Key Laboratory of Sugarcane Biology and Genetic Breeding, Ministry of Agriculture, Fujian Agriculture and Forestry University, Fuzhou, 350002, China.
Plant Cell Rep. 2019 May;38(5):637-655. doi: 10.1007/s00299-019-02394-1. Epub 2019 Feb 12.
Six Sec14-like PITP genes from sugarcane were identified, two of them were cloned, and their biological functions were characterized indicating their involvement in plant defense against biotic and abiotic stresses. Sec14, a phosphatidylinositol transfer protein (PITP) is widely present in eukaryotes. In this study, the structure and expression patterns of six Sec14-like PITP genes (ScSEC14-1, ScSEC14p, ScSFH1, ScSFH2, ScPATL1, and ScPATL2) from sugarcane were analyzed, and two of them (ScSEC14-1 and ScSEC14p) were cloned and functionally verified. Phylogenetic analysis divided these genes into four groups, including group I (ScSFH1 and ScSFH2), group II (ScPATL1 and ScPATL2), Group III (ScSEC14p), and group V (ScSEC14-1). qRT-PCR analysis showed tissue-specific expression of these genes, primarily in the root, leaf, and bud tissues. They responded differently to SA, MeJA, and ABA stresses. ScSEC14-1, ScSEC14p, and ScSFH2 were upregulated by CuCl and CdCl, while ScSEC14-1, ScSFH1, ScSFH2, and ScPATL1 were upregulated by PEG and NaCl. When infected by Sporisorium scitamineum, the transcripts of ScSFH1, ScSFH2, ScPATL1, and ScPATL2 were upregulated in the resistant genotype Yacheng 05-179, while those of ScSEC14-1 and ScSEC14p were upregulated in the susceptible genotype ROC22. Subcellular localization showed that ScSEC14-1 and ScSEC14p were mainly localized in the plasma membrane and cytoplasm. Enhanced growth of Escherichia coli BL21 cells expressing ScSEC14-1 and ScSEC14p showed high tolerance to NaCl and mannitol stresses. The transient overexpression of ScSEC14-1 and ScSEC14p in Nicotiana benthamiana leaves enhanced its resistance to the infection of tobacco pathogens Ralstonia solanacearum and Fusarium solani var. coeruleum. We can conclude the involvement of ScSEC14-1 and ScSEC14p in the defense against biotic and abiotic stresses, which should facilitate further research on Sec14-like PITP gene family, especially its regulatory mechanisms in sugarcane.
从甘蔗中鉴定出 6 个 Sec14 样 PITP 基因，克隆了其中 2 个，并对其生物学功能进行了表征，表明它们参与了植物对生物和非生物胁迫的防御。Sec14 是一种磷酸肌醇转移蛋白 (PITP)，广泛存在于真核生物中。在这项研究中，分析了来自甘蔗的 6 个 Sec14 样 PITP 基因（ScSEC14-1、ScSEC14p、ScSFH1、ScSFH2、ScPATL1 和 ScPATL2）的结构和表达模式，并克隆和功能验证了其中 2 个（ScSEC14-1 和 ScSEC14p）。系统发育分析将这些基因分为 4 组，包括第 I 组（ScSFH1 和 ScSFH2）、第 II 组（ScPATL1 和 ScPATL2）、第 III 组（ScSEC14p）和第 V 组（ScSEC14-1）。qRT-PCR 分析显示这些基因在组织中具有特异性表达，主要在根、叶和芽组织中表达。它们对 SA、MeJA 和 ABA 胁迫的反应不同。CuCl 和 CdCl 上调 ScSEC14-1、ScSEC14p 和 ScSFH2，PEG 和 NaCl 上调 ScSEC14-1、ScSFH1、ScSFH2 和 ScPATL1。感染香蕉穿孔线虫后，在抗源品种 Yacheng 05-179 中，ScSFH1、ScSFH2、ScPATL1 和 ScPATL2 的转录本上调，而在感病品种 ROC22 中，ScSEC14-1 和 ScSEC14p 的转录本上调。亚细胞定位表明 ScSEC14-1 和 ScSEC14p 主要定位于质膜和细胞质。表达 ScSEC14-1 和 ScSEC14p 的大肠杆菌 BL21 细胞的生长增强表明其对 NaCl 和甘露醇胁迫具有高耐受性。ScSEC14-1 和 ScSEC14p 在烟草原生质体叶片中的瞬时过表达增强了其对烟草病原菌青枯菌和尖孢镰刀菌的抗性。我们可以得出结论，ScSEC14-1 和 ScSEC14p 参与了对生物和非生物胁迫的防御，这将有助于进一步研究 Sec14 样 PITP 基因家族，特别是其在甘蔗中的调控机制。
