基于DDRT-PCR研究茶树对茶尺蠖取食诱导的基因表达谱差异

Differential Gene Expression Profiles Analysis of Tea Plant Induced by Tea Looper (Ectropic oblique) Attack Using DDRT-PCR

利用DDRT-PCR技术初步研究了茶树被害虫茶尺蠖取食后基因表达谱差异。结果表明:接头引物A3、A4、A6、A7和A8分别与随机引物R1-R8,R12组合扩增后,总共得到222条差异片段,占总条带数的32.8%。在所鉴定的20个差异片段中,有8个片段序列没有发现同源序列;有5个片段序列与未知功能蛋白相关;其余7个差异表达的片段序列可分为6类,即分别与光合系统II色素蛋白(C15)、非生物抗性诱导蛋白(D22)、糖苷代谢酶(A45)、核酸和蛋白转录调控因子(A12,D63)、生长素调节蛋白(E94)和营养性抗虫蛋白(D73)。其中差异表达片段C15、A45、D22、D73和E94在植物与害虫相互作用相关分子机制研究中首次被发现。

DDRT-PCR technique was used to explore the differential gene expression profiles analysis of tea plant induced by tea looper （Ectropic oblique） attack. Results showed that 222 differential expression fragments were obtained, which was 32.8% of total fragments, by amplified in anchor primers A3, A4, A6, A7, A8 and random primers R1-R8, R12, respectively. Twenty fragments of which were re-amplified by RT-PCR. Based on the BLASTx search in GenBank, it were found that eight of the fragments shared no homology, five of the fragments shared homology with the unknown proteins, and seven of the fragments shared higher score homology with the known proteins, which can be divided into six groups related to photosynthetic pigment protein in photosystem Ⅱ （C15）, abiotic resistance induced protein（D22）, glucoside metabolism enzyme（A45）, nucleic acid and protein transcription and regulation factors（A12, D63）, auxin-regulated protein（E94） and VIP2 protein（D73）. The fragments of C15, A45, D22, D73 and E94 were firstly found in the studies of molecular mechanism of plant-insect interaction, up to day.