灰葡萄孢菌诱导的百合转录辅激活因子LlMBF1a基因的克隆及其特征分析

doi:10.3969/j.issn.1004-1524.2018.12.06

浙江农业学报 ›› 2018, Vol. 30 ›› Issue (12): 2024-2030.DOI: 10.3969/j.issn.1004-1524.2018.12.06

灰葡萄孢菌诱导的百合转录辅激活因子LlMBF1a基因的克隆及其特征分析

曹兴¹, 张秀省¹, 侯栋², 隋娟娟³, 穆红梅¹, 高祥斌¹, 吕福堂¹, 郭尚敬¹, 王桂清^1,*

1.聊城大学农学院,山东聊城 252059;
2.甘肃省农业科学院蔬菜研究所,甘肃兰州 730070;
3.阜阳师范学院生物与食品工程学院,安徽阜阳 236037

收稿日期:2018-03-12 出版日期:2018-12-25 发布日期:2018-12-28
通讯作者: 王桂清,E-mail: wangguiqing@lcu.edu.cn
作者简介:曹兴(1984—),男,山东滕州人,博士,讲师,主要从事园艺作物病害防治研究。E-mail: caoxinglina@163.com
基金资助:
国家自然科学基金(31601788);山东省自然科学基金(ZR2016CB36);聊城大学博士科研启动基金(318051312)

Cloning and characterization of LlMBF1a in lily induced by gray mold pathogen

CAO Xing¹, ZHANG Xiusheng¹, HOU Dong², SUI Juanjuan³, MU Hongmei¹, GAO Xiangbin¹, LYU Futang¹, GUO Shangjing¹, WANG Guiqing^1,*

1. College of Agriculture, Liaocheng University, Liaocheng 252059, China;
2. Vegetables Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou 730070, China;
3. College of Biology and Food Engineering, Fuyang Normal College, Fuyang 236037, China

Received:2018-03-12 Online:2018-12-25 Published:2018-12-28

摘要/Abstract

摘要： 转录辅激活因子MBF1a参与了植物对病原真菌的防御反应。利用电子克隆结合RT-PCR技术从百合中分离了LlMBF1a基因,其开放阅读框长429 bp,编码142个氨基酸。LlMBF1a分子量为15.53 ku,理论等电点为10.12,为稳定的水溶性非分泌蛋白,含有典型的MBF1结构域和α螺旋-转角-α螺旋模序。亚细胞定位分析表明,LlMBF1a主要在细胞核中表达。LlMBF1a在根、茎、鳞茎、叶中的表达量无显著差异,受灰葡萄孢菌侵染诱导表达且在相对抗病品种中持续高水平表达,表明LlMBF1a可能参与了百合对灰霉病的抗性反应。

关键词: 百合, MBF1, 灰葡萄孢菌, 基因克隆, 亚细胞定位, 基因表达

Abstract: Previous studies revealed that MBF1a was involved in host defense response. The MBF1a homologous gene was isolated from lily cultivar White Heaven by silico cloning and RT-PCR approaches. The open reading frame of LlMBF1a gene was 429 bp, encoding a protein of 142 amino acid residues, and the molecular weight of LlMBF1a protein was 15.53 ku with a theoretical isoelectric point of 10.12. Bioinformatics analysis predicted that LlMBF1a was a stable soluble non-secreted protein, containing a typical MBF1 motif and a helix-turn-helix motif. Subcellular localization assay indicated LlMBF1a expressed in nucleus. The expression of LlMBF1a was at a similar level in leaf, stem, bulb and root, and was induced by Botrytis cinerea. The up-regulation of LlMBF1a in resistant variety was higher and more persistent than that in susceptible variety, suggesting LlMBF1a might be correlated to the resistance of lily against gray mold.

Key words: lily, MBF1, Botrytis cinerea, gene cloning, subcellular localization, gene expression

中图分类号:

S682.2⁺65

曹兴, 张秀省, 侯栋, 隋娟娟, 穆红梅, 高祥斌, 吕福堂, 郭尚敬, 王桂清. 灰葡萄孢菌诱导的百合转录辅激活因子LlMBF1a基因的克隆及其特征分析[J]. 浙江农业学报, 2018, 30(12): 2024-2030.

CAO Xing, ZHANG Xiusheng, HOU Dong, SUI Juanjuan, MU Hongmei, GAO Xiangbin, LYU Futang, GUO Shangjing, WANG Guiqing. Cloning and characterization of LlMBF1a in lily induced by gray mold pathogen[J]. , 2018, 30(12): 2024-2030.

参考文献

[1] 瞿友均, 徐源辉. 百合灰霉病的发生特点和综合防治方法[J]. 中国植保导刊, 2010, 30(4): 26-27.
QU Y J, XU Y H.Lily gray mould occurrence and control[J]. China Plant Protection, 2010, 30(4): 26-27. (in Chinese with English abstract)
[2] OZAKI J, TAKEMARU K I, IKEGAMI T, et al.Identification of the core domain and the secondary structure of the transcriptional coactivator MBF1[J]. Genes Cells, 1999, 4(7): 415-424.
[3] 张毅, 张岗, 董艳玲, 等. 条锈菌诱导的小麦MBF1转录辅激活因子基因的克隆及其特征分析[J]. 作物学报, 2009, 35(1): 11-17.
ZHANG Y, ZHANG G, DONG Y L, et al.Cloning and characterization of a MBF1 transcriptional coactivator factor in wheat induced by stripe rust pathogen[J]. Acta Agronomica Sinica, 2009, 35(1): 11-17. (in Chinese with English abstract)
[4] KIM M J, LIM G H, KIM E S, et al.Abiotic and biotic stress tolerance in Arabidopsis overexpressing the multiprotein bridging factor 1a (MBF1a) transcriptional coactivator gene[J]. Biochemical & Biophysical Research Communications, 2007, 354(2): 440-446.
[5] GODAY A V, ZANETTI M E, SAN SEGUNDO B, et al.Identification of a putative Solanum tuberosum transcriptional coativator up-regulated in potato tubers by Fasarium solanif sp. Eumartii infection and wounding[J]. Physiologia Plantarum, 2001, 112(2): 217-222.
[6] 隋娟娟, 李晓昕, 杨秋燕, 等. 重瓣百合LiSEP3基因克隆与表达分析[J]. 南京林业大学学报(自然科学版), 2017, 41(1): 42-48.
SUI J J, LI X X, YANG Q Y, et al.Cloning and expression analysis of LiSEP3 in double lily[J]. Journal of Nanjing Forestry University (Nature Sciences Edition), 2017, 41(1): 42-48. (in Chinese with English abstract)
[7] GONG B H, YI J, WU J, et al.LlHSFA1, a novel heat stress transcription factor in lily (Lilium longiflorum), can interact with LlHSFA2 and enhance the thermotolerance of transgenic Arabidopsis thaliana[J]. Plant Cell Reports, 2014, 33(9): 1519-1533.
[8] SCHMITTGEN T D, LIVAK K J.Analyzing real-time PCR data by the comparative CT method[J]. Nature Protocols, 2008, 3(6): 1101-1108.
[9] 朱丽梅, 侯建文, 罗凤霞, 等. 百合灰霉病的诊断及其病原物的分离纯化[J]. 金陵科技学院学报, 2009, 25(3): 59-63.
ZHU L M, HOU J W, LUO F X, et al.Diagnosis of lily gray mould and isolation and purification of its pathogen[J]. Journal of Jinling Institute of Technology, 2009, 25(3): 59-63. (in Chinese with English abstract)
[10] CAO X Q, SHI S, ZHANG Z.First report of Botrytis leaf blight on lily (Lilium longiflorum) caused by Botrytis cinerea in Beijing, China[J]. Plant Disease, 2017, 102(5):1033.
[11] 朱丽梅, 张长青, 陈浩, 等. 百合灰霉病高效杀菌剂的筛选和联合作用研究[J]. 金陵科技学院学报, 2011, 27(1): 51-54.
ZHU L M, ZHANG C Q, CHEN H, et al.Screening and combined action of efficient fungicides for controlling lily Botrytis cinerea[J]. Journal of Jinling Institute of Technology, 2011, 27(1): 51-54. (in Chinese with English abstract)
[12] 梁巧兰, 张娜, 魏列新, 等. 深绿木霉蛋白质TraT2A诱导兰州百合抗灰霉病的作用[J]. 中国生物防治学报, 2017, 33(4): 545-551.
LIANG Q L, ZHANG N, WEI L X, et al.Effect of Trichoderma atroviride proteinaceous TraT2A induced Lanzhou lily resistant to gray mold caused by Botrytis cinerea[J]. Chinese Journal of Biological Control, 2017, 33(4): 545-551. (in Chinese with English abstract)
[13] 朱丽梅, 胡凤荣, 罗凤霞. 不同百合品种对百合灰霉病的抗病性鉴定[J]. 植物保护, 2010, 36(3): 148-151.
ZHU L M, HU F R, LUO F X.Identification of disease resistance of Lilium species to lily gray mould[J]. Plant Protection, 2010, 36(3): 148-151. (in Chinese with English abstract)
[14] 杜运鹏. 我国百合属植物资源评价及抗病基因同源序列(RGA)的研究[D]. 北京: 北京林业大学, 2014.
DU Y P.Collection and evaluation of Lilium spp. in China (Liliaceae) and the research on resistance gene analog (RGA) [D]. Beijing: Beijing Forestry University, 2014. (in Chinese with English abstract)
[15] 宋翔宇. 椭圆灰葡萄孢(Botrytis cinerea)胁迫下亚洲百合的生理响应及调节基因MAPK的克隆[D]. 沈阳: 沈阳农业大学, 2016.
SONG X Y.Physiological responses of Lilium asiatic hybrid to Botrytis cinerea stress and cloning of regulatory gene-MAPK[D]. Shenyang: Shenyang Agriculture University, 2016. (in Chinese with English abstract)
[16] 韩青, 杨野, 陈瑞, 等. 超表达岷江百合类萌发素蛋白基因LrGLP2增强烟草对几种病原真菌的抗性[J]. 植物生理学报, 2015, 51(12): 2223-2230.
HAN Q, YANG Y, CHEN R, et al.Overexpression of germin-like protein gene from Lilium regale enhance resistance to several pathogenic fungi in transgenic tobacco plants[J]. Plant Physiology Journal, 2015, 51(12): 2223-2230. (in Chinese with English abstract)
[17] TSUDA K, YAMAZAKI K I.Structure and expression analysis of three subtypes of Arabidopsis MBF1 genes[J]. Biochimica et Biophysica Acta-Gene Structure and Expression, 2004, 1680(1): 1-10.
[18] SUGIKAWA Y, EBIHARA S, TSUDA K, et al.Transcriptional coactivator MBF1s from Arabidopsis predominantly localize in nucleolus[J]. Journal of Plant Research, 2005, 118(6): 431-437.
[19] 韩泽刚, 赵曾强, 何兰兰, 等. 枯萎病菌诱导感、抗陆地棉品种的转录因子表达变化[J]. 作物学报, 2015, 41(2): 228-239.
HAN Z G, ZHAO Z Q, HE L L, et al.Expression changes of transcription factors in susceptible and resistant upland cotton (Gossypium hirsutum L.) cultivars in response to Fusarium wilt[J]. Acta Agronomica Sinica, 2015, 41(2): 228-239. (in Chinese with English abstract)
[20] 唐平华, 陈国平, 潘宇番, 等. 番茄多蛋白桥梁因子基因LeMBF1的克隆及抗病性分析[J]. 生命科学研究, 2012, 16(2): 138-143.
TANG P H, CHEN G P, PAN Y F, et al.Cloning and analysis of pathogen resistance of multiprotein bridging factor gene LeMBF1 in tomato[J]. Life Science Research, 2012, 16(2): 138-143. (in Chinese with English abstract)
[21] 杨科府, 陈瑜, 王慧中, 等. RNA沉默机制及其介导的植物抗病毒基因工程研究进展[J]. 浙江农业学报, 2009, 21(6): 654-659.
YANG K F, CHEN Y, WANG H Z, et al.Review on RNA silencing and its mediated-genetic engineering of plant virus-resistance[J]. Acta Agriculturae Zhejiangensis, 2009, 21(6): 654-659. (in Chinese with English abstract)

灰葡萄孢菌诱导的百合转录辅激活因子LlMBF1a基因的克隆及其特征分析

Cloning and characterization of LlMBF1a in lily induced by gray mold pathogen

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	黄咏明, 宋放, 王策, 姚京磊, 王志静, 何利刚, 吴黎明, 蒋迎春. 根系修剪对枳生长及相关基因表达的影响[J]. 浙江农业学报, 2021, 33(2): 270-277.
[2]	何佳琦, 翟莹, 张军, 邱爽, 李铭杨, 赵艳, 张梅娟, 马天意. 大豆转录因子GmDof1.5的克隆与非生物胁迫诱导表达[J]. 浙江农业学报, 2021, 33(1): 1-7.
[3]	苏学思, 张玉宝, 王若愚, 王亚军, 唐国亮, 金卫杰. 车前草花叶病毒衣壳蛋白的原核表达和多克隆抗体制备[J]. 浙江农业学报, 2021, 33(1): 104-111.
[4]	徐秀红, 刘金亮, 李栋成, 刘仁祥. 不同类型烟草种质的烟碱含量变化与相关基因表达水平[J]. 浙江农业学报, 2020, 32(9): 1555-1563.
[5]	刘坤举, 张小辉, 庞有志, 赵淑娟, 祁艳霞, 王乾昆. 朝鲜鹌鹑GNAS基因表达、克隆及其多态性与羽色的相关性[J]. 浙江农业学报, 2020, 32(8): 1369-1377.
[6]	郭丽娜, 赵慧婷, 任有蛇, 徐兵, 姜玉锁. 中华蜜蜂AcerOr2基因昆虫表达载体pIB/V5-His的构建[J]. 浙江农业学报, 2020, 32(6): 994-999.
[7]	柯义强, 郭鹏辉, 马洪鑫, 杨许花, 高丹丹, 刘湘君, 马忠仁, 丁功涛. 兰州百合组培快繁体系的构建[J]. 浙江农业学报, 2020, 32(6): 1000-1008.
[8]	李秋玲, 齐颖, 王琛, 张一名, 王新妤, 尚校兰, 贾永红, 李美茹, 储明星. 热应激对中国荷斯坦牛乳腺组织基因表达及信号通路的影响[J]. 浙江农业学报, 2020, 32(5): 770-778.
[9]	卢祎, 高有领, 王水涛, 何盛盛. MicroRNA-499对中华鳖脂类代谢相关基因表达的影响[J]. 浙江农业学报, 2020, 32(5): 798-803.
[10]	李维芳, 王春蕾, 王妮, 邓雨正, 姚彦东, 魏丽娟, 廖伟彪. 一氧化氮影响植物不定根发生的研究进展[J]. 浙江农业学报, 2020, 32(4): 742-752.
[11]	秦玲, 张鑫, 荣春笑, 莫传园, 范露, 闫婕, 孟莹, 张满让. 苹果多胺氧化酶(PAO)基因家族鉴定与表达分析[J]. 浙江农业学报, 2020, 32(2): 262-273.
[12]	张政, 王晓荣, 钱虹, 张兰, 颜鹏, 张丽平, 张新富, 李鑫, 韩文炎. 炭疽病对茶树叶片光合特性的影响[J]. 浙江农业学报, 2020, 32(11): 2020-2026.
[13]	王伟科, 宋吉玲, 陆娜, 袁卫东, 闫静, 陈观平. 秀珍菇原基形成相关基因PpFBD1的克隆与表达研究[J]. 浙江农业学报, 2020, 32(1): 93-97.
[14]	张爱菊, 刘士力, 刘金殿, 张根芳, 周志明. 一种三角帆蚌肌浆网Ca²⁺-ATP酶基因cDNA的全长克隆与表达分析[J]. 浙江农业学报, 2019, 31(4): 545-555.
[15]	陈洪强, 夏惠, 王进, 邓群仙, 梁东, 吕秀兰, 唐礼平. 葡萄STS基因家族的鉴定与表达分析[J]. 浙江农业学报, 2019, 31(3): 401-407.