Genome-Wide Identification and Expression Analysis of nsLTP Gene Family in Rapeseed (Brassica napus) Reveals Their Critical Roles in Biotic and Abiotic Stress Responses

Non-specific lipid transfer proteins (nsLTPs) are small cysteine-rich basic proteins which play essential roles in plant growth, development and abiotic/biotic stress response. However, there is limited information about the nsLTP gene (BnLTP) family in rapeseed (Brassica napus). In this study, 283 BnLTP genes were identified in rapeseed, which were distributed randomly in 19 chromosomes of rapeseed. Phylogenetic analysis showed that BnLTP proteins were divided into seven groups. Exon/intron structure and MEME motifs both remained highly conserved in each BnLTP group. Segmental duplication and hybridization of rapeseed’s two sub-genomes mainly contributed to the expansion of the BnLTP gene family. Various potential cis-elements that respond to plant growth, development, biotic/abiotic stresses, and phytohormone signals existed in BnLTP gene promoters. Transcriptome analysis showed that BnLTP genes were expressed in various tissues/organs with different levels and were also involved in the response to heat, drought, NaCl, cold, IAA and ABA stresses, as well as the treatment of fungal pathogens (Sclerotinia sclerotiorum and Leptosphaeria maculans). The qRT-PCR assay validated the results of RNA-seq expression analysis of two top Sclerotinia-responsive BnLTP genes, BnLTP129 and BnLTP161. Moreover, batches of BnLTPs might be regulated by BnTT1 and BnbZIP67 to play roles in the development, metabolism or adaptability of the seed coat and embryo in rapeseed. This work provides an important basis for further functional study of the BnLTP genes in rapeseed quality improvement and stress resistance.

[1]  Peijian Cao,et al.  Systematic analysis of the non-specific lipid transfer protein gene family in Nicotiana tabacum reveal its potential roles in stress responses. , 2022, Plant physiology and biochemistry : PPB.

[2]  Ruijun Duan,et al.  Genome-wide identification and expression profile under abiotic stress of the barley non-specific lipid transfer protein gene family and its Qingke Orthologues , 2021, BMC genomics.

[3]  Guiyin Zhang,et al.  Tissue-specific expression of GhnsLTPs identified via GWAS sophisticatedly coordinates disease- and insect-resistance by regulating metabolic flux redirection in cotton. , 2021, The Plant journal : for cell and molecular biology.

[4]  Kede Liu,et al.  DELLA proteins BnaA6.RGA and BnaC7.RGA negatively regulate fatty acid biosynthesis by interacting with BnaLEC1s in Brassica napus , 2021, Plant biotechnology journal.

[5]  Linhai Wang,et al.  Genome-Wide Analysis of nsLTP Gene Family and Identification of SiLTPs Contributing to High Oil Accumulation in Sesame (Sesamum indicum L.) , 2021, International journal of molecular sciences.

[6]  Sixue Chen,et al.  Guard cell redox proteomics reveals a role of lipid transfer protein in plant defense. , 2021, Journal of proteomics.

[7]  F. Brini,et al.  Pathogenesis Related Proteins (PRs): From Cellular Mechanisms to Plant Defense. , 2020, Current protein & peptide science.

[8]  Qingyong Yang,et al.  Genome- and transcriptome-wide association studies provide insights into the genetic basis of natural variation of seed oil content in Brassica napus. , 2020, Molecular plant.

[9]  Chenchen Wang,et al.  A nonspecific lipid transfer protein, StLTP10, mediates resistance to Phytophthora infestans in potato , 2020, Molecular plant pathology.

[10]  A. Raza.  Eco-physiological and Biochemical Responses of Rapeseed (Brassica napus L.) to Abiotic Stresses: Consequences and Mitigation Strategies , 2020, Journal of Plant Growth Regulation.

[11]  R. Dill‐Macky,et al.  A lipid transfer protein has antifungal and antioxidant activity and suppresses Fusarium head blight disease and DON accumulation in transgenic wheat. , 2020, Phytopathology.

[12]  Xinhua Zhao,et al.  An Advanced Lipid Metabolism System Revealed by Transcriptomic and Lipidomic Analyses Plays a Central Role in Peanut Cold Tolerance , 2020, Frontiers in Plant Science.

[13]  Y. Chai,et al.  Genome-wide mining and comparative analysis of fatty acid elongase gene family in Brassica napus and its progenitors. , 2020, Gene.

[14]  Margaret H. Frank,et al.  TBtools - an integrative toolkit developed for interactive analyses of big biological data. , 2020, Molecular plant.

[15]  Shuai Li,et al.  Genome-wide identification and characterization of nonspecific lipid transfer protein (nsLTP) genes in Arachis duranensis. , 2020, Genomics.

[16]  Yaoguang Liu,et al.  A lipid transfer protein variant with a mutant eight-cysteine motif causes photoperiod- and thermo-sensitive dwarfism in rice , 2019, Journal of experimental botany.

[17]  A. Kasianov,et al.  Non-Specific Lipid Transfer Proteins in Triticum kiharae Dorof. et Migush.: Identification, Characterization and Expression Profiling in Response to Pathogens and Resistance Inducers , 2019, Pathogens.

[18]  Shanshan Wang,et al.  The lipid transfer protein OsLTPL159 is involved in cold tolerance at the early seedling stage in rice , 2019, Plant biotechnology journal.

[19]  Shengting Li,et al.  Identification of candidate genes controlling oil content by combination of genome-wide association and transcriptome analysis in the oilseed crop Brassica napus , 2019, Biotechnology for biofuels.

[20]  Haoli Ma,et al.  Genome-wide identification, characterization and expression analysis of the non-specific lipid transfer proteins in potato , 2019, BMC Genomics.

[21]  S. M. Monti,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 , 2019, Scientific Reports.

[22]  A. Diévart,et al.  Genome-wide analysis of the barley non-specific lipid transfer protein gene family , 2019, The Crop Journal.

[23]  N. Buhot,et al.  Involvement of lipid transfer proteins in resistance against a non-host powdery mildew in Arabidopsis thaliana. , 2018, Molecular plant pathology.

[24]  U. Baumann,et al.  Genome-wide identification and analysis of non-specific Lipid Transfer Proteins in hexaploid wheat , 2018, Scientific Reports.

[25]  Z. Fang,et al.  Genome-wide identification and characterization of non-specific lipid transfer proteins in cabbage , 2018, PeerJ.

[26]  Lifei Zhu,et al.  NtLTP4, a lipid transfer protein that enhances salt and drought stresses tolerance in Nicotiana tabacum , 2018, Scientific Reports.

[27]  T. Salminen,et al.  Plant lipid transfer proteins: are we finally closing in on the roles of these enigmatic proteins? , 2018, Journal of Lipid Research.

[28]  Y. Chai,et al.  Genome-Wide Survey and Characterization of Fatty Acid Desaturase Gene Family in Brassica napus and Its Parental Species , 2018, Applied Biochemistry and Biotechnology.

[29]  Uwe Scholz,et al.  MISA-web: a web server for microsatellite prediction , 2017, Bioinform..

[30]  B. Payre,et al.  The Arabidopsis Lipid Transfer Protein 2 (AtLTP2) Is Involved in Cuticle-Cell Wall Interface Integrity and in Etiolated Hypocotyl Permeability , 2017, Front. Plant Sci..

[31]  Xuede Wang,et al.  Genomic Identification and Comparative Expansion Analysis of the Non-Specific Lipid Transfer Protein Gene Family in Gossypium , 2016, Scientific Reports.

[32]  Jingjuan Yu,et al.  A Non-specific Setaria italica Lipid Transfer Protein Gene Plays a Critical Role under Abiotic Stress , 2016, Frontiers in plant science.

[33]  T. Salminen,et al.  Lipid transfer proteins: classification, nomenclature, structure, and function , 2016, Planta.

[34]  R. Prasad,et al.  Enhanced Tolerance of Transgenic Potato Plants Over-Expressing Non-specific Lipid Transfer Protein-1 (StnsLTP1) against Multiple Abiotic Stresses , 2016, Front. Plant Sci..

[35]  Kun Lu,et al.  Genome-wide association analysis and differential expression analysis of resistance to Sclerotinia stem rot in Brassica napus. , 2016, Plant biotechnology journal.

[36]  T. Ovchinnikova,et al.  Lipid Transfer Proteins As Components of the Plant Innate Immune System: Structure, Functions, and Applications , 2016, Acta naturae.

[37]  Sudhir Kumar,et al.  MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. , 2016, Molecular biology and evolution.

[38]  Qingyong Yang,et al.  A genome-wide association study reveals novel elite allelic variations in seed oil content of Brassica napus , 2016, Theoretical and Applied Genetics.

[39]  X. Wang,et al.  A lipid transfer protein, OsLTPL36, is essential for seed development and seed quality in rice. , 2015, Plant science : an international journal of experimental plant biology.

[40]  F. Liu,et al.  Non-specific lipid transfer proteins in plants: presenting new advances and an integrated functional analysis. , 2015, Journal of experimental botany.

[41]  K. Masmoudi,et al.  A wheat lipid transfer protein (TdLTP4) promotes tolerance to abiotic and biotic stress in Arabidopsis thaliana. , 2015, Plant physiology and biochemistry : PPB.

[42]  K. Wei,et al.  Non-specific lipid transfer proteins in maize , 2014, BMC Plant Biology.

[43]  Monika M. Edstam,et al.  Involvement of GPI-anchored lipid transfer proteins in the development of seed coats and pollen in Arabidopsis thaliana. , 2014, Physiologia plantarum.

[44]  Käthe M. Dahlström,et al.  Characterization of the GPI-anchored lipid transfer proteins in the moss Physcomitrella patens. , 2014, Plant physiology and biochemistry : PPB.

[45]  Kun Xu,et al.  Genome-Wide Survey and Expression Analysis of the Putative Non-Specific Lipid Transfer Proteins in Brassica rapa L , 2014, PloS one.

[46]  S. Powers,et al.  bZIP67 Regulates the Omega-3 Fatty Acid Content of Arabidopsis Seed Oil by Activating FATTY ACID DESATURASE3[W][OPEN] , 2013, Plant Cell.

[47]  K. Blomqvist,et al.  Coexpression patterns indicate that GPI-anchored non-specific lipid transfer proteins are involved in accumulation of cuticular wax, suberin and sporopollenin , 2013, Plant Molecular Biology.

[48]  Xiaochun Ge,et al.  The rice OsDIL gene plays a role in drought tolerance at vegetative and reproductive stages , 2013, Plant Molecular Biology.

[49]  Y. Wang,et al.  Transformation of LTP gene into Brassica napus to enhance its resistance to Sclerotinia sclerotiorum , 2013, Russian Journal of Genetics.

[50]  X. Fu,et al.  Overexpression of an nsLTPs-like antimicrobial protein gene (LJAMP2) from motherwort (Leonurus japonicus) enhances resistance to Sclerotinia sclerotiorum in oilseed rape (Brassica napus) , 2013 .

[51]  Xiaoyan Zhang,et al.  Lipid transfer protein 3 as a target of MYB96 mediates freezing and drought stress in Arabidopsis , 2013, Journal of experimental botany.

[52]  K. Katoh,et al.  MAFFT Multiple Sequence Alignment Software Version 7: Improvements in Performance and Usability , 2013, Molecular biology and evolution.

[53]  S. Powers,et al.  bZIP 67 Regulates the Omega-3 Fatty Acid Content of Arabidopsis Seed Oil by Activating FATTY ACID DESATURASE 3 , 2013 .

[54]  I. Hwang,et al.  Characterization of glycosylphosphatidylinositol-anchored lipid transfer protein 2 (LTPG2) and overlapping function between LTPG/LTPG1 and LTPG2 in cuticular wax export or accumulation in Arabidopsis thaliana. , 2012, Plant & cell physiology.

[55]  Zhao Li,et al.  Overexpression of wheat lipid transfer protein gene TaLTP5 increases resistances to Cochliobolus sativus and Fusarium graminearum in transgenic wheat , 2012, Functional & Integrative Genomics.

[56]  T. Salminen,et al.  Evolutionary history of the non-specific lipid transfer proteins. , 2011, Molecular plant.

[57]  B. Weisshaar,et al.  TRANSPARENT TESTA1 interacts with R2R3-MYB factors and affects early and late steps of flavonoid biosynthesis in the endothelium of Arabidopsis thaliana seeds. , 2011, The Plant journal : for cell and molecular biology.

[58]  Xingyong Yang,et al.  Enhanced resistance to fungal pathogens in transgenic Populus tomentosa Carr. by overexpression of an nsLTP-like antimicrobial protein gene from motherwort (Leonurus japonicus). , 2010, Tree physiology.

[59]  R. Weselake,et al.  A survey of quantitative real-time polymerase chain reaction internal reference genes for expression studies in Brassica napus. , 2010, Analytical biochemistry.

[60]  C. Kieslich,et al.  A multifaceted study of stigma/style cysteine-rich adhesin (SCA)-like Arabidopsis lipid transfer proteins (LTPs) suggests diversified roles for these LTPs in plant growth and reproduction , 2010, Journal of experimental botany.

[61]  Dabing Zhang,et al.  OsC6, Encoding a Lipid Transfer Protein, Is Required for Postmeiotic Anther Development In Rice1[W][OA] , 2010, Plant Physiology.

[62]  C. Jang,et al.  Evolution of non-specific lipid transfer protein (nsLTP) genes in the Poaceae family: their duplication and diversity , 2008, Molecular Genetics and Genomics.

[63]  J. Rose,et al.  The biochemistry and biology of extracellular plant lipid‐transfer proteins (LTPs) , 2008, Protein science : a publication of the Protein Society.

[64]  B. Kleinhenz,et al.  A crop loss-related forecasting model for sclerotinia stem rot in winter oilseed rape. , 2007, Phytopathology.

[65]  A. Carvalho,et al.  Role of plant lipid transfer proteins in plant cell physiology—A concise review , 2007, Peptides.

[66]  P. Porter,et al.  Impact of Sclerotinia Stem Rot on Yield of Canola. , 2007, Plant disease.

[67]  M. Gautier,et al.  Genome-wide analysis of the rice and arabidopsis non-specific lipid transfer protein (nsLtp) gene families and identification of wheat nsLtp genes by EST data mining , 2008, BMC Genomics.

[68]  M. Teece,et al.  Increased Accumulation of Cuticular Wax and Expression of Lipid Transfer Protein in Response to Periodic Drying Events in Leaves of Tree Tobacco1[W] , 2005, Plant Physiology.

[69]  M. Barbetti,et al.  World-Wide Importance of Phoma Stem Canker (Leptosphaeria maculans and L. biglobosa) on Oilseed Rape (Brassica napus) , 2005, European Journal of Plant Pathology.

[70]  B. Hwang,et al.  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 , 2005, Planta.

[71]  R. Shin,et al.  Induction of pepper cDNA encoding a lipid transfer protein during the resistance response to tobacco mosaic virus , 2002, Plant Molecular Biology.

[72]  D. Marion,et al.  From elicitins to lipid-transfer proteins: a new insight in cell signalling involved in plant defence mechanisms. , 2002, Trends in plant science.

[73]  F Beaudoin,et al.  Plant protein families and their relationships to food allergy. , 2001, Biochemical Society transactions.

[74]  M. Sawaya,et al.  Structural basis of non-specific lipid binding in maize lipid-transfer protein complexes revealed by high-resolution X-ray crystallography. , 2001, Journal of molecular biology.

[75]  L. C. Loon,et al.  The families of pathogenesis-related proteins, their activities, and comparative analysis of PR-1 type proteins , 1999 .

[76]  J. Kader.  Lipid-transfer proteins: a puzzling family of plant proteins , 1997 .

[77]  J. Kader.  LIPID-TRANSFER PROTEINS IN PLANTS. , 1996, Annual review of plant physiology and plant molecular biology.

[78]  M. Yamada.  Lipid Transfer Proteins in Plants and Microorganisms , 1992 .

[79]  P. Shewry,et al.  Molecular evolution of the seed storage proteins of barley, rye and wheat. , 1985, Journal of molecular biology.