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Expression, tissue distribution and subcellular localization of dehydrin TAS14 in salt-stressed tomato plants

机译:脱水素TAS14在盐胁迫番茄中的表达,组织分布和亚细胞定位

摘要

We previously isolated and characterized TAS14, an mRNA that is induced in tomato upon osmotic stress or abscisic acid (ABA) treatment and that shares expression and sequence characteristics with other dehydrin genes in different species. Affinity-purified antibodies against TAS14 protein were used to study the expression of TAS14 protein, both in seedlings and mature plants, its tissue distribution and its subcellular localization. TAS14 protein was not detected in 4-day-old seedlings but accumulated after ABA, NaCl or mannitol treatments. In NaCl-treated seedlings, some protein was detectable after 6 h of treatment and reached maximal levels between 24 and 48 h. Concentrations ranging from 5 to 12.5 g/l NaCl induced the protein to similar levels. In salt-stressed mature plants, TAS14 was expressed abundantly and continuously in aerial parts, but only slightly and transiently in roots. Immunocytochemical analysis of salt-treated plants showed TAS14 accumulated in adventitious root primordia and associated to the provascular and vascular tissues in stems and leaves. Immunogold electron microscopy localized TAS14 protein both in the cytosol and in the nucleus, associated to the nucleolus and euchromatin. Since TAS14 is a phosphoprotein in vivo, the classes of protein kinases potentially responsible for its in vivo phosphorylation were tested in in vitro phosphorylation assays. TAS14 protein was phosphorylated in vitro by both casein kinase II and cAMP-dependent protein kinase.
机译:我们之前已经分离并鉴定了TAS14,一种在渗透胁迫或脱落酸(ABA)处理下在番茄中诱导的mRNA,并且与不同物种中的其他脱水素基因共享表达和序列特征。使用亲和纯化的TAS14蛋白抗体来研究TAS14蛋白在幼苗和成熟植物中的表达,其组织分布和亚细胞定位。 TAS14蛋白未在4天大的幼苗中检测到,但在ABA,NaCl或甘露醇处理后积累。在用NaCl处理的幼苗中,处理6小时后可检测到一些蛋白质,并在24至48小时之间达到最大水平。 NaCl浓度范围从5到12.5 g / l诱导蛋白质达到相似水平。在盐胁迫的成熟植物中,TAS14在地上部分大量且连续表达,但在根部仅短暂而短暂地表达。盐处理过的植物的免疫细胞化学分析显示,TAS14积累在不定根原基中,并与茎和叶中的前血管和维管组织有关。免疫金电子显微镜在细胞质和细胞核中均定位了TAS14蛋白,与核仁和常染色质相关。由于TAS14是体内的磷蛋白,因此在体外磷酸化测定中测试了可能导致其体内磷酸化的蛋白激酶类别。 TAS14蛋白在体外被酪蛋白激酶II和cAMP依赖性蛋白激酶磷酸化。

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