Laboratory of Fruit Genetic Engineering, The Institute of Enology and Viticulture, University of Yamanashi, Kofu, Yamanashi 400-0005, Japan.
J Plant Physiol. 2010 Jul 1;167(10):812-9. doi: 10.1016/j.jplph.2009.12.005. Epub 2010 Jan 22.
We report the cloning and characterization of heat shock-induced genes in grapevine (Vitis vinifera L.). Using the cDNA subtraction method, four heat shock-induced genes were identified in heat shock-treated Pinot noir grapevine. The four genes were immediately induced and upregulated in leaves and berry skins by heat treatment at 45 degrees C. The recovery treatment at 26 degrees C reduced the upregulated transcription of the heat shock-induced genes to near basal levels. The predicted amino acid sequences of three genes, HSG4, HSG14, and HSG19, showed high homologies to those of known small heat shock proteins of other plants. The predicted amino acid sequence of the fourth gene, HSG1, has two conserved motifs, the IQ motif and the Bcl-2-associated athanogene (BAG) domain, suggesting that HSG1 may be a member of the BAG family in grapevine. Although no morphological changes were observed, the overexpression Arabidopsis lines of HSG1, HSG4, HSG14, or HSG19 exhibited faster growth including floral transition than the control line, suggesting that the constitutive expression of HSG1, HSG4, HSG14, or HSG19 protein resulted in increased growth rates without any detectable harm. The thermotolerance test demonstrated that all of the overexpression lines remained viable and noticeably healthy compared with the control line even after exposure to severe heat shock, suggesting that HSG1, HSG4, HSG14, or HSG19 protein might be related to the acquisition of thermotolerance in grapevine.
我们报告了葡萄（Vitis vinifera L.）中热激诱导基因的克隆和特性。使用 cDNA 消减法，在热激处理的黑比诺葡萄中鉴定出 4 个热激诱导基因。这 4 个基因在 45°C 热处理下立即在叶片和果皮中被诱导并上调。26°C 的恢复处理将热激诱导基因的上调转录降低到接近基础水平。三个基因 HSG4、HSG14 和 HSG19 的预测氨基酸序列与其他植物的已知小热休克蛋白具有高度同源性。第四个基因 HSG1 的预测氨基酸序列有两个保守基序，即 IQ 基序和 Bcl-2 相关抗凋亡基因（BAG）结构域，表明 HSG1 可能是葡萄中 BAG 家族的一员。尽管没有观察到形态变化，但 HSG1、HSG4、HSG14 或 HSG19 的过表达拟南芥系比对照系表现出更快的生长速度，包括花转变，这表明 HSG1、HSG4、HSG14 或 HSG19 蛋白的组成型表达导致生长速度增加，而没有任何可检测到的伤害。耐热性测试表明，与对照系相比，所有过表达系在暴露于剧烈热激后仍保持存活且明显健康，这表明 HSG1、HSG4、HSG14 或 HSG19 蛋白可能与葡萄获得耐热性有关。
