9个拟南芥凝集素类受体激酶基因功能的初步研究
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摘要
植物凝集素类受体激酶(LecRLK)在植物防御、应答多种生物与非生物胁迫、参与激素调控和生长发育中具有重要的作用。但是该家族大部分成员的功能尚不明了。本文对LecRLK家族中的9个成员进行了生物信息学和相关分子遗传学的初步研究。启动子元件分析结果表明,9个成员均具有多个光和逆境响应元件,暗示其可能参与光信号以及非生物胁迫信号途径的调控。基因芯片及实时定量PCR结果显示,9个LecRLK基因具有明显的时空表达特异性,说明其可能在植物不同生长发育阶段及组织部位发挥着重要调控作用。利用三引物法对9个基因的突变体进行了筛选,最终得到14个纯合缺失突变体材料。上述结果为后续深入开展这些基因的生理功能研究提供参考数据。
Lectin receptor like kinases( LecRLK) play an important role in plant defense,various kinds of responses to biological and abiotic stress,involving in hormones signaling,regulation of growth and development. However,the function of most members of LecRLK family remains unclear. In this paper,we carried out a preliminary study on the bioinformatics and molecular genetics of nine LecRLK members. The analysis of Cis-elements in promoter showed that the nine genes had multiple light and stress responsive elements,suggesting that they could be involved in the light signal transduction and abiotic stress response. The gene chip analysis and real-time quantitative PCR results indicated that the nine genes showed significant differences in the temporal-spacial expression,revealing that they may be implicated in various growth periods and diverse plant tissues. Using three primers verification arrays,we got 14 homozygous knockout mutant lines as materials. Our work could contributed to the further studies on the physiological functions of these genes.
Lectin receptor like kinases( LecRLK) play an important role in plant defense,various kinds of responses to biological and abiotic stress,involving in hormones signaling,regulation of growth and development. However,the function of most members of LecRLK family remains unclear. In this paper,we carried out a preliminary study on the bioinformatics and molecular genetics of nine LecRLK members. The analysis of Cis-elements in promoter showed that the nine genes had multiple light and stress responsive elements,suggesting that they could be involved in the light signal transduction and abiotic stress response. The gene chip analysis and real-time quantitative PCR results indicated that the nine genes showed significant differences in the temporal-spacial expression,revealing that they may be implicated in various growth periods and diverse plant tissues. Using three primers verification arrays,we got 14 homozygous knockout mutant lines as materials. Our work could contributed to the further studies on the physiological functions of these genes.
引文
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[2]BOUWMEESTER K,GOVERS F.Arabidopsis L-type lectin receptor kinases:phylogeny,classification,and expression profiles[J].Journal of Experimental Botany,2009,60(15):4383-4396.
[3]VAID N,PANDEY P K,TUTEJA N.Genome-wide analysis of lectin receptor-like kinase family from Arabidopsis and rice[J].Plant Molecular Biology,2012,80(4-5):365-388.
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[8]BOUWMEESTER K,DE SAIN M,WEIDE R,et al.The lectin receptor kinase LecRK-I.9 is a novel phytophthora resistance component and a potential host target for a RXLR effector[J].PLo S Pathogens,2011,7:e1001327.
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[14]HUANG P,JU H W,MIN J H,et al.Overexpression of Ltype lectin-like protein kinase 1 confers pathogen resistance and regulates salinity response in Arabidopsis thaliana[J].Plant Science,2013,203-204:98-106.
[15]SUN X L,YU Q Y,TANG L L,et al.Gs SRK,a G-type lectin S-receptor-like serine/threonine protein kinase,is a positive regulator of plant tolerance to salt stress[J].Journal of Plant Physiology,2013,170(5):505-515.
[16]HE X J,ZHANG Z G,YAN D Q,et al.A salt-responsive receptor-like kinase gene regulated by the ethylene signaling pathway encodes a plasma membrane serine/threonine kinase[J].Theoretical and Applied Genetics,2004,109(2):377-383.
[17]BOUWMEESTER K,DE SAIN M,WEIDE R,et al.The lectin receptor kinase LecRK-I.9 is a novel Phytophthora resistance component and a potential host target for a RXLR effector[J].PLo S Pathogens,2011,7(3):e1001327.
[18]LYOU S H,PARK H J,JUNG C,et al.The Arabidopsis At LEC gene encoding a lectin-like protein is up-regulated by multiple stimuli including developmental signal,wounding,jasmonate,ethylene,and chitin elicitor[J].Molecules and Cells,2009,27(1):75-81.
[19]JOSHI A,DANG H Q,VAID N,et al.Pea lectin receptorlike kinase promotes high salinity stress tolerance in bacteria and expresses in response to stress in planta[J].Glycoconjugate Journal,2010,27(1):133-150.
[20]XIN Z,WANG A,YANG G,et al.The Arabidopsis A4 subfamily of lectin receptor kinases negatively regulates abscisic acid response in seed germination[J].Plant Physiology,2009,149(1):434-444.
[21]WAN J,PATEL A,MATHIEU M,et al.A lectin receptorlike kinase is required for pollen development in Arabidopsis[J].Plant Molecular Biology,2008,67(5):469-482.
[22]RIOU C,HERVE C,PACQUIT V,et al.Expression of an Arabidopsis lectin kinase receptor gene,lecRK-a1,is induced during senescence,wounding and in response to oligogalacturonic acids[J].Plant Physiology and Biochemistry,2002,40(5):431-438.
[23]ZUO K,ZHAN J,WANG J,et al.Molecular cloning and characterization of GhlecRK,a novel kinase gene with lectinlike domain from Gossypium hirsutum[J].DNA Sequence,2004,15(1):58-65.
[2]BOUWMEESTER K,GOVERS F.Arabidopsis L-type lectin receptor kinases:phylogeny,classification,and expression profiles[J].Journal of Experimental Botany,2009,60(15):4383-4396.
[3]VAID N,PANDEY P K,TUTEJA N.Genome-wide analysis of lectin receptor-like kinase family from Arabidopsis and rice[J].Plant Molecular Biology,2012,80(4-5):365-388.
[4]TAKASAKI T,HATAKEYAMA K,SUZUKI G,et al.The S receptor kinase determines self-incompatibility in Brassica stigma[J].Nature,2000,403(6772):913-916.
[5]CAMBI A,KOOPMAN M,FIGDOR C G.How C-type lectins detect pathogens[J].Cellular Microbiology,2005,7(4):481-488.
[6]HERVE C,SERRES J,DABOS P,et al.Characterization of the Arabidopsis lecRK-a genes:members of a superfamily encoding putative receptors with an extracellular domain homologous to legume lectins[J].Plant Molecular Biology,1999,39(4):671-682.
[7]VAID N,MACOVEI A,TUTEJA N.Knights in action:lectin receptor-like kinases in plant development and stress responses[J].Molecular Plant,2013,6(5):1405-1418.
[8]BOUWMEESTER K,DE SAIN M,WEIDE R,et al.The lectin receptor kinase LecRK-I.9 is a novel phytophthora resistance component and a potential host target for a RXLR effector[J].PLo S Pathogens,2011,7:e1001327.
[9]MELLERSH D G,HEATH M C.Plasma membrane-cell wall adhesion is required for expression of plant defense responses during fungal penetration[J].Plant Cell,2001,13(2):413-424.
[10]MELOTTO M,UNDERWOOD W,KOCZAN J,et al.Plant stomata function in innate immunity against bacterial invasion[J].Cell,2006,126(5):969-980.
[11]MELOTTO M,UNDERWOOD W,HE S Y.Role of stomata in plant innate immunity and foliar bacterial diseases[J].Annual Review of Phytopathology,2008,46:101-122.
[12]DESCLOS-THEVENIAU M,ARNAUD D,HUANG T Y,et al.The Arabidopsis lectin receptor kinase LecRK-V.5represses stomatal immunity induced by Pseudomonas syringae pv.tomato DC3000[J].PLo S Pathogens,2012,8(2):e1002513.
[13]NAVARRO-GOCHICOA M T,CAMUT S,TIMMERS A C,et al.Characterization of four lectin-like receptor kinases expressed in roots of Medicago truncatula.structure,location,regulation of expression,and potential role in the symbiosis with Sinorhizobium meliloti[J].Plant Physiology,2003,133(4):1893-1910.
[14]HUANG P,JU H W,MIN J H,et al.Overexpression of Ltype lectin-like protein kinase 1 confers pathogen resistance and regulates salinity response in Arabidopsis thaliana[J].Plant Science,2013,203-204:98-106.
[15]SUN X L,YU Q Y,TANG L L,et al.Gs SRK,a G-type lectin S-receptor-like serine/threonine protein kinase,is a positive regulator of plant tolerance to salt stress[J].Journal of Plant Physiology,2013,170(5):505-515.
[16]HE X J,ZHANG Z G,YAN D Q,et al.A salt-responsive receptor-like kinase gene regulated by the ethylene signaling pathway encodes a plasma membrane serine/threonine kinase[J].Theoretical and Applied Genetics,2004,109(2):377-383.
[17]BOUWMEESTER K,DE SAIN M,WEIDE R,et al.The lectin receptor kinase LecRK-I.9 is a novel Phytophthora resistance component and a potential host target for a RXLR effector[J].PLo S Pathogens,2011,7(3):e1001327.
[18]LYOU S H,PARK H J,JUNG C,et al.The Arabidopsis At LEC gene encoding a lectin-like protein is up-regulated by multiple stimuli including developmental signal,wounding,jasmonate,ethylene,and chitin elicitor[J].Molecules and Cells,2009,27(1):75-81.
[19]JOSHI A,DANG H Q,VAID N,et al.Pea lectin receptorlike kinase promotes high salinity stress tolerance in bacteria and expresses in response to stress in planta[J].Glycoconjugate Journal,2010,27(1):133-150.
[20]XIN Z,WANG A,YANG G,et al.The Arabidopsis A4 subfamily of lectin receptor kinases negatively regulates abscisic acid response in seed germination[J].Plant Physiology,2009,149(1):434-444.
[21]WAN J,PATEL A,MATHIEU M,et al.A lectin receptorlike kinase is required for pollen development in Arabidopsis[J].Plant Molecular Biology,2008,67(5):469-482.
[22]RIOU C,HERVE C,PACQUIT V,et al.Expression of an Arabidopsis lectin kinase receptor gene,lecRK-a1,is induced during senescence,wounding and in response to oligogalacturonic acids[J].Plant Physiology and Biochemistry,2002,40(5):431-438.
[23]ZUO K,ZHAN J,WANG J,et al.Molecular cloning and characterization of GhlecRK,a novel kinase gene with lectinlike domain from Gossypium hirsutum[J].DNA Sequence,2004,15(1):58-65.