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棉花学报  2011 

棉花黄萎病菌插入突变体库的构建及致病相关基因DVK1的克隆与鉴定

DOI: 1002-7807(2011)01-0064-05, PP. 64-68

Keywords: T-DNA插入,突变体文库,基因克隆,棉花黄萎病,功能鉴定

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Abstract:

利用ATMT(Agrobacteriumtumefaciens-mediatedtransformation)技术自建Verticilliumdahliae强致病力落叶型菌株T-DNA插入突变体库,共获得5000个转化子;随机挑选1000个转化子进行致病力鉴定,筛选获得5株致病力衰退的突变体。挑选致病力下降最为明显的突变体d1,通过TAIL-PCR技术最终获得一段长3237bp的DVK1全长cDNA序列,编码1079个氨基酸的蛋白。基于DNA序列比对发现,DVK1基因含有2个内含子,分别为52bp和36bp。进一步比对发现T-DNA插入到DVK1基因启始密码上游740bp的启动子中。通过遗传互补实验d1恢复了致病性,进一步证明DVK1基因与黄萎菌致病性相关。

References

[1]  YANG Chang, Guo Wang-zhen, Li Guo-ying, et al. QTLs mapping for Verticillium wilt resistance at seedling and maturity stages in Gossypium barbadense L. [J]. Plant Science, 2008, 174: 290-298.
[2]  RAUYAREE P, Ospina-Giraldo M D, Kang S, et al. Mutations in VMK1, a mitogen-activated protein kinase gene, affect microsclerotia formation and pathogenicity in Verticillium dahliae[J]. Curr Genet, 2005, 48: 109-116.
[3]  KLIMES A, Dobinson K F. A hydrophobin gene, VDH1, is involved in microsclerotial development and spore viability in the plant pathogen Verticillium dahliae[J]. Fungal Genetics and Biology, 2006, 43:283-294.
[4]  KLIMES A, Amyotte S G, Grant S, et al. Microsclerotia development in Verticillium dahliae: regulation and differential expression of the hydrophobin gene VDH1[J]. Genetics and Biology, 2008, 45: 1525-1532.
[5]  TALBOT N J, Ebbole D J, Hamer J E. Identification and characterization of MPG1, a gene involved in pathogenicity from the rice blast fungus Magnaporthe grisea[J]. The Plant Cell, 1993, 5: 1575-1590.
[6]  SAMBROOK J, Fritsch E F, Maniatis T. Molecular cloning: a laboratory manual[M]. 2nd ed. Coldspring Harbor, NY, USA: Cold Spring Harbor Laboratory Press, 1989.
[7]  彭 姗,吕学莲,高 峰,等.一种新的棉花黄、枯萎病快速接种方法的研究[J].棉花学报,2008,20(3):174-178.PENG Shan, Lü Xue-lian, Gao Feng, et al. Study on a new rapid inoculation method for Verticillium wilt and Fusarium wilt of cotton[J]. Cotton Science, 2008, 20(3): 174-178.
[8]  LIU Yao-Guang, Mitsukawa N, Oosumi, et a1. Efficient isolation and mapping of Arabidopsis thaliana T-DNA insert junctions by thermal asymmetric interlaced PCR[J]. Plant J, 1995, 8: 457-463.
[9]  MULLINS E D,Chen X, Romaine C P, et al. Agrobacterium-mediated transformation of Fusarium oxysporum: an efficient tool for insertional mutagenesis and gene transfer[J]. Phytopathology, 2001, 91: 173-180.
[10]  HUANG H C, Hanna M R. An efficient method to evaluate alfalfa cultivars for resistance to Verticillium wilt[J]. Canadian Journal of Plant Science, 1991, 71(3): 871-875.
[11]  KENNETH C S, William E G. A root-injection method to assess Verticillium wilt resistance of peppermint and its use in identifying resistant somaclones of cv. Black Mitcham[J]. Euphytica, 1999, 106(3): 223-230.
[12]  马 平,Huang H C,李社增,等.一种新的棉花黄萎病快速接种技术及其在病原菌致病力和寄主抗病性鉴定上的应用[J].植物病理学报,2004,34(6):536-541.MA Ping, Huang H C, Li She-zeng, et a1. A new inoculation method for Verticillium wilt on cotton and its application in evaluating pathogenesis and host resistance[J]. Acta Phytopathologica Sinica, 2004, 34(6): 536-541.
[13]  王省芬,马峙英.一种新的棉花黄萎病抗性鉴定方法[J].棉花学报,2002,14(4):231-233.WANG Xing-feng, Ma Zhi-ying. A new method for identification of cotton Verticillium wilt resistance[J]. Cotton Science, 2002, 14(4): 231-233.
[14]  DENKIN S M, Nelson D R. Regulation of Vibrio anguillarum empA metalloprotease expression and its role in virulence[J]. Appl Environ Microbiol, 2004, 70: 4193-4204.
[15]  CORBETT C R, Burtnick M N, Kooi T C, et a1. An extracellular zinc metalloprotease gene of Burkholderia cenocepacia[J]. Microbiology, 2003, 149: 2263-2271.

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