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Comprehensive studies on endogenous PR-3 chitinases expression in rive plant

河流植物内源PR-3几丁质酶表达的综合研究

基本信息

批准号：
14560004
负责人：
NAKAZAKI Tetsuya
金额：
$ 1.66万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14560004/
关键词：
PR-3 locus of chitinase gene expression character GST fusion protein expression system

项目摘要

Chitinases are PR-3 (pathogenesis-related protein-3) proteins, hydrolyzing the B -1,4-linkage in N-acetyl-D-glucosamine residues of chitin, the main component of fungus cell walls. Chitinases so far have been considered to play an important role in the disease-defense mechanisms of plants. The objective of this study was to elucidate the number of chitinase genes in rice, and their allelic relationships, chromosomal locations and expression patterns. The enzymatic characters of proteins encoded by the genes were also analyzed.Database analyses of nucleotide sequences in rice showed that there were 24 non-redundant PR-3 chitinase sequences. They were formed into 12 groups, each of which was found to correspond to one locus. Thus a total of 12 chitnase loci (Cht1-Cht12) were identified. Through the analysis of the catalytic domain structure we proposed a new classification system of chitinases. A GST fusion protein expression system of rice chitinases was also established. Using chitinase proteins obtained by this system, enzymatic characters and antifungal activities of rice PR-3 chitinases were evaluated. These chitinases showed different patterns in gene expression strength and time. The fruits obtained in this study would be available for elucidating biological roles and functions of PR-3 chitinases in plants.

几丁质酶是PR-3（病程相关蛋白-3）蛋白，水解真菌细胞壁的主要成分几丁质的N-乙酰基-D-葡萄糖胺残基中的B-1，4-键。几丁质酶在植物抗病机制中起着重要作用。本研究的目的是阐明水稻几丁质酶基因的数量、等位基因关系、染色体定位和表达模式。对水稻PR-3几丁质酶基因的核苷酸序列进行数据库分析，发现该基因编码的蛋白质有24个非冗余序列。他们被分成12组，每组被发现对应于一个位点。因此，共鉴定出12个几丁质酶基因座（Cht 1-Cht 12）。通过对几丁质酶催化结构域的分析，提出了一个新的几丁质酶分类体系。建立了水稻几丁质酶GST融合蛋白表达系统。利用该系统获得的几丁质酶蛋白，对水稻PR-3几丁质酶的酶学性质和抑菌活性进行了研究。这些几丁质酶在基因表达强度和时间上表现出不同的模式。本研究结果为进一步阐明PR-3几丁质酶在植物中的生物学作用和功能奠定了基础。