ZHAO Fengzhi,WANG Xueying,WANG Xinlei,et al.Cloning and Expression Analysis of BoPGIP2 Gene From Brassica oleracea[J].Northern Horticulture,2016,40(07):77-84.[doi:10.11937/bfyy.201607021]
花椰菜BoPGIP2基因的克隆与表达水平分析
- Title:
- Cloning and Expression Analysis of BoPGIP2 Gene From Brassica oleracea
- 文献标志码:
- A
- 摘要:
- 以3个品系花椰菜为试材,采用同源克隆的方法,克隆了花椰菜BoPGIP2基因,并对表达水平进行分析。结果表明:该基因与油菜的PGIP2基因序列有较高的相似性;ExPASy分析发现BoPGIP2理论分子量为37.02 kDa,等电点为8.51,分子式为C1672H2604N436O487S13,不稳定系数为34.95,为稳定蛋白,脂肪族指数为92.48,GRAVY值为-0.096,表明其在一级结构上的亲水性、疏水性差异不显著。亚细胞定位预测分析表明,此蛋白属于分泌途径信号肽蛋白;qRT-PCR分析发现BoPGIP2基因在花椰菜高抗菌核病品系、中抗菌核病品系中均是茎和叶中相对表达量较高,而在易感菌核病品系中则是根中相对表达量最高,叶和茎中相对表达量较低,这与抗菌核病强弱具有一致性,推测此基因很可能在调节花椰菜的抗菌核病中起着重要作用。
- Abstract:
- aking 3 strains Brassica napus as materials,using homologous cloning method,the PGIP2 gene of Brassica oleracea was cloned and the expression level was analysed.The results showed that BoPGIP2 had high similarity with PGIP2 gene of Brassica napus.The ExPASy analysis showed that molecular weight of BoPGIP2 was 37.02 kDa,theoretical pI was 8.51.The instability index (II) was 34.95,and this showed the protein was stable.Grand average of hydropathicity was -0.096,so it was not a hydrophilic protein.The subcellular localization analysis indicated that BoPGIP2 was a signal peptide of secretory pathway.qRT-PCR found that BoPGIP2 has high expression in stems and leaves in both high resistant varieties and resistant varieties,whereas BoPGIP2 expressed highly in root in susceptible varieties,it was consistent with their resistance to Sclerotinia sclerotiorum.Therefore,it might play an important role in the regulation of resistance to Sclerotinia sclerotiorum of Brassica oleracea.
参考文献/References:
[1]李素文,赵前程,孙德岭,等.国内外花椰菜种植面积及产量变化趋势[J].中国蔬菜,2005(3):36-37. [2]张凯,徐艳辉.花椰菜的分子遗传与育种研究进展概述[J].辽宁农业科学,2003(1):31-34. [3]王燕,朱隆静,柳李旺,等.花椰菜生物技术育种研究进展[J].分子植物育种,2008(3):549-554. [4]BOLAND G,HALL R.Index of plant hosts of Sclerotinia sclerotiorum[J].Can J Plant Pathol,1994(16):93-108. [5]PURDY L.Sclerotinia sclerotiorum:history diseases and symptomatology,host range,geographic distribution and impac[J].Phytopathology,1979,69:875-890. [6]BOLTON M D T B.Sclerotinia sclerotiorum (Lib.) de Bary:biology and molecular traits of a cosmopolitan pathogen[J].Molecular Plant Pathology,2006(1):1-16. [7]JAMAUX J,GELIE B,LAMARQUE C.Early stages of infection of rapeseed petals and leaves by Sclerotinia sclerotiorum revealed by scanning electron microscopy[J].Plant Pathol,1995,44:22-30. [8]庞超,赵卫,谢永江.油菜菌核病的危害症状及防治对策[J].现代农业科技,2015(9):137-142. [9]杨子祥,苏银玲,沙毓沧.花椰菜菌核病的发生与防治技术[J].长江蔬菜,2012(21):46-47. [10]王忠华,贾育林,夏英武.植物抗病分子机制研究进展[J].植物学通报,2004(5):521-530. [11]HAMMOND-KOSACK K E,JONES J D J.Plant disease resistance genes[J].Annu Rev Plant Physiol Plant Mol Biol,1997,48(4):575-607. [12]STASKAWICZ B J,AUSUBEL F M,BAKER B J,et al.Molecular genetics of plant disease resistance[J].Science,1995,268(5211):661-667. [13]李文娴.PGIP的基因克隆及抗真菌活性研究[D].昆明:昆明理工大学,2010. [14]HUANGFU H Y,GUAN C Y,JIN F R,et al.Prokaryotic expression and protein function of Brassica napus PGIP2 and its genetic transformation[J].Plant Biotechnology Reports,2014,8(2):171-181. [15]桂枝,高建明,袁庆华.多聚半乳糖醛酸酶抑制蛋白(PGIP)的研究进展概述[J].天津农学院学报,2011(3):36-41. [16]罗娅,汤浩茹,张勇,等.草莓多聚半乳糖醛酸酶抑制蛋白基因(PGIP)的克隆及组织特异性表达[J].农业生物技术学报,2010(3):424-430. [17]陈夕军,张磊,陈羽,等.油菜Bnpgip2-1基因的克隆表达与生物信息学分析[J].中国油料作物学报,2014(6):701-706. [18]陈夕军,刘晓维,左示敏,等.水稻多聚半乳糖醛酸酶抑制蛋白基因(Ospgip1)原核表达及编码产物生物信息学分析[J].中国水稻科学,2011(2):136-142. [19]符聪慧,王建平,张冲,等.苹果PGIP基因部分家族成员启动子的克隆与功能分析[J].园艺学报,2014,41(11):2169-2178. [20]王秀菊.辣椒多聚半乳糖醛酸酶抑制蛋白基因克隆及功能研究[D].泰安:山东农业大学,2011. [21]刘颖,吴剑锋,卢海宇,等.普通白菜PGIP基因BcPGIP分子特征研究[J].核农学报,2010(2):281-286. [22]李广平,乔玉山,陶建敏,等.中国李PGIP基因的克隆及序列分析[J].西北植物学报,2006(9):1870-1873. [23]李宽.花椰菜菌核病抗性评价体系的建立及抗性相关基因的表达研究[D].天津:南开大学,2014. [24]HENRIK N,JACOB E,SREN B,et al.Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites[J].Protein Engineering,1997(10):1-6. [25]OLOF E,HENRIK N,SREN B,et al.Predicting subcellular localization of proteins based on their N-terminal amino acid sequence[J].J Mol Biol,2000,300:1005-1016. [26]MARCO B,STEFAN B,ANDREW W,et al.SWISS-MODEL:modelling protein tertiary and quaternary structure using evolutionary information[J].Nucleic Acids Research,2014,42:252-258. [27]ARNOLD K,BORDOLI L,KOPP J,et al.The SWISS-MODEL workspace:a web-based environment for protein structure homology modelling[J].Bioinformatics,2006(22):195-201. [28]BENKERT P,BIASINI M,SCHWEDE T.Toward the estimation of the absolute quality of individual protein structure models[J].Bioinformatics,2011(27):343-350. [29]MILAGROS F M,FLORENCIA P O,GUSTAVO R D,et al.Isolation and characterization of a polygalacturonase-inhibiting protein from potato leaves:Accumulation in response tosalicylic acid,wounding and infection[J].Plant Physiology and Biochemistry,2001(2):129-136.
相似文献/References:
[1]黄俊轩,刘艳军,李建科,等.以花椰菜花球为外植体的离体再生体系的建立[J].北方园艺,2014,38(05):88.
HUANG Jun-xuan,LIU Yan-jun,LI Jian-ke,et al.Regeneration of Cauliflower by in vitro Flower Bulb[J].Northern Horticulture,2014,38(07):88.
[2]李天星.乙酰甲胺磷对花椰菜种子萌发及其幼苗生长的影响[J].北方园艺,2013,37(08):127.
LI Tian-xing.Effects of Different Concentrations of Acephate Stress on Seed Germination and Growth of Brassica oleracea var. botrytis Seedlings [J].Northern Horticulture,2013,37(07):127.
[3]李天星.氯化钠对乙酰甲胺磷胁迫下花椰菜种子萌发及幼苗生长的影响[J].北方园艺,2013,37(19):45.
LI Tian-xing.Effects of NaCl on Seed Germination and Growth of Brassica oleracea var. botrytis L.Seedlings Under Acephate Stress[J].Northern Horticulture,2013,37(07):45.
[4]郭晓芹,仪泽会,惠麦侠,等.大白菜与花椰菜种间杂交结实性调查[J].北方园艺,2012,36(17):8.
GUO Xiao-qin,YI Ze-hui,HUI Mai-xia,et al.Study of Seed Characteristics from InterspecificHybridization between Chinese Cabbage and Cauliflower[J].Northern Horticulture,2012,36(07):8.
[5]钟建明.高山条件下间作蚕豆对花椰菜产量、品质、生境的影响[J].北方园艺,2015,39(03):34.[doi:10.11937/bfyy.201503010]
ZHONG Jian-ming.Effect of Intercropping Broad Bean on the Yields,Quality and Habitat of Broccoli Under High Mountain Conditions[J].Northern Horticulture,2015,39(07):34.[doi:10.11937/bfyy.201503010]
[6]朱 惠 霞,胡 立 敏,陶 兴 林.两个花椰菜品种再生体系的研究[J].北方园艺,2010,34(09):0.[doi:10.11937/bfyy.201009053]
ZHU Hui-xia,HU Li -min,TAO Xing -lin.Study on Regeneration System of 2 Cauliflowers ( Brassica oleracea var. botrytis)[J].Northern Horticulture,2010,34(07):0.[doi:10.11937/bfyy.201009053]
[7]李 元 万,王 晓 巍,张 玉 鑫,等.农大哥和 AM 生物菌肥对花椰菜产量和效益的影响[J].北方园艺,2010,34(09):0.[doi:10.11937/bfyy.201009012]
LI Yuan -wan,W ANG Xiao -wei,ZHANG Yu -xin,et al.Effect of Nongdage and AM Bacterial Manure on Yield and Profit of Cauliflower[J].Northern Horticulture,2010,34(07):0.[doi:10.11937/bfyy.201009012]
[8]王帘里,翟国亮.地下滴灌增温对盆栽花椰菜生长、产量和营养品质的影响[J].北方园艺,2016,40(06):14.[doi:10.11937/bfyy.201606004]
WANG Lianli,ZHAI Guoliang.Effect of Temperature-increased Subsurface Drip Irrigation on Potted Cauliflower Growth,Yield and Nutrition Quality[J].Northern Horticulture,2016,40(07):14.[doi:10.11937/bfyy.201606004]
[9]任喜波,袁进成,戴希尧,等.花椰菜种质资源遗传完整性变化的生理生化特性研究[J].北方园艺,2016,40(10):19.[doi:10.11937/bfyy.201610005]
REN Xibo,YUAN Jincheng,DAI Xiyao,et al.Study on Physiological and Biochemical Characteristics and Genetic Integrity in Cauliflower[J].Northern Horticulture,2016,40(07):19.[doi:10.11937/bfyy.201610005]
[10]蔡 绵 聪,等.花 椰 菜 氮 磷 钾 施 肥 效 应 研 究[J].北方园艺,2009,33(01):0.[doi:10.11937/bfyy.200901001]
CAI Mian -cong,LI Shu-yi,CHEN Zhen -yuan,et al.The Effect of N, P and K Fertilizers on Yield and Q uality of Brassia oleracea var. botrytis[J].Northern Horticulture,2009,33(07):0.[doi:10.11937/bfyy.200901001]
备注/Memo
第一作者简介:赵风治(1990-),男,河北邯郸人,硕士研究生,研究方向为分子遗传学。E-mail:zhaofengzhi145@163.com.责任作者:陈成彬(1972-),男,河北冀州人,博士,副教授,研究方向为细胞分子遗传学。E-mail:chencb@nankai.edu.cn.基金项目:天津市自然科学基金重点基金资助项目(13JCZDJC29000);天津市应用基础与前沿技术研究计划一般资助项目(14JCYBJC31000)。