| 程晓欣,翟许玲,闫晋强,等.冬瓜ARF4基因克隆及表达特性分析[J].广东农业科学,2024,(11-12):- |
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冬瓜ARF4基因克隆及表达特性分析 |
| The Cloning and Expression Characteristics of BhARF4 Gene in Wax Gourd |
| 投稿时间：2024-09-03 修订日期：2024-10-18 |
| DOI： |
| 中文关键词: 冬瓜 ARF 果实发育 |
| 英文关键词: wax gourd ARF fruit development |
| 基金项目:国家自然科学基金（32202504）；广西科技计划项目（桂科AA22068086）；江门市科技计划项目（江科〔2022〕191 号） |
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| 摘要点击次数: 50 |
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| 中文摘要: |
| 【目的】克隆冬瓜ARF4基因,开展生物信息分析,并研究其在不同组织的表达模式,为进一步深入探究BhARF4在冬瓜果实发育中的功能提供了参考。【方法】以冬瓜B227高代自交系开花当天的果实cDNA为模板,根据参考基因中BhARF4的CDS序列设计特异性克隆引物进行克隆。在结果期,取B227不同器官(根、茎、叶、开花当天的雌花和雄花样本,以及授粉后不同发育阶段的果实(0d,5d,10d,15d,20d和30d)样本,通过qRT-PCR分析BhARF4在不同组织的表达模式。【结果】BhARF4含有12个外显子和2个内含子,CDS序列长2412bp；motif和保守结构域分析显示,BhARF4蛋白含有9个motif并含有保守的ARF、AUX_IAA和B3结构域；系统进化分析显示,BhARF4蛋白与黄瓜、甜瓜的ARF4蛋白亲缘关系最近；基因表达模式分析显示,BhARF4在冬瓜果实中表达较高,尤其是当天授粉的果实。【结论】明确了BhARF4的基因结构,BhARF4蛋白的保守结构域等生物学信息特征,以及基因表达模式,推测BhARF4在冬瓜果实生长阶段,尤其是果实生长初期发挥重要作用。 |
| 英文摘要: |
| 【Objective】The ARF4 gene was cloned and its expression patterns in different tissues were studied, thereby providing a reference for further exploration of the function of BhARF4 in the development of wax gourd fruits. 【Method】Specific cloning primers were designed according to the CDS sequence of BhARF4 in the reference gene,using the cDNA of the fruits of B227 on the day of flowering as the template. At the fruiting stage, samples of B227 were taken from different organs (root, stem, leaf, female and male flowers on the day of flowering, as well as fruits on the different day of after pollination(DAP)(0 DAP, 5 DAP, 10 DAP, 15 DAP, 20 DAP and 30 DAP).The expression patterns of BhARF4 in different tissues and different stages of fruits were analyzed by qRT-PCR.【Result】The BhARF4 gene comprises 12 exons and 2 introns, with its CDS sequence length of 2412 bp. Motif and conserved domain analysis revealed that the BhARF4 protein contains 9 motifs and conserved ARF, AUX_IAA, and B3 domains. A phylogenetic analysis revealed that the BhARF4 protein exhibited the highest degree of similarity to the ARF4 protein derived from cucumber and melon. The gene expression pattern analysis revealed that BhARF4 exhibited high expression levels in wax gourd fruits, particularly those on 0 DAP.【Conclusion】The gene structure of BhARF4, the conserved domain of the BhARF4 protein, and other biological information characteristics, as well as the gene expression pattern, were elucidated. It is hypothesized that BhARF4 plays a significant role in the growth of wax gourd fruit, particularly during the initial stages of development. |
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