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

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

Abstract

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

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.

References

[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.