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本研究以拟南芥多氨氧化酶(AtPAO2)为探针,在雷蒙德氏棉基因组数据库中获得同源基因并设计引物,利用RT-PCR技术克隆PAO基因,分离一个新的陆地棉PAO基因命名为GhPAO2。该基因cDNA全长为2 237 bp,具有一个1473 bp的开放阅读框,编码490个氨基酸残基,预测该基因蛋白质分子量为54.41 kD,等电点为5.69。利用Real-time PCR对该基因在不同组织和多种非生物胁迫处理进行表达分析,结果表明:GhPAO2基因主要在棉花花瓣和叶中表达;不同胁迫处理下GhPAO2基因表达量均发生显著变化,其中低温处理和20%PEG处理后基因表达量迅速上升并在1 h时表达量达到最高,200 mmol/L NaCl和1 mmol/L ABA处理则在24 h时表达量达到最高,表明GhPAO2基因受低温、PEG、Na Cl和ABA诱导表达,可能在棉花抵御非生物胁迫过程中发挥重要作用。
In this study, Arabidopsis thaliana polyamine oxidase (AtPAO2) as a probe, access to homologous genes in the Redmond cotton genome database and design of primers, cloning PAO gene using RT-PCR technology to isolate a new upland cotton PAO The gene is named GhPAO2. The full-length cDNA of this gene was 2 237 bp in length with a 1473 bp open reading frame encoding 490 amino acid residues. The molecular weight of this gene was predicted to be 54.41 kD with an isoelectric point of 5.69. Real-time PCR was used to analyze the expression of GhPAO2 in different tissues and abiotic stresses. The results showed that GhPAO2 gene was mainly expressed in petals and leaves of cotton; GhPAO2 gene expression changed significantly under different stress treatments, The expression of GhPAO2 increased rapidly at 1 h after treatment with low temperature and 20% PEG treatment, and reached the peak at 1 h. The expression of GhPAO2 gene was highest at 24 h after treatment with 200 mmol / L NaCl and 1 mmol / L ABA. , PEG, NaCl and ABA induced cotton may play an important role in the defense against abiotic stress.