Scientia Agricultura Sinica ›› 2017, Vol. 50 ›› Issue (13): 2488-2497.doi: 10.3864/j.issn.0578-1752.2017.13.008

• PLANT PROTECTION • Previous Articles     Next Articles

Gene Cloning and Expression Analysis of Canker-Related Transcription Factor CsBZIP40 in Citrus

JIA RuiRui, ZHOU PengFei, BAI XiaoJing, CHEN ShanChun, XU LanZhen, PENG AiHong, LEI TianGang, YAO LiXiao, CHEN Min, HE YongRui, LI Qiang   

  1. Citrus Research Institute, Southwest University/Citrus Research Institute, Chinese Academy of Agricultural Sciences/National Citrus Engineering Research Center, Chongqing 400712
  • Received:2017-01-09 Online:2017-07-01 Published:2017-07-01

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Abstract: 【Objective】 The objective of this study is to annotate the BZIP family and clone the citrus canker related transcription factor CsBZIP40. It is also aimed to confirm the subcellular localization and the expression profile reduced by exogenous hormone and mechanical damage and the relations between CsBZIP40 and Xanthomonas citri subsp. citri (Xcc) infection. 【Method】Based on the public genome databases, the BZIP gene family was expertly and comprehensively annotated and named based on the chromosomal localization of all the members of BZIP; the motifs of the BZIPs were analyzed by MEME online tool. the phylogenetic tree of BZIPs in Citrus and Arabidopsis thaliana was constructed using software Mega 6.0 based on which the category of BZIP family was obtained. Canker-related transitional factor CsBZIP40 obtained from transcriptome data was also detected by qRT-PCR. Elements in the promoter and the nuclear localization signal were analyzed with database plantCARE and online tool cNLSmapper, respectively. And then the subcellular localization was confirmed by GFP fusion experiments in onion to confirm the prediction of nuclear localization analyzed with softwares. Expression profiles induced by salicylic acid (SA), jasmonic acid methyl ester (MeJA), ethylene (ET) and mechanical damage of CsBZIP40 were checked with qRT-PCR. 【Result】A total of 47 BZIP genes were annotated from the whole genome of Citrus sinensis and all the BZIPs are located on every chromosome except the 9th one. The BZIP concentration on chromosome 3 are 4.5×10-7/Mb which is the highest while chromosome 2 is the lowest, contains only 2% of all BZIPs in citrus. There were fewer gene duplication events detected from BZIP family of citrus compared with other plants, such as Arabidopsis, grapevine and so on. That is why citrus has a smaller BZIP family size. The full-length of CsBZIP40 is 5 756 bp with a 1 530 bp open reading frame which codes a protein containing 509 amino acids. It is closely related to AT1g08320 in A. thaliana based on the evolutionary analysis. In citrus, the BZIPs have been annotated which can be divided into 10 different sub-families. CsBZIP40 belongs to sub-family D, which is always take part in the pathogen resistance in plants. The gene promoter contains multiple cis involved in plant adversity or hormone response, such as Box-W1, HSE, ERE and so on. Subcellular localization results confirmed the prediction of protein localization in nucleus. Based on the qPCR data, the exogenous salicylic acid cannot induce the different expression of CsBZIP40, in contrast, jasmonic acid methyl ester, mechanical damage and ethylene can induce significant differences in gene expression level. Xcc attack can significantly increase the expression level of CsBZIP40 in Calamondin but no difference in Newhall navel orange. 【Conclusion】CsBZIP40 would be an important transcription factor which is closely associated with the resistance of citrus canker. This gene should be a potential candidate in the molecular breeding to improve the canker resistance of citrus. 

Key words: citrus canker, BZIP, transcription factors, subcellular localization

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