不同水稻品种基因表达对干旱胁迫的应答
摘要
本文以中旱3号和矮仔占为材料,分析了10%聚乙二醇(PEG6000)模拟干旱胁迫对上述两水稻品种叶片含水量的影响,应用基因芯片技术分析了中旱3号和矮仔占叶片基因表达对干旱胁迫的应答,并应用RT-PCR对部分干旱胁迫诱导基因的转录水平进行了验证。进而探讨了水稻对干旱胁迫的应答机制,其主要结果如下:
1.干旱胁迫对水稻幼苗含水量的影响
PEG6000模拟干旱对两种供试水稻幼苗叶片含水量均有明显影响。供试水稻幼苗叶片的相对含水量随着干旱处理时间的延长而降低,并且中旱3号相对含水量下降的幅度较小,矮仔占相对含水量下降幅度较大。这表明,中旱3号为耐旱品种,矮仔占为干旱胁迫敏感型品种。
2.水稻基因表达对干旱胁迫应答的基因芯片分析
PEG6000模拟干旱胁迫对两供试水稻品种幼苗叶片的基因表达有明显的影响。中旱3号幼苗叶片中受到PEG6000模拟干旱胁迫诱导基因有信号受体相关基因、信号转导相关基因、转录因子及转录调控相关基因、抗脱水特异性蛋白相关基因、解毒相关基因、次生物和抗生素合成相关基因、分子构型和大分子修饰相关基因、物质运输与代谢相关基因几大类型。其中,中旱3号叶片中信号转导、转录调控、抗脱水特异性蛋白以及解毒相关基因受到干旱胁迫大量诱导,尤其是干旱胁迫诱导中旱3号叶片中木菠萝凝集素、水通道蛋白强烈表达,而在矮仔占叶片中未检测到其诱导作用。这可能是中旱3号耐干旱胁迫主要机制。
相反,矮仔占叶片中受干旱胁迫所诱导基因的数量不仅很少,而且受诱导基因的上调倍数也较低,尤其是受诱导的抗脱水特异性蛋白相关基因数量很少。这可能是矮仔占对干旱胁迫敏感的主要机制。
3.干旱胁迫诱导基因的RT-PCR验证
RT-PCR结果表明干旱胁迫对中旱3号叶片中与活性甲基循环、转移相关基因的表达既有抑制作用又有诱导作用。而对矮仔占叶片中与活性甲基循环、转移相关基因的表达具有抑制作用,这与基因芯片的检测结果相一致。
In this work, the responses of rice (Zhonghan3 and Aizaizhan) to drought stress (simulated by 10%PEG6000) were investigated with gene-chip analysis and RT-PCR technology. Results were showed as follow:
1. The effects drought stress on relative water content in rice seedling leaves
The relative water content in both rice seedling leaves were significantly affected by drought stress. The relative water content in rice seedlings decreased along with time change under the drought stress and it decreased lightly in zhonghan3, but more severe in aizaizhan. The results suggested that the zhonghan3 was more tolerant to water stress, whereas aizaizhan sensitive.
2. Gene expression analysis of rice using gene-chip under drought stress
The gene expression in both rice seedling leaves were significantly affected by drought stress. Receptor gene, signal transduction gene, transcription factors and transcriptional regulation gene, anti-dehydration-specific gene, detoxication gene, synthesis of secondary metabolites and antibiotics gene, modification of molecular structure and macromolecular, transportation and metabolic gene all were induced by drought stress in zhonghan3 leaves. Among them, signal transduction gene, transcription factors and transcriptional regulation gene, anti-dehydration-specific gene and detoxication gene were largely induced by drought stress. In particular, jacalin and aquaporin were strongly expressed in zhonghan3 seedling leaves, but not detected in aizaizhan. This may be the main mechanism for tolerance to drought stress of zhonghan3.
Contrary,the mumber of genes, induced by drought stress was small and the magnitude up-regulated was also small in aizaizhan leaves. Especially the mumber of anti-dehydration-specific gene was very small. This may be mainly contributed to the sensitivity to drought stress of aizaizhan.
3. Identifing drought stress-induced gene in rice seedling leaves with RT-PCR technology
The result showed that active methyl cycle and transfer genes would be inhibited or inducted in zhonghan3 leaves by drought stress, but only be inhibited in aizaizhan leaves. The results were consistent with the result of gene-chip.
1.干旱胁迫对水稻幼苗含水量的影响
PEG6000模拟干旱对两种供试水稻幼苗叶片含水量均有明显影响。供试水稻幼苗叶片的相对含水量随着干旱处理时间的延长而降低,并且中旱3号相对含水量下降的幅度较小,矮仔占相对含水量下降幅度较大。这表明,中旱3号为耐旱品种,矮仔占为干旱胁迫敏感型品种。
2.水稻基因表达对干旱胁迫应答的基因芯片分析
PEG6000模拟干旱胁迫对两供试水稻品种幼苗叶片的基因表达有明显的影响。中旱3号幼苗叶片中受到PEG6000模拟干旱胁迫诱导基因有信号受体相关基因、信号转导相关基因、转录因子及转录调控相关基因、抗脱水特异性蛋白相关基因、解毒相关基因、次生物和抗生素合成相关基因、分子构型和大分子修饰相关基因、物质运输与代谢相关基因几大类型。其中,中旱3号叶片中信号转导、转录调控、抗脱水特异性蛋白以及解毒相关基因受到干旱胁迫大量诱导,尤其是干旱胁迫诱导中旱3号叶片中木菠萝凝集素、水通道蛋白强烈表达,而在矮仔占叶片中未检测到其诱导作用。这可能是中旱3号耐干旱胁迫主要机制。
相反,矮仔占叶片中受干旱胁迫所诱导基因的数量不仅很少,而且受诱导基因的上调倍数也较低,尤其是受诱导的抗脱水特异性蛋白相关基因数量很少。这可能是矮仔占对干旱胁迫敏感的主要机制。
3.干旱胁迫诱导基因的RT-PCR验证
RT-PCR结果表明干旱胁迫对中旱3号叶片中与活性甲基循环、转移相关基因的表达既有抑制作用又有诱导作用。而对矮仔占叶片中与活性甲基循环、转移相关基因的表达具有抑制作用,这与基因芯片的检测结果相一致。
In this work, the responses of rice (Zhonghan3 and Aizaizhan) to drought stress (simulated by 10%PEG6000) were investigated with gene-chip analysis and RT-PCR technology. Results were showed as follow:
1. The effects drought stress on relative water content in rice seedling leaves
The relative water content in both rice seedling leaves were significantly affected by drought stress. The relative water content in rice seedlings decreased along with time change under the drought stress and it decreased lightly in zhonghan3, but more severe in aizaizhan. The results suggested that the zhonghan3 was more tolerant to water stress, whereas aizaizhan sensitive.
2. Gene expression analysis of rice using gene-chip under drought stress
The gene expression in both rice seedling leaves were significantly affected by drought stress. Receptor gene, signal transduction gene, transcription factors and transcriptional regulation gene, anti-dehydration-specific gene, detoxication gene, synthesis of secondary metabolites and antibiotics gene, modification of molecular structure and macromolecular, transportation and metabolic gene all were induced by drought stress in zhonghan3 leaves. Among them, signal transduction gene, transcription factors and transcriptional regulation gene, anti-dehydration-specific gene and detoxication gene were largely induced by drought stress. In particular, jacalin and aquaporin were strongly expressed in zhonghan3 seedling leaves, but not detected in aizaizhan. This may be the main mechanism for tolerance to drought stress of zhonghan3.
Contrary,the mumber of genes, induced by drought stress was small and the magnitude up-regulated was also small in aizaizhan leaves. Especially the mumber of anti-dehydration-specific gene was very small. This may be mainly contributed to the sensitivity to drought stress of aizaizhan.
3. Identifing drought stress-induced gene in rice seedling leaves with RT-PCR technology
The result showed that active methyl cycle and transfer genes would be inhibited or inducted in zhonghan3 leaves by drought stress, but only be inhibited in aizaizhan leaves. The results were consistent with the result of gene-chip.
引文
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