Transgenic Rice Plants Harboring Genomic DNA from Zizania latifolia Confer Bacterial Blight Resistance

doi:10.3969/j.issn.1001-7216.2010.05.001

Chinese Journal of Rice Science ›› 2010, Vol. 24 ›› Issue (5): 447-452 .DOI: 10.3969/j.issn.1001-7216.2010.05.001

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Transgenic Rice Plants Harboring Genomic DNA from Zizania latifolia Confer Bacterial Blight Resistance

SHEN Wei-wei^1,3;SONG Cheng-li¹;CHEN Jie²;FU Ya-ping²; WU Jian-li^2,*; JIANG Shao-mei^3,*

¹College of Life Sciences, Jiangxi Normal University, Nanchang 330022, China; ² China National Rice Research Institute, Hangzhou 310006, China; ³Jiangxi University of Finance and Economics, Nanchang 330013, China; *Corresponding authors, E-mail: beishangd@163.com; shmjiang111@yahoo.com.cn

Received:1900-01-01 Revised:1900-01-01 Online:2010-09-10 Published:2010-09-10

转菰候选基因克隆获得抗白叶枯病水稻植株

沈玮玮^1,3;宋成丽¹;陈洁²;付亚萍²;吴建利^2,*_;江绍玫^3,*

¹江西师范大学生命科学学院, 江西南昌 330022; ²中国水稻研究所, 浙江杭州 310006; ³江西财经大学, 江西南昌 330013; *通讯联系人, E-mail: beishangd@163.com; shmjiang111@yahoo.com.cn

Abstract

Abstract: Based on the sequence of resistance gene analog FZ14 derived from Zizania latifolia(Griseb.), a pair of specific PCR primers FZ14P1/FZ14P2 was designed to isolate candidate disease resistance gene. The pooledPCR approach was adapted using the primer pairs to screen a genomic transformationcompetent artificial chromosome(TAC) library derived from Zizania latifolia. A positive TAC clone (ZR1) was obtained and confirmed by sequence analysis. The results indicated that ZR1 consisted of conserved motifs similar to Ploop (kinase 1a), kinase 2, kinase 3a and GLPL(Gly-Leu-Pro-Leu), indicating that it could be a portion of NBSLRR type of resistance gene. Using Agrobacterium tumafaciensmediated transformation of Nipponbare mature embryo, a total of 48 independent transgenic T0 plants were obtained. Among them, 36 plants were highly resistant to the virulent bacterial strain PXO71. The results indicated that ZR1 contains at least one bacterial blight resistance gene.

摘要： 以菰NBS-LRR类抗病基因同源序列FZ14（GenBank登录号：DQ239432）为模板设计特异性引物FZ14P1/P2 ，通过克隆池PCR法经分级筛选，从菰基因组TAC文库中获得1个阳性克隆（ZR1），序列分析比对证实该阳性克隆为含有菰抗病基因同源序列FZ14的抗病基因候选克隆。同时，ZR1具有植物NBSLRR类型抗性基因中的Ploop (kinase 1a )、 kinase 2、kinase 3a和GLPL(Gly-Leu-Pro-Leu)等保守基序，可能为抗性基因的部分序列。通过农杆菌介导转化水稻品种日本晴，获得36个对白叶枯病菌PXO71具有明显抗性的独立转化子，结果表明，菰ZR1克隆中至少含有1个白叶枯病抗性基因。

SHEN Wei-wei,SONG Cheng-li,CHEN Jie,FU Ya-ping,WU Jian-li,JIANG Shao-mei. Transgenic Rice Plants Harboring Genomic DNA from Zizania latifolia Confer Bacterial Blight Resistance[J]. Chinese Journal of Rice Science, 2010, 24(5): 447-452 .

沈玮玮,宋成丽,陈洁,付亚萍,吴建利,江绍玫,. 转菰候选基因克隆获得抗白叶枯病水稻植株 [J]. 中国水稻科学, 2010, 24(5): 447-452 .

References

[1]Liu Y G, Shirano Y, Fukaki H, et al. Complementation of plant mutants with large genomic DNA fragments by a transformation-competent artificial chromosome vector accelerates positional cloning. Proc Natl Acad Sci USA, 1999, 96: 6535-6540.
[2]Kong F N, Jiang S M, Shi X L, et al. Construction and characterization of a transformation-competent artificial chromosome (TAC) library of Zizania latifolia (Griseb.). Plant Mol Biol Rep, 2006, 24: 219-227.
[3]Liu Y G, Nagaki K, Fujita M, et al. Development of an efficient maintenance and screening system for large-insert genomic DNA libraries of hexaploid wheat in a transformation-competent artificial chromosome (TAC) vector. Plant J, 2000, 23: 687-695.
[4]秦跟基，陈佩度，刘耀光，等. 用克隆池PCR法筛选小麦簇-毛麦易位系6VS/6AL基因组TAC 文库获得抗病基因候选克隆. 生物工程学报, 2002, 18: 313-317.
[5]Fenillet C, Schachermayr G, Keller B. Molecular cloning of a new receptor-like kinase gene encoded at the Lr10 disease resistance locus of wheat. Plant J, 1997, 11: 45-52.
[6]Bendahmane A, Querci M, Kanyuka K, et al. Agrobacterium transient expression system as a tool for the isolation of disease resistance genes: Application to the Rx2 locus in potato. Plant J, 2000, 21: 73-81.
[7]Staskawicz B J, Ausubel F M, Baker B J, et al. Molecular genetics of plant disease resistance. Science, 1995, 268: 661-667.
[8]Chen Y, Long L, Lin X, et al. Isolation and characterization of a set of disease resistance-gene analogs (RGAs) from wild rice, Zizania latifolia (Griseb):Ⅰ. Introgression, copy number lability, sequence change, and DNA methylation alteration in several rice-Zizania introgression lines. Genome, 2006, 49: 150-158.
[9]朱正歌, 肖晗, 付亚萍, 等. 水稻转座子突变体库的构建及突变类型的遗传分析. 生物工程学报, 2001, 17(3): 288-292.
[10]Kauffman, H E, Reddy A P K, Merca S D, et al. An improved technique for evaluating resistance to rice varieties of Xanthomonas oryzae pv. oryzea. Plant Dis Rep, 1973, 57: 537-541.
[11] 卢扬江, 郑康乐. 提取水稻DNA的一种简易方法. 中国水稻科学, 1992, 6(1): 47-48.
[12]Dangl J L, Jones J D. Plant pathogens and integrated defense responses to infection. Nature, 2001, 411: 826-833.
[13]Merers B C, Dickerman A W, Michelmore R W, et al. Plant disease resistance genes encode members of an ancient and diverse protein family within the nucleotide-binding superfamily. Plant J, 1999, 20: 317-332.
[14] 徐兵强, 杜中军, 黄俊生. RGA法克隆候选抗病基因的研究进展. 分子植物育种, 2004, 3: 421-428.
[15]Ramalingam J, Vera Cruz C M, Kukreja K, et al. Candidate resistance genes from rice, barley, and maize and their association with qualitative and quantitative resistance in rice. Mol Plant Microbe Int, 2003, 16: 14-24.
[16]韩德俊, 曹莉, 陈耀锋, 等. 植物抗病基因与病原菌无毒基因互作的分子基础. 遗传学报, 2005, 32: 1319-1326.
[17] Liu X Q, Lin F, Wang L, et al. The in silico map-based cloning of Pi36, a rice CC-NBS-LRR gene which confers race-specific resistance to the blast fungus. Genetics, 2007, 176: 2541-2549.
[18] Wang Z X, Yano M, Yamanouchi U, et al. The Pi b gene for rice blast resistance belongs to the nucleotide binding and leucine-rich repeat class of plant disease resistance genes. Plant J, 1999, 19: 55-64.
[19] Bryan G, Wu K S, Farrall L, et al. A single amino acid difference distinguishes resistance and susceptible alleles of the rice blast resistance gene Pita. Plant Cell, 2000, 12: 2033-2045.
[20] Zhou B, Qu S H, Liu G F, et al. The eight amino-acid differences within three leucine-rich repeats between Pi2 and Pizt resistance proteins determine the resistance specificity to Magnaporthe grisea. Mol Plant Microbe Int, 2006, 19: 1216-1228.
[21] Qu S H, Liu G F, Zhou B, et al. The broad-spectrum blast resistance gene Pi9 encodes a nucleotide-binding site-leucine-rich repeat protein and is a member of a multigene family in rice. Genetics, 2006, 172: 1901-1914.
[22] Lin F, Chen S, Que Z Q, et al. The blast resistance gene Pi37 encodes an NBS-LRR protein and is a member of a resistance gene cluster on rice chromosome 1. Genetics, 2007, 177: 1871-1880.
[23]Yoshimura S, Yamanouchi U, Katayose Y, et al. Expression of Xa1, a bacterial blight-resistance gene in rice, is induced by bacterial inoculation. Proc Natl Acad Sci USA, 1998, 95: 1663-1668.
[24] 金旭炜, 王春莲, 杨清, 等. 水稻抗白叶枯病近等基因系CBB30的培育及Xa30(t)的初步定位. 中国农业科学, 2007, 40(6): 1094-1100.
[25] 郑崇珂, 王春莲, 于元杰, 等. 水稻抗白叶枯病新基因Xa32(t)的鉴定和初步定位. 作物学报， 2009, 35(7): 1173-1180.
[26] 于彦春, 张光恒, 郭龙彪, 等. CecropinB与Xa21基因共表达提高转基因水稻白叶枯病抗性. 中国水稻科学, 2006, 20(1): 105-108.

Transgenic Rice Plants Harboring Genomic DNA from Zizania latifolia Confer Bacterial Blight Resistance

转菰候选基因克隆获得抗白叶枯病水稻植株

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