Effects of Pyramiding of Two Rice Blast Resistance Genes on Resistance of Rice Cultivar ‘Yunjing 37’

doi:10.11924/j.issn.1000-6850.casb2024-0486

Abstract

Abstract:

To improve the resistance of rice cultivar ‘Yunjing 37’ to Manaporthe oryzae, the rice cultivar ‘Yunjing 37’ was used as the recurrent parent. By employing a combination of conventional hybridization, backcrossing, selfing, and marker-assisted selection (MAS) strategies, the blast resistance genes Pi57 and Pi69 were pyramided into the japonica cultivar ‘Yunjing 37’. M. oryzae strains virulent to ‘Yunjing 37’ were used to evaluate the blast resistance of BC₂F₃ lines containing homozygous Pi57+Pi69, Pi57 or Pi69, as well as their parental materials, through artificial spray inoculation at the seedling stage. Results showed that plants possessing homozygous Pi57+Pi69 were resistant to all blast strains used in this study, while those lines containing only homozygous Pi57 or Pi69 exhibited resistance or susceptible reactions consistent with their donor parents. These results indicated that pyramiding of blast resistance genes can significantly enhance the resistance of rice varieties against M. oryzae. The newly developed materials will provide valuable germplasm resources for blast-resistant rice breeding.

Key words: Oryza sativa L., Magnaporthe oryzae, resistance genes, Pi57, Pi69, gene-pyramiding, evaluation

DONG Liying, XIANG Jinqian, LIU Pei, LIU Shufang, LI Qicheng, CHEN Jianchen, YANG Qinzhong. Effects of Pyramiding of Two Rice Blast Resistance Genes on Resistance of Rice Cultivar ‘Yunjing 37’[J]. Chinese Agricultural Science Bulletin, 2025, 41(24): 98-103.

Figures/Tables 5

References 29

[1]	COUCH B C, HOHN L M. A multilocus gene genealogy concordant with host preference indicates segregation of a new species, Magnaporthe oryzae, from M. grisea[J]. Mycologia, 2002,94:683-693.
[2]	ASHKANI S, YUSOP M R, SHABANIMOFRAD M, et al. Allele mining strategies: principles and utilisation for blast resistance genes in rice (Oryza sativa L.)[J]. Current issues in molecular biology, 2015,17:57-74.
[3]	WILSON R A, TALBOT N J. Under pressure: investigating the biology of plant infection by Magnaporthe oryzae[J]. Nature reviews microbiology, 2009, 7:185-195.
[4]	KALIA S, RATHOUR R. Current status on mapping of genes for resistance to leaf and neck-blast disease in rice[J]. 3 Biotech, 2019,9:209.
[5]	ARUNAKUMARI K, DURGARANI C V, SATTURU V, et al. Marker-assisted pyramiding of genes conferring resistance against bacterial blight and blast diseases into Indian rice variety MTU1010[J]. Rice science, 2016, 23(6):306-316. doi: 10.1016/j.rsci.2016.04.005
[6]	WING R A, PURUGGANAN M D, ZHANG Q. The rice genome revolution: from an ancient grain to green super rice[J]. Nature reviews genetics, 2018, 19:505-517. doi: 10.1038/s41576-018-0024-z pmid: 29872215
[7]	KHUSH G S, JENA K K. Current status and future prospects for research on blast resistance in rice (Oryza sativa L.). In: WANG G L, VALENT B (eds) Advances in genetics, genomics and control of rice blast disease[M]. Dordrecht: Springer, 2009:1-10.
[8]	DENG Y, ZHAI K, XIE Z, et al. Epigenetic regulation of antagonistic receptors confers rice blast resistance with yield balance[J]. Science, 2017,355:962-965.
[9]	杨瑰丽, 张瑞祥, 王慧, 等. 利用分子标记辅助选择改良水稻保持系香味和稻瘟病抗性[J]. 华南农业大学学报, 2022, 43(3):9-17.
[10]	HITTALMANI S, PARCO A, MEW T V, et al. Fine mapping and DNA marker-assisted pyramiding of the three major genes for blast resistance in rice[J]. Theoretical and applied genetics, 2000, 100(7):1121-1128.
[11]	陈学伟, 李仕贵, 马玉清, 等. 水稻抗稻瘟病基因Pi-d(t)¹、Pi-b、Pi-ta²的聚合及分子标记选择[J]. 生物工程学报, 2004, 20(5):708-714.
[12]	陈建民, 付志英, 权宝权, 等. 分子标记辅助培育双抗稻瘟病和白叶枯病杂交稻恢复系[J]. 分子植物育种, 2009, 7(3):465-470.
[13]	田红刚, 陈红旗, 胡江, 等. 分子标记转育3个基因改良空育131稻瘟病抗性[J]. 核农学报, 2016, 30(11):2096-2103. doi: 10.11869/j.issn.100-8551.2016.11.2096
[14]	廖常青, 陈英, 向聪, 等. 分子标记选择改良水稻两系不育系创5S稻瘟病抗性[J]. 作物研究, 2018, 32(4):271-275,282.
[15]	白玉路, 王平, 张琼, 等. 水稻骨干恢复系川恢907抗稻瘟病基因聚合与精准改良[J]. 西南农业学报, 2019, 32(5):947-951.
[16]	王雨, 孙全翌, 杜海波, 等. 利用抗稻瘟病基因Pigm和抗纹枯病数量性状基因qSB-9^TQ、qSB-11^HJX改良南粳9108的抗性[J]. 中国水稻科学, 2023, 37(2):125-132. doi: 10.16819/j.1001-7216.2023.220413
[17]	李全衡, 陈洁, 郑智, 等. 云南省水稻品种科研育种及应用[J]. 中国种业, 2021,12:27-30.
[18]	SALEH D, MILAZZO J, ADREIT H, et al. South-East Asian is the center of origin, diversity and dispersion of the rice blast fungus, Magnaporthe oryzae[J]. New phytologist, 2014,201:1440-1456.
[19]	陈越, 陈玲, 张敦宇, 等. 云南水稻种质资源的遗传多样性及群体结构分析[J]. 西北农业学报, 2019, 28(11):1778-1789.
[20]	董丽英, 王群, 刘树芳, 等. 云南省稻瘟病菌群体对稻瘟病抗性单基因系的致病性分析[J]. 西南农业学报, 2012,25:467-473.
[21]	董丽英, 李国生, 赵秀兰, 等. 云南粳稻品种(系)对稻瘟病的抗性鉴定与评价[J]. 西南农业学报, 2018, 31(12):2458-246.
[22]	周伍民, 董丽英, 刘树芳, 等. 抗稻瘟病基因特异分子标记开发及在云南粳稻中的应用[EB/OL]. 分子植物育种, 2021-09-29, https://kns.cnki.net/kcms/detail/46.1068.s.20210928.0151.008.html.
[23]	董丽英, 杨勤忠, 刘树芳, 等. 一种稻瘟病抗性基因Pi57及其编码蛋白与应用[P].中国专利: 201811221584.X, 2019-06-11.
[24]	杨勤忠, 董丽英, 刘树芳, 等. 一种抗稻瘟病基因Pi69及其编码蛋白及应用[P]. 中国专利: 202211195864.4, 2023-06-23.
[25]	WARUDE D, CHAVAN P, JOSHI K, et al. DNA isolation from fresh, dry plant samples with highly acidic tissue extracts[J]. Plant molecular biology reporter, 2003, 21(4):467.
[26]	HOSPITAL F, CHEVALET C, MULSANT P. Using markers in gene introgression breeding programs[J]. Genetics, 1992,132:1199-1210.
[27]	马作斌, 赵家铭, 崔月峰, 等. 分子标记辅助选育抗稻瘟病水稻新品种‘铁粳16’[J]. 分子植物育种, 2021, 19(2):512-517.
[28]	于苗苗, 戴正元, 潘存红, 等. 广谱稻瘟病抗性基因Pigm和Pi2的抗谱差异及与Pi1的互作效应[J]. 作物学报, 2013, 39(11):1927-1934. doi: 10.3724/SP.J.1006.2013.01927
[29]	唐清杰, 韩义胜, 严小微. 多抗基因聚合的特种稻海丰黑糯2号抗病虫鉴定和分子标记检测[J]. 分子植物育种, 2018, 16(2):466-471.

目的基因	标记名称	引物序列(5'-3')	片段大小/bp	退火温度/℃	标记类型	参考文献
Pi57	Pi57F	TCTTCCTTGCTTGAACTCGA	143^a/185^b	58	InDel	[23]
Pi57	Pi57R	TTTAAAAGGATCTGCATGCC	143^a/185^b	58	InDel	[23]
Pi69	Pi69F	TCGTCACCTTCTGGTCATAA	70^a/76^b	60	InDel	[24]
Pi69	Pi69R	CCTCTCCTCTTAGTTGCTCC	70^a/76^b	60	InDel	[24]

目的基因	标记名称	引物序列(5'-3')	片段大小/bp	退火温度/℃	标记类型	参考文献
Pi57	Pi57F	TCTTCCTTGCTTGAACTCGA	143^a/185^b	58	InDel	[23]
Pi57	Pi57R	TTTAAAAGGATCTGCATGCC	143^a/185^b	58	InDel	[23]
Pi69	Pi69F	TCGTCACCTTCTGGTCATAA	70^a/76^b	60	InDel	[24]
Pi69	Pi69R	CCTCTCCTCTTAGTTGCTCC	70^a/76^b	60	InDel	[24]

序号	水稻材料编号	菌株编号
序号	水稻材料编号	CH1603	08-1-1a-2	2011-1-1-39-1
1	云粳37	S	S	S
2	IL-E1454	R	R	R
3	IL106	S	S	R
4	云粳37+Pi57	R	R	R
5	云粳37+Pi69	S	S	R
6	云粳37+Pi57+Pi69	R	R	R
7	LTH（感病对照）	S	S	S

序号	水稻材料编号	菌株编号
序号	水稻材料编号	CH1603	08-1-1a-2	2011-1-1-39-1
1	云粳37	S	S	S
2	IL-E1454	R	R	R
3	IL106	S	S	R
4	云粳37+Pi57	R	R	R
5	云粳37+Pi69	S	S	R
6	云粳37+Pi57+Pi69	R	R	R
7	LTH（感病对照）	S	S	S