Rice Blast Resistance of Transgenic Rice Plants with Pi-d2 Gene

doi:10.3969/j.issn.1001-7216.2010.01.06

Abstract

Abstract: Transgenic rice lines harboring Pid2 gene under the control of different promoters from three expression vector pCB6.3kb, pCB53kb and pZH01272kb were tested for resistance to Magnaporthe grisea. The results showed that 9 stably transgenic lines had different resistances to 39 strains of M. grisea from Sichuan Province, China, and the highest diseaseresistance frequency was 917%. Four early homozygous transgenic lines of Pid2 gene exhibited resistance to more than 8148% of 58 strains from Chinese Academy of Agricultural Sciences, which indicates that the transgenic rice lines possessed widespectrum resistance against various rice blast races. The results of transgenic embryonic callus selected by crude toxin showed that the induction rate of embryonic callus reduced by increased concentration of toxin in medium. When the toxin concentration was 40% in medium, the induction rate of embryonic callus of transgenic plant was 4933%, while that of the receptor control was 5%. The disease incidence of neck blast were 0% to 50% in the transgenic lines under induction of rice blast disease in fields, showing the higher resistance to rice blast than that of receptor control.

Key words: transgenic rice, resistance gene, rice blast, resistance spectrum, crude toxin

摘要： 对由pCB63kb、pCB53kb和pZH01272kb三个不同表达载体转化的转稻瘟病抗性基因Pid2水稻株系进行稻瘟病抗性分析。结果表明，转Pid2基因水稻的9个高代株系对来自四川的39个稻瘟病菌株表现不同的抗性，抗病频率最高达917%；4个转基因早代纯合株系对来自中国农业科学院的58个菌株中的8148%以上菌株表现抗性，具有广谱抗性特点。稻瘟病菌粗毒素筛选结果表明，来自转基因植株的幼胚愈伤组织诱导率随培养基中粗毒素浓度提高而降低，粗毒素浓度达到40%时，幼胚愈伤诱导率为4933%，受体对照幼胚愈伤组织诱导率为5%。田间诱发条件下，转基因株系在大田的穗瘟发病率为0%~50%, 抗性较受体对照大幅度提高。

关键词: 转基因水稻, 抗病基因, 稻瘟病, 抗谱, 粗毒素

CHEN De-xi,CHEN Xue-wei,LEI Cai-lin,MA Bing-tian,WANG Yu-ping,LI Shi-gui . Rice Blast Resistance of Transgenic Rice Plants with Pi-d2 Gene [J]. Chinese Journal of Rice Science, DOI: 10.3969/j.issn.1001-7216.2010.01.06 .

陈德西,陈学伟,雷财林, 马炳田,王玉平,李仕贵,. 转Pi-d2基因水稻对稻瘟病的抗性分析[J]. 中国水稻科学, DOI: 10.3969/j.issn.1001-7216.2010.01.06 .

References

[1]Qu S H. Rice diseases. 2nd edn. Kew, Surrey：Commonwealth Mycological Institute, CABI, 1985： 109-201.

[2]Nishizawa Y, Nishio Z, Nakazono K, et al. Enhanced resistance to blast (Magnaporthe grisea) in transgenic japonica rice by constitutive expression of rice chitinase.Theor Appl Genet, 1999, 99: 383-390.

[3]Starklorenzen P, Nelke B, Han G, et al. Transfer of a grapevine synthase gene to rice (Oryza sativa L.). Plant Cell Rep, 1997, 16: 668-673.

[4]Feng D R, Xu X P, Wei J W, et al. Enhancement of rice disease resistance by two antifungal protein genes.Acta Bot Sin, 1999, 41(11): 1187-1191.

[5]Ming X T, Wang L J, An C C, et al. Resistance to rice blast (Pyricularia oryzae) caused by the expression of trichosanthin gene in transgenic rice plants transferred through Agrobacterium method. Chin Sci Bull, 2000, 19: 1774-1778.

[6]Kanzaki H, Nirasawa S, Saitoh H, et al. Overexpression of the wasabi defensin gene confers enhanced resistance to blast fungus(Magnaporthe grisea) in transgenic rice. Theor Appl Genet, 2002, 105: 809-814.

[7]Wang Z X, Yano M, Yamanouchi U, et al. The Pib 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.

[8]Bryan G T, Wu K S, Farrall L, et al. A single amino acid difference distinguishes resistant and susceptible alleles of the rice blast resistance gene Pita. Plant Cell, 2000, 12: 2033-2045.

[9]Qu S H, Liu G F, Zhou B, et al. The broadspectrum blast resistance gene Pi9 encodes an NBSLRR protein and is a member of a multigene family in rice. Genetics, 2006, 172: 1901-1914.

[10]Zhou B, Qu S H, Liu G F, et al. The eight amino acid differences within three leucinerich repeats between Pi2 and Pizt resistance proteins determine the resistance specificity to Magnaporthe grisea. Mol PlantMicrobe Interact, 2006, 19: 1216-1228.

[11] Chen X W, Shang J J, Chen D X, et al. A Blectin receptor kinase gene conferring rice blast resistance. Plant J, 2006, 46: 794-804.

[12] Lin F, Chen S, Que Z Q, et al. Molecular cloning and characterization of the rice blast resistance gene Pi37 in the wellknown cultivar St. No. 1//Abstracts of the 4th International Rice Blast Conference. Changsha, China, October 9 14, 2007. Changsha:[s.n.], 2007.

[13]刘文萍, 刘丽艳, 吕晓波, 等. 稻瘟病菌粗毒素的制备及其致病力的测定. 黑龙江农业科学, 1997（6）：14-17.

[14]许明辉, 李成云, 李进斌，等. 转溶菌基因水稻稻瘟病抗谱分析. 中国农业科学, 2003, 36(4): 387-392.

[15] 许明辉，李成云，李进斌，等. 转几丁质酶葡聚糖基因水稻稻瘟病抗谱分析. 中国水稻科学， 2003， 17(4): 307-310.

[16]Cao M X, Huang J Q， Wei Z M, et al. Agrobacteriummediated multiple gene transformation in rice using a single vector. J Integr Plant Biol, 2005, 47(2): 233-242.

[17]陈红旗, 陈宗祥, 倪深，等. 利用分子标记技术聚合3个稻瘟病基因改良金23B的稻瘟病抗性. 中国水稻科学， 2008，12（1）： 23-27.