光滑鳖甲Anatolica polita抗冻蛋白基因的发育阶段性表达以及低温对其表达的影响  

Developmental and Cold Stress Regulation of Antifreeze Protein Genes in Anatolica polita borealis

作　　者：王晶[1] 邱立明[1] 王岩[1] 马纪[1] 

机构地区：[1]新疆大学生命科学与技术学院/新疆生物资源与基因工程重点实验室,乌鲁木齐830046

出　　处：《新疆农业科学》2010年第9期1697-1702,共6页Xinjiang Agricultural Sciences

基　　金：国家自然科学基金项目"新疆荒漠昆虫准噶尔小胸鳖甲抗冻蛋白基因表达特性研究"(30760056);新疆生物资源基因工程重点实验室开放课题"荒漠昆虫光滑鳖甲抗冻机理研究"(XJDX0201-2010-05);新疆大学博士启动基金"环境因素对光滑鳖甲抗冻蛋白基因表达的影响"(BS090129)

摘　　要：【目的】光滑鳖甲是广泛分布于新疆荒漠地区的一种拟步甲科(Tenebrionidae)昆虫,它能够通过抗冻蛋白的表达从而在长时间低温和剧烈的温度变化等环境条件下生存。研究光滑鳖甲抗冻蛋白基因(Apafp)在不同发育阶段表达差异和冷胁迫对其表达的影响。【方法】通过实时定量PCR方法对Apafp752和Apafp914两种抗冻蛋白基因的mRNA水平变化规律进行了研究。【结果】随着幼虫龄期的增大,Apafps mRNA水平逐渐增加,至8～9龄幼虫阶段达到最高约为小龄幼虫的7～8倍,蛹期时则显著降低。血淋巴液的渗透浓度值也呈现相同趋势。【结论】发育阶段的变化是对光滑鳖甲抗冻蛋白基因表达的重要调节因素。在低温胁迫下4～6龄幼虫afps的表达显著提高,说明冷胁迫能促进幼虫afps的累积。结合Apafps基因表达和低温存活率可以推测光滑鳖甲大龄幼虫可能是一种过冬虫态。【Objective】Anatolica polita,a beetle in Tenebrionidae,is widely distributed in the desert of Xinjiang.It can survive the cold and huge temperature fluctuation by producing antifreeze proteins（AFPs）.To investigate the expression of antifreeze protein genes in different developmental stages,and influence of cold stress on the developmental expression of antifreeze protein genes.【Method】mRNA levels of two AFP genes Apafp752 and Apafp914 were tested by using Real-Time PCR.【Result】The results showed that mRNA levels of Apafp752 and Apafp914 in the 8th and 9th larvae were the highest which were 7-8 times of the small instars,but decreased in pupa.The osmolality displayed similar trends with the larva development.【Conclusion】This result implied that development stage is a primary factor in the regulation of expression of Apafps.Under cold stress,Apafps expression in 4-6 instars was significantly increased,indicating that expression of larval Apafps could be stimulated by cold stress.It could be predicted that old larvae might be in one of the overwintering stages in Anatolica polita considering the expression pattern in the immature stages and the survival rate of the larvae under cold stress.

关 键 词：光滑鳖甲 抗冻蛋白(AFPs) REAL-TIME PCR 发育阶段 冷胁迫 

分 类 号：S188[农业科学—农业基础科学]