稻曲病菌基因组DNA提取方法比较与小文库构建

doi:10.13560/j.cnki.biotech.bull.1985.2017-0686

生物技术通报 ›› 2018, Vol. 34 ›› Issue (4): 102-106.doi: 10.13560/j.cnki.biotech.bull.1985.2017-0686

稻曲病菌基因组DNA提取方法比较与小文库构建

伏荣桃¹, 王剑¹, 陈诚¹, 龚学书¹, 卢代华¹, 罗曦¹, 郑爱萍²

1. 四川省农业科学院植物保护研究所农业部西南作物有害生物综合治理重点实验室,成都 610066;
2. 四川农业大学水稻研究所,成都 611130

收稿日期:2017-08-21 出版日期:2018-04-20 发布日期:2018-05-04
作者简介:伏荣桃,女,博士研究生,研究方向：水稻真菌病害;E-mail：furongtao@126.com
基金资助:
四川省财政创新能力提升工程项目（2016GYSH-014）; 四川省农业科学院青年基金项目（2015JSCX-017）

Comparing the Methods of Isolating Genomic DNA and Construction of Its Small Genomic Library of Ustilaginoidea virens

FU Rong-tao¹, WANG Jian¹, CHEN Cheng¹, GONG Xue-shu¹, LU Dai-hua¹, LUO Xi¹, ZHENG Ai-ping²

1. Key Laboratory of Integrated Pest Management on Crops in Southwest,Ministry of Agriculture Science,Institute of Plant Protection,Sichuan Academy of Agriculture Science,Chengdu 610066;
2. Rice Research Institute,Sichuan Agricultural University,Chengdu 611130

Received:2017-08-21 Published:2018-04-20 Online:2018-05-04

摘要/Abstract

摘要： 水稻稻曲病是由稻曲病菌引起的真菌病害,现已成为水稻重要的病害之一。旨在寻找一种最佳的稻曲病菌基因组DNA提取方法,并构建基因组小文库。采用CTAB、SDS和真菌DNA试剂盒3种提取方法提取稻曲病菌基因组DNA,并用Illumina系统测序分析,构建基因组小文库。结果显示,CTAB提取法能得到高质量的基因组DNA,小文库测序组装共得29 350 288碱基（bp）和17 908 Scaffold,总碱基的GC含量为49.79%。CTAB提取法是稻曲病菌基因组DNA最佳的提取方法,基因组小文库成功的构建为稻曲病菌的基因组gap的填补和致病相关基因的鉴定提供了数据资源。

关键词: 稻曲病菌, CTAB, SDS, 基因组小文库

Abstract: Rice false smut caused by Ustilaginoidea virens is one of important fungal diseases for rice. This experiment aimed to select an optimal method of isolating genomic DNA,and to construct small genomic library of U. virens. The genomic DNA of U. virens was extracted by 3 methods of CTAB,SDS,and fungal DNA isolation kit. The DNA sample was sequenced using Illumina system,and small genomic library was constructed. The results showed that the high-quality genomic DNA was obtained by CTAB. Meanwhile,the small genomic library of U. virens was contained 29 350 268 bases and 17 904 Scaffold,and the GC content was 49.79% of total bases. We found that CTAB was the best method of isolating genomic DNA of U. virens. The successful construction of the small genomic library provided data resource for filling the genome gap and identifying pathogenetic gene.

Key words: Ustilaginoidea virens, CTAB, SDS, small genomic library

伏荣桃, 王剑, 陈诚, 龚学书, 卢代华, 罗曦, 郑爱萍. 稻曲病菌基因组DNA提取方法比较与小文库构建[J]. 生物技术通报, 2018, 34(4): 102-106.

FU Rong-tao, WANG Jian, CHEN Cheng, GONG Xue-shu, LU Dai-hua, LUO Xi, ZHENG Ai-ping. Comparing the Methods of Isolating Genomic DNA and Construction of Its Small Genomic Library of Ustilaginoidea virens[J]. Biotechnology Bulletin, 2018, 34(4): 102-106.

参考文献

[1] Tanaka E, Ashizawa T, Sonoda R, et al.Villosiclava virens gen. nov. , comb. nov. , teleomorph of Ustilaginoidea virens, the causal agent of rice false smut[J]. Mycotaxon, 2008, 106(1):491-501.
[2] Takahashi Y. On Ustilago virens Cooke and a new species of Tilletia parasitic on rice plant[J]. Botanical Magazine Tokyo, 1896, 10:16-20.
[3] 伏荣桃, 王剑, 卢代华, 等. 国内外水稻稻曲病研究进展[J]. 中国农学通报, 2016, 32(12):189-194.
[4] Lu S, Tian J, Sun W, et al.Bis-naphtho-γ-pyrones from fungi and their bioactivities[J]. Molecules, 2014, 19:7169-7188.
[5] Nakamura K, Izumiyama N, Ohtsubo K, et al.‘Lupinosis’ - Like lesions in mice caused by Ustiloxin, produced by Ustilaginoidea virens:a morphological study[J]. Natural Toxins, 1994, 2:22-28.
[6] Fu RT, Ding L, Zhu J, et al.Morphological structure of propagules and electrophoretic karyotype analysis of false smut Villosiclava virens in rice[J]. The Journal of Microbiology, 2012, 50(2):263-269.
[7] Tang XY, Jin J, Hu DW, et al.Elucidation of the infection process of Ustilaginoidea virens(teleomorph:Villosiclava virens)in rice spikelets[J]. Plant Pathology, 2012, 62(1):1-8.
[8] Zhang Y, Zhang K, Fang AF, et al.Specific adaption of Ustilaginoidea virens in occupying host florets revealed by comparative and functional genomics[J]. Nature Communication, 2014, 5:3849.
[9] Fan J, Guo XY, Huang F, et al.Epiphytic colonization of Ustilaginoidea virens on biotic and abiotic surfaces implies the widespread presence of primary inoculums for rice false smut disease[J]. Plant Pathology, 2014, 63:937-945.
[10] Hu ML, Luo LX, Wang S, et al.Infection process of Ustilaginoidea virens during artificial inoculation of rice panicles[J]. European Journal of Plant Pathology, 2014, 139:67-77.
[11] Fu RT, Yin CC, Deng QM, et al.Mating type loci analysis indicates that Villosiclava virens, the casual agent of false smut disease of rice, is a homothallic fungus[J]. Tropical Plant Pathology, 2014, 39(3):203-209.
[12] Singh RA, Dubey KS.Sclerotial germination and ascospore formation of claviceps oryzae-sative in India[J]. India Phytopathology, 1984, 37:168-170.
[13] 刘庆丽, 王晓明, 王革娇, 等. 稻曲病菌UV-2 菌株细菌人工染色体文库构建及分析[J]. 微生物学通报, 2013, 40(9):1715-1722.
[14] 张震, 王教瑜, 杜新法, 等. 稻曲病菌cDNA文库的构建[J]. 植物病理学报, 2008, 38(5):462-467.
[15] 韩冰, 蔺瑞明, 曹远银, 等. 小麦条锈菌 DNA 提取方法的比较研究[J]. 中国农学通报, 2006, 22(4):81-83.
[16] 冮洁, 杜连祥, 路福平, 等. 基因工程菌里氏木霉染色体 DNA的提取方法[J]. 生物技术, 2004, 14(2):24-26.
[17] 张宝俊, 李志岗, 张家榕, 等. 4种镰刀菌基因组DNA提取方法的比较研究[J]. 安徽农业科学, 2008, 38(31):13559-13560, 13563.
[18] 贺羽, 李鑫, 王有福, 等. 苹果球壳孢腐烂病菌DNA不同提取方法的比较[J]. 江苏农业科学, 2012, 40(6):38-40.
[19] 秦培钢. 水稻纹枯病菌Rhizoctonia solani AG1 IA Fosmid 基因组文库的构建及线粒体全基因组组装分析[D]. 成都, 四川农业大学, 2012.
[20] 张晓晖, 郭春华, 江晓霞, 等. 康氏木霉基因组DNA提取方法的比较研究[J]. 生物技术通报, 2007(5):128-130.

稻曲病菌基因组DNA提取方法比较与小文库构建

Comparing the Methods of Isolating Genomic DNA and Construction of Its Small Genomic Library of Ustilaginoidea virens

PDF (PC)

可视化

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

本文评价

[1]	张婷婷, 滕丽, 蔡小姚, 龙慧, 李升愉, 刘红美. 稻曲病菌菌株GZ-14-07中携带的全病毒基因组特征研究[J]. 生物技术通报, 2020, 36(10): 80-87.
[2]	杨云, 刘红昌, 夏金兰, 马亚龙, 聂珍媛. 极端嗜酸热古菌Acidianus manzaensis膜蛋白提取方法的建立及应用[J]. 生物技术通报, 2016, 32(11): 130-136.
[3]	关静, 孙仁强, 宋佳雯, 黄迪, 成楠, 岑院汉凤, 张扬, 陈曦乐, ,赵怡姗, 朱晨旦, 郑烨, 康立, 高红亮. 微生物蛋白质谷氨酰胺酶的初步分离纯化[J]. 生物技术通报, 2014, 0(2): 161-165.
[4]	范晶 ,李仲芳, 高明辉 ,伏秦超. 一种适用于RAPD分析的微量山茶DNA提取方法的建立[J]. 生物技术通报, 2014, 0(12): 67-72.
[5]	魏晓丽;金一帮;朱明;李亮;温浩;. 小鼠beta防御素2原核表达与抗菌作用的初步研究[J]. , 2012, 0(10): 95-99.
[6]	蔡翠霞;肖维威;康文杰;周琳华;吴永彬;郑远金;马文丽;. 改良Chelex-100法快速提取转基因农产品DNA[J]. , 2011, 0(10): 210-214.
[7]	刘胜洪;刘文;刘明峰;杨凤婷;刘庆生;梁红;. 一种高效提取猕猴桃DNA和RNA的方法[J]. , 2011, 0(09): 171-175.
[8]	王大威;张健;姜伟;张凤久;. 一种快速分离检测产脂肽类生物表面活性剂枯草芽孢杆菌的方法[J]. , 2011, 0(09): 142-146.
[9]	向小聪;孟庆石;鲍锦库;潘映红;. 小麦叶片类囊体膜蛋白复合体的分离和分析技术研究[J]. , 2011, 0(07): 148-153.
[10]	王冠明;韩烈保;马秀杰;程晓霞;. 麦冬基因组DNA提取方法研究[J]. , 2010, 0(06): 100-106.
[11]	李苗;高丽美;李永锋;韩榕;. 增强UV-B辐射对小麦幼苗基因组DNA的影响及RAPD分析体系的建立[J]. , 2010, 0(05): 87-92.
[12]	孙孝德;孙国华;杨建敏;吉成龙;王卫军;宋志乐;. 刺参基因组DNA提取的探讨[J]. , 2010, 0(03): 149-153.
[13]	赵宏宇;蔡禄;赵秀娟;王晶妍;李晶;. 三种快速制备含重复序列质粒PCR模板的方法[J]. , 2010, 0(02): 119-122.
[14]	吴转娣;昝逢刚;赵丽宏;罗遵喜;张树珍;. 甘蔗基因组DNA小量提取与大量提取方法研究[J]. , 2009, 0(S1): 172-175.
[15]	施维属;潘腾飞;钟凤林;潘东明;李开拓;王江波;郭志雄;刘天亮;. 柑橘基因组DNA快速提取及ISSR-PCR扩增体系优化[J]. , 2009, 0(10): 109-113.