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Identification of key factor fur cell wall disassembly in fruit by proteome analysis and transgenic technology

蛋白质组分析和转基因技术鉴定水果毛皮细胞壁解体关键因素

基本信息

批准号：
17380024
负责人：
KUBO Yasutaka
金额：
$ 10.85万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17380024/
关键词：
Cell wall protein Fruit softening YSST 2-D gel electophoreisis Pear Melon Chinese pear YSST解析細胞壁 DNAアレイ 1-MCP プロテオーム解析形質転換エチレン

项目摘要

Cell wall is main factor for physical structure of plant and the main limiting factors limiting their storage, transport and marketability. Previous studies indicated importance of plant hormone ethylene for fruit ripening and softening. However, key factor (s) in cell wall disassembly have not been identified. 'La France' European pear fruit and melon fruit undergo dramatic softening during ripening in an ethylene-dependent manner, whereas Chinese pear does not exhibit softening in spite of its massive ethylene production during ripening. To broad-scale and comprehensive screening for factors involved in fruit softening, in addition to gene expression analysis of polygalacturonase, α-arabinofuranosidase and β-galactosidase, the two-dimensional gel electrophoresis profiles and YSST analysis of cell wall proteins were employed. The accumulation levels of 200 protein spots were isolated in the ripening 'La France' pear. When softening pears of this variety were treated with 1-MCP, a strong ethylene inhibitor, fruit softening was retarded. On the comparison between treated and non-treated fruit, 3 protein spots were selected as candidates for key factors of fruit softening, since their accumulation showed specific pattern for softening.On YSST analysis, 170 clones (70 contigs and 100 singletons) encoding secreted proteins including clones which have not been previously reported were isolated. Based on the macroarray analysis, 28 clones showed elevated or down-regulated expression with pear fruit development and ripening. On comparison in 'La France' and the non-softening 'Yali' pear fruit, 11 potential new cell wall modifying proteins among 28 clones were selected for candidates for key factor of fruit softening.

细胞壁是决定植物物理结构的主要因素，也是限制植物贮藏、运输和商品化的主要限制因素。以前的研究表明，植物激素乙烯对果实成熟和软化具有重要作用。然而，细胞壁分解的关键因子(S)还没有确定。‘La France’欧洲梨和甜瓜果实在成熟过程中会以一种依赖乙烯的方式发生戏剧性的软化，而中国的梨并没有表现出软化，尽管它在成熟过程中会产生大量的乙烯。为了更大范围、更全面地筛选果实软化相关因子，除对多聚半乳糖醛酸酶、α-阿拉伯呋喃糖苷酶和β-半乳糖苷酶的基因表达进行分析外，还采用了双向凝胶电泳法和细胞壁蛋白质双向凝胶电泳法。在成熟的‘La France’梨中分离到200个蛋白质点的积累水平。用1-MCP处理该品种的软化梨时，果实软化有一定的延缓作用。通过对处理和未处理果实的比较，筛选出3个蛋白质点作为果实软化的关键因子，它们的积累表现出特定的软化模式。在YSST分析中，分离到170个编码分泌蛋白的克隆(70个重叠群和100个单态)，其中包括以前未见报道的克隆。根据大阵列分析，28个无性系在梨果实发育和成熟过程中表达上调或下调。通过对‘La France’和未软化的‘鸭梨’果实的比较，从28个克隆中筛选出11个潜在的新的细胞壁修饰蛋白作为果实软化的关键因子。

项目成果

期刊论文数量（0）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

セイヨウナシ果実におけるYSSTを用いた細胞外分泌タンパク質をコードするcDNAのクローニング

利用 YSST 在梨果实中克隆编码细胞外分泌蛋白的 cDNA

DOI：
发表时间：
2006
期刊：
影响因子：
0
作者：
Mwaniki;Mercy W.牛島幸一郎・山根久代・Jocelyn K.C. Rose.稲葉昭次・久保康隆
通讯作者：
Mercy W.牛島幸一郎・山根久代・Jocelyn K.C. Rose.稲葉昭次・久保康隆

β-Galactiosidase and α-L-arabinofuranosidase activities and gene expression in European and Chinese pear fruit during ripening

欧洲和中国梨果实成熟过程中β-半乳糖苷酶和α-L-阿拉伯呋喃糖苷酶活性及基因表达

DOI：
发表时间：
2007
期刊：
J.Japan.Soc.Hort.Sci. 76
影响因子：
0
作者：
Mwaniki M.W;F.M.Mathookob;M.MatsuzakiK.Hiwasac;A.Tateishi;K.Ushijimae;R.Nakanoe;A.Inabae;Y.Kubo
通讯作者：
Y.Kubo

メロン果実の成熟に伴う軟化関連酵素遺伝子の発現解析

甜瓜果实成熟过程中软化相关酶基因的表达分析