Chi Wen-Tzu, Fung Raymond W M, Liu Hsiang-Chin, Hsu Ching-Chi, Charng Yee-Yung
Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan.
Plant Cell Environ. 2009 Jul;32(7):917-27. doi: 10.1111/j.1365-3040.2009.01972.x. Epub 2009 Mar 3.
Plant temperature-induced lipocalins (TILs) have been shown to be responsive to heat stress (HS), but the nature of this response was unknown. In this study, a reverse genetic approach was taken to elucidate the role of Arabidopsis TIL1 (At5g58070) in thermotolerance. A T-DNA knock-out line of TIL1 (til1-1) showed severe defects in basal (BT) and acquired thermotolerance (AT), which could be complemented by introducing the wild-type gene. However, over-expression of TIL1 did not significantly enhance thermotolerance in transgenic plants. TIL1 is peripherally associated with plasma membrane. Transcriptomic analysis showed that the heat shock response in til1-1 seedlings was about the same as in the wild-type plants except the expression of TIL1. The level of TIL1 did not affect the temperature threshold for heat shock protein induction. Ion leakage analysis revealed no significant difference in membrane stability between the wild-type and til1-1 seedlings. These results suggest that TIL1 is not involved in regulating membrane fluidity or stability. Nevertheless, the mutant plants were also more sensitive than the wild type to tert-butyl hydroperoxide, a reagent that induces lipid peroxidation. Taken together, these data indicate that TIL1 is an essential component for thermotolerance and probably functions by acting against lipid peroxidation induced by severe HS.
植物温度诱导脂钙蛋白(TILs)已被证明对热胁迫(HS)有响应,但其响应的本质尚不清楚。在本研究中,采用反向遗传学方法来阐明拟南芥TIL1(At5g58070)在耐热性中的作用。TIL1的T-DNA敲除系(til1-1)在基础耐热性(BT)和获得性耐热性(AT)方面表现出严重缺陷,通过导入野生型基因可对其进行互补。然而,TIL1的过表达并未显著提高转基因植物的耐热性。TIL1与质膜外周相关。转录组分析表明,除了TIL1的表达外,til1-1幼苗中的热休克反应与野生型植物大致相同。TIL1的水平不影响热休克蛋白诱导的温度阈值。离子渗漏分析显示野生型和til1-1幼苗之间的膜稳定性没有显著差异。这些结果表明,TIL1不参与调节膜流动性或稳定性。然而,突变体植物也比野生型对叔丁基过氧化氢更敏感,叔丁基过氧化氢是一种诱导脂质过氧化的试剂。综上所述,这些数据表明TIL1是耐热性的重要组成部分,可能通过对抗严重热胁迫诱导的脂质过氧化发挥作用。