College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian, China.
Mid-Florida Research and Education Center, Department of Environmental Horticulture, Institute of Food and Agricultural Sciences, University of Florida, Apopka, FL 32703, USA.
Int J Mol Sci. 2021 Jan 8;22(2):568. doi: 10.3390/ijms22020568.
14-3-3 proteins (14-3-3s) are among the most important phosphorylated molecules playing crucial roles in regulating plant development and defense responses to environmental constraints. No report thus far has documented the gene family of in and their roles in response to stresses. In this study, nine genes, designated as ( through ) were identified from the latest genome. Phylogenetic analysis classified them into ε-like and non-ε groups, which were supported by gene structure analysis. The nine were located on five chromosomes, and none had duplication. Publicly available RNA-Seq raw data and microarray databases were mined for 14-3-3 expression profiles in different organs of citrus and in response to biotic and abiotic stresses. RT-qPCR was used for further examining spatial expression patterns of in citrus and their temporal expressions in one-year-old "Xuegan" plants after being exposed to different biotic and abiotic stresses. The nine were expressed in eight different organs with some isoforms displayed tissue-specific expression patterns. Six of the positively responded to citrus canker infection ( pv. ). The showed expressional divergence after phytohormone application and abiotic stress treatments, suggesting that 14-3-3 proteins are ubiquitous regulators in . Using the yeast two-hybrid assay, CitGF14a, b, c, d, g, and h were found to interact with CitGF14i proteins to form a heterodimer, while CitGF14i interacted with itself to form a homodimer. Further analysis of co-expression and potential interactors established a 14-3-3s protein interaction network. The established network identified genes and several candidate clients which may play an important role in developmental regulation and stress responses in this important fruit crop. This is the first study of 14-3-3s in citrus, and the established network may help further investigation of the roles of 14-3-3s in response to abiotic and biotic constraints.
14-3-3 蛋白(14-3-3s)是最重要的磷酸化分子之一,在调控植物发育和对环境胁迫的防御反应中起着至关重要的作用。迄今为止,尚无报道记录 在 中的基因家族及其在应对胁迫时的作用。在这项研究中,从最新的 基因组中鉴定了 9 个 基因,分别命名为 (通过 )。系统发育分析将它们分为 ε 样和非-ε 组,这得到了基因结构分析的支持。这 9 个 位于 5 条染色体上,没有重复。从柑橘不同器官和生物及非生物胁迫下的公共可用 RNA-Seq 原始数据和微阵列数据库中挖掘 14-3-3 表达谱。使用 RT-qPCR 进一步检测柑橘中 的空间表达模式及其在一年生“血橙”植物暴露于不同生物和非生物胁迫后的时间表达。这 9 个 在 8 个不同的器官中表达,有些同工型表现出组织特异性表达模式。其中 6 个 对柑橘溃疡病( pv. )感染呈正响应。 在植物激素处理和非生物胁迫处理后表现出表达差异,表明 14-3-3 蛋白是 中的普遍调节因子。利用酵母双杂交试验,发现 CitGF14a、b、c、d、g 和 h 与 CitGF14i 蛋白相互作用形成异二聚体,而 CitGF14i 相互作用形成同源二聚体。进一步的共表达分析和潜在相互作用物分析建立了 14-3-3s 蛋白相互作用网络。建立的网络确定了 基因和几个候选客户,它们可能在这个重要的水果作物的发育调控和应激反应中发挥重要作用。这是柑橘 14-3-3s 的首次研究,建立的网络可能有助于进一步研究 14-3-3s 在应对生物和非生物胁迫中的作用。