Lily ASR protein-conferred cold and freezing resistance in Arabidopsis
Lily ASR protein-conferred cold and freezing resistance in Arabidopsis
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DOI：
10.1016/j.plaphy.2011.07.002
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发表时间：
2011-09-01
影响因子：
6.5
通讯作者：
Wang, Co-Shine
中科院分区：
文献类型：
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作者：
Hsu, Yi-Feng;Yu, Shu-Chuan;Wang, Co-Shine
百合LLA 23蛋白是脱落酸、胁迫和成熟诱导（ASR）蛋白家族的成员。在花椰菜花叶病毒35 S启动子下组成型过表达LLA 23赋予拟南芥耐寒和抗冻性。两株转基因35 S：：LLA 23植株的表型生长和存活率表现出比野生型（WT）植株更高的抗寒性和抗冻性。WT叶中的电解质渗漏在-2 ° C下相对于在22 ° C下增加约四倍，而两种转基因叶在相同条件下显示出很少的离子渗漏。在正常生长条件下过表达LLA 23的转基因系35 S：：LLA 23 E的微阵列分析先前由Yang等人进行（Protoplasma，2008，233：241-254）。基因芯片分析表明，百合ASR诱导12个冷应答基因表达上调，25个冷应答基因表达下调。许多ASR调节基因编码的蛋白质参与防御/应激相关的，转录和代谢的类别。定量聚合酶链反应分析证实了在微阵列分析中观察到的mRNA水平的变化。因此，我们的研究结果提供了体内证据，表明LLA 23介导冷/冷冻应激反应信号。为了进一步了解LLA 23蛋白的功能，使用了体外酶保护测定，其中乳酸脱氢酶和苹果酸脱氢酶经受不利条件。该测定显示，加入LLA 23后，两种酶活性均显著保留，其上级于海藻糖或BSA，表明LLA 23蛋白可以保护酶活性免受冻融循环的影响。35 S：：LLA 23幼苗在-4 ℃下也表现出比WT上级的酶活性。这些结果表明，LLA 23可能作为一种植物保护剂以及一个转录因子赋予35 S：：LLA 23植物增强的抗寒性和抗冻性。(C)2011年Elsevier Masson SAS。All rights reserved.
The lily LLA23 protein is a member of the abscisic acid, stress and ripening-induced (ASR) protein family. Constitutive overexpression of LLA23 under the cauliflower mosaic virus 35S promoter confers cold and freezing tolerance in Arabidopsis. The phenotypical growth and survival percentage of the two transgenic 35S::LLA23 plants showed higher resistance to cold and freezing conditions than those of wild-type (WT) plants. The electrolyte leakage in WT leaves increased by approximately fourfold at -2 degrees C relative to that at 22 degrees C whereas both transgenic leaves showed little ion leakage under the same conditions. A microarray analysis of LLA23-overexpressing transgenic line, 35S::LLA23E, under normal growing conditions was previously conducted by Yang et al. (Protoplasma, 2008, 233:241-254). Microarray analysis showed that 12 cold-responsive genes are upregulated and 25 cold-responsive genes are downregulated by lily ASR. Many ASR-regulated genes encode proteins involved in the classes of defense/stress-related, transcription, and metabolism. Quantitative polymerase chain reaction analysis confirms the changes in mRNA levels observed in the microarray analysis. Thus, our results provide in vivo evidence implying that LLA23 mediates cold/freezing stress-responsive signaling. To gain further insight into the functions of LLA23 protein, an in vitro enzyme protection assay was used in which lactate dehydrogenase and malate dehydrogenase were subjected to unfavorable conditions. The assay revealed that both enzyme activities were significantly retained with the addition of LLA23, which was superior to either trehalose or BSA, suggesting that the LLA23 protein can protect enzymatic activities against freeze-thaw cycles. The 35S::LLA23 seedlings also exhibited enzyme activity superior to WT at -4 degrees C. These results suggest that LLA23 may act as an osmoprotectant as well as a transcription factor to confer 35S::LLA23 plants enhanced cold and freezing resistance. (C) 2011 Elsevier Masson SAS. All rights reserved.