细叶百合LpPEX7基因克隆及盐胁迫下的表达特性分析

doi:10.7525/j.issn.1673-5102.2020.02.015

植物研究 ›› 2020, Vol. 40 ›› Issue (2): 274-283.doi: 10.7525/j.issn.1673-5102.2020.02.015

细叶百合LpPEX7基因克隆及盐胁迫下的表达特性分析

何好, 朱国庆, 陈诗雅, 徐畅, 金淑梅

东北盐碱植被恢复与重建教育部重点实验室, 生命科学学院, 东北林业大学, 哈尔滨 150040

收稿日期:2019-10-10 出版日期:2020-03-05 发布日期:2020-03-06
通讯作者: 金淑梅,E-mail:jinshumei1972@163.com E-mail:jinshumei1972@163.com
作者简介:何好(1995-),男,硕士研究生,主要从事分子植物育种研究。
基金资助:
黑龙江省自然科学基金联合引导项目（LH2019C011）；东北油田盐碱植被恢复与重建教育部重点实验室开放基金（SAVER1701）；国家自然科学基金面上项目（31070616，31500317）

Cloning of LpPEX7 Gene from Lilium pumilum and Its Expression Characteristics under Salt Stress

HE Hao, ZHU Guo-Qing, CHEN Shi-Ya, XU Chang, JIN Shu-Mei

Key Laboratory of Saline-alkali Vegetation Ecology Restoration, Ministry of Education, College of Life Sciences, Northeast Forestry University, Harbin 150040

Received:2019-10-10 Online:2020-03-05 Published:2020-03-06
Supported by:
Nature Scientific Foundation of Heilongjiang Province project(LH2019C011);Open Fund Key of Laboratory of Saline-alkali Vegetation Ecology Restoration in Oil Field(SAVER1701);National Natural Science Foundation of China(31070616,31500317)

摘要/Abstract

摘要： 从细叶百合的鳞茎中克隆出过氧化物酶体生物合成蛋白基因（LpPEX7），该基因ORF全长957 bp，编码318个氨基酸。LpPEX7蛋白序列包含6个WD40保守结构域，通过同源蛋白序列比对和进化树分析，发现LpPEX7与其他植物的PEX7蛋白具有较高的同源性。LpPEX7基因在细叶百合种子，叶片和鳞茎中的表达量比较高，在根和花中表达量比较低，在H₂O₂，NaCl，NaHCO₃不同逆境处理条件下，LpPEX7基因的表达量都发生了改变。在盐碱和氧化胁迫处理下，LpPEX7过表达拟南芥株系种子的萌发要早于野生型种子的萌发，这些研究结果表明LpPEX7基因与盐碱、氧化逆境有一定的应答关系，为细叶百合的耐盐碱性分子机理研究提供一个非常重要的候选基因。

关键词: 细叶百合, 过氧化物酶体生物合成蛋白, 基因表达, 拟南芥

Abstract: The peroxidasome biosynthetic gene(LpPEX7) of Lilium pumilum DC was cloned, and its open reading frame length is 957 bp, encoding 318 amino acids. The deduced amino acid sequence of LpPEX7 contains six WD40 conserved domains, through sequence alignment analysis of homologous proteins and evolutionary tree analysis, it was found that LpPEX7 had higher homology with some PEX7 from other plants. The expression of LpPEX7 was higher in seeds, leaves and bulbs, but lower in roots and flowers of L.pumilum. The expression of LpPEX7 changed under H₂O₂, NaCl and NaHCO₃ stress conditions. Subsequently, the seeds germination of LpPEX7 over-expression Arabidopsis thaliana lines as earlier than that of wild seeds under the treatment of salinity and oxidative stress. These results show that the LpPEX7 gene has a certain response relationship with salinity and oxidative adversity. It provides a very important candidate gene for the study of salt-tolerant alkaline molecular mechanism of Lily.

Key words: Lilium pumilum DC, peroxidasome biosynthetic gene, gene expression, Arabidopsis thaliana

中图分类号:

Q949.71+8.23

何好, 朱国庆, 陈诗雅, 徐畅, 金淑梅. 细叶百合LpPEX7基因克隆及盐胁迫下的表达特性分析[J]. 植物研究, 2020, 40(2): 274-283.

HE Hao, ZHU Guo-Qing, CHEN Shi-Ya, XU Chang, JIN Shu-Mei. Cloning of LpPEX7 Gene from Lilium pumilum and Its Expression Characteristics under Salt Stress[J]. Bulletin of Botanical Research, 2020, 40(2): 274-283.

参考文献 28

[1]	史海水,王少卿,廖祥儒.植物体中的过氧化物酶体[J].生物学通报,2005,40(2):18-19.Shi H S,Wang S Q,Liao X R.Peroxisomes in plants[J].Bulletin of Biology,2005,40(2):18-19.
[2]	崔慧萍,周薇,郭长虹.植物过氧化物酶体在活性氧信号网络中的作用[J].中国生物化学与分子生物学报,2017,33(3):220-226.Cui H P,Zhou W,Guo C H.The role of plant peroxisomes in ROS signalling network[J].Chinese Journal of Biochemistry and Molecular Biology,2017,33(3):220-226.
[3]	Hu J P,Aguirre M,Peto C,et al.A role for peroxisomes in photomorphogenesis and development of Arabidopsis[J].Science,2002,297(5580):405-409.
[4]	Islinger M,Grille S,Fahimi H D,et al.The peroxisome:an update on mysteries[J].Histochemistry and Cell Biology,2012,137(5):547-574.
[5]	姚琨,练从龙,王菁菁,等.胡杨PePEX11基因参与调节盐胁迫下拟南芥的抗氧化能力[J].北京林业大学学报,2018,40(5):19-28.Yao K,Lian C L,Wang J J,et al.PePEX11 functions in regulating antioxidant capacity of Arabidopsis thaliana under salt stress[J].Journal of Beijing Forestry University,2018,40(5):19-28.
[6]	赵亚虹.蜡梅过氧化物酶体生成蛋白基因CpPEX22的克隆及功能初步分析[D].重庆:西南大学,2013.Zhao Y H.Cloning and functional analysis of the peroxisome generate protein gene CpPEX22 from Chimonanthus praecox[D].Chongqing:Southwest University,2013.
[7]	孙艳,孙雪培,姜玲玲,等.过氧化物酶体生物发生研究进展[J].生物学杂志,2015,32(2):83-86.Sun Y,Sun X P,Jiang L L,et al.Research progress in peroxisome biogenesis[J].Journal of Biology,2015,32(2):83-86.
[8]	Kiel J A K W,Van Der Berg M,Bovenberg R A L,et al.Penicillium chrysogenum Pex5p mediates differential sorting of PTS1 proteins to microbodies of the methylotrophic yeast Hansenula polymorpha[J].Fungal Genetics and Biology,2004,41(7):708-720.
[9]	Purdue P E,Yang X D,Lazarow P B.Pex18p and Pex21p,a novel pair of related peroxins essential for peroxisomal targeting by the PTS2 pathway[J].The Journal of Cell Biology,1998,143(7):1859-1869.
[10]	An C J,Gao Y F,Li J Y,et al.Alternative splicing affects the targeting sequence of peroxisome proteins in Arabidopsis[J].Plant Cell Reports,2017,36(7):1027-1036.
[11]	Corpas F J,Barroso J B.Peroxisomal plant nitric oxide synthase(NOS) protein is imported by peroxisomal targeting signal type 2(PTS2) in a process that depends on the cytosolic receptor PEX7 and calmodulin[J].FEBS Letters,2016,588(12):2049-2054.
[12]	Motley A M,Hettema E H,Ketting R,et al.Caenorhabditis elegans has a single pathway to target matrix proteins to peroxisomes[J].EMBO Reports,2000,1(1):40-46.
[13]	Singh T,Hayashi M,Mano S,et al.Molecular components required for the targeting of PEX7 to peroxisomes in Arabidopsis thaliana[J].The Plant Journal,2010,60(3):488-498.
[14]	Pilar A V C,Strasser R,Mclean J,et al.Analysis of the Leishmania peroxin 7 interactions with peroxin 5,peroxin 14 and PTS2 ligands[J].Biochemical Journal,2014,460(2):273-282.
[15]	Ramón N M,Bartel B.Interdependence of the peroxisome-targeting receptors in Arabidopsis thaliana:PEX7 facilitates PEX5 accumulation and import of PTS1 cargo into peroxisomes[J].Molecular Biology of the Cell,2010,21(7):1263-1271.
[16]	Cross L L,Ebeed H T,Baker A.Peroxisome biogenesis,protein targeting mechanisms and PEX gene functions in plants[J].Biochimica et Biophysica Acta(BBA)-Molecular Cell Research,2016,1863(5):850-862.
[17]	Orth T,Reumann S,Zhang X C,et al.The Peroxin11 protein family controls peroxisome proliferation in Arabidopsis[J].The Plant Cell,2007,19(1):333-350.
[18]	Rodríguez-Serrano M,Romero-Puertas M C,Sanz-Fernández M,et al.Peroxisomes extend peroxules in a fast response to stress via a reactive oxygen species-mediated induction of the peroxin PEX11a[J].Plant Physiology,2016,171(3):1665-1674,doi:10.1104/pp.16.00648.
[19]	杜运鹏,李双,何恒斌,等.百合鳞片总RNA提取方法的比较[J].分子植物育种,2010,8(4):832-836.Du Y P,Li S,He H B,et al.Comparative of methods for RNA extraction from lily bulb scales[J].Molecular Plant Breeding,2010,8(4):832-836.
[20]	Zhang X R,Henriques R,Lin S S,et al.Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method[J].Nature Protocols,2006,1(2):641-646,doi:10.1038/nprot.2006.97.
[21]	Cui S K,Mano S,Yamada K,et al.Novel proteins interacting with peroxisomal protein receptor PEX7 in Arabidopsis thaliana[J].Plant Signaling & Behavior,2013,8(10):e26829,doi:10.4161/psb.26829.
[22]	Stirnimann C U,Petsalaki E,Russell R B,et al.WD40 proteins propel cellular networks[J].Trends in Biochemical Sciences,2010,35(10):565-574.
[23]	Hou Y F,Huang J,Yu S L,et al.The 6-phosphogluconate dehydrogenase genes are responsive to abiotic stresses in rice[J].Journal of Integrative Plant Biology,2007,49(5):655-663.
[24]	冷春玲,宋洁.吡咯喹啉醌生物学功能研究进展[J].辽东学院学报:自然科学版,2014,21(2):103-108.Leng C L,Song J.Biological function of pyrroloquinoline quinine:research progress[J].Journal of Liaodong University:Natural Sciences,2014,21(2):103-108.
[25]	Fang Y J,Wu H,Zhang T W,et al.A complete sequence and transcriptomic analyses of date palm(Phoenix dactylifera L.) mitochondrial genome[J].PLoS One,2012,7(5):e37164.
[26]	Zhang G Q,Xu Q,Bian C,et al.The Dendrobium catenatum Lindl.genome sequence provides insights into polysaccharide synthase,floral development and adaptive evolution[J].Scientific Reports,2016,6:19029.
[27]	Al-Shanfari A B,Abdullah S N A,Saud H M,et al.Differential gene expression identified by suppression subtractive hybridization during late ripening of fruit in oil palm(Elaeis guineensis Jacq.)[J].Plant Molecular Biology Reporter,2012,30(3):768-779.
[28]	Yang K,Tian Z X,Chen C H,et al.Genome sequencing of adzuki bean(Vigna angularis) provides insight into high starch and low fat accumulation and domestication[J].Proceedings of the National Academy of Sciences of the United States of America,2015,112(43):13213-13218.

细叶百合LpPEX7基因克隆及盐胁迫下的表达特性分析

Cloning of LpPEX7 Gene from Lilium pumilum and Its Expression Characteristics under Salt Stress

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献 28

相关文章 15

编辑推荐

Metrics

本文评价

[1]	江转转, 龚莉, 宋亚玲. 拟南芥叶绿体分裂蛋白PARC6影响子叶与真叶的生长[J]. 植物研究, 2023, 43(5): 700-710.
[2]	宋煜, 林文昊, 荆一博, 董懿, 金淑梅. 细叶百合Catalase基因的克隆及表达分析[J]. 植物研究, 2023, 43(5): 756-767.
[3]	郑晟, 高海霞, 苏敏, 卢尚欢, 张腾国, 武国凡. 外源蔗糖影响AtKEA1和AtKEA2调节拟南芥幼苗根的生长[J]. 植物研究, 2023, 43(4): 562-571.
[4]	徐磊, 胥晓, 刘沁松. 外源水杨酸对盐胁迫下珙桐幼苗抗氧化系统和基因表达的影响[J]. 植物研究, 2023, 43(4): 572-581.
[5]	裘喻平, 王益川, 郭红卫. 植物根毛发育调控机制的研究进展[J]. 植物研究, 2023, 43(3): 321-332.
[6]	蔡圆圆, 夏季奔奔, 应文涵, 王洁瑶, 谢涛, 邢孔丫, 冯宣军, 华学军. 拟南芥线粒体蛋白突变体ssr1-2表型的详细鉴定与分析[J]. 植物研究, 2023, 43(3): 421-431.
[7]	杨洪, 王立丰, 代龙军, 郭冰冰. 死皮对橡胶树树皮线粒体超微结构及活性氧代谢的影响[J]. 植物研究, 2023, 43(1): 69-75.
[8]	李梦烁, 刘莹泽, 鲁焕, 强胜. 基因转录介导同质园条件下拟南芥不同地理种群的光合能力分化[J]. 植物研究, 2023, 43(1): 90-99.
[9]	覃碧, 刘明洋, 王萌, 王立丰, 黄飞. 橡胶树DELLA基因HbRGL1的克隆与表达分析[J]. 植物研究, 2022, 42(6): 997-1004.
[10]	代龙军, 刘明洋, 阳江华, 周凯, 郭冰冰, 杨洪, 王立丰. 巴西橡胶树胶乳中DUF1262结构域蛋白结构与基因表达分析[J]. 植物研究, 2022, 42(5): 802-810.
[11]	陈思思, 谢牧洪, 崔茂凯, 李文凯, 徐正刚, 贾彩霞, 杨桂燕. 构树转录因子BpbZIP1的鉴定及镉胁迫响应分析[J]. 植物研究, 2022, 42(3): 394-402.
[12]	王梦姣, 曹钰雪, 徐永盛, 丁风鹅, 苏乔. 过表达海洋微生物宏基因组MbCSP提高转基因拟南芥的抗旱和耐寒性[J]. 植物研究, 2022, 42(2): 243-251.
[13]	马凯恒, 何佳凝, 谢牧洪, 任斌, 黄倩雪, 杨桂燕. 核桃JrNPR1基因的克隆与抗病响应潜力[J]. 植物研究, 2022, 42(1): 39-46.
[14]	陈华峰, 樊松乐, 王立丰. 巴西橡胶树赤霉素信号转导途径HbRGA1结构和功能分析[J]. 植物研究, 2021, 41(4): 531-539.
[15]	武国凡, 成宏斌, 吴玉俊, 沈娟, 吴旺泽. CRISPR/Cas9介导靶向敲除拟南芥BRI1突变体的鉴定[J]. 植物研究, 2021, 41(3): 362-371.