Isolation of a cDNA clone and characterization of expression of the highly abundant, cold acclimation-associated 14 kDa dehydrin of blueberry
Isolation of a cDNA clone and characterization of expression of the highly abundant, cold acclimation-associated 14 kDa dehydrin of blueberry
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DOI：
10.1016/j.plantsci.2004.11.007
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发表时间：
2005-04-01
期刊：
影响因子：
5.2
通讯作者：
Rowland, LJ
中科院分区：
文献类型：
--
作者：
Dhanaraj, AL;Slovin, JP;Rowland, LJ
脱水蛋白是由脱水胁迫诱导的植物蛋白家族，所述脱水胁迫例如由干旱、盐度和低温/冷冻温度引起的那些。一些植物蛋白的诱导可能也对阿坝和/或短光周期有反应。以前，我们报道，在蓝莓中的一个家庭的65，60和14 kDa积累在花芽在冬季，这些蛋白质的水平与耐寒性。随着暴露于温暖的温度，14 kDa的葡萄糖蛋白水平的下降也与抗寒性的丧失或去适应密切相关。在本研究中，我们确定和测序的cDNA克隆从蓝莓花芽RNA编码的14 kDa的tuberculin。通过比较来自蛋白质的部分肽序列与来自cDNA的推导的蛋白质序列来确认克隆的身份。该cDNA全长653 bp，由55 bp的5 '非翻译区、303 bp的开放阅读框、213 bp的3'非翻译区和82 bp的polyA尾组成。该cDNA有2个K盒，表明是β-胡萝卜素，没有Y或S片段;因此它被分类为K（2）型β-胡萝卜素。在转录和蛋白质水平的表达的14 kDa的拟南芥蛋白在茎和叶组织诱导的寒冷和干旱胁迫下，在两个基因型，'Bluecrop'和'总理'，这在他们的耐寒性和耐旱性方面不同的研究。使用多克隆抗体监测蛋白质的表达，所述多克隆抗体针对蓝莓甜蛋白的共有K盒的合成肽产生，发现所述合成肽与许多蓝莓甜蛋白交叉反应。14 kDa的拟南芥蛋白，像其他K（n）型拟南芥蛋白，强烈诱导冷胁迫和干旱胁迫在较小程度上。一个以前未表征的16 kDa的蛋白质在蛋白质印迹上显示出类似的诱导，然而，它似乎随着时间的推移增加在南方品种'总理'的茎的实验过程中。与其他一些树皮蛋白和树皮贮藏蛋白一样，16 kDa的树皮蛋白的表达可能对短光周期有反应。在蛋白质水平上的14 kDa的tetrain的表达模式与在RNA水平上的模式非常一致，并且在更耐寒哈代和耐旱的基因型“Bluecrop”中，比在“Premier”中，14 kDa的tetrain信息被诱导至更高的水平。出版社：Elsevier爱尔兰Ltd.
Dehydrins are a family of plant proteins that are induced by dehydrative stresses such as those caused by drought, salinity, and low/freezing temperature. Induction of some dehydrins may be responsive to ABA and/or short photoperiod as well. Previously, we reported that in blueberry a family of dehydrins of 65, 60, and 14 kDa accumulates in floral buds during the winter, and the levels of these proteins correlate with cold tolerance. Decline in level of the 14 kDa dehydrin with exposure to warm temperatures correlates well with loss of hardiness or deacclimation also. In the present study, we identified and sequenced a cDNA clone from blueberry floral bud RNA that encodes the 14 kDa dehydrin. The identity of the clone was confirmed by comparing partial peptide sequences from the protein with deduced protein sequence from the cDNA. The cDNA was found to be a full-length 653 bp clone, comprised of a 55 bp 5'UTR, an ORF of 303 bp, a 213 bp 3'UTR, and an 82 bp polyA tail. The cDNA has 2 K boxes, indicative of dehydrins, and no Y or S segments; thus it was classified as a K(2) type dehydrin. Expression of the 14 kDa dehydrin was studied at the transcript and protein levels in stem and leaf tissues under induced cold and drought stress in two genotypes, 'Bluecrop' and 'Premier', which differ in terms of their cold and drought tolerances. Expression of the protein was monitored using a polyclonal antibody raised against a synthetic peptide of the consensus K box of blueberry dehydrins, which was found to cross react with many blueberry dehydrins. The 14 kDa dehydrin, like other K(n) type dehydrins, was strongly induced by cold stress and to a lesser extent by drought stress. A previously uncharacterized 16 kDa dehydrin showed similar induction on Western blots; however, it appeared to increase over time during the course of the experiments in stems of the southern variety 'Premier'. Like some other dehydrins and bark storage proteins, expression of the 16 kDa dehydrin may be responsive to short photoperiods. Patterns of expression of the 14 kDa dehydrin at the protein level were very consistent with patterns at the RNA level, and the 14 kDa dehydrin message was induced to higher levels in the more cold hardy and drought tolerant genotype, 'Bluecrop', than in 'Premier'. Published by Elsevier Ireland Ltd.