The Quantitative Research Of Transcription Levels Of BmGST Genes In Bombyx Mori

Glutathione S-transferases (GSTs) are a multifunctional supergene family and some play an important role in insecticide resistance. In order to explore the roles of Bombyx mori glutathione S-transferase genes (BmGSTs), we investigated the transcription levels of some BmGST genes in Bombyx mori and Bombyx mandarina with the dual spike-in qPCR method. The main results are as follows:1. Induced expression and quantitative analysis of BmGSTd1We used a real-time fluorescent quantitative PCR method to detect the transcription levels of BmGSTd1 in different tissues of the 5th instar larvae feeding on both normal mulberry leaves and sodium fluoride treated mulberry leaves. The detection results were normalized by using 3 Bombyx mori internal reference genes namely Actin3, GAPDH and 28S rRNA. The results indicated that the transcription levels of BmGSTd1 were different in various tissues of the 5th instar larvae. They were also varied while different internal reference genes were used. It is thus concluded that, for detection of gene transcription levels, it is very important to choose adequate internal reference genes so as to ensure the reliability of the detection results.2. Transcription levels of BmGST genes in varieties of silkwormWe have determined the transcription levels of BmGSTs in five varieties of Bombyx mori and Bombyx mandarina with the dual-spike-in qPCR method. The results indicated that the transcription levels of BmGSTs shows differences between different varieties. To sum up, the transcription levels of BmGSTs in Jingsong and Haoyue are highest, those of L11 are following, and the transcription levels of BmGSTs in Dazao and Bombyx mandarina are lowest. And the transcription levels of BmGSTs in different tissues are also different.3. Insecticides-induced transcription levels of BmGST genes We have determined the transcription levels of BmGSTs induced by Dichlorvos and Deltamethrin in silkworm L11 and Bombyx mandarina with the dual-spike-in qPCR method. The results indicated that the transcription levels of BmGSTs induced by Dichlorvos and Deltamethrin in midgut of L11 are up-regulated most, the following are fat body. In Bombyx mandarina, the transcription levels of BmGSTs induced by Deltamethrin are significantly up-regulated, but those induced by Dichlorvos can not be detected significantly. The transcription levels of BmGSTe5 in midgut and BmGSTs1 in fatbody are up-regulated most, so we predict that these two genes are related with resistance of silkworm.4. NaF-induced transcription levels of BmGST genesWe have determined the transcription levels of BmGSTs induced by NaF in silkworm Haoyue with dual-spike-in qPCR method. The results indicated that the transcription of some BmGSTs such as BmGSTe5 and BmGSTs1 in the midgut as well as BmGSTo3 and BmGSTs1 in the fat body can be induced by NaF, and they can be inhibited by NaF, such as the BmGSTs in the Malpighian tubule.The production of this research illuminated the basic information of distributing of BmGSTs in various tissues of Bombyx mori, and it can provide data for studying the function of BmGSTs in the development of resistance and developing new pesticide resistant silkworm varieties.