昆虫学报 ›› 2019, Vol. 62 ›› Issue (8): 930-936.doi: 10.16380/j.kcxb.2019.08.005

• 研究论文 • 上一篇    下一篇

家蚕微孢子虫感染对家蚕海龟蛋白(Bmtutl)基因表达水平的影响

朱峰, 高建华, 张永红, 肖圣燕, 邵榆岚, 唐芬芬, 白兴荣*   

  1. (云南省农业科学院蚕桑蜜蜂研究所, 云南蒙自 661101)
  • 出版日期:2019-08-20 发布日期:2019-08-29

Effect of Nosema bombycis infection on the expression level of Bombyx Turtle (Bmtutl) gene in Bombyx mori

ZHU Feng, GAO Jian-Hua, ZHANG Yong-Hong, XIAO Sheng-Yan, SHAO Yu-Lan, TANG Fen-Fen, BAI Xing-Rong*   

  1.  (Institute of Sericulture and Apiculture, Yunnan Academy of Agricultural Sciences, Mengzi, Yunnan 661101, China)
  • Online:2019-08-20 Published:2019-08-29

摘要:

【目的】本研究旨在阐明家蚕微孢子虫Nosema bombycis感染不同时间对家蚕Bombyx mori幼虫不同组织中家蚕海龟蛋白(Bombyx Turtle, Bmtutl)基因表达水平的影响,为揭示家蚕微孢子虫的侵染机制奠定基础。【方法】利用生物信息学方法对家蚕海龟蛋白3种亚型Bmtutl-464, Bmtutl-519和Bmtutl-810的序列结构特征进行了分析;利用qPCR检测家蚕微孢子虫感染后12, 24, 48, 72, 96和120 h,家蚕幼虫中肠、血淋巴与脂肪体组织中Bmtutl-464, Bmtutl-519和Bmtutl-810基因表达水平的变化情况。【结果】家蚕海龟蛋白3种亚型的二级结构均主要由无规则卷曲、α螺旋、β转角和延伸链组成,其中无规则卷曲所占比例最高。但是PredictProtein分析发现,Bmtutl-464, Bmtutl-519和Bmtutl-810之间的蛋白/多核苷酸结合位点存在较大差异。qPCR结果表明,感染家蚕微孢子虫后,家蚕幼虫中肠、血淋巴与脂肪体组织中Bmtutl-464, Bmtutl-519和Bmtutl-810基因的整体表达处于被抑制状态,尤其在脂肪体中最为明显:Bmtutl-519和Bmtutl-810基因的表达在家蚕微孢子虫感染家蚕后的72 h开始受到显著抑制,特别是Bmtutl-519基因,其相对表达水平均不到对照的5.0%。【结论】家蚕海龟蛋白这3种亚型的序列结构特征存在较大差异,家蚕微孢子虫感染在一定程度抑制了家蚕幼虫中肠、血淋巴与脂肪体组织中Bmtutl-464, Bmtutl-519和Bmtutl-810基因尤其是Bmtutl-519的表达。结果说明,与其他两种家蚕海龟蛋白亚型相比,Bmtutl-519蛋白可能在家蚕微孢子虫侵染宿主的过程中起主要作用。

关键词: 家蚕, 家蚕微孢子虫, 家蚕海龟蛋白, 序列分析, 时相表达

Abstract: 【Aim】 The aim of this study is to elucidate the effect of Nosema bombycis infection on the expression level of Bombyx Turtle (Bmtutl) gene in different tissues of larvae of the silkworm (Bombyx mori) at different time post infection, so as to lay the foundation for revealing the infection mechanism of N. bombycis. 【Methods】 The sequence characteristics of three isoforms of Bmtutl (Bmtutl-464, Bmtutl-519 and Bmtutl-810) of B. mori were analyzed by bioinformatics methods. The expression levels of Bmtutl-464, Bmtutl-519 and Bmtutl-810 in the midgut, hemolymph, and fat body of B.mori larvae infected by N. bombycis at 12, 24, 48, 72, 96 and 120 h post inoculation, respectively, were detected by qPCR. 【Results】 The secondary structures of the three Bmtutl isoforms are mainly composed of random coil, α-helix, β-turn and extended strand, and the proportion of random coil is the highest. However, PredictProtein analysis indicated that there was a large difference in protein/polynucleotide binding sites between Bmtutl-464, Bmtutl-519 and Bmtutl-810. qPCR analysis showed that the expression of Bmtutl-464, Bmtutl-519 and Bmtutl-810 in the midgut, hemolymph and fat body of B. mori larvae was inhibited after N. bombycis infection, and most significantly inhibited in the fat body. The expression of Bmtutl-519 and Bmtutl-810 in the fat body of B. mori larvae infected by N. bombycis was strongly inhibited since 72 h post inoculation, and especially the relative expression level of Bmtutl-519 in the fat body in the treatment group was less than 5.0% of that in the control. 【Conclusion】 The three isoforms of Bmtutl have different sequence characteristics. N. bombycis infection inhibits the expression of Bmtutl-464, Bmtutl-519 and Bmtutl-810 in the midgut, hemolymph and fat body of B. mori larvae, especially the expression of Bmtutl-519. These results suggest that Bmtutl-519 protein may play a major role in the process of N. bombycis infecting the silkworms as compared with the other two Bmtutl isoforms.

Key words:  Bombyx mori, Nosema bombycis, Bombyx mori Turtle protein, sequence analysis, phase expression