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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，也是制约植物贮藏、运输和销售的主要制约因素。以往的研究表明植物激素乙烯对果实成熟和软化的重要性。然而，细胞壁分解的关键因素尚未确定。“La France”欧洲梨果和甜瓜果实在乙烯依赖的成熟过程中经历了戏剧性的软化，而中国梨尽管在成熟过程中产生了大量的乙烯，但却没有表现出软化。为了广泛、全面地筛选果实软化的影响因素，除了对多半乳糖醛酸酶、α-阿拉伯糖葡糖苷酶和β-半乳糖苷酶进行基因表达分析外，还采用了二维凝胶电泳图谱和细胞壁蛋白的YSST分析。从成熟的‘La France’梨中分离出200个蛋白质斑点的积累水平。用1-MCP（一种强乙烯抑制剂）处理该品种的软化梨，可延缓果实软化。通过对处理和未处理果实的比较，选择了3个蛋白质斑点作为果实软化的关键因子候选，因为它们的积累具有特定的软化模式。在YSST分析中，分离到170个编码分泌蛋白的克隆（70个contigs和100个singleton），其中包括以前未报道的克隆。经macroarray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‘Yali’梨果实的比较，从28个无性系中筛选出11个潜在的细胞壁修饰蛋白作为果实软化关键因子的候选。

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