College of Life Sciences, Shandong Normal University, Jinan 250014, China.
Plant Sci. 2006 Sep;171(3):398-407. doi: 10.1016/j.plantsci.2006.04.011. Epub 2006 May 26.
Small heat shock proteins (sHSPs) are the major family of HSP induced by heat stress in plants. In this report, an approximately 1.9kb of Lehsp23.8 5'-flanking sequence was isolated from tomato genome. By using the β-glucuronidase (GUS) reporter gene system, the developmental and tissue specific expression of the gus gene controlled by the Lehsp23.8 promoter was characterized in transgenic tomato plants. Strong GUS staining was detected in the roots, leaves, flowers, fruits and germinated seeds after heat shock. The heat-induced GUS activity was different in the floral tissues at various developmental stages. Fluorometric GUS assay showed that the heat-induced GUS activity was higher in the pericarp than in the placenta, and it was the lowest in the locular gel. The heat-shock induction of the Lehsp23.8 promoter depended on the different stages of fruit development. The optimal heat-shock temperatures leading to the maximal GUS activity in the pericarp of green, breaker, pink and red fruits were 42, 36, 39 and 39°C, respectively. The heat-induced GUS activity in tomato fruits increased gradually within 48h of treatment and weakened during tomato fruit ripening. Obvious GUS activities under cold, exogenous ABA and heavy metal (Cd(2+), Cu(2+), Pb(2+) or Zn(2+)) stress conditions were also detected. These results show that the Lehsp23.8 promoter is characterized as strongly heat-inducible and multiple-stress responsive.
小分子热休克蛋白(sHSPs)是植物受热胁迫时诱导产生的 HSP 主要家族。本研究从番茄基因组中分离得到一个约 1.9kb 的 Lehsp23.8 5'侧翼序列。利用β-葡萄糖醛酸酶(GUS)报告基因系统,研究了 Lehsp23.8 启动子调控的 gus 基因在转基因番茄植株中的发育和组织特异性表达。经热激处理后,根、叶、花、果实和萌发种子中均检测到强烈的 GUS 染色。在不同发育阶段的花组织中,热诱导的 GUS 活性存在差异。荧光 GUS 分析显示,热诱导的 GUS 活性在果皮中最高,在胎座中最低,在心室凝胶中最低。Lehsp23.8 启动子的热激诱导依赖于果实发育的不同阶段。导致绿熟、裂果、粉红和红熟果实果皮中 GUS 活性达到最大值的最佳热激温度分别为 42、36、39 和 39°C。处理后 48h 内,番茄果实中热诱导的 GUS 活性逐渐增加,在番茄果实成熟过程中减弱。在冷胁迫、外源 ABA 和重金属(Cd(2+)、Cu(2+)、Pb(2+)或 Zn(2+))胁迫条件下也检测到明显的 GUS 活性。这些结果表明,Lehsp23.8 启动子具有较强的热诱导和多重胁迫响应特性。