不同移虫日龄蜂王卵巢中hexamerin110、hexamerin70b的差异表达分析
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摘要
为探究不同移虫日龄的西方蜜蜂蜂王卵巢中hexamerin110、hexamerin70b的表达及其调控机理,以β-actin为内参基因,运用q RT-PCR技术检测不同移虫日龄蜂王卵巢中hexamerin110、hexamerin70b m RNA的表达水平,用Western blot方法检测hexamerin110、hexamerin70b蛋白的表达。结果显示,hexamerin110、hexamerin70无论是在m RNA水平还是蛋白水平,随着移虫日龄的增加,其表达量均呈降低的趋势。1日龄移虫培育的蜂王卵巢hexamerin110基因表达量显著高于其他组(P<0.05),1日龄移虫培育的蜂王卵巢的hexamerin70b基因的表达量极显著高于其他组(P<0.01)。Hexamerin110、hexamerin70b在1日龄移虫发育的蜂王卵巢中的表达水平最高,推测其可能与蜂王生殖发育以及级型分化有关。
To explore the expression and molecular regulation mechanism of hexamerin110 and hexamerin70 b in the ovaries of queens reared from different instar of the grafted larvae,m RNA and protein expression of hexamerin110 and hexamerin70 b were assessed by q RT-PCR and Western blot. The results showed that the expression levels of the hexamerin110 and hexamerin70 b decreased with the increasing instar of the grafted larvae,the expression levels of the hexamerin110 in ovaries of queens reared from 1 day instar of the grafted larvae were significantly higher than others( P < 0. 05).Meanwhile,the expression levels of the hexamerin70 b in ovaries of queens reared from 1 day instar of the grafted larvae were extremely significantly higher than others( P < 0. 01). The expression levels of the hexamerin110 and hexamerin70 b of 1 day instar of the grafted larvae are the highest,it is speculated that hexamerin110 and hexamerin70 b may be associated with queen reproductive development and caste differentiation.
To explore the expression and molecular regulation mechanism of hexamerin110 and hexamerin70 b in the ovaries of queens reared from different instar of the grafted larvae,m RNA and protein expression of hexamerin110 and hexamerin70 b were assessed by q RT-PCR and Western blot. The results showed that the expression levels of the hexamerin110 and hexamerin70 b decreased with the increasing instar of the grafted larvae,the expression levels of the hexamerin110 in ovaries of queens reared from 1 day instar of the grafted larvae were significantly higher than others( P < 0. 05).Meanwhile,the expression levels of the hexamerin70 b in ovaries of queens reared from 1 day instar of the grafted larvae were extremely significantly higher than others( P < 0. 01). The expression levels of the hexamerin110 and hexamerin70 b of 1 day instar of the grafted larvae are the highest,it is speculated that hexamerin110 and hexamerin70 b may be associated with queen reproductive development and caste differentiation.
引文
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Wheeler DE,Buck NA.Storage proteins in ants during development and colony founding[J].Journal of Insect Physiological,1995,41:885-894.
Wheeler DE,Tuchinskaya I,Buck NA,et al.Hexameric storage proteins during metamorphosis and egg production in the diamondback moth,Plutella xylostella(Lepidoptera)[J].Journal of Insect Physiology,2000,46:951-958.
Wirtz P,Beetsma J.Induction of caste differentiation in the honeybee(Apis mellifera)by juvenile hormone[J].Entomologia Experimentalis et Applicata,1972,(15):517-520.
Bitondi MMG,Nascimento AM,Cunha AD,et al.Characterization and expression of the Hex 110 gene encoding a glutamine-rich hexamerin in the honey bee,Apis mellifera[J].Archives of Insect Biochemistry and Physiology,2006,63(2):57-72.
Burmester T,Scheller K.Ligands and receptors:Common theme in insect storage protein transport[J].Naturwissenschaften,1999,86:468-474.
Chen X,Hu FL.Molecular determinants of caste differentiation in honey bee[J].Journal of Bee,2011,4:1003-1091.[陈璇,胡福良.雌性蜜蜂级型决定的分子机制[J].蜜蜂杂志,2011,4:1003-1091]
Cunha AD,Nascimento AM,Guidugli KR,et al.Molecular cloning and expression of a hexamerin cD NA from the honey bee,Apis mellifera[J].Journal of Insect Physiology,2005,51:1135-1147.
Hartfelder K,Engels W.2 social insect polymorphism:Hormonal regulation of plasticity in development and reproduction in the honeybee[J].Current Topics in Developmental Biology,1998,40:45-77.
Martins JR,Nunes FMF,Cristino AS,et al.The four hexamerin genes in the honey bee:Structure,molecular evolution and function deduced from expression patterns in queens,workers and drones[J].BMC Molecular Biology,2010,11:23.
Martins JR,Nunes FMF,Sim7es ZLP,et al.A honeybee storage protein gene,hex70a,expressed in developing gonads and nutritionally regulated in adult fat body[J].Journal of Insect Physiology,2008,54:867-877.
Page RE,Peng CYS.Aging and development in social insects with emphasis on the honey bee,Apis mellifera L.[J].Experimental Gerontology,2001,36(4):695-711.
Pan ML,Telfer WH.Methionine-rich hexamerin and arylphorin as precursor reservoirs for reproduction and metamorphosis in female luna moths[J].Archives of Insect Biochemistry and Physiology,1996,33:149-162.
Pan ML,Telfer WH.Storage hexamer utilization in two lepidopterans:Differences correlated with the timing of egg formation[J].Journal of Insect Science,2001,1:1-9.
Peng WJ.About the royal jelly[J].Apiculture of China,2007,58(12):46.[彭文君.关于蜂王浆[J].中国蜂业,2007,58(12):46]
Riddiford LM,Truman JW,Mirth CK,et al.A role for juvenile hormone in the prepupal development of Drosophila melanogaster[J].Development,2010,137(7):1117-1126.
Seo SJ,Kang YJ,Cheon HM,et al.Distribution and accumulation of storage protein-1 in ovary of Hyphantria cunea Drury[J].Archives of Insect Biochemistry and Physiology,1998,37:115-128.
Shen F.Research on the Effect of the Instar of the Grafted Larvae on Growth and Development and Ovary Proteomic of Queen[D].Yangzhou University,2015.[沈芳.移虫日龄对蜂王生长发育及蜂王卵巢蛋白质组学的影响[D].扬州大学,2015]
Shi YY.Influence of Nutritional and Spatial Factors in the Development of Females Honeybees(Apis mellifera,Apidae)[D].Jiangxi Agricultural University,2011.[石元元.营养和空间因素对雌性蜜蜂发育的影响[D].江西农业大学,2011]
Wheeler DE,Buck NA.Storage proteins in ants during development and colony founding[J].Journal of Insect Physiological,1995,41:885-894.
Wheeler DE,Tuchinskaya I,Buck NA,et al.Hexameric storage proteins during metamorphosis and egg production in the diamondback moth,Plutella xylostella(Lepidoptera)[J].Journal of Insect Physiology,2000,46:951-958.
Wirtz P,Beetsma J.Induction of caste differentiation in the honeybee(Apis mellifera)by juvenile hormone[J].Entomologia Experimentalis et Applicata,1972,(15):517-520.