response to high-salt stress in Halophyte cells

盐生植物细胞对高盐胁迫的反应

• 批准号: 09660013
• 负责人: YASUDA Takeshi
• 金额: $ 2.11万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09660013/
• 关键词: salt stress; Atriplex; Halophyte; cultured cell; germin-like protein; 塩性ストレス; 塩性植物; 特異的タンパク; cDNA

Halophyte plants are able to grow under high saline environments. For examining cellular response of Halophyte cells to salt stress, cultured cells from 3 Atriplex species ; 1 entiformis (AL), halimus (AH) and halimus (AH) were treated with and without 0.2M NaCl, and polypeptide patterns were analyzed by 2-D electrophoresis. Significant increase of polypeptide spot were detected in insoluble protein fractions of salt treated Atriplex cells on NEPHGE/SDS electrophoresis. A polypeptide named SP1 was with a molecular mass of 27 kDa with pI over 8.5. The polypeptide was glycoprotein with Con A binding activity. Eluted proteins from electrophoretic spot of LH were used for antibody preparation by mice and N-terminal amino acid sequencing. mRNA were isolated from salt-treated LH cells and cDNA library was prepared, and library screening were done using a probe, which amplified by PCR on primers constructed from N-terminal amino-acid sequences. We have determined the nucleotide sequence of a full length cDNA for SP1 and submitted on the DDBJ/EMBL/GenBank databases (AB024338). Reduced amino acid sequences reveals that SP1 is 224-AA, 24 kDa and pI=9.72. A search of the GenBank data base for SP1 amino acid sequence showed similarity to Germin and oxalate-oxidase.

卤素植物能够在高盐水环境下生长。为了检查卤素细胞对盐胁迫的细胞反应，来自3种阿特莱克斯物种的培养细胞； 1 Entiformis（Al），Halimus（AH）和Halimus（AH）用NaCl和0.2M NaCl处理，并通过2-D电泳分析多肽模式。在Nephge/SDS电泳上的盐处理的Atriplex细胞的不溶性蛋白质部分中检测到多肽斑点的显着增加。一个名为SP1的多肽的分子量为27 kDa，PI超过8.5。多肽是具有结合活性的糖蛋白。 LH电泳位点洗脱的蛋白质用于小鼠和N末端氨基酸测序的抗体制备。从盐处理的LH细胞中分离出mRNA并制备cDNA文库，并使用探针进行筛选，该探针通过PCR对由N末端氨基酸序列构建的引物进行扩增。我们已经确定了SP1全长cDNA的核苷酸序列，并在DDBJ/EMBL/GenBank数据库（AB024338）上提交。减少的氨基酸序列表明SP1为224-AA，24 kDa，PI = 9.72。对SP1氨基酸序列的GenBank数据库的搜索显示与生殖和草酸盐氧化酶相似。