茶树MEP途径中HDS与HDR基因的cDNA全长克隆、功能分析与表达特征分析-茶学专业论文.docxVIP

I II 显低于对照，而后者与对照没有明显的差异，但是两者各自的最大表达量随着 激素种类的不同而存在差异；经 ABA 处理后，CsHDS 在各时间点表达量均显 著低于对照，而 CsHDR 除了处理后 6 h 以外，其它时间点也表现类似规律； 经推荐浓度的吡虫啉喷施处理后 4 d，CsHDS 的表达量显著上调，而 CsHDR 则相反。 关键词：茶树；CsHDS；CsHDR；茶尺蠖；基因克隆；表达特征 I III Abstract Terpenoids are a class of compounds which constructed from their basic structural unit- isoprene. Terpenoids is an important part of secondary metabolites in tea plant, and their also plays an important role in indirect defense. 4-hydroxy-3-methylbut-2-enyl diphosphate synthase (HDS), 4-hydroxy-3-methyl- but-2-enyl diphosphate reductase (HDR), the two above-mentioned participate in the last two steps of the MEP pathways which is upstream biosynthesis pathway in terpenes metabolism. The HDR is also a critical rate-limiting enzyme of MEP pathway. However, the research around HDS and HDR and its function in tea plant is still unknown. In this paper, we mainly described the cDNA cloning, identification and expression analysis of the HDS, HDR in different organs of Camellis sinensis. The preliminary results are as follows: First, the two EST sequences which have the high homology with the HDS, HDR were isolated from SSH library after Ectropic oblique attack in the leaves of Camellis sinensis. Then we get HDS, HDR full-length cDNA sequences by using the RACE and named for CsHDS, CsHDR. The full length of CsHDS is 2603bp with a open reading frame of 2226bp, encoding 741 amino acids; The full length of CsHDR is 1716 bp with a open reading frame of 1380 bp, encoding 459 amino acids. The two gene have been submitted to GenBank and the accession numbers are JQ014629, JQ014628. Bioinformatics analysis revealed CsHDS had two conservative GcpE functional domains and three active site of conserved cysteine residues like other HDS in plant; The CsHDR had a conservative LYTB functional domains and four active site of conserved cysteine residues The two genes all contained chloroplast transit peptides. Tissue specific expression analysis showed that the CsHDS had the high