Population Genetic Structure Analysis of Insect Fungal Pathogen Causing Silkworm Grey Muscardine

doi:10.16409/j.cnki.2095-039x.2016.01.009

Abstract

Abstract: Fungal pathogen Isaria javanica can cause a low rate of silkworm grey muscardine. In this study, two I. javanica populations collected from Qianshan and Tongxiang were fingerprinting analyzed using ISSR markers. In the assayed 19 isolates, 120 discernible DNA bands ranging in size from 200 to 2000 bp were generated by 12 ISSR primers, and 93 of the bands (70.45%) were polymorphic. Nei's gene diversity (h) and Shannon's Information index (I_s) showed a similar trend with the percentage of polymorphism at the population level. At the point of genetic similarity coefficient 0.76, the strains could be divided into 2 groups which correlated with the geographic origin of sampling. The result of 3-D principal coordinate analyses (PCA) based on ISSR data shared high similarity with UPGMA cluster analysis. The gene flow (N_m) between the two populations was 0.5678, and the genetic differentiation coefficient (G_st) was 0.3287, which indicates higher genetic diversity between populations than within populations. Single population clustering results showed that Qianshan population had an evident dominant group at the genetic similarity coefficient of 0.8, while Tongxiang population did not have a predominant lineage. Based on the analyses above, we can conclude that pathogen populations causing enzootic grey muscardine are geographically heterogenous, which may be an important factor for the maintenance of enzootic populations.

Key words: Bombyx mori, grey muscardine, Isaria javanica, enzootic, ISSR

CLC Number:

S476.12

ZHANG ShengLi, CHEN Xue, PU Shunchang, XU Yanping, HE Lingmin, LI Zengzhi, LUAN Fenggang. Population Genetic Structure Analysis of Insect Fungal Pathogen Causing Silkworm Grey Muscardine[J]. journal1, 2016, 32(1): 55-61.

References

[1] 孙黎峰, 韩益飞, 杨佩华, 等. 2008年江苏省秋季蚕病疫情调查分析[J]. 江苏蚕业, 2009, (3):7-11
[2] 李佳丽, 蔡悦, 栾丰刚, 等. 皖南与皖西南蚕区球孢白僵菌种群遗传结构的ISSR分析[J]. 应用生态学报, 2010, 21(12):3239-3247.
[3] 穆莉. 家蚕的真菌病//中国虫生真菌研究与应用(第三卷)[M]. 北京:中国农业科学技术出版社, 1993, 50-53.
[4] 朱方容, 邱全立, 何彬, 等. 寄生桑粉虱的拟青霉菌对家蚕的致病性研究[J]. 蚕业科学, 1997, 22(3):137-141.
[5] 黄勃, 王四宝, 金伟, 等. 家蚕灰僵病病原的鉴定[J]. 蚕业科学, 2008, 34(2):257-261.
[6] 朱衡, 瞿峰, 朱立煌. 利用氯化苄提取适于分子生物学分析的真菌DNA[J]. 菌物系统, 1994, 13(1):34-40.
[7] Wang S, Miao X, Zhao W, et al. Genetic diversity and population structure among strains of the entomopathogenic fungus, Beauveria bassiana, as revealed by inter-simple sequence repeats (ISSR)[J]. Mycological Research, 2005, 109(12):1364-1372.
[8] Hall T A. BioEdit:a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT[J]. Nucleic Acids Symposium Series, 1999, 41:95-98.
[9] Tamura K, Dudley J, Nei M, et al. MEGA4:Molecular evolutionary genetics analysis (MEGA) software version 4.0[J]. Molecular Biology and Evolution, 2007, 24(8):1596-1599.
[10] 王勇, 陈克平, 姚勤. 系统发生分析软件PAUP和TreePuzzle使用方法介绍[J]. 安徽农业科学, 2008, 36(2):818-821, 829.
[11] Swofford D L. PAUP*. Phylogenetic Analysis Using Parsimony (*and Other Methods). Version 4[M]. Massachusetts:Sinauer Associates, 2003.
[12] Felsenstein J. Confidence limits on phylogenies:an approach using the bootstrap[J]. Evolution, 1985, 39(4):783-791.
[13] Ronquist F, Huelsenbeck J P. MrBayes 3:Bayesian phylogenetic inference under mixed models[J]. Bioinformatics, 2003, 19(12):1572-1574.
[14] 王勇, 陈克平, 姚勤. 系统发生分析程序MrBayes 3.1使用方法介绍[J]. 安徽农业科学, 2009, 37(33):16665-16669.
[15] Yeh F C, Yang R C, Boyle T B J, et al. POPGENE, the user-friendly shareware for population genetic analysis//Molecular Biology and Biotechnology Centre[M]. University of Alberta, 1997.
[16] Rohlf F J. NTSYS-pc:Numerical Taxonomy and Multivariate Analysis System, Version 2.1, User Guide[M]. New York:Exeter Software, 2000.
[17] 朱方容. 广西部分地区桑蚕僵病流行的原因及防控对策[J]. 广西蚕业, 2008, 45(3):29-32.
[18] 孙始威, 孙振兴, 葛宜和, 等. 基于ISSR标记的扁玉螺(Neverita didyma)自然居群遗传结构[J]. 生态学报, 2008, 28(11):5499-5505.
[19] 杜广祖, 王晶晶, 陈斌. 云南球孢白僵菌的遗传多样性研究[J]. 云南农业大学学报, 2012, 27(3):327-333.
[20] 栾丰刚. 引起昆虫自然流行病的昆虫病原真菌种群遗传结构研究[D]. 合肥:安徽农业大学, 2011.
[21] Tigano-Milani M S, Honeycutt R J, Lacey L A, et al. Genetic variability of Paecilomyces fumosoroseus isolates revealed by molecular markers[J]. Journal of Invertebrate Pathology, 1995, 65(3):274-282.
[22] Estrada M E, Camacho M V, Benito C. The molecular diversity of different isolates of Beauveria bassiana (Bals.) Vuill. as assessed using intermicrosatellites (ISSRs)[J]. Cellular and Molecular Biology Letters, 2007, 12(2):240-252.
[23] Glare T R, Inwood A J. Morphological and genetic characterization of Beuvaria spp. from New Zealand[J]. Mycological Research, 1998, 102(2):250-256.
[24] Fernandes E K K, Moraes A M L, Pacheco R S, et al. Genetic diversity among Brazilian isolates of Beauveria bassiana:comparisons with non-Brazilian isolates and other Beauveria species[J]. Journal of Applied Microbiology, 2009, 107(3):760-774.
[25] 胡晓磊, 何玲敏, 陈雪, 等. 中国南方球孢白僵菌的遗传多样性和种群遗传结构[J]. 中国生物防治学报, 2013, 29(1):31-41.
[26] 何玲敏, 胡晓磊, 陈雪, 等. 中国北方球孢白僵菌的遗传多样性和种群遗传结构[J]. 应用生态学报, 2012, 23(11):3087-3095.
[27] Shimazu M, Takatsuka J. Isaria javanica (anamorphic Cordycipitaceae) isolated from gypsy moth larvae, Lymantria dispar (Lepidoptera:Lymantriidae), in Japan[J]. Applied Entomology and Zoology, 2010, 45(3):497-504.
[28] Specht A, Azevedo J L, de Luna E A, et al. Occurrence of the entomopathogenic fungus Isaria javanica (Frieder. & Bally) Samson & Hywell-Jones (Fungi, Sordariomycetes) infecting Lonomia obliqua Walker (Lepidoptera, Saturniidae, Hemileucinae) caterpillars[J]. Revista Brasileira De Entomologia, 2009, 53(3):493-494.
[29] Scorsetti A C, Humber R A, de Gregorio C, et al. New records of entomopathogenic fungi infecting Bemisia tabaci and Trialeurodes vaporariorum, pests of horticultural crops, in Argentina[J]. BioControl, 2008, 53(5):787-796.
[30] 展茂魁, 何玲敏, 陈名君, 等. 防治假眼小绿叶蝉的虫生真菌高毒菌株的筛选及田间防治效果[J]. 中国生物防治学报, 2012, 28(1):41-46.