Plant non-specific lipid transfer proteins （nsLtps） have been reported to be involved in plant defense activity against bacterial and fungal pathogens. In this study, we identified 135 （122 putative and 13 previous...Plant non-specific lipid transfer proteins （nsLtps） have been reported to be involved in plant defense activity against bacterial and fungal pathogens. In this study, we identified 135 （122 putative and 13 previously identified） Solanaceae nsLtps, which are clustered into 8 different groups. By comparing with Boutrot’s nsLtp classification, we classified these eight groups into five types （I, II, IV, IX and X）. We compared Solanaceae nsLtps with Arabidopsis and Gramineae nsLtps and found that （1） Types I, II and IV are shared by Solanaceae, Gramineae and Arabidopsis; （2） Types III, V, VI and VIII are shared by Gramineae and Arabidopsis but not detected in Solanaceae so far; （3） Type VII is only found in Gramineae whereas type IX is present only in Arabidopsis and Solanaceae; （4） Type X is a new type that accounts for 52.59% Solanaceae nsLtps in our data, and has not been reported in any other plant so far. We further built and compared the three-dimensional structures of the eight groups, and found that the major functional diversification within the nsLtp family could be predated to the monocot/dicot divergence, and many gene duplications and sequence variations had happened in the nsLtp family after the monocot/dicot divergence, especially in Solanaceae.展开更多
以芹菜(Apium graveolens)‘六合黄心芹’、‘津南实芹’和‘美国西芹’为试验材料,采用RT-PCR技术分别获得其cDNA序列。序列分析表明:来源于3个芹菜品种的非特异性脂转移蛋白(Non-specific lipid transfer protein,nsLTP)基因核苷酸序...以芹菜(Apium graveolens)‘六合黄心芹’、‘津南实芹’和‘美国西芹’为试验材料,采用RT-PCR技术分别获得其cDNA序列。序列分析表明:来源于3个芹菜品种的非特异性脂转移蛋白(Non-specific lipid transfer protein,nsLTP)基因核苷酸序列高度保守,全长357bp,编码118个氨基酸,起始密码子ATG之后含有27个氨基酸残基的信号肽序列,推测其成熟的蛋白含91个氨基酸残基,预测其蛋白质分子量为11.75kD,pI值为9.36。芹菜的nsLTP蛋白主要由α-螺旋和随机卷曲组成。空间结构上分析显示,芹菜nsLTP蛋白中H1区域明显分为H1a和H1b两个亚区域,而模板碧桃中H1区域为一个连续的螺旋结构,存在明显的差异。进化分析显示,芹菜nsLTP与香石竹、大洋洲滨藜等植物的nsLTP相似性较高,在保守位置具有8个半胱氨酸残基。实时定量PCR表达分析表明,该基因主要在芹菜的茎以及茎尖生长活跃中心表达,具有明显的组织特异性。展开更多
Non-specific lipid transfer proteins(nsLTPs/LTPs) that can transport various phospholipids across the membrane in vitro are widespread in the plant kingdom, and they play important roles in many biological processes t...Non-specific lipid transfer proteins(nsLTPs/LTPs) that can transport various phospholipids across the membrane in vitro are widespread in the plant kingdom, and they play important roles in many biological processes that are closely related to plant growth and development. Recently, nsLTPs have been shown to respond to different forms of abiotic stresses. Despite the vital roles of nsLTPs in many plants, little is known about the nsLTPs in wheat. In this study, 330 nsLTP proteins were identified in wheat and they clustered into five types(1, 2, c, d, and g) by phylogenetic analysis with the nsLTPs from maize, Arabidopsis, and rice. The wheat nsLTPs of type d included three subtypes(d1, d2, and d3) and type g included seven subtypes(g1–g7). Genetic structure and motif pattern analyses showed that members of each type had similar structural composition. Moreover, GPI-anchors were found to exist in non-g type members from wheat for the first time. Chromosome mapping revealed that all five types were unevenly and unequally distributed on 21 chromosomes. Furthermore, gene duplication events contributed to the proliferation of the nsLTP genes. Large-scale data mining of RNA-seq data covering multiple growth stages and numerous stress treatments showed that the transcript levels of some of the nsLTP genes could be strongly induced by abiotic stresses, including drought and salinity, indicating their potential roles in mediating the responses of the wheat plants to these abiotic stress conditions. These findings provide comprehensive insights into the nsLTP family members in wheat, and offer candidate nsLTP genes for further studies on their roles in stress resistance and potential for improving wheat breeding programs.展开更多
Plant nonspecific lipid transfer proteins (nsLTPs) are widely distributed through plant kingdom and are characterized by the presence of a central hydrophobic cavity, suitable for binding various hydrophobic molecules...Plant nonspecific lipid transfer proteins (nsLTPs) are widely distributed through plant kingdom and are characterized by the presence of a central hydrophobic cavity, suitable for binding various hydrophobic molecules. Despite extensive research on nsLTP in different plant species, mostly angiosperm, and the great diversity of physiological processes in which they seem to be involved, their exact functions still remain unclear. Also, very limited experimental data are available on nsLTP in gymnosperm. In this study, we report for the first time on the molecular cloning of nsLTP, from Pinus sylvestris L.(PsLTP1, GenBank accession JN980402.1) and the expression pattern of PsLTP1 during ontogenesis and in response to environmental stress conditions. Total RNA from roots of 7-day old pine seedlings was used to isolate the cDNA clone, corresponding to Scots pine lipid transfer protein. The open reading frame of PsLTP1 consists of 372 bp encoding a protein of 123 amino acids. Amino acid sequence alignment revealed that mature PsLTP1 shares high level of similarity with nsLTP from other conifers and with well-studied nsLTPs from angiosperms. The PsLTP1 contains a 27-amino-acid N-terminal signal sequence and presents all the features of a plant nsLTP. Amino acid comparison analysis and 3D structure prediction showed that PsLTP1 is a type 1 nsLTP. The results of the expression analysis of Scots pine PsLTP1 gene revealed that its transcripts accumulate in actively growing tissues. Furthermore, transcription of PsLTP1 was upregulated in response to cold and salt treatments, and downregulated during acidic, osmotic and water stresses.展开更多
TDF1(transcription-drived fragment) was homologous to the predicted S. lycopersicum nonspecific lipid-transfer protein,nsLTP 2-like(91%), and it was significantly upregulated in response to C. fulvum(cladosporium fulv...TDF1(transcription-drived fragment) was homologous to the predicted S. lycopersicum nonspecific lipid-transfer protein,nsLTP 2-like(91%), and it was significantly upregulated in response to C. fulvum(cladosporium fulvum) infection in tomato plants.In this experiment, the full-length cDNA of nsLTP 2-like was cloned using RACE technology based on the sequence of TDF1(GenBank: JZ717725). A full-length, 625 bp(GenBank: KU366289), cDNA sequence, which with 98% similarity to nsLTP 2-like gene(GenBank: XM015233692) was obtained. This cDNA contains an ORF(open reading frame) with full-length of 345 bp, coding of 114 amino acids, including 12.3% Ala and Gly. Protein molecular weight was 11.51 ku, the isoelectric point(pI) was 8.99, and average overall hydrophilicity was 0.412, with one phosphorylation sites, belonging to volatile acidic nuclear protein. Secondary structure prediction showed that α-Helix accounts for 30.7%, extension chain for 12.28%, β-corner for 9.65%, and random coil for 47.37%. Through comparative analysis of the homology among species, it was found that the amino acid sequence of tomato nsLTP 2-like protein had a high similarity with other plants, and with a specific conserved sequence which might related features in nsLTP 2-like protein. It also be analyzed the gene expression pattern of tomato in different parts and under different stress conditions.The results showed that nsLTP 2-like gene was up-regulated in varying degrees, under the condition of cold stress, exogenous hormone spraying and cladosporium fulvum infection. Therefore, it was speculated that the gene played a role in response to abiotic and biotic stress in tomato.展开更多
This study is to investigate the role of lipid transfer protein （LTP1） gene of potato （Solanum tuberosum） in bacterial wilt （Ralstonia solanacearum） resistance. A novel cDNA clone encoding nsLTP was isolated fro...This study is to investigate the role of lipid transfer protein （LTP1） gene of potato （Solanum tuberosum） in bacterial wilt （Ralstonia solanacearum） resistance. A novel cDNA clone encoding nsLTP was isolated from cultivated potato （Solanum tuberosum） infected with R. solanacearum by 5＇-rapid amplification of cDNA ends （RACE）. The temporal and spatial expression of StLTPbl was studied during the early stages of potato-R, solanacearum interaction by reverse transcriptase PCR （RT-PCR） and Northern blotting. The sequence analysis of the cloned cDNA, named StLTPbl, showed 691 bp which encoded a type 1 nsLTP of 91 amino acids. Construction of a phylogenic tree showed that StLTPbl is well conserved in the coding region with high identity at the amino acid level with other Solanaceae nsLTPs. The temporal and spatial expression of StLTPbl was studied during the early stages of potato-R, solanacearum interaction. StLTPbl transcription is induced faster and transcripts accumulate to higher concentrations in resistant compared with susceptible genotypes by the pathogen. Dominant differences in the pathogen-induced gene expression pattern between the upper and lower leaves and stems were observed within the same genotypes. In situ hybridization results showed that the StLTPbl mRNA was localized in phloem cells of vascular tissues in potato leaf and stem tissues after pathogen infection. Salicylic acid, methyl jasmonate and abscisic acid could induce StLTPbl gene expression without significant difference between the upper and lower tissues. These abiotic elicitors could produce a long-lastingeffect on the StLTPbl during early stages of potato-R, solanacearum interaction. Differential expression of StLTPbl gene between resistance and susceptible potato genotypes in response to R. solanacearum suggests that this gene plays a key role in plant defense mechanisms.展开更多
TaMs1 encodes a non-specific lipid transfer protein(nsLTP) and is required for pollen development in wheat. Although MS1 is a Poaceae-specific gene, the roles of MS1 genes in other Poaceae plants are unknown, especial...TaMs1 encodes a non-specific lipid transfer protein(nsLTP) and is required for pollen development in wheat. Although MS1 is a Poaceae-specific gene, the roles of MS1 genes in other Poaceae plants are unknown, especially in rice and maize. Here, we identified one ortholog in rice(OsLTPg29) and two orthologs in maize(ZmLTPg11 and ZmLTPx2). Similar to TaMs1, both OsLTPg29 and ZmLTPg11 genes are specifically expressed in the microsporocytes, and both OsLTPg29 and ZmLTPg11 proteins showed lipid-binding ability to phosphatidic acid and several phosphoinositides. To determine their roles in pollen development, we created osltpg29 mutants and zmltpg11 zmltpx2 double mutants by CRISPR/Cas9.osltpg29, not zmltpg11 zmltpx2, is defective in pollen development, and only OsLTPg29, not ZmLTPg11,can rescue the male sterility of tams1 mutant. Our results demonstrate that the biological function of MS1 in pollen development differs in the evolution of Poaceae plants.展开更多
【目的】AsE246是我们首次报道的紫云英根瘤特异表达的非特异性转脂蛋白(nsLTP1:non specificlipid transfer protein 1)编码基因。本实验旨在筛选和鉴定与AsE246相互作用的宿主植物靶蛋白,并分析靶基因在共生和胁迫条件下的表达特征。...【目的】AsE246是我们首次报道的紫云英根瘤特异表达的非特异性转脂蛋白(nsLTP1:non specificlipid transfer protein 1)编码基因。本实验旨在筛选和鉴定与AsE246相互作用的宿主植物靶蛋白,并分析靶基因在共生和胁迫条件下的表达特征。【方法】利用酵母双杂交技术、小范围杂交技术及实时荧光定量PCR,筛选与AsE246的相互作用蛋白,并定量分析靶基因在结瘤与固氮过程中的时空表达特性。【结果】获取一个阳性克隆,其cDNA序列经Blast分析表明:候选靶蛋白是一个DnaJ-like蛋白,该蛋白相应基因命名为AsDJL1。AsE246与AsDJL1在酵母体内确实相互作用。AsDJL1在固氮根瘤中特异性增强表达,在NaCl胁迫下表达水平显著提高,在(NH4)2SO4胁迫下表达水平显著下降。【结论】本实验是筛选与LTP相互作用蛋白的首次报道。获得了直接的实验证据表明互作基因AsDJL1与AsE246具有高度相似的表达特征和功能,为深入研究二者的相互作用及其在共生固氮和应答环境胁迫中的调控机制,提供了一定的工作基础和理论依据。展开更多
基金supported by the National Natural Science Foundation of China (Grant No. 30900831)
文摘Plant non-specific lipid transfer proteins （nsLtps） have been reported to be involved in plant defense activity against bacterial and fungal pathogens. In this study, we identified 135 （122 putative and 13 previously identified） Solanaceae nsLtps, which are clustered into 8 different groups. By comparing with Boutrot’s nsLtp classification, we classified these eight groups into five types （I, II, IV, IX and X）. We compared Solanaceae nsLtps with Arabidopsis and Gramineae nsLtps and found that （1） Types I, II and IV are shared by Solanaceae, Gramineae and Arabidopsis; （2） Types III, V, VI and VIII are shared by Gramineae and Arabidopsis but not detected in Solanaceae so far; （3） Type VII is only found in Gramineae whereas type IX is present only in Arabidopsis and Solanaceae; （4） Type X is a new type that accounts for 52.59% Solanaceae nsLtps in our data, and has not been reported in any other plant so far. We further built and compared the three-dimensional structures of the eight groups, and found that the major functional diversification within the nsLtp family could be predated to the monocot/dicot divergence, and many gene duplications and sequence variations had happened in the nsLtp family after the monocot/dicot divergence, especially in Solanaceae.
文摘以芹菜(Apium graveolens)‘六合黄心芹’、‘津南实芹’和‘美国西芹’为试验材料,采用RT-PCR技术分别获得其cDNA序列。序列分析表明:来源于3个芹菜品种的非特异性脂转移蛋白(Non-specific lipid transfer protein,nsLTP)基因核苷酸序列高度保守,全长357bp,编码118个氨基酸,起始密码子ATG之后含有27个氨基酸残基的信号肽序列,推测其成熟的蛋白含91个氨基酸残基,预测其蛋白质分子量为11.75kD,pI值为9.36。芹菜的nsLTP蛋白主要由α-螺旋和随机卷曲组成。空间结构上分析显示,芹菜nsLTP蛋白中H1区域明显分为H1a和H1b两个亚区域,而模板碧桃中H1区域为一个连续的螺旋结构,存在明显的差异。进化分析显示,芹菜nsLTP与香石竹、大洋洲滨藜等植物的nsLTP相似性较高,在保守位置具有8个半胱氨酸残基。实时定量PCR表达分析表明,该基因主要在芹菜的茎以及茎尖生长活跃中心表达,具有明显的组织特异性。
基金supported by the National Transgenic Key Project of the Ministry of Agriculture of China(2018ZX0800909B)the Major Program of Technological Innovation of Hubei Province,China(2018ABA085)+1 种基金the Open Project Program of Engineering Research Center of Ecology and Agricultural Use of Wetland,Ministry of Education,China(KF201802)the Southwest Agricultural Crop Pest Management Key Laboratory Open Fund of Ministry of Agriculture,China(2018-XNZD-01)。
文摘Non-specific lipid transfer proteins(nsLTPs/LTPs) that can transport various phospholipids across the membrane in vitro are widespread in the plant kingdom, and they play important roles in many biological processes that are closely related to plant growth and development. Recently, nsLTPs have been shown to respond to different forms of abiotic stresses. Despite the vital roles of nsLTPs in many plants, little is known about the nsLTPs in wheat. In this study, 330 nsLTP proteins were identified in wheat and they clustered into five types(1, 2, c, d, and g) by phylogenetic analysis with the nsLTPs from maize, Arabidopsis, and rice. The wheat nsLTPs of type d included three subtypes(d1, d2, and d3) and type g included seven subtypes(g1–g7). Genetic structure and motif pattern analyses showed that members of each type had similar structural composition. Moreover, GPI-anchors were found to exist in non-g type members from wheat for the first time. Chromosome mapping revealed that all five types were unevenly and unequally distributed on 21 chromosomes. Furthermore, gene duplication events contributed to the proliferation of the nsLTP genes. Large-scale data mining of RNA-seq data covering multiple growth stages and numerous stress treatments showed that the transcript levels of some of the nsLTP genes could be strongly induced by abiotic stresses, including drought and salinity, indicating their potential roles in mediating the responses of the wheat plants to these abiotic stress conditions. These findings provide comprehensive insights into the nsLTP family members in wheat, and offer candidate nsLTP genes for further studies on their roles in stress resistance and potential for improving wheat breeding programs.
基金supported by grants from the Ministry of Education and Science of Ukraine(0116U003593)grant from cieA3(Campus de Excelencia Internacional Agroalimentario)-UCO,Spain
文摘Plant nonspecific lipid transfer proteins (nsLTPs) are widely distributed through plant kingdom and are characterized by the presence of a central hydrophobic cavity, suitable for binding various hydrophobic molecules. Despite extensive research on nsLTP in different plant species, mostly angiosperm, and the great diversity of physiological processes in which they seem to be involved, their exact functions still remain unclear. Also, very limited experimental data are available on nsLTP in gymnosperm. In this study, we report for the first time on the molecular cloning of nsLTP, from Pinus sylvestris L.(PsLTP1, GenBank accession JN980402.1) and the expression pattern of PsLTP1 during ontogenesis and in response to environmental stress conditions. Total RNA from roots of 7-day old pine seedlings was used to isolate the cDNA clone, corresponding to Scots pine lipid transfer protein. The open reading frame of PsLTP1 consists of 372 bp encoding a protein of 123 amino acids. Amino acid sequence alignment revealed that mature PsLTP1 shares high level of similarity with nsLTP from other conifers and with well-studied nsLTPs from angiosperms. The PsLTP1 contains a 27-amino-acid N-terminal signal sequence and presents all the features of a plant nsLTP. Amino acid comparison analysis and 3D structure prediction showed that PsLTP1 is a type 1 nsLTP. The results of the expression analysis of Scots pine PsLTP1 gene revealed that its transcripts accumulate in actively growing tissues. Furthermore, transcription of PsLTP1 was upregulated in response to cold and salt treatments, and downregulated during acidic, osmotic and water stresses.
基金Supported by the National Key R&D Program of China(2017YFD0101900)China Agriculture Research System(CARS-23-A-16)the Science Foundation of Heilongjiang Province(C2017024)
文摘TDF1(transcription-drived fragment) was homologous to the predicted S. lycopersicum nonspecific lipid-transfer protein,nsLTP 2-like(91%), and it was significantly upregulated in response to C. fulvum(cladosporium fulvum) infection in tomato plants.In this experiment, the full-length cDNA of nsLTP 2-like was cloned using RACE technology based on the sequence of TDF1(GenBank: JZ717725). A full-length, 625 bp(GenBank: KU366289), cDNA sequence, which with 98% similarity to nsLTP 2-like gene(GenBank: XM015233692) was obtained. This cDNA contains an ORF(open reading frame) with full-length of 345 bp, coding of 114 amino acids, including 12.3% Ala and Gly. Protein molecular weight was 11.51 ku, the isoelectric point(pI) was 8.99, and average overall hydrophilicity was 0.412, with one phosphorylation sites, belonging to volatile acidic nuclear protein. Secondary structure prediction showed that α-Helix accounts for 30.7%, extension chain for 12.28%, β-corner for 9.65%, and random coil for 47.37%. Through comparative analysis of the homology among species, it was found that the amino acid sequence of tomato nsLTP 2-like protein had a high similarity with other plants, and with a specific conserved sequence which might related features in nsLTP 2-like protein. It also be analyzed the gene expression pattern of tomato in different parts and under different stress conditions.The results showed that nsLTP 2-like gene was up-regulated in varying degrees, under the condition of cold stress, exogenous hormone spraying and cladosporium fulvum infection. Therefore, it was speculated that the gene played a role in response to abiotic and biotic stress in tomato.
基金grateful to Ren Caihong(College of Life Science,Shanxi Normal University of China)for technical assistance.This work was supported by National 863 Program(2003AA207130)Natural Science Foundation of Shanxi Province of China(20051042).
文摘This study is to investigate the role of lipid transfer protein （LTP1） gene of potato （Solanum tuberosum） in bacterial wilt （Ralstonia solanacearum） resistance. A novel cDNA clone encoding nsLTP was isolated from cultivated potato （Solanum tuberosum） infected with R. solanacearum by 5＇-rapid amplification of cDNA ends （RACE）. The temporal and spatial expression of StLTPbl was studied during the early stages of potato-R, solanacearum interaction by reverse transcriptase PCR （RT-PCR） and Northern blotting. The sequence analysis of the cloned cDNA, named StLTPbl, showed 691 bp which encoded a type 1 nsLTP of 91 amino acids. Construction of a phylogenic tree showed that StLTPbl is well conserved in the coding region with high identity at the amino acid level with other Solanaceae nsLTPs. The temporal and spatial expression of StLTPbl was studied during the early stages of potato-R, solanacearum interaction. StLTPbl transcription is induced faster and transcripts accumulate to higher concentrations in resistant compared with susceptible genotypes by the pathogen. Dominant differences in the pathogen-induced gene expression pattern between the upper and lower leaves and stems were observed within the same genotypes. In situ hybridization results showed that the StLTPbl mRNA was localized in phloem cells of vascular tissues in potato leaf and stem tissues after pathogen infection. Salicylic acid, methyl jasmonate and abscisic acid could induce StLTPbl gene expression without significant difference between the upper and lower tissues. These abiotic elicitors could produce a long-lastingeffect on the StLTPbl during early stages of potato-R, solanacearum interaction. Differential expression of StLTPbl gene between resistance and susceptible potato genotypes in response to R. solanacearum suggests that this gene plays a key role in plant defense mechanisms.
基金supported by Peking University Institute of Advanced Agricultural Sciences, and Beijing Municipal Government Science Foundation (IDHT20170513)。
文摘TaMs1 encodes a non-specific lipid transfer protein(nsLTP) and is required for pollen development in wheat. Although MS1 is a Poaceae-specific gene, the roles of MS1 genes in other Poaceae plants are unknown, especially in rice and maize. Here, we identified one ortholog in rice(OsLTPg29) and two orthologs in maize(ZmLTPg11 and ZmLTPx2). Similar to TaMs1, both OsLTPg29 and ZmLTPg11 genes are specifically expressed in the microsporocytes, and both OsLTPg29 and ZmLTPg11 proteins showed lipid-binding ability to phosphatidic acid and several phosphoinositides. To determine their roles in pollen development, we created osltpg29 mutants and zmltpg11 zmltpx2 double mutants by CRISPR/Cas9.osltpg29, not zmltpg11 zmltpx2, is defective in pollen development, and only OsLTPg29, not ZmLTPg11,can rescue the male sterility of tams1 mutant. Our results demonstrate that the biological function of MS1 in pollen development differs in the evolution of Poaceae plants.
文摘【目的】AsE246是我们首次报道的紫云英根瘤特异表达的非特异性转脂蛋白(nsLTP1:non specificlipid transfer protein 1)编码基因。本实验旨在筛选和鉴定与AsE246相互作用的宿主植物靶蛋白,并分析靶基因在共生和胁迫条件下的表达特征。【方法】利用酵母双杂交技术、小范围杂交技术及实时荧光定量PCR,筛选与AsE246的相互作用蛋白,并定量分析靶基因在结瘤与固氮过程中的时空表达特性。【结果】获取一个阳性克隆,其cDNA序列经Blast分析表明:候选靶蛋白是一个DnaJ-like蛋白,该蛋白相应基因命名为AsDJL1。AsE246与AsDJL1在酵母体内确实相互作用。AsDJL1在固氮根瘤中特异性增强表达,在NaCl胁迫下表达水平显著提高,在(NH4)2SO4胁迫下表达水平显著下降。【结论】本实验是筛选与LTP相互作用蛋白的首次报道。获得了直接的实验证据表明互作基因AsDJL1与AsE246具有高度相似的表达特征和功能,为深入研究二者的相互作用及其在共生固氮和应答环境胁迫中的调控机制,提供了一定的工作基础和理论依据。
基金supported by grants from the National Science and Technology Pillar Program of China(2007BAD78B03)the “Eleventh-Five” Key Project of Sichuan ProvinceChina(07SG111-003-1)
