苹果HD-ZipⅠ家族基因序列特征及其在果实中的激素响应

doi:10.3864/j.issn.0578-1752.2015.21.011

摘要/Abstract

摘要： 【目的】对苹果基因组中挖掘得到的HD-Zip I（Homeodomain leucine zipper I）家族基因进行生物信息预测，结合生理生化数据为苹果果实发育的激素调控网络研究提供最佳候选基因。【方法】应用ExPASy进行HD-Zip I家族蛋白基本理化性质分析，采用MEME和PLACE软件预测各成员启动子含有的顺式作用元件及其功能，利用GSDS绘制基因内含子/外显子结构示意图；通过内源乙烯浓度、可溶性固形物、可滴定酸、硬度等品质指标检测‘皇家嘎啦’苹果果实在不同激素处理条件下的后熟进程；运用半定量RT-PCR分析HD-Zip I家族各成员对外源激素的响应。【结果】鉴定完整的22个HD-Zip I基因可以细分为5个亚类，同一亚类基因的内含子/外显子结构相似，但仍有差异；各基因起始密码子上游-600 bp区域内广泛分布多个响应赤霉素、细胞分裂素、茉莉酸、乙烯和脱落酸的顺式作用元件，以（GA/TC）8重复序列最为保守和丰富。乙烯和脱落酸两种激素处理后，果实内源乙烯浓度及硬度等各指标变化表现出极高的相似度，均加快了‘皇家嘎啦’果实的后熟进程；不同于其他各家族成员，MdHZ1和MdHZ17两个基因的转录水平在乙烯和脱落酸处理后48 h均被不同程度的上调，而在有延缓果实衰老效果的腐胺（PUT）和赤霉素（GA4）处理组中表现为下调，表明MdHZ1和MdHZ17可能与果实的后熟进程有较为紧密的联系。MdHZ16在成熟果实组织中未检测到表达,其他家族成员的激素响应特性未表现出一定的规律性，表明同一家族内即使结构相似的基因也并不一定呈现相似的表达模式，家族基因上游调控机制有一定的复杂性。【结论】通过苹果基因组扫描获得22个HD-Zip I基因，这些基因编码的蛋白结构相似度较高；基因启动子区域含有多个参与不同激素交互作用的顺式作用元件，这或许为家族成员差异响应各外源激素处理的原因；MdHZ1和MdHZ17两个基因与果实的后熟进程表现出较为紧密的联系，可以作为候选基因进行果实后熟相关方面的深入研究。

关键词: 苹果, HD-Zip I家族, 激素, 后熟, 差异表达基因

Abstract: 【Objective】By integrating bioinformatics and analyses of physiological features, the functions of homeodomain leucine zipper I (HD-Zip I) in apple genome were studied. This study may aid in the selection of appropriate candidate genes for apple ripening research.【Method】ExPASy was employed to analyze the family members’ basic physical and chemical properties, MEME and PLACE were used to predict cis-elements in the promoters and their functions, and GSDS was used to draw the stucture map of intron/extron. The ripening characters such as internal ethylene concentration, soluble solids, hardness and titratable acid after various hormone-treatments were tested. Semi-quantitative PCR was used to detect gene responses to different plant hormones.【Result】A total of 22 HD-Zip I genes were identified in Royal Gala, and they are further divided into ?ve clades with phylogenetic analysis. The intron/exon structures showed a partly conservation in structure from the same cluster. Cis-elements related to hormone responses (such as gibberellin, cytokinins, jasmonic acid, ethylene and abscisic acid) are widely distributed in the -600 bp upsteam region from ATG, and the most conserved and abundant one is (GA/TC)8 repeated sequence. The treatments of ethylene and abscisic acid similarly increased the inner ethylene content and promoted the maturation process. Differing from other members, the transcriptional levels of MdHZ1 and MdHZ17 were both up-regulated on 48 h after treatments, while their levels were down-regulated by PUT and GA4, which could delay the ripening process of apples in the experiments. These data showed a close connection between genes MdHZ1, MdHZ17 and fruit ripening. At the same time, it was also detected that the family members answered the hormone treatments differently and MdHZ16 was the undetectable one. Therefore, the up-regulate network of this family is complex and even the similar structure genes within a same family do not present alike expression patterns.【Conclusion】There are 22 HD-Zip I genes identified from apple and they possess similar gene structures. Promoter regions contain lots of cis-elements involved in hormone interaction response, and this may be a reason for the differently answer to exogenous hormone treatments. MdHZ1 and MdHZ17 could be the main candidate genes participating in ripening progress.

Key words: apple, HD-Zip I gene family, hormone, ripening, significant genes

温小红，姜永华，亢键，王豪杰，任小林. 苹果HD-ZipⅠ家族基因序列特征及其在果实中的激素响应[J]. 中国农业科学, 2015, 48(21): 4314-4322.

WEN Xiao-hong, JIANG Yong-hua, KANG Jian, WANG Hao-jie,REN Xiao-lin. A Genome-Wide Survey of HD-ZipⅠ Genes and Their Responses to Hormone in Apple[J]. Scientia Agricultura Sinica, 2015, 48(21): 4314-4322.

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