规程[S]. 北京: 中华人民共和国农业部,2014. |
| [21] | MURRAY M G, THOMPSON W F. Rapid isolation of high molecular weight plant DNA[J]. Nucleic Acids Research, 1980, 8(19): 4 321-4 326. |
| [22] | MUDALKAR S, GOLLA R, GHATTY S, et al. De novo transcriptome analysis of an imminent biofuel crop, Camelina sativa, L. using Illumina GAIIX sequencing platform and identification of SSR markers[J]. Plant Molecular Biology, 2014, 84(1/2): 159-171. |
| [23] | International Rice Genome Sequencing Project, SASAKI T. The map-based sequence of the rice genome[J]. Nature, 2005, 436: 793-800. |
| [24] | ALTSCHUL S F, GISH W R, MILLER W B, et al. Basic local alignment search tool (BLAST)[J]. Journal of Molecular Biology, 2012, 215(3): 403-410. |
| [25] | LI H, DUR B R. Fast and accurate short read alignment with Burrows-Wheeler transform[J]. Bioinformatics, 2009, 25(14): 1 754-1 760. |
| [26] | TAKAGI H, ABE A, YOSHIDA K, et al. QTL-seq: Rapid mapping of quantitative trait loci in rice by whole genome resequencing of DNA from two bulked populations[J]. Plant Journal, 2013, 74: 174-183. |
| [27] | RYM F, HIROKI T, MULUNEH T, et al. MutMap+: Genetic mapping and mutant identification without crossing in rice[J]. PLoS One, 2013, 8(7): e68529. |
| [28] | HAN Y C, LV P, HOU S L, et al. Combining next generation sequencing with bulked segregant analysis to fine map a stem moisture locus in sorghum (Sorghum bicolor L. Moench)[J]. PLoS One, 2015, 10: e0127065. |
| [29] | LUO X D, LIU J, ZHAO J, et al. Rapid mapping of candidate genes for cold tolerance in Oryza rufipogon Griff.by QTL-seq of seedlings[J]. Journal of Integrative Agriculture, 2018, 17(2): 265-275. |
| [30] | CHEN S, WANG L, QUE Z Q, et al. Genetic and physical mapping of Pi37(t), a new gene conferring resistance to rice blast in the famous cultivar St. No.1[J]. Theoretical and Applied Genetics, 2005, 111(8): 1 563-1 570. |
| [31] | YOU Q Y, ZHAI K R, YANG D L, et al. An E3 ubiquitin ligase-BAG protein module controls plant innate immunity and broad-spectrum disease resistance[J]. Cell Host Microbe, 2016, 20(6): 758-769. |
| [32] | LIU D F, CHEN X J, LIU J Q, et al. The rice ERF transcription factor OsERF922 negatively regulates resistance to Magnaporthe oryzae and salt tolerance[J]. Journal of Experimental Botany, 2012, 63(10) : 3 899-3 911. |
| [33] | QIU D Y, XIAO J, DING X H, et al. OsWRKY13 mediates rice disease resistance by regulating defense-related genes in salicylate- and jasmonate-dependent signaling[J]. Molecular Plant-Microbe Interactions, 2007, 20(5): 492-499. |
| [34] | ABBRUSCATO P, NEPUSZ T, MIZZI L, et al. OsWRKY22, a monocot WRKY gene, plays a role in the resistance response to blast[J]. Molecular Plant Pathology, 2012, 13(8): 828-41. |
| [35] | LIN F, CHEN S, QUE Z Q, et al. The blast resistance gene Pi37 encodes a nucleotide binding site-leucine-rich repeat protein and is a member of a resistance gene cluster on rice chromosome 1[J]. Genetics, 2007, 177(3): 1 871-1 880. |
| [36] | TAKAHASHI A, HAYASHI N, MIYAO A, et al. Unique features of the rice blast resistance Pish locus revealed by large scale retrotransposon-tagging[J]. BMC Plant Biology, 2010,10: 175. |
| [37] | LIU X L, WANG L, WANG X W, et al. Mutation of the chloroplast-localized phosphate transporter OsPHT2;1 reduces flavonoid accumulation and UV tolerance in rice[J]. The Plant Journal, 2020, 102(1): 53-67. |
| [38] | HE S L, JIANG J Z, CHEN B H, et al. Overexpression of a constitutively active truncated form of OsCDPK1 confers disease resistance by affecting OsPR10a expression in rice[J]. Scientific Reports, 2018, 8: 403. |
/
| 〈 |
|
〉 |