立枯丝核菌不同融合群菌株GPD基因的克隆及序列特征分析

doi:10.3969/j.issn.1001-7216.2013.06.011

中国水稻科学 ›› 2013, Vol. 27 ›› Issue (6): 639-646.DOI: 10.3969/j.issn.1001-7216.2013.06.011

立枯丝核菌不同融合群菌株GPD基因的克隆及序列特征分析

王艳丽,任衍春,张震,王教瑜,毛雪琴,姜华,邱海萍,柴荣耀,杜新法,孙国昌^*

浙江省植物有害生物防控重点实验室省部共建国家重点实验室培育基地/浙江省农业科学院植物保护与微生物研究所杭州 310021；

收稿日期:2013-07-10 修回日期:2013-08-10 出版日期:2013-11-10 发布日期:2013-11-10
通讯作者: 孙国昌*
基金资助:
国家自然科学基金资助项目(31071644)；农业部转基因专项（2011ZX08009003001）；浙江省公益性重大项目(2011C129011，2011C129012，2011C12022)；浙江省农科院国际合作项目（2009）。

Characterization of Glyceraldehyde3phosphate Dehydrogenase Gene Sequences of Different Anastomosis Group Rhizoctonia solani

WANG Yanli, REN Yanchun, ZHANG Zhen, WANG Jiaoyu, MAO Xueqin, JIANG Hua, QIU Haiping, CHAI Rongyao, DU Xinfa, SUN Guochang^*

State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Institute of Plant Protection Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;

Received:2013-07-10 Revised:2013-08-10 Online:2013-11-10 Published:2013-11-10
Contact: SUN Guochang*

摘要/Abstract

摘要： 利用同源克隆的方法对11个立枯丝核菌标准融合群菌株的GPD基因进行了分离。分析发现不同融合群立枯丝核菌GPD基因在外显子数目、编码蛋白长度及内含子剪切方式等方面存在差异，如AG22ⅢB、AG22Ⅳ、AG8三个融合群的GPD基因的外显子个数为10个，其他融合群均为11个；AG8融合群的GPD基因的编码蛋白长度为267个氨基酸，其余融合群的GPD基因的编码蛋白长度约为340个氨基酸。AG22ⅢB、AG22Ⅳ、AG8三个融合群的GPD基因在98、98、272氨基酸位点的内含子发生了缺失。基于蛋白序列的进化分析表明不同融合群之间GPD基因编码蛋白存在明显差异，可以有效地将不同融合群菌株区分开来；同时，不同物种之间GPD基因在进化上仍具有一定保守性，立枯丝核菌自身的GPD基因蛋白序列归为一类且与担子菌门同源性最高。这些结果表明进行保守基因序列差异分析是进行立枯丝核菌分类分群研究的新思路。

关键词: 立枯丝核菌, GPD 基因, 克隆, 进化分析, 融合群

Abstract: Full length cDNA sequences of glyceraldehyde3phosphate dehydrogenase (GPD) gene from 11 anastomosis groups (AGs) of Rhizoctonia solani were isolated by PCRbased strategies. The open reading frames (ORFs) of the GPD genes(from ATG to TGA) showed some differences in exon numbers, splicing modes and lengths of coding sequences. GPD genes from AG22ⅢB, AG22Ⅳand AG8 have 10 exons, while those from the other AGs have 11 exons; GPD gene from AG8 encodes a protein of 267 amino acids, those from the other AGs encode proteins with about 340 amino acids. Meanwhile, intron deletions were found in the GPD genes from AG22ⅢB, AG22Ⅳ and AG8 at amino acid 98, 98 and 272, respectively. Phylogenetic analysis show that the GPD genes can effectively distinguish the different AGs. Further, the GPD genes from different AGs can differentiate all the AGs of R. solani from other fungi, implying the GPD gene is a useful alternative to the 5.8S rDNAITS gene for determination of phylogenetic relationships among different AGs and between Rhizoctonia and other fungi.

Key words: Rhizoctonia solani, GPD, gene clone, phylogenetic analysis, anastomosis group

中图分类号:

王艳丽,任衍春,张震,王教瑜,毛雪琴,姜华,邱海萍,柴荣耀,杜新法,孙国昌*. 立枯丝核菌不同融合群菌株GPD基因的克隆及序列特征分析[J]. 中国水稻科学, 2013, 27(6): 639-646.

WANG Yanli, REN Yanchun, ZHANG Zhen, WANG Jiaoyu, MAO Xueqin, JIANG Hua, QIU Haiping, CHAI Rongyao, DU Xinfa, SUN Guochang*. Characterization of Glyceraldehyde3phosphate Dehydrogenase Gene Sequences of Different Anastomosis Group Rhizoctonia solani [J]. Chinese Journal of Rice Science, 2013, 27(6): 639-646.

参考文献

\[1\]刘力, 葛起新. 华东地区立枯丝核菌融合群鉴定. 浙江农业大学学报, 1987, 13(3): 227233.

\[2\]Anderson N A. The genetics and pathology of Rhizoctonia solani. Ann Rev Phytopathol, 1982(20): 329347.

\[3\]张天晓, 张志光. 立枯丝核菌 Rhizoctonia solani Kühn 的研究. 湖南师范大学：自然科学学报, 1986, 9(1): 7682.

\[4\]Carling D E, Kuninaga S, Brainard K A. Hyphal anastomosis reactions, rDNAinternal transcribed spacer sequences, and virulence levels among subsets of Rhizoctonia solani anastomosis group2 (AG2) and AGBI. Phytopathology, 2002, 92: 4350.

\[5\]李华荣. 丝核菌的菌丝融合群及其遗传多样性研究的新进展. 菌物系统, 1999, 18(2): 100107.

\[6\]Redkar R J, Herzog R W, Singh N K. Transcriptional activation of the Aspergillus nidulans gpdA promoter by osmotic signals. Appl Environ Microbiol, 1998, 64: 22292231.

\[7\]Jeong M J, Park S C, Kwon H B, et al. Isolation and characterization of the gene encoding glyceraldehyde3phosphate dehydrogenase. Biochem Biophysic Res Commun,2000, 278: 192196.

\[8\]Schaeffer H J, Forstheoefel N R, Cushman J C. Identification of enchancer and silencer regions involved in saltresponsive expression of Crassulacean acid metabolism (CAM) genes in the facultative halophyte Mesembryanthemum crystallinum. Plant Mol Biol, 1995, 28: 205218.

\[9\]Vernon D M, Ostrem J A, Bohnert H J. Stress perception and response in a facultative halophyte: The regulation of salinity induced genes in Mesembryanthemum crystallinum. Plant Cell Environ, 1993, 16: 437444.

\[10\]曾娟，王源超，申贵，等. 大豆疫霉甘油醛3磷酸脱氢酶基因在病原物植物互作中诱导表达及其抗氧化作用在酵母遗传互补系统中的功能验证. 科学通报, 2006, 51(10): 11751181.

\[11\]肖川, 刘晓斐, 詹树萱, 等. 水稻甘油醛3磷酸脱氢酶cDNA结构分析和分子进化. 自然科学进展, 1998, 8(4): 411419.

\[12\]宋时英, 郭剑, 李军, 等. 甘油醛3磷酸脱氢酶结构的保守性. 生物物理学报, 1998, 14(3): 401406.

\[13\]Ogoshi A. Ecology and pathogenicity of anastomosis and intraspecific groups of Rhizoctonia solani Kühn. Ann Rev Phytopathol, 1987, 25: 125143.

\[14\]Ogoshi A. Studies on the taxonomy of the genus Rhizoctonia. Annu Phytopathol Soc Japan, 1984, 50: 307309.

\[15\]Parmeter J R, Sherwood R T, Platt W D, et al. Anastomosis grouping among isolates of Thanatephorus cucumeris. Phytopathology,1969, 59(1): 12701278.

\[16\]方中达. 常用培养基的配制. 植病研究方法. 北京: 农业出版社, 1982: 8387.

\[17\]朱旭芬. 基因工程实验指导. 北京: 高等教育出版社, 2006: 1825.

\[18\]Rosewich U L, Pettway R, McDonald B, et al. High levels of gene flow and heterozygote excess characterize Rhizoctonia solani AG1 IA (Thanatephorus cucumeris) from texas. Fungal Genet Biol,1999, 28(3): 148159.

\[19\]Duncan S, Barton J E, Obrien P A. Analysis of variation in isolates of Rhizoctoniasolani by random amplified polymorphic DNA assay. Mycolog Res,1993, 97: 10751082.

\[20\]Ceresini P C, Shew H D, Vilgalys R J, et al. Genetic structure of populations of Rhizoctonia solani AG3 on potato in eastern North Carolina. Mycologia, 2002, 94(3): 450460.

\[21\]PadashtDehkaei F, Ceresini P, Zala M, et al. Population genetic evidence that basidiospores play an important role in the disease cycle of riceinfecting populations of Rhizoctonia solani AG1 IA in Iran. Plant Pathol, 2013, 62: 4958.

\[22\]Sharon M, Kuninaga S, Hyakumachi M, et al. The advancing identification and classification of Rhizoctonia spp. using molecular and biotechnological methods compared with the classical anastomosis grouping. Mycoscience, 2006, 47(6): 299316.

\[23\]Kuninaga S, Natsuaki T, Takeuchi T,et al. Sequence variation of the rDNA ITS regions within and between anastomosis groups in Rhizoctonia solani. Cur Genet, 1997, 32(3): 237243.

\[24\]陈涛，张震，柴荣耀，等．浙江省水稻纹枯病菌的遗传分化与致病力研究．中国水稻科学, 2010, 24(1): 6772

\[25\]Takeda K, Kang Y, Yazawa K, et al. Phylogenetic studies of Nocardia species based on gyrB gene analyses. J Med Microbiol, 2010, 59: 165171.

\[26\]Kwok A Y. and Chow A W. Phylogenetic study of Staphylococcus and Macrococcus species based on partial hsp60 gene sequences. Int J Syst Evol Microbiol, 2003, 53(1): 8792.

\[27\]梁日深，周爱国，陈金涛，等. 基于RAG2 基因序列分析仿石鲈科鱼类及其相关种类系统进化关系.水产学报, 2013, 37(5): 651660.

\[28\]Zheng A, Lin R, Zhang D, et al. The evolution and pathogenic mechanisms of the rice sheath blight pathogen. Nat Commun, 2013, 4: 1424.

[1]	廉院训, 韦子芸, 张强, 李清, 任德勇, 胡江, 朱丽, 高振宇, 张光恒, 郭龙彪, 曾大力, 钱前, 沈兰. 水稻斑马叶突变体zl7的鉴定与基因的精细定位[J]. 中国水稻科学, 2023, 37(2): 113-124.
[2]	曹跃炫, 严绘景, 王克剑, 刘朝雷. 苗期快速分选水稻人工无融合生殖克隆种子[J]. 中国水稻科学, 2022, 36(6): 656-662.
[3]	裘霖琳, 刘窍, 付亚萍, 刘文真, 胡国成, 翟玉凤, 庞礴, 汪得凯. 水稻矮化小穗基因DSP2的鉴定与克隆[J]. 中国水稻科学, 2022, 36(2): 150-158.
[4]	张涛荟, 王海宇, 万华, 张莉萍, 谢振威, 陈可毅, 何晓栋, 赵志刚, 万建民. 水稻雌雄不育突变体Osfma2的细胞学观察及基因图位克隆[J]. 中国水稻科学, 2022, 36(1): 13-26.
[5]	吴先美, 李三峰, 胡萍, 何瑞, 焦然, 毛一剑, 鲁草林, 胡娟, 林晗, 吴荣梁, 朱旭东, 饶玉春, 王跃星. 水稻分蘖调控基因HTD3的克隆与功能研究[J]. 中国水稻科学, 2021, 35(6): 535-542.
[6]	朱玉君, 左紫薇, 张振华, 樊叶杨. 一种水稻微效QTL精细定位和克隆新途径[J]. 中国水稻科学, 2021, 35(4): 407-414.
[7]	杜溢墨潘天田云录刘世家刘喜江玲张文伟王益华* 万建民. 水稻粉质皱缩胚乳突变体fse4的表型分析与基因克隆[J]. 中国水稻科学, 2019, 33(6): 499-512.
[8]	冯辉, 范亚磊, 张金凤, 朱红利, 魏利辉. 谷氧还蛋白(AbGrx-1)对水稻干尖线虫在氧化胁迫下的保护作用[J]. 中国水稻科学, 2019, 33(6): 565-574.
[9]	何婧, 刘寒, 郭留明, 李静, 张恒木. 水稻小热休克蛋白OsHSP20抗体特性鉴定及应用[J]. 中国水稻科学, 2019, 33(3): 235-240.
[10]	吕凤, 唐倩莹, 王启明, 郑海, 尤世民, 柏文婷, 肖晏嘉, 赵志刚, 万建民. 水稻雌性败育基因FA的图位克隆[J]. 中国水稻科学, 2018, 32(6): 519-528.
[11]	江绍锋, 王陈骄子, 舒灿伟, 周而勋. 水稻纹枯病菌RsPhm基因的克隆及其表达分析[J]. 中国水稻科学, 2018, 1(1): 111-118.
[12]	张习春, 鲁菲菲, 吕育松, 罗荣剑, 焦桂爱, 邬亚文, 唐绍清, 胡培松, 魏祥进. 两个垩白突变体的鉴定及突变基因的图位克隆[J]. 中国水稻科学, 2017, 31(6): 568-579.
[13]	赵晨星, 俞叶微, 许益鹏, 俞晓平. 褐飞虱两个dynamin-1-like基因的克隆、多克隆抗体制备及表达定位[J]. 中国水稻科学, 2017, 31(4): 345-354.
[14]	吕育松, 谢耘丰, 圣忠华, 邬亚文, 唐绍清, 胡培松, 魏祥进. 矮秆小粒水稻潇湘矮的形态学与分子遗传学分析[J]. 中国水稻科学, 2017, 31(3): 238-246.
[15]	井文, 章文华. 水稻耐盐基因定位与克隆及品种耐盐性分子标记辅助选择改良研究进展[J]. 中国水稻科学, 2017, 31(2): 111-123.

立枯丝核菌不同融合群菌株GPD基因的克隆及序列特征分析

Characterization of Glyceraldehyde3phosphate Dehydrogenase Gene Sequences of Different Anastomosis Group Rhizoctonia solani

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics