Molecular Cloning of the PsiERF Gene and Its Expression in Korla Fragrant Pears

doi:10.6048/j.issn.1001-4330.2018.09.006

Abstract

Abstract: 【Objective】 The objective of this project is to compare the transcriptome expression in the Korla fragrant pears' flowers with either a president or deciduous calyx in the hope of screening differently expressed genes related to sepals.【Method】One nucleotide sequence which contained a AP2/ERF domain shared by the DEGs between transcriptome and DEG sequencing was selected from the Korla fragrant pears' flowers transcriptome database and named as PsiERF, and was analyzed by bioinformatics. Real-time quantitative PCR was used to compare temporal changes in the expression of PsiERF in floral organs with either persistent or deciduous calyx. 【Result】The full length cDNA of PsiERF is 1,195 bp which contained a 123 bp 5'-UTR(untranslated region), a 277 bp 3'-UTR and a 795 bp ORF encoding 264 amino acids. According to the analysis with real-time RT-PCR, the expression of PsiERF in flowers with persistent calyx was significantly (P<0.01) higher than that in flowers with deciduous calyx at the early and late bloom stages. And at the late bloom stage, persistent calyx was significantly (P<0.01) higher than deciduous calyx.【Conclusion】The present work has screened, cloned and identified a new PsiERF gene in Korla fragrant pear, which belongs to the ERF subtribe of the superfamily ethylene responsive factor (AP2/ERF) and our results indicated that PsiERF is closely related to the calyx persistence.

Key words: Korla fragrant pear; ERF; persistent and deciduous calyx; bioinformatics ; ;

摘要： 【目的】筛选克隆库尔勒香梨萼片脱落与宿存的相关基因,研究其在脱萼样品与宿萼样品中表达。【方法】以库尔勒香梨脱萼和宿萼花器官转录组测序数据为基础,从Unigenes中选出一条转录组和数字表达谱测序结果共有的与萼片脱落宿存相关的具有AP2/ERF结构域的差异基因,命名为PsiERF,对其进行生物信息学分析,通过实时荧光定量PCR分析其在花期不同阶段不同器官中的表达量。【结果】PsiERF cDNA序列的长度是1 195 bp,包含一个编码264个氨基酸长度为795 bp的开放阅读框(ORF),一个123 bp的5'端非编码区和一个277 bp的3'端非编码区。库尔勒香梨的PsiERF的核酸与氨基酸序列与其他植物的ERF109基因具有高度的同源性。实时荧光定量结果显示,其在全花样本中,宿萼组全花样品表达量在所有时期均高于脱萼组全花样品,而在宿萼组样品末花期萼片表达量极显著高于子房。【结论】筛选、克隆并鉴定一个新的库尔勒香梨PsiERF基因,该基因属于AP2/ERF家族中的ERF亚家族,与库尔勒香梨的萼片宿存脱落存在密切关系。

关键词: 库尔勒香梨, ERF, 萼片脱落于宿存, 生物信息学

CLC Number:

S611.2

XU Hang, QUAN Shao-wen, MA Li, ZHOU Li, NIU Jian-xin. Molecular Cloning of the PsiERF Gene and Its Expression in Korla Fragrant Pears[J]. Xinjiang Agricultural Sciences, 2018, 55(9): 1616-1625.

徐航, 全绍文, 马丽, 周丽, 牛建新. 库尔勒香梨PsiERF基因的克隆和表达分析[J]. 新疆农业科学, 2018, 55(9): 1616-1625.

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