家蚕中肠丝氨酸蛋白酶基因对家蚕微孢子虫入侵的响应模式  

Response pattern of serine protease genes of Bombyx mori midguts infected with Nosema bombycis

作　　者：王钰[1] 党晓群[1] 李治[1] 庞敏[2] 周泽扬[1] 王林玲[1] 

机构地区：[1]重庆师范大学动物生物学重点实验室,重庆400047 [2]重庆文理学院林学与生命科学学院,重庆402168

出　　处：《安徽农业大学学报》2013年第2期175-179,共5页Journal of Anhui Agricultural University

基　　金：重庆市自然科学基金(cstc2011jjA80003);重庆市教委科学技术研究项目(KJ120606);重庆市研究生教育教学改革研究项目(yjg12311);重庆文理学院科研项目(Z2009SKl2)共同资助

摘　　要：为探究接种家蚕微孢子虫(Nosema bombycis)24与48 h后,家蚕中肠6种丝氨酸蛋白酶基因(serineprotease gene,sps)的表达变化趋势,采用实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)方法对正常组和微孢子虫感染试验组的6个家蚕中肠丝氨酸蛋白酶基因(编号分别为sp1、sp3、sp5、sp8、sp11和sp12)进行定量分析。结果显示,与正常家蚕中肠内丝氨酸蛋白酶基因的转录水平相比,sp1、sp3、sp5和sp12在接种家蚕微孢子虫24与48 h后均上调表达,其中sp5上调最明显,接种家蚕微孢子虫24与48 h的表达量分别是对照的1.9和1.2倍;sp8和sp11在受微孢子虫感染24 h后表达上调,而48 h后表达量下降。结果表明,丝氨酸蛋白酶(serineprotease,SP)参与了家蚕中肠对微孢子虫入侵的免疫反应;不同丝氨酸蛋白酶上调倍数不同,说明不同丝氨酸蛋白酶参与家蚕免疫反应的程度不同;家蚕微孢子虫侵染48 h后,sp8和sp11出现了下调,推测微孢子虫的成功入侵抑制了sp8和sp11的表达。家蚕中肠不同丝氨酸蛋白酶在微孢子虫感染的不同时间的表达变化趋势为进一步研究丝氨酸蛋白酶家族在家蚕微孢子虫与宿主家蚕互作中的作用奠定了基础。To investigate the expression trends of six serine protease genes sps of silkworm Bombyx mori midguts induced by Nosema bombycis（N.bombycis）at post-infection（p.i.） 24 and 48 h,quantitative real-time PCR（qRT-PCR） was adopted to analyze the different mRNA levels of 6 sps（sp1,sp3,sp5,sp8,sp11 and sp12） between the N.bombycis-infected groups and the normal group.Compared to the normal group,the expression quantities of sp1,sp3,sp5 and sp12 were up-regulated at p.i.24 and p.i.48 h;it was notable that the transcriptional level of sp5 was increased 1.9 and 1.2 folds,respectively;the expression levels of sp8 and sp11 were up-regulated after 24 h,but down-regulated after 48 h.In conclusion,the transcriptional levels of the six sps of B.mori were all up-regulated after infected with N.bombycis for 24 h,suggesting that sps is involved in the defense of N.bombycis for 24 h,and sps is involved in the defense of N.bombycis.The different transcriptional levels of the genes also implied that they would play different roles in the response to the pathogen.sp5 might make a special function in the immune process and the infection of N.bombycis might be the areason to cause the down-regulated expression of sp8 and sp11.The expression trend of the six serine protease genes of silkworm B.mori L midguts at different time points would provide a new clue to discover the molecular mechanism of serine proteases in the interaction between B.mori and N.bombycis.

关 键 词：家蚕微孢子虫 家蚕中肠 丝氨酸蛋白酶 实时荧光定量PCR 

分 类 号：S884.21[农业科学—特种经济动物饲养]