Molecular cloning and expression of a novel eukaryotes elongation factor1A gene (ZjeEF-1α) from Chinese jujube in response to phytoplasma infection. (December 2016)

• Record Type: Journal Article
• Authors: Xie, Nannan; Yuan, Zan; Zhang, Liman; Zhao, Jin; Liu, Mengjun
• Is Part Of: Physiological and molecular plant pathology. Volume 96(2016:Oct.)
• Page Start: 101
• Page End: 108
• Publication Date: 2016-12
• Subjects: ZjeEF-1α -- Chinese jujube -- Phytoplasma -- RT-qPCR -- Western blotting; Plant diseases -- Periodicals; Diseased plants -- Physiology -- Periodicals; Phytopathogenic microorganisms -- Host plants -- Periodicals; 632
• DOI: 10.1016/j.pmpp.2016.10.004
• Languages: English
• ISSNs: 0885-5765

Abstract

Abstract: Eukaryotic translation elongation factor 1A (eEF1A) in plant is a multifunctional protein. In our previous study, we identified a 651-bp EST sequence of eEF-1α from a suppression subtractive hybridization (SSH) library of Chinese jujube ( Ziziphus jujuba Mill.), which was established under jujube witches' broom (JWB) phytoplasma stress. Here we isolated the eEF1A gene ( ZjeEF-1α, JF488063) from Chinese jujube by single oligonucleotide nested (SON)-PCR and rapid amplification of cDNA ends (RACE). ZjeEF-1α contains an open reading frame (ORF) of 1344 bp, encoding a polypeptide of 447 amino acids with an estimated molecular mass of 49.29 kDa and a theoretical pI of 9.10. ZjeEF-1α shared over 92% similarity with eEF-1α proteins from other 24 eukaryotes including plants and animals. Search Pfam predicted that ZjeEF-1A contains three typical eEF1A functional domains. Moreover, the corresponding genomic sequence of ZjeEF-1α was also sequenced, in which a 115-bp intervening sequence with splicing site GT-AG was found. Real-time quantitative PCR (RT-qPCR) and western blotting assays revealed that the expression of ZjeEF-1α was enhanced in resistant cultivar 'Xingguang' and decreased in sensitive cultivar 'Zanhuangdazao' at early stages of phytoplasma infection. It is proved that ZjeEF-1A expression is early response to phytoplasma infection and its possible functions in resistant process to JWB phytoplasma are discussed. Highlights: ZjeEF-1α was firstly isolated from Chinese jujube. ZjeEF-1α expression was enhanced in resistant cultivar at early stages of phytoplasma infection. ZjeEF-1α is earlier response to phytoplasma infection.