龙眼ERF家族成员鉴定及其在体胚发生早期的表达
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国家自然科学基金(31572088,31672127);


Identification of ERF Gene Family and Their Expression Analysis during Early Somatic Embryogenesis in Dimocarpus longan
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    摘要:

    为了解龙眼ERF家族的基本性质以及在体胚发生早期的表达规律,该研究对龙眼基因组鉴定出108个ERF家族成员进行基本理化性质分析与进化树构建,并结合龙眼转录组及miRNA、lncRNA数据库对DlERF家族成员与lncRNA、miRNA之间的关系预测与表达模式分析。结果显示:(1)理化性质及进化树分析表明,AP2/ERF结构域在龙眼中相对最为保守,DlERF对龙眼的抗胁迫能力以及对病菌的防御能力可能起着重要作用。(2)RNASeq中的表达量分析可知,95个ERF基因在转录组中检测到表达,在愈伤组织(EC)、不完全胚性紧实结构(ICpEC)与球形胚(GE)阶段中分别存在31、11与53个ERF基因高表达。(3)qRTPCR结果显示,在龙眼体胚发生早期显著表达5个ERF基因中,Dlo_008317.1ERF152、Dlo_022310.1ERF11与Dlo_009939.1ERF98在GE阶段表达量显著高于EC与ICpEC阶段,Dlo_009070.1ERF1063与Dlo_022634.1ERF221则分别在EC与ICpEC阶段高表达。(4)DlERF家族成员与相关lncRNA、miRNA表达量分析显示,LTCONS_00013739对靶基因Dlo_008317.1ERF152为正调控关系;Dlo_miR413与Dlo_miR1510a共同靶向Dlo_009070.1ERF1063,从EC到GE阶段均表现出显著负调控关系,而Dlo_miR413与Dlo_008317.1ERF152的表达量表现出正相关,推测相比于Dlo_008317.1ERF152,Dlo_miR413更倾向于调控Dlo_009070.1ERF1063;调控Dlo_009939.1ERF98相关的Dlo_miR408与Dlo_miR774b,从表达趋势来看,Dlo_miR774b在龙眼体胚发生早期过程能够负调控Dlo_009939.1ERF98;Dlo_miR399c与Dlo_022310.1ERF11在EC到GE阶段可能存在负调控关系。(5)不同梯度浓度的乙烯处理使得DlERF基因表达显著下调。这些发现提供了有关龙眼体胚发生早期DlERF的重要见解,从而为将来对龙眼体胚发生早期过程中DlERF的功能分析奠定了基础。

    Abstract:

    In order to understand the basic properties of the longan ERF family and the expression of early embryogenesis, this study identified 108 ERF family members for basic physicochemical properties and phylogenetic tree construction in the longan genome. The relationship between DlERF family members and lncRNA and miRNA was predicted and expressed in combination with longan transcriptome, miRNA and lncRNA database. The results showed that: (1) the physicochemical properties and phylogenetic tree analysis of the family indicated that the AP2/ERF domain is relatively the most conservative in longan, and DlERF may play an important role in the longterm stress resistance and defense ability of the pathogen. (2) Analysis of expression levels in RNASeq revealed that 95 ERF genes were detected in the transcriptome, and there were 31, 11 and 53 ERF genes highly expressed in the callus, incomplete embryonic compact structure and globular embryo stage, respectively. (3) qRTPCR results showed that among the five ERF genes that were significantly expressed early in the longan somatic embryo, the expression levels of Dlo_008317.1ERF152, Dlo_022310.1ERF11 and Dlo_009939.1ERF98 in GE stage were significantly higher than those of EC and ICpEC, Dlo_009070.1ERF1063 and Dlo_022634.1ERF221 were highly expressed in the EC and ICpEC phases, respectively. (4) Analysis of DlERF family members and related lncRNA and miRNA expression levels showed that LTCONS_00013739 has a positive regulatory relationship with the target gene Dlo_008317.1ERF152; Dlo_miR413 and Dlo_miR1510a target Dlo_009070.1ERF1063 together, showing a significant negative regulatory relationship from EC to GE, while Dlo_miR413 and Dlo_008317.1ERF152 expression were positively correlated, presumably compared to Dlo_008317.1ERF152 Dlo_miR413 is more inclined to regulate Dlo_009070.1ERF1063; Dlo_miR408 and Dlo_miR774b regulate Dlo_009939.1ERF98. From the expression trend, Dlo_miR774b can negatively regulate Dlo_009939.1ERF98 in the early stage of longan somatic embryogenesis; Dlo_miR399c and Dlo_022310.1ERF11 may have negative regulation relationship in EC to GE stage. (5) Ethylene treatment with different gradient concentrations significantly downregulated DlERF gene expression. These findings provide important insights into DlERF during somatic embryogenesis in longan, and lay a foundation for further functional analysis of DlERF during somatic embryogenesis in longan.

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陈 燕,吕科良,厉 雪,等.龙眼ERF家族成员鉴定及其在体胚发生早期的表达[J].西北植物学报,2018,38(11):1986-1999

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  • 在线发布日期: 2018-12-20
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