广西师范大学学报（自然科学版） ›› 2016, Vol. 34 ›› Issue (3): 131-137.doi: 10.16088/j.issn.1001-6600.2016.03.019

• • 上一篇    下一篇

沙田柚WRKY转录因子的克隆与序列分析

张渝1,2, 刘玉洁1,2, 郭丹妮1,2, 覃信梅1,2, 韩愈1,2, 李惠敏1,2, 秦新民1,2   

  1. 1.广西师范大学生命科学学院,广西桂林541006;
    2.广西师范大学珍稀濒危动植物生态与环境保护教育部重点实验室,广西桂林541006
  • 收稿日期:2015-11-27 出版日期:2016-09-30 发布日期:2018-09-17
  • 通讯作者: 秦新民(1956—),男,广西灵川人,广西师范大学教授,博士。E-mail:xmqin@mailbox.gxnu.edu.cn
  • 基金资助:
    国家自然科学基金资助项目(31360477);广西教育厅项目基金资助(2013YB036)

Cloning and Sequence Analysis of WRKY Transcription Factorfrom Citrus grandis var. Shatinyu Hort

ZHANG Yu1,2, LIU Yujie1,2, GUO Danni1,2, QIN Xinmei1,2, HAN Yu1,2, LI Huimin1,2, QIN Xinmin1,2   

  1. 1.College of Life Science, Guangxi Normal University, Guilin Guangxi 541006,China;
    2.Key Laboratory of Ecology of Rare and Endangered Species and Environmental Protection(Ministry of Education),Guangxi Normal University, Guilin Guangxi 541006,China
  • Received:2015-11-27 Online:2016-09-30 Published:2018-09-17

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摘要： 利用高通量测序技术对沙田柚自交和异交花柱进行转录组测序,差异分析得到沙田柚WRKY转录因子的序列。该基因的全长序列为1 178 bp (GenBank accession No. KU173833),含有993 bp的开放阅读框(ORF)可编码330个氨基酸,编码蛋白的相对分子质量为37.00 ku,理论等电点为5.79。与未授粉的花柱相比,沙田柚异花授粉1、2、3 d花柱中WRKY转录因子基因的表达量(RPKM)分别为5.67、26.04、17.08;而自花授粉1、2、3 d花柱中WRKY转录因子基因的表达量(RPKM)分别为15.67、14.96、3.89。氨基酸序列分析表明,该氨基酸序列与甜橙、金桔的同源性分别为98%、97%。系统进化分析发现沙田柚WRKY转录因子与甜橙、金桔的亲缘关系很近,属于一个分支。

关键词: 沙田柚, WRKY, 转录因子, 自交不亲和性

Abstract: In this study, the transcriptome of the self-pollinated styles and cross-pollinated styles of Citrus grandis var. Shatinyu Hort were sequenced by high-throughput sequencing technology. WRKY transcription factor gene of Citrus grandis var. Shatinyu Hort was obtained through variance analysis method. It is 1 178 bp in length with an open reading frame (ORF) of 993 bp, encoding 330 amino acids with deduced molecular weight of 37.00 ku, and theoretical pI value of 5.79. Compared with un-pollinated styles, the expression (RPKM) of WRKY transcription factor gene is respectively 5.67, 26.04 and 17.08 in 1, 2, 3 days cross-pollinated styles, and expression (RPKM) of the gene is respectively 15.67, 14.96 and 3.89 in 1, 2, 3 days self-pollinated styles. The homology analysis of amino acid sequence indicated that the WRKY transcription factor protein shared higher homology with that of Citrus sinensis (98%) and Citrus japonica (97%). Phylogenetic analysis revealed that WRKY transcription factor gene showed closer kinship with that of Citrus sinensis and Citrus_japonica,indicating that they belong to the same evolutionary branch.

Key words: Citrus grandis var. Shatinyu Hort, WRKY, transcription factor, self-incompatibility

中图分类号: 

  • Q946
[1] PAETSCH M, MAYLAND-QUELLHORST S, NEUFFER B. Evolution of the self-incompatibility system in the Brassicaceae, identification of S-locus receptor kinase(SRK) in self-incompatible Capsella grandiflora[J]. Heredity, 2006,97:283-290.
[2] SHIBA H, PARK J I, SUZUKI G, et al. Duplicated SP11genes produce alternative transcripts in the S15 haplotype of Brassica oleracea[J]. Genes and Systems, 2004,79:87-93.
[3] SHIMOSATO H, YOKOTA N, SHIBA H, et al. Characterization of the SP11/SCR high-affinity binding site involved in self/nonself-recognition in brassica self-incompatibility[J]. The Plant Cell, 2007,19:107-117.
[4] ANDERSON M A, CORNISH E C, MAU S L, et al. Cloning of cDNA for a stylar glycoprotein associated with expression of self-incompatibility in Naicotiana alal[J]. Nature,1986,321:38-44.
[5] HUA Z, MENG X, KAO T H. Comparison of Petunis inflata S-locus F-box protein(Pi SLF) with Pi SLFlike proteins reveals its unique function in S-RNase based self-incompatibility[J]. The Plant Cell, 2007:19:3593-3609.
[6] VAUGHAN S P, RAMANE H, WATARI A, et al. The S haplotype-specific F-box protein gene, SFB, is defective in self-compatible haplotypes of Prunus avium and P.mume[J]. The Plant Journal, 2004,39:573-586.
[7] 秦新民,张渝,刘玉洁,等.沙田柚S-RNase基因的克隆及序列分析[J].广西师范大学学报(自然科学版),2015,33(1):139-145.
[8] DE GRAFF B H, VATOVEC S, JUAREZ-DIAZ J A, et al. The Papaver self-incompatibility pollen S-determinant, PrpS, function in Arabidopsis thaliana[J]. Current Biology, 2012,22:154-159.
[9] MCCLURE B, MOU B, CANEVASCINI S, et al. A small asparagine-rich protein required for S-allele-specific pollen rejection in Nicotiana[J]. Proc Natl Acad Sci USA ,199,96:13548-13553.
[10] GOLDRAIJ A, KONDO K, LEE C B, et al. Compartmentalization of S-RNase and HT-B degradation in self-incompatible Nicotiana[J]. Nature, 2006,439:805-810.
[11] NATHAN HANCOCK C, KENT L, MCCLURE B A. The stylar120 kDa glycoprotein is required for S-specific pollen rejection in Nicotiana[J]. Plant J, 2005,43:716-723.
[12] DISTEFANO G, CARUSO M, LA MALFA S, et al. Histological and molecular analysis of pollen-pistil interaction in Clementine[J]. Plant Cell Rep, 2009,281:439-1451.
[13] ISHIGURO S, NAKAMURA K. Characterization of a cDNA encoding a novel DNA-binding protein, SPF1, that recognizes SP8 sequences in the 5’upstream regions of genes coding for sporamin and b-amylase from sweet potato[J]. Mol Gen Genet, 1994,244:563-571.
[14] RUSHTON P J,MACDONALD H, HUTTLY A K, et al. Members of a new family of DNA-binding proteins bind to a conserved cis-element in the promoters of a-Amy2 genes[J]. Plant Mol Biol, 1995,29:691-702.
[15] RUSHTON P J, TORRES J M, WERNERT P, et al. Interaction of elicitor-induced DNA binding proteins with elicitor response elements in the promoters of parsley PR1 genes[J]. EMBO J, 1996,15:5690-5700.
[16] DE PATER S, GRECO V, PHAM K, et al. Characterization of a zinc-dependent transcriptional activator from Arabidopsis[J]. Nucleic Acids Res, 1996,24:4624-4631.
[17] BAKKHI M, OELMULLER R. WRKY transcription factors: Jack of many trades in plants[J]. Plant Signaling and Behavior, 2014, 9(2): e27700.
[18] 宋钰,荆邵娟,余迪求.水稻WRKY转录调控因子基因功能研究进展[J]. 中国水稻科学,2009,23(5):447-455.
[19] ZHOU X, JIANG Y, YU D. WRKY22 transcription factor mediates dark-induced leaf senescence in Arabidopsis[J]. Molecules and Cells, 2011, 31: 303-313.
[20] BARIOLA P A, HOWARD C J, TAYLOR C B, et al. The Arabidopsis ribonuclease gene RNS1 is tightly controlled in response to phosphate limitation[J]. The Plant Journal, 1994, 6: 673-685.
[21] ZHANG S W, HUANG G X, DING F, et al. Mechanism of seedlessness in a new lemon cultivar‘Xiangshui’ [Citrus limon (L.) Burm. F][J]. Sex Plant Reprod,2012, 25:337-345.
[22] ZHANG S, DING F, HE X, et al. Characterization of the ‘Xiangshui’lemon transcriptome by de novo assembly to discover genes associated with self-incompatibility[J]. Molecular Genetics and Genomics, 2015, 290: 365-375.
[23] AGARWAL P, REDDY M P, CHIKARA J. WRKY: its structure, evolutionary relationship, DNA-binding selectivity, role in stress tolerance and development of plants[J]. Molecular biology reports, 2011, 38: 3883-3896.
[24] RUSHTON P J, SOMSSICH I E, RINGLER P, et al. WRKY transcription factors[J]. Trends in Plant Science, 2010, 15: 247-258.
[25] GUAN Y, MENG X, KHANNA R, et al. Phosphorylation of a WRKY transcription factor by MAPKs is required for pollen development and function in Arabidopsis[J]. Plos Genetics, 2014, 10:1-12.
[26] LUO M, DENNIS E S, BERGER F, et al. MINISEED3 (MINI3), a WRKY family gene, and HAIKU2 (IKU2), a leucine-rich repeat (LRR) KINASE gene, are regulators of seed size in Arabidopsis[J]. Proceedings of the National Academy of Sciences of the United States of America, 2005, 102: 17531-17536.
[27] ROBATZEK S, SOMSSICH I E. A new member of the Arabidopsis WRKY transcripyion factor family, AtWRKY6, is associated with both senescence-and defence-related processes[J]. The Plant Journal, 2001, 2:123-133.
[28] 李欢,何炎森,李科,等. 多花水仙WRKY转录因子的克隆与序列分析[J]. 热带作物学报,2014,35(12):2378-2383.
[1] 秦新民, 张渝, 刘玉洁, 郭丹妮, 李惠敏. 沙田柚S-RNase基因的克隆及序列分析[J]. 广西师范大学学报（自然科学版）, 2015, 33(1): 139-145.
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