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Cloning and expression analysis of phenylalanine ammonia-lyase in relation to chilling tolerance in harvested banana fruit.

机译:香蕉果实中苯丙氨酸解氨酶的克隆和表达分析与耐寒性的关系。

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摘要

Bananas are highly susceptible to chilling injury (CI) and phenylalanine ammonia-lyase (PAL), as a key enzyme involved in plant phenylpropanoid metabolism, has been associated with low temperature stress in plant tissues. However, little is known about the role of PAL (including PAL activity, gene and protein expression) in postharvest chilling tolerance of banana fruit. Two partial cDNAs sequences (MaPAL1 and MaPAL2) with about 760 bp were cloned from banana pulp by RT-PCR. Western and northern hybridizations were used to investigate expression of PAL protein and PAL genes in fruit stored for 10 days at 7 degrees C (chilling temperature) and then transferred to 22 degrees C (room temperature). The effects of propylene (a functional ethylene analogue) on their expression in relation to CI were also examined. Northern and western blot analyses revealed that mRNA transcripts of MaPAL1 and MaPAL2 and PAL protein levels in banana fruit during storage increased, reaching a peak at about day 8, and finally decreased at chilling temperature. Prior to low temperature storage, pretreatment with propylene could alleviate CI and enhance PAL activity, protein amount and mRNA transcripts of MaPAL1 and MaPAL2. Moreover, changes in PAL activity, protein amount and accumulation of MaPAL1 and MaPAL2 exhibited almost the same patterns. The results suggest that the induction of PAL in banana fruit during low temperature storage is regulated at transcriptional and translational levels, and is related to reduction in CI symptoms..
机译:香蕉极易受到冷害(CI)的影响,而苯丙氨酸氨解酶(PAL)作为植物苯丙氨酸代谢的关键酶,已与植物组织的低温胁迫相关。然而,人们对PAL在香蕉果实采后耐寒性中的作用(包括PAL活性,基因和蛋白质表达)了解甚少。通过RT-PCR从香蕉果肉中克隆了两个约760 bp的部分cDNA序列(MaPAL1和MaPAL2)。 Western和Northern杂交用于研究PAL蛋白和PAL基因在7摄氏度(冷藏温度)保存10天,然后转移至22摄氏度(室温)的水果中的表达。还检查了丙烯(功能性乙烯类似物)对其与CI有关的表达的影响。 Northern和Western印迹分析表明,在贮藏期间香蕉果实中的MaPAL1和MaPAL2以及PAL蛋白的mRNA转录物增加,在第8天达到峰值,并在冷藏温度下最终降低。在低温储存之前,用丙烯进行预处理可以减轻CI并增强PAL活性,MaPAL1和MaPAL2的蛋白质含量以及mRNA转录本。此外,MaPAL1和MaPAL2的PAL活性,蛋白质含量和积累的变化表现出几乎相同的模式。结果表明,低温贮藏期间香蕉果实中PAL的诱导受转录和翻译水平的调节,并且与CI症状的减轻有关。

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