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孟氏隐唇瓢虫气味结合蛋白ComOBP1基因的克隆和时空表达
  • ISSN号:1674-0858
  • 期刊名称:《环境昆虫学报》
  • 时间:0
  • 分类:Q963[生物学—昆虫学] S476[农业科学—农业昆虫与害虫防治;农业科学—植物保护]
  • 作者机构:[1]有害生物控制与资源利用国家重点实验室,生物多样性演化与保护广东普通高校重点实验室,中山大学生命科学学院,广州510275, [2]广东省昆虫研究所,广东省农业害虫综合治理重点实验室,广东省野生动物保护与利用公共实验室,广州510260, [3]根特大学生物工程学院植物保护系,比利时根特市B-9000
  • 相关基金:国家重点基础研究发展计划(973计划)项目(2013CB127600); 国家自然科学基金面上项目(31171899,31572052);国家自然科学基金项目(31301720)
中文摘要:

采用RT-PCR和RACE技术,从孟氏隐唇瓢虫Cryptolaemus montrouzieri Mulsant中成功克隆出气味结合蛋白(Com OBP1)基因的全序列(Genbank登陆号:KU170686)。Com OBP1基因全长922 bp,包括5'端长为40 bp的非编码区域(UTR),及3'端长为462 bp的UTR,开放阅读框ORF长为420 bp,编码139个氨基酸,预测的分子量为15.516 k Da,等电点p I为6.57,存在AATAAA加尾信号。N-末端疏水区包含由20个氨基酸构成的信号肽,无跨膜结构,有4个保守的半胱氨酸,属于Minus-C OBP,也是在孟氏隐唇瓢虫中发现的第一个Minus-C OBP。氨基酸序列中有且仅有一个N-糖基化位点为62 NLSA,并存在2个潜在的磷酸化位点。与其它昆虫的Minus-C OBPs进行同源性比较并构建系统发育树,发现与同为鞘翅目Coleoptera昆虫的同源性较高。利用Real-time PCR、RT-PCR技术对Cmon OBP1基因在孟氏隐唇瓢虫不同发育阶段,不同组织,不同营养条件及不同食性下的表达水平进行了测定,结果显示,Cmon OBP1基因在整个发育阶段均有表达,雄性成虫期具有最高表达量,且多在成虫的头部及翅部表达。当营养条件发生变化时,表达丰度不会发生变化,当猎物由天然猎物柑橘粉蚧Planococcus citri Risso变成碗豆修尾蚜Megoura japonica Matsumura时,表达量会明显下降。该结果表明,孟氏隐唇瓢虫的不同发育阶段、不同组织及猎物种类会影响Cmon OBP1基因的表达,从而进一步影响其嗅觉行为。同时,Cmon OBP1基因可能在雄虫相关的信息素感受过程中发挥着重要作用。

英文摘要:

The odorant binding protein gene Cmon OBP1( Genbank number: KU170686) from Cryptolaemus montrouzieri Mulsant was cloned by technology of RT-PCR and RACE. The full length was922 bp,containing a 5' UTR( untranslated region) of 40 bp and a 3' UTR of 462 bp. ORF( Open Reading Frame) was 420 bp which encoded a polypeptide of 139 amino acids with an estimated molecular weight of15. 516 k Da and p I of 6. 57. Besides,the sequence had an AATAAA tailing signal. The predicted Nterminal hydrophobic region containing 20 amino residues displayed the characteristic features of a signal peptide,and no transmembrane domain was predicted. Four conservative cysteine residues classified Cmon OBP1 as the first Minus-C OBP which was found in C. montrouzieri. The amino acid sequence had only one N-glycosylation site 62 NLSA,and two potential phosphorylation sites. Compared with other insect Minus-C OBPs,Cmon OPB1 was much identitier to the other Coleoptera. By Real-time PCR and RT-PCR methods,we determined the expression pattern of Cmon OBP1 in different developmental stages,various adult tissues,different larval food regimes and other food. Spatio-temporal expression result showed that Cmon OBP1 expressed in every stage and the male adult had the highest level. In addition,Cmon OPB1 had a higher expression level in adult head and wing than that of other adult tissues. When larval diet changed either poor or rich,the expression level wouldn' t be influenced markedly,while when the food changed from natural prey Planococcus citri Risso to Megoura japonica Matsumura,the expression level decreased significantly. The result suggested that the expression level of Cmon OBP1 gene was development,tissues and prey type dependent,indicating a distinct sensitivity in olfactory. In addition,Cmon OBP1 gene might play an important role in the process of male adult feeling related pheromone.

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期刊信息
  • 《环境昆虫学报》
  • 中国科技核心期刊
  • 主管单位:广东省科学技术协会
  • 主办单位:广东省昆虫学会 中国昆虫学会
  • 主编:庞义
  • 地址:广州新港西路105号
  • 邮编:510260
  • 邮箱:J.Env.Ent@gdei.gd.cn
  • 电话:020-84456131
  • 国际标准刊号:ISSN:1674-0858
  • 国内统一刊号:ISSN:44-1640/Q
  • 邮发代号:46-18
  • 获奖情况:
  • 国内外数据库收录:
  • 美国剑桥科学文摘,英国动物学记录,中国中国科技核心期刊,中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:2216