Oct. 22-25, 2012
Screening on Genetic Markers Relating to Trait of High Royal Jelly Production Using Gene Chip
Jiao Pan1, Wenfeng Li, Zhiguo Li, Limin Yan, Shenglu Chen, Songkun Su,*
College of Animal Sciences, Zhejiang University, Hangzhou, China
College of Bee Science, Fujian Agriculture and Forestry University, Fuzhou, China
Royal jelly, a principal food of the honeybee queen and larva under 3 days old, is secreted by the hypopharyngeal and mandibular glands of nursing bees. Royal jelly has peculiar effects on health care owning to its highly active nutrients. Through years of directional selection and group closed propagation breeding, we have successfully cultivated a bee breed of high royal jelly yield, which enable China's both royal jelly production and export volume rank first in the world. So, the researches on genes related to high royal jelly producing traits and their molecular mechanisms have important theoretical significance and promising prospects in practice.
In this study, we cultivated four colonies of high royal jelly yielding bees(HRJB) and low royal jelly yielding bees (LRJB) respectively (F1,16= 66.04, P = 0.0002) by directional selection breeding. A new gene chip Agilent of honeybee covering 11689 genes (9792 genes retrieved from NCBI, 1722 newly tested genes and 175 genes which might be closely related to the royal jelly secretion) was designed and prepared, and hybridized with cDNA of nursing bees collected when they were feeding the larva in queen cells. In total, 369 differentially expressed genes between HRJB and LRJB were detected, and 201 genes were up-regulated (1.70%) in HRJB, while 168 genes were down-regulated (1.43%) in HRJB.
The cluster analysis, gene ontology (GO) and pathway of 369 differently expressed genes between HRJB and LRJB were analyzed using bio-informatics software and database. GO analyses showed that the differently expressed genes mainly participated in four biological processes including developmental regulation of organic tissue, synthesis and deposit of nutrients, oxidoreduction coenzyme and glucose metabolism, biosynthesis and metabolism of organic acid. They mainly have the following molecular functions: neuropeptide and its' receptor activity, metabolism enzymes activity of various nutrients. Pathway analyses revealed that the differently expressed genes mainly belong to 46 pathways, which can be classified into signal pathway, synthesis and metabolism of nutrients pathway, processing and modification of protein pathway.
Furthermore, 10 different colonies of bee samples were collected from royal jelly producing colonies in field, 5 of which belonged to HRJB, and another 5 were LRJB. Ten of the differentially expressed genes obtained above were further confirmed using qRT-PCR and correlation analysis, 3 genes were successfully verified to be the molecular markers relating to high royal jelly production trait. This study provided genetic basis for further studies on molecular mechanisms of high royal jelly production.
The research was supported by earmarked funds for Modern Agro-industry Technology Research System (No. CARS-45-KXJ3) and Nature and Science Foundation Commission of Zhejiang Province (R3080306) to SKS.
1. 浙江大学动物科学学院 杭州 中国
2. 福建农林大学蜂学学院 福州 中国
本研究通过定向选育手段，在同一蜂种中分别培育了王浆高产和王浆低产蜜蜂(x vs y, F1,16= 66.04, P = 0.0002)。根据从NCBI上检索的9792条已注释的蜜蜂基因、本实验室在蜜蜂幼虫中新检测的1722条基因和175条与王浆分泌可能紧密相关的基因，设计和制备了蜜蜂基因芯片（Agilent），分别与王浆高产蜜蜂（HRJB）及王浆低产蜜蜂(LRJB)中正在吐浆的哺育蜂的头部cDNA进行杂交，初步筛选出369条差异表达的基因，其中在王浆高产蜜蜂中上调表达的基因有201个（1.70%），下调表达168个（1.43%）。
关键词：蜜蜂(Apis mellifera)；王浆产量性状；基因芯片；qRT-PCR 分子标记