毛果杨nsLTP基因家族全基因组水平鉴定及其表达特性分析

程赫; 田双慧; 张洋; 刘聪; 夏德安; 魏志刚

毛果杨nsLTP基因家族全基因组水平鉴定及其表达特性分析

程赫¹,
田双慧¹,
张洋¹,
刘聪¹,
夏德安¹,
魏志刚^2, ,

1. 林木遗传育种国家重点实验室东北林业大学;
2. 国家林业和草原局盐碱地研究中心中国林业科学研究院

基金项目:

国家自然科学基金项目(31770640)

国家重点研发计划子课题项目(2017YFC0504102-04)~~

详细信息

作者简介:
程赫(1996—),女,硕士研究生,主要从事林木抗性育种方面的研究

通讯作者:
魏志刚 E-mail:zhigangwei1973@163.com

中图分类号: S792.11
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出版历程
- 收稿日期: 2021-03-27
- 刊出日期: 2022-05-19

Genome-wide Identification and Expression Analysis of nsLTP Gene Family in Populus trichocarpa

1. State Key Laboratory of Forest Tree Genetics and Breeding,Northeast Forestry University;
2. Saline Soil Research Center,National Forestry and Grassland Administration,Chinese Academy of Forestry

摘要

摘要: 植物非特异性脂质转移蛋白(non-specific lipid transfer proteins,nsLTP)是一类多基因家族编码碱性蛋白，负责脂肪酸体外结和与膜之间的磷脂转移，在植物生长发育和逆境胁迫响应中扮演着重要角色。目前为止，尚无模式植物毛果杨(Populus trichocarpa)nsLTP家族的研究报导。本研究从全基因组水平对PtrnsLTP家族成员的基因数量、亲缘关系、基因结构、编码蛋白保守基序等特性进行了分析，结果表明：PtrnsLTP家族共由39个基因组成，进化成5个亚家族，其中A亚族含有6个基因、B亚族含有2个、C亚族含有13个、D亚族含有3个、E亚族含有15个。PtrnsLTP家族包含7对旁系同源基因，其中1对大于1,6对Ka/Ks均远小于1，且这6对基因均处于同一个大的进化分支上，进化压力的不同导致基因间的功能出现了分化，编码蛋白均含有Motif 1和Motif 2保守基序。利用qRT-PCR技术并结合杨树转录组数据对PtrnsLTP的组织表达与盐胁迫响应特性研究发现：各家族成员在毛果杨根、茎和叶中均有表达且经qRT-PCR技术验证后与网站预测结果基本吻合，有11、15和13个成员分别在根、茎和叶中有较高的表达，表明该基因家族参与了杨树不同组织的生长发育；NaCl胁迫下，该家族39个基因中分别有26个成员在根部、14个成员在叶部表达量随着胁迫时间的增加而升高，而32个基因在茎部表现为先升高后降低的趋势。本研究结果对于PtrnsLTP家族基因生物学功能的鉴定与盐胁迫响应基因资源的工作有着积极的推动作用。
- 毛果杨 /
- nsLTP /
- 生物信息学分析 /
- 盐胁迫
Abstract: Non-specific lipid transfer proteins（nsLTP）,a polygene family of basic lipid transfer proteins,are responsible for the transfer of phospholipids between fatty acids and membrane in vitro,and play an important role in plant growth and development and the response to stress.There have been no reports on the nsLTP family of Populus trichocarpa.In this study,the number of genes,genetic relationship,gene structure and conserved motif of coding proteins of PtrnSLTP family members were analyzed at the genome-wide level.The results showed that the PtrnsLTP family was composed of 39 genes and evolved into five subfamilies,among which six genes in subfamily A,two genes in subfamily B,13 genes in subfamily C,three genes in subfamily D,and 15genes in subfamily E.PtrnsLTP family contained seven pairs of paragenetic homologous genes,among which one pair was greater than one,and the six pairs of Ka/Ks were all far less than one.Moreover,these six pairs of genes were all on the same large evolutionary branch,and different evolutionary pressures led to functional differentiation among genes.Both encoded proteins contained conserved motifs in Motif 1 and Motif 2.Using qRT-PCR technology combined with P.trichocarpa data to study the tissue expression of PtrnsLTP and the response characteristics of salt stress,it was found that all family members expressed in the roots,stems and leaves of P.trichocarpa.The results were basically consistent with the prediction results by the bioinformatics analysis of the website.There were 11,15 and 13 members respectively having higher expression in roots,stems and leaves,indicated that this gene family was involved in the growth and development of different tissues of P.trichocarpa.Under NaCl stress,among the 39 genes in the family,26 genes expressed in the root and 14genes expressed in the leaves increased with the increase of stress time,while 32 genes expressed in the stem first increased and then decreased.The results have a positive role in promoting the identification of the biological function of PtrnsLTP family genes and the work of salt stress response gene resources.
- Populus trichocarpa /
- nsLTP /
- bioinformatics analysis /
- salt stress

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参考文献(38)

[1]	DENG W J,LI R Q,XU Y W,et al.A lipid transfer protein variant with a mutant eight-cysteine motif causes photoperiod-and-thermo-sensitive dwarfism in rice[J].Journal of Experimental Botany,2019,71(4):1294-1305.
[2]	JOSÉ-ESTANYOL M,GOMIS-RÜTH F X,PUIG-DOMÈNECH R.The eight-cysteine motif,a versatile structure in plant proteins[J].Plant Physiology and Biochemistry,2004,42(5):355-365.
[3]	LERCHE M H,KRAGELUND B B,BECH L M,et al.Barley lipid-transfer protein complexed with palmitoyl Co A:the structure reveals a hydrophobic binding site that can expand to fit both large and small lipid-like ligands[J].Structure,1997,5(2):291-306.
[4]	KADER J C.Proteins and the intracellular exchange of lipids:I.stimulation of phospholipid exchange between mitochondria and microsomal fractions by proteins isolated from potato tuber[J].Biochimica et Biophysica Acta(BBA)-Lipids and Lipid Metabolism,1975,380(1):31-44.
[5]	王洪.油菜非特异性脂转运蛋白基因家族的全基因组水平鉴定和表达分析[D].杨凌:西北农林科技大学,2015.WANG H.Genome-wide identification and expression analysis of brnsltps gene family in brassica rapa[D].Yangling:Northwest Agriculture and Forestry University,2015.
[6]	孟令波,刘关君,李淑敏,等.黄瓜非特异性脂质转移蛋白基因的序列分析及细菌性角斑病菌浸染下的表达[J].东北农业大学学报,2013,44(1):55-60. MENG L B,LIU G J,LI S M,et al.Sequence analyses of non-specific transfer protein from cucumber and expression after challenging by Pseudomonas syringae pv.Lachry-mans[J].Journal of Northeast Agricultural University,2013,44(1):55-60.
[7]	KINLAW C S,GERTTULA S M,CARTER M C.Lipid transfer protein genes of loblolly pine are members of a complex gene family[J].Plant Molecular Biology,1994,26(4):1213-1216.
[8]	BLUNDELL T L,SIBANDA B L,STERNBERG M J E,et al.Knowledge-based prediction of protein structures and the design of novel molecules[J].Nature,1987,326(6111):347-352.
[9]	DOULIEZ J P,MICHON T,ELMORJANI K,et al.Mini Review:structure,biological and technological functions of lipid transfer proteins and indolines,the major lipid binding proteins from cereal kernels[J].Journal of Cereal Science,2000,32(1):1-20.
[10]	BOUTROT F,CHANTRET N,GAUTIER M F.Genomewide analysis of the rice and arabidopsis non-specific lipid transfer protein(ns Ltp)gene families and identification of wheat ns Ltp genes by EST data mining[J].BMCGenomics,2008,9(1):86.
[11]	SAMUEL D,LIU Y J,CHENG C S,et al.Solution structure of plant nonspecific lipid transfer protein-2 from rice(Oryza sativa)[J].Journal of Biological Chemistry,2002,277(38):35267-35273.
[12]	KADER J C.Lipid-transfer proteins in plants[J].Annual Review of Plant Physiology and Plant Molecular Biology,1996,47:627-654.
[13]	ZHANG D S,LIANG W Q,YIN C S,et al.Os C6,encoding a lipid transfer protein,is required for postmeiotic anther development in rice[J].Plant Physiology,2010,154(1):149-162.
[14]	XIE W Q,ZHAO L Q,BAI W Y,et al.The effects of calmodulin on the lipid-binding activity of Ca M-binding protein-10 and maize non-specific lipid transfer protein[J].Journal of Plant Physiology and Molecular Biology,2006,32(6):679-684.
[15]	MOLINA A,SEGURA A,GARCÍA-OLMEDO F.Lipid transfer proteins(ns LTPs)from barley and maize leaves are potent inhibitors of bacterial and fungal plant pathogens[J].FEBS Letters,1993,316(2):119-122.
[16]	GARCÍA-OLMEDO F,MOLINA A,SEGURA A,et al.The defensive role of nonspecific lipid-transfer proteins in plants[J].Trends in Microbiology,1995,3(2):72-74.
[17]	强晓晶.小盐芥Th PIP1基因的水稻遗传转化及耐盐机理研究[D].北京:中国农业科学院,2015. QIANG X J.Thellungiella halophila Th PIPl gene transferring rice and mechanism of salt stress tolerance[D].Beijing:Chinese Academy of Agricultural Sciences,2015.
[18]	ANDREA P,SNEHA D,HELENE P.Salt stress in Arabidopsis:lipid transfer protein AZI1 and its control by mitogen-activated protein kinase MPK3[J].Molecular Plant,2014,7(4):722-738.
[19]	MCLAUGHLIN J E,BIN-UMERM A,WIDIEZ T,et al.A lipid transfer protein increases the glutathione content and enhances Arabidopsis resistance to a trichothecene mycotoxin[J].PLo S One,2015,10(6):e0130204.
[20]	JUNG H W,KIM K D,HWANG B K.Identification of pathogen-responsive regions in the promoter of a pepper lipid transfer protein gene(CALTPI)and the enhanced resistance of the CALTPI transgenic Arabidopsis against pathogen and environmental stresses[J].Planta,2005,221(3):361-373.
[21]	郜刚,任彩虹,金黎平,等.马铃薯非特异性脂质转移蛋白基因St LTPa1的克隆和表达[J].作物学报,2008,34(9):1510-1517. GAO G,REN C H,JIN L P,et al.Cloning,expression and characterization of a non-specific lipid transfer protein gene from potato[J].Acta Agronomica Sinica,2008,34(9):1510-1517.
[22]	FANG Z W,HE Y Q,LIU Y K,et al.Bioinformatic identification and analyses of the non-specific lipid transfer proteins in wheat[J].Journal of Integrative Agriculture,2020,19(5):1170-1185.
[23]	TAMURA K,STECHER G,PETERSON D,et al.MEGA6:molecular evolutionary genetics analysis version 6.0[J].Molecular Biology and Evolution,2013,30(12):2725-2729.
[24]	LETUNIC I,BORK P.Interactive Tree of Life(i TOL)v4:recent updates and new developments[J].Nucleic Acids Research,2019,47(W1):W256-W259.
[25]	ZHANG Z,LI J,ZHAO Z Q,et al.Ka Ks_Calculator:calculating Ka and Ks through model selection and model averaging[J].Genomics,Proteomics&Bioinformatics,2006,4(4):259-263.
[26]	LI J,GAO G Z,XU K,et al.Genome-wide survey and expression analysis of the putative non-specific lipid transfer proteins in Brassica rapa L.[J].PLos One,2014,9(1):e84556.
[27]	BLANC G,WOLFE K H.Widespread paleopolyploidy in model plant species inferred from age distributions of duplicate genes[J].The Plant Cell,2004,16(7):1667-1678.
[28]	LIU W F,HUANG D W,LIU K,et al.Discovery,identification and comparative analysis of non-specific lipid transfer protein(ns Ltp)family in solanaceae[J].Genomics,Proteomics&Bioinformatics,2010,8(4):229-237.
[29]	TSUBOI S,OSAFUNE T,TSUGEKI R,et al.Nonspecific lipid transfer protein in castor bean cotyledon cells:subcellular localization and a possible role in lipid metabolism[J].Journal of Biochemistry,1992,111(4):500-508.
[30]	ARONDEL V,VERGNOLLE C,CANTREL C,et al.Lipid transfer proteins are encoded by a small multigene family in Arabidopsis thaliana[J].Plant Science,2000,157(1):1-12.
[31]	HINCHA D K,NEUKAMM B,SROR H A M,et al.Cabbage cryoprotectin is a member of the nonspecific plant lipid transfer protein gene family[J].Plant Physiology,2001,125(2):835-846.
[32]	TREVIN-O M B,O’CONNELL M A.Three drought-responsive members of the nonspecific lipid-transfer protein gene family in Lycopersicon pennellii show different developmental patterns of expression[J].Plant Physiology,1998,116(4):1461-1468.
[33]	JI J L,LÜH H,YANG L M,et al.Genome-wide identification and characterization of non-specific lipid transfer proteins in cabbage[J].Peer J,2018,6:e5379.
[34]	SAKHARKAR M K,CHOW V T K,KANGUEANE P.Distributions of exons and introns in the human genome[J].In Silico Biology,2004,4(4):387-393.
[35]	JAIN M,KHURANA P,TYAGI A K,et al.Genome-wide analysis of intronless genes in rice and Arabidopsis[J].Functional&Integrative Genomics,2008,8(1):69-78.
[36]	D’AGOSTINO N,BUONANNO M,AYOUB J,et al.Identification of non-specific Lipid Transfer Protein gene family members in Solanum lycopersicum and insights into the features of Sola l 3 protein[J].Scientific Reports,2019,9(1):1607.
[37]	LIANG W J,MA X L,WAN P,et al.Plant salt-tolerance mechanism:a review[J].Biochemical and Biophysical Research Communications,2018,495(1):286-291.
[38]	COLCOMBET J,HIRT H.Arabidopsis MAPKs:a complex signalling network involved in multiple biological processes[J].Biochemical Journal,2008,413(2):217-226.