中国米虾bursicon基因的功能及作用机制  

THE FUNCTION AND MECHANISM OF BURSICON GENE IN CARIDINA

作　　者：任丽琦 翁洁羊 王鑫 孙金生 李冉 REN Li-Qi, WENG Jie-Yang, WANG Xin, SUN Jin-Sheng , LI Ran(Tianjin Key Laboratory of Animal and Plant Resistance, College of Life Science, Tianjin Normal University, Tianjin 300387, Chin)

机构地区：[1]天津师范大学生命科学学院天津市动植物抗性重点实验室,天津300387

出　　处：《水生生物学报》2018年第3期503-511,共9页Acta Hydrobiologica Sinica

基　　金：国家自然科学基金青年基金(31302168);天津市应用基础与前沿技术研究计划(14JCYBJC30700);天津师范大学博士基金(52XB1303)资助~~

摘　　要：为探讨米虾鞣化激素在其蜕皮周期及表皮角质层形成过程中的作用,采用PCR技术克隆得到了米虾鞣化激素两个亚基基因的开放阅读框(ORF)序列。bursicon-αORF全长441 bp,共编码146个氨基酸;bursicon-βORF全长411 bp,共编码136个氨基酸。利用实时荧光定量PCR分析米虾整个蜕皮周期中鞣化激素2个亚基基因的表达特征,结果发现,鞣化激素bursicon-α和bursicon-β在米虾蜕皮周期的各个阶段的相对表达量存在差异,在蜕皮前期(D期)相对表达量开始上升,到D3期时相对表达量最高,蜕皮期E期相对表达量最低。RNA干扰(RNA interference,RNAi)介导bursicon-α和bursicon-β基因沉默后,发现米虾的蜕皮周期延长,表皮角质层明显变薄。结果提示,鞣化激素(Bursicon)与新形成的外骨骼中角质层的加厚与硬化密切相关,进而影响蜕皮时间。To study the function of bursicon gene in Caridina during molting and cuticle formation, its open reading frame（ORF） containing two subunits genes were amplified by PCR respectively. The ORF of bursicon-α is 441 bp in length, encoding a protein with 146 amino acid residues. The ORF of bursicon-β is 411 bp in length, encoding a protein with 136 amino acid residues. The dynamic changes in the expression of both bursicon subunit genes during the molt cycle in Caridina have been analyzed by the real-time quantitative PCR. Relative expression of both bursicon-αand bursicon-β had different levels in different stages of the molt cycle, which increased through premolt stage（D stage） and reached their peak level at the D3 stage. The m RNA accumulation decreased to its lowest level at the molt stage（E stage）. RNA interference-mediated knockdown of bursicon-α and bursicon-β retard the molting process and ecdysis behavior of Caridina. The cuticle of ds RNA-treated Caridina was thinner than that in the control group. These findings demonstrate that bursicon is involved in the thickening and hardening of cuticle in newly formed exoskeleton.Moreover, bursicon plays an important role in molt cycle.

关 键 词：米虾 鞣化激素 表皮鞣化 蜕皮周期 实时荧光定量PCR RNA干扰 

分 类 号：Q344.1[生物学—遗传学]