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利用矮牵牛(Petunia hybrida)基因表达谱芯片筛选出应答低温胁迫的相关基因,从中发现1个B-box型锌指蛋白基因Ph BBX8。通过RT-PCR克隆得到该基因的编码区序列(CDS),为1 242 bp,预测其编码氨基酸413 aa,N端含有2个B-box盒,C端含有1个CCT结构域。系统进化树分析发现,Ph BBX8与拟南芥At BBX8等聚类为一支。利用半定量RT-PCR检测其在根、茎、叶和花中的表达特性,结果表明该基因在花中表达量较高,其次为根,而在茎和叶中表达较弱;利用实时定量PCR检测了在低温、干旱、ABA、Me JA、高盐和高渗胁迫处理下矮牵牛叶片中Ph BBX8的诱导表达情况,结果表明,Ph BBX8表现出不同程度的上调,其中除干旱胁迫外,其他胁迫下上调倍数都较高,初步推测该基因与矮牵牛应答低温等非生物逆境相关。
The genes related to low temperature stress were screened by Petunia hybrida cDNA microarray. One B-box zinc finger protein gene Ph BBX8 was found. The coding region (CDS) of this gene was 1 242 bp by RT-PCR. The predicted amino acid sequence was 413 aa. There are two B-box boxes at the N-terminal and one CCT domain at the C-terminal. Phylogenetic tree analysis found that Ph BBX8 and At BBX8 Arabidopsis clustered into one. The results of semi-quantitative RT-PCR showed that the gene was expressed in roots, stems, leaves and flowers. The results showed that the gene was highly expressed in roots, followed by roots and weakly expressed in stems and leaves. PCR was used to detect Ph BBX8 expression in petunia leaves under low temperature, drought, ABA, Me JA, high salt and high osmotic stress. The results showed that Ph BBX8 showed up-regulated in different degrees, except for drought stress , And the up-regulation times were higher under other stress. It was preliminarily presumed that this gene was related to the abiotic stress such as low temperature response of petunia.