香蕉枯萎病菌4号生理小种中CHS基因家族的克隆与蛋白序列分析

生物技术通报 ›› 2014, Vol. 0 ›› Issue (3): 79-85.

香蕉枯萎病菌4号生理小种中CHS基因家族的克隆与蛋白序列分析

毛超^1，2, 杨腊英², 戴青冬^1，2, 黄俊生²

(1.海南大学，海口 570228；2.中国热带农业科学院环境与植物保护研究所农业部热带农林有害生物入侵监测与控制重点实验室海南省热带农业有害生物检测与控制重点实验室，海口 571101)

收稿日期:2013-09-14 出版日期:2014-03-29 发布日期:2014-03-31
作者简介:毛超，男，硕士研究生，研究方向:微生物功能基因组学；E-mail:mc111666@163.com
基金资助:
国家公益性行业(农业)科研专项(200903049)，中央级公益性科研院所基本科研业务费专项(2011HZS1J022)，海南大学“211工程”建设项目

Cloning and Protein Sequence Analysis of CHS Gene Family in Fusarium oxysporumf.sp. cubense Race 4

Mao Chao^1，2, Yang Laying², Dai Qingdong^1，2, Huang Junsheng²

(1. Hainan University, Haikou 570228；2. China Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Science. Key Laboratory of Monitoring and Control of Tropical Agricultural and Forest Invasive Alien Pests, Ministry of Agriculture. Hainan Key Laboratory for Monitoring and Control of Triopical Agricultural Pests, Haikou 571101)

Received:2013-09-14 Published:2014-03-29 Online:2014-03-31

摘要/Abstract

摘要： 为了解香蕉枯萎病菌4号生理小种(FOC4)中的几丁质合酶(Chitin synthase，CHS)基因的特点，从而为进一步研究相关基因功能奠定基础。根据丝状真菌中CHS基因设计简并引物，采用RT-PCR的方法扩增得到FOC4中全长cDNA，测序后对该基因家族编码的蛋白进行分析。结果表明，FOC4共有8个CHS基因，根据蛋白聚类分析表明可其可划分为2个家族，并可根据氨基酸序列特征进一步划分为7个小类，该基因家族成员在不同种镰刀菌中保守程度较高。

关键词: FOC4, 几丁质合酶, 序列分析

Abstract: In order to know the characteristics of chitin synthase genes in Fusarium oxysporum f. sp. cubenes race 4 and lay an information foundation for further study of their functions, the degenerate primers were designed according to the sequence of several filamentous fungi’s CHS genes. The FOC4 CHS genes were amplified by PCR and RT-PCR methods. Their cDNAs were sequenced and protein sequences were analyzed. The results showed eight CHS genes were identified in FOC4. Through phylogenetic analysis, the FOC4 CHS proteins can be classified into two families. They also can be classified into seven classes through characteristics of the amino acid sequences analysis. The FOC4 CHS amino acid sequences showed high identity with other Fusarium species by homology camparison analysis.

Key words: FOC4, Cchitin synthase, Sequence analysis

毛超, 杨腊英, 戴青冬, 黄俊生. 香蕉枯萎病菌4号生理小种中CHS基因家族的克隆与蛋白序列分析[J]. 生物技术通报, 2014, 0(3): 79-85.

Mao Chao, Yang Laying, Dai Qingdong, Huang Junsheng. Cloning and Protein Sequence Analysis of CHS Gene Family in Fusarium oxysporumf.sp. cubense Race 4[J]. Biotechnology Bulletin, 2014, 0(3): 79-85.

参考文献

[1] Leong SK, Latiffah Z, Baharuddin S, et al. Molecular characterization of Fusarium oxysporum f. sp. cubense of banana[J]. American Journal of Applied Sciences, 2009, 6(7):1301-1307.
[2] 冯贻安, 崔志峰. 真菌几丁质合酶的研究进展[J]. 微生物学通报, 2008, 35(2):267-271.
[3] Schmidt M. Survival and cytokinesis of Saccharomyces cerevisiae in the absence of chitin[J]. Microbiology, 2004, 150(10):3253-3260.
[4] 刘向勇, 张小华, 王跃嗣, 等. 酿酒酵母CHS3基因缺失对热耐受性的影响[J]. 中国酿造, 2010(5):127-129.
[5] Weber I, Assmann D, Thines E, et al. Polar localizing class V myosin chitin synthases are essential during early plant infection in the plant pathogenic fungus Ustilago maydis[J]. The Plant Cell, 2006, 18(1):225-242.
[6] Larson TM, Kendra DF, Busman M, et al. Fusarium verticillioides chitin synthases CHS5 and CHS7 are required for normal growth and pathogenicity[J]. Curr Genet, 2011(57):177-189.
[7] 李秀峰, 庄佩君, 唐振华, 等. 真菌中的几丁质合成酶[J]. 世界农药, 2000, 22(5):33-38.
[8] 冀颖, 李梅, 田云龙, 等. 哈茨木霉几丁质合酶基因ThChsC的克隆及序列分析[J]. 中国农业科学, 2011, 44(17):3537-3546.
[9] Bulawa CE. Genetics and molecular biology of chitin synthesis in fungi[J]. Annu Rev Microbiol, 1993(47):505-534.
[10] 阳江华, 黄德宝, 刘术金, 等. 巴西橡胶树六个蔗糖转运蛋白基因的克隆与序列分析[J]. 热带作物学报, 2007, 28(4):32-38.
[11] Cohen E. Chitin synthesis and degradation as targets for pesticide action[J]. Arch Insect Biochem Physiol, 1993(22):245-261.
[12] Kim J, Lee H, Lee J, et al. Gibberella zeae chitin synthase genes, GzCHS5 and GzCHS7, are required for hyphal growth, perithecia formation, and pathogenicity[J]. Curr Genet, 2009(55):449-549.
[13] 石峰, 秦云霞, 唐朝荣, 等. 巴西橡胶树6个小G蛋白基因cDNA的克隆与序列分析[J]. 热带作物学报, 2009, 30(3):320-326.
[14] Munro CA, Gow NA. Chitin synthesis in human pathogenic fungi[J]. Med Mycol, 2001(39):41-53.
[15] Ni?o-Vega GA, Carrero L, San BG, et al. Isolation of the CHS4 gene of Paracoccidioides brasiliensis and its accommodation in a new class of chitin synthases[J]. Med Mycol, 2004(42):51-57.
[16] 漆艳香, 张欣, 蒲金基, 等. 巴西橡胶树棒孢霉落叶病菌Slt2类MAPK同源基因CMP1的克隆与序列分析[J]. 热带作物学报, 2010, 31(11):1951-1958.
[17] Proctor RH, Desjardins AE, Plattner RD, et al. A polyketide synthase gene required for biosynthesis of fumonisin mycotoxins in Gibberella fujikuroi mating population A[J]. Fungal Genet Biol, 1998(27):100-112.
[18] Muller C, Hjort CM, Hansen K, et al. Altering the expression of two chitin synthase genes differentially affects the growth and morphology of Aspergillus oryzae[J]. Microbiology, 2002, 148(12):4025-4033.
[19] Garcia RL J, Trilla JA, Castro C, et al. Characterization of the chitin biosynthesis process as a compensatory mechanism in the fks1 mutant of Saccharomyces cerevisiae[J]. FEBS Letters, 2000, 478(12):84-88.
[20] Munro CA, Whitton RK, Hughes HB, et al. CHS8-a fourth chitin synthase gene of Candida albicans contributes to in vitro chitin synthase activity, but is dispensable for growth[J]. Fungal Genetics and Biology, 2003, 40(2):146-158.
[21] Takeshita N, Yamashita S, Ohta A, et al . Aspergillus nidulans class V and VI chitin synthases CsmA and CsmB, each with a myosin motor-like domain, perform compensatory functions that are essential for hyphal tip growth[J]. Molecular Microbiology, 2006, 59(5):1380-1394.
[22] Lee JI, Choi JH, Park BC, et al. Differential expression ofthe chitin synthase genes of Aspergillus nidulans, chsA, chsB, and chsC, in response to developmental status and environmental factors[J]. Fungal Genetics and Biology, 2004, 41(6):635-646.
[23] Mellado E, Dubreucq G, Mol P, et al. Cell wall biogenesis in a double chitin synthase mutant(chsG-/chsE-)of Aspergillus fumigatus[J]. Fungal Genetics and Biology, 2003, 38(1):98-109.
[24] Mellado E, Aufauvre-Brown A, Gow NAR, et al. The Aspergillus fumigatus chsC and chsG genes encode Class III chitin synthases with different functions[J]. Mol Microbiol, 1996, 20(3):667-679.