Nucleotide-binding site leucine-rich repeat receptors (NBS-LRR/NLRs) are crucial intracellular immune proteins in plants. Previous article reported a novel NLR protein SUT1 (SUPPRESSORS OF TOPP4-1, 1), which is involv...Nucleotide-binding site leucine-rich repeat receptors (NBS-LRR/NLRs) are crucial intracellular immune proteins in plants. Previous article reported a novel NLR protein SUT1 (SUPPRESSORS OF TOPP4-1, 1), which is involved in autoimmunity initiated by type one protein phosphatase 4 mutation (topp4-1) in Arabidopsis, however, its role in planta is still unclear. This study employed Nicotiana benthamiana, a model platform, to conduct an overall structural and functional analysis of SUT1 protein. The transient expression results revealed that SUT1 is a typical CNL (CC-NBS-LRR) receptor, both fluorescence data and biochemical results showed the protein is mainly anchored on the plasma membrane due to its N-terminal acylation site. Further truncation experiments announced that its CC (coiled-coil) domain possessed cell-death-inducing activity. The outcomes of point mutations analysis revealed that not only the CC domain, but also the full-length SUT1 protein, whose function and subcellular localization are influenced by highly conserved hydrophobic residues. These research outcomes provided favorable clues for elucidating the activation mechanism of SUT1.展开更多
目的为解析党参NBS-LRR(Nucleotide-binding site and leucine-rich repeat)抗病基因家族,探究党参抗根腐病机制,从而解决党参根腐病害难题,促进党参育种及产业发展。方法基于党参响应根腐病病原菌的转录组数据,通过运用生物信息学方法...目的为解析党参NBS-LRR(Nucleotide-binding site and leucine-rich repeat)抗病基因家族,探究党参抗根腐病机制,从而解决党参根腐病害难题,促进党参育种及产业发展。方法基于党参响应根腐病病原菌的转录组数据,通过运用生物信息学方法对党参NBS-LRR家族基因进行理化性质、基因结构、系统发育、表达模式及互作网络分析。结果成功鉴定到88个党参NBS-LRR家族基因,包括N、NL、CN、CNL、TN、TNL、PN共7种类型,分别有50、14、1、14、4、3、2个基因。结果表明,党参CNL及TNL类基因结构比较保守;党参CNL亚家族基因在进化过程中发生扩增;党参NBS-LRR家族基因在尖孢镰刀菌(Fusarium oxysporum)侵染条件下存在时间表达模式差异,且侵染前期(6-24 h)高表达的基因DN64786c1g6、DN64786c1g5、DN48234c0g2、DN54844c1g2、DN59747c0g3、DN56071c1g8、DN64591c1g1、DN48464c1g1、DN59886c0g1在调控党参抗病过程中发挥重要作用。其中党参的抗病蛋白DN54844c1g2可能与GLR家族互作,进而通过调节Ca2+内流参与免疫调控;DN64786c1g5可能与CYTC-1和CYTC-2互作,进而通过参与氧化还原反应参与党参响应根腐病过程;DN59747c0g3可能与MPK3互作,进而通过参与MAP信号级联、磷酸化WRKY转录因子以及参与超敏反应(HR),在党参响应根腐病过程中发挥重要作用。结论党参NBSLRR家族基因的鉴定及表达分析对于探究党参抗根腐病机制、发掘基因功能具有重要意义。展开更多
根据抗病基因核苷酸结合位点(Nuc leotide b ind ing site,NBS)设计简并性引物,从陆地棉cDNA中进行RT-PCR扩增。获得含NBS保守域的EST,进一步用RACE技术和TAIL PCR技术获得其中1个EST的全长基因序列,并获得GHNBS基因的5′调控序列。此...根据抗病基因核苷酸结合位点(Nuc leotide b ind ing site,NBS)设计简并性引物,从陆地棉cDNA中进行RT-PCR扩增。获得含NBS保守域的EST,进一步用RACE技术和TAIL PCR技术获得其中1个EST的全长基因序列,并获得GHNBS基因的5′调控序列。此基因被命名为GHNBS。该基因的编码区长2 583 bp,编码861个氨基酸,GHNBS编码的氨基酸序列与拟南芥R基因具有28%的同源性。GHNBS与拟南芥的其他几个NBS-LRR基因比较发现,它们在保守区外的相似性相当低。Southern杂交和网上数据库搜索分析都表明GHNBS是1个寡拷贝基因。通过半定量RT-PCR分析发现GHNBS在棉花的蕾、花瓣、韧皮部、根及叶中均有表达且在根、叶中表达量比其它部位强,在木质部基本不表达。展开更多
NBS-LRR(nucleotide-binding site and leucine-rich-repeat)是植物中最大类抗病基因家族之一。番茄基因组测序完成为全基因组水平上分析NBS-LRR抗病基因家族提供了机遇。利用生物信息学方法对番茄NBS-LRR抗病基因家族成员数目进行鉴定...NBS-LRR(nucleotide-binding site and leucine-rich-repeat)是植物中最大类抗病基因家族之一。番茄基因组测序完成为全基因组水平上分析NBS-LRR抗病基因家族提供了机遇。利用生物信息学方法对番茄NBS-LRR抗病基因家族成员数目进行鉴定,并对其染色体定位和系统发育关系进行了分析。随后,将番茄和马铃薯NBS-LRR抗病基因进行了比较基因组学分析。结果表明:番茄基因组共包括252个NBS-LRR抗病基因,分布于番茄12条染色体上;63.5%的基因成簇存在,大部分为串联重复;系统发育关系分析表明番茄CC-NBS-LRR(CNL)亚家族较其他亚家族扩展程度大;同线性分析发现番茄中共79个NBS-LRR抗病基因与马铃薯基因具有同源关系。结果将为番茄NBS-LRR抗病基因家族的深入研究提供依据,同时也为利用番茄NBS-LRR基因进行基因定位以及相关抗病基因克隆等奠定基础。展开更多
文摘Nucleotide-binding site leucine-rich repeat receptors (NBS-LRR/NLRs) are crucial intracellular immune proteins in plants. Previous article reported a novel NLR protein SUT1 (SUPPRESSORS OF TOPP4-1, 1), which is involved in autoimmunity initiated by type one protein phosphatase 4 mutation (topp4-1) in Arabidopsis, however, its role in planta is still unclear. This study employed Nicotiana benthamiana, a model platform, to conduct an overall structural and functional analysis of SUT1 protein. The transient expression results revealed that SUT1 is a typical CNL (CC-NBS-LRR) receptor, both fluorescence data and biochemical results showed the protein is mainly anchored on the plasma membrane due to its N-terminal acylation site. Further truncation experiments announced that its CC (coiled-coil) domain possessed cell-death-inducing activity. The outcomes of point mutations analysis revealed that not only the CC domain, but also the full-length SUT1 protein, whose function and subcellular localization are influenced by highly conserved hydrophobic residues. These research outcomes provided favorable clues for elucidating the activation mechanism of SUT1.
文摘目的为解析党参NBS-LRR(Nucleotide-binding site and leucine-rich repeat)抗病基因家族,探究党参抗根腐病机制,从而解决党参根腐病害难题,促进党参育种及产业发展。方法基于党参响应根腐病病原菌的转录组数据,通过运用生物信息学方法对党参NBS-LRR家族基因进行理化性质、基因结构、系统发育、表达模式及互作网络分析。结果成功鉴定到88个党参NBS-LRR家族基因,包括N、NL、CN、CNL、TN、TNL、PN共7种类型,分别有50、14、1、14、4、3、2个基因。结果表明,党参CNL及TNL类基因结构比较保守;党参CNL亚家族基因在进化过程中发生扩增;党参NBS-LRR家族基因在尖孢镰刀菌(Fusarium oxysporum)侵染条件下存在时间表达模式差异,且侵染前期(6-24 h)高表达的基因DN64786c1g6、DN64786c1g5、DN48234c0g2、DN54844c1g2、DN59747c0g3、DN56071c1g8、DN64591c1g1、DN48464c1g1、DN59886c0g1在调控党参抗病过程中发挥重要作用。其中党参的抗病蛋白DN54844c1g2可能与GLR家族互作,进而通过调节Ca2+内流参与免疫调控;DN64786c1g5可能与CYTC-1和CYTC-2互作,进而通过参与氧化还原反应参与党参响应根腐病过程;DN59747c0g3可能与MPK3互作,进而通过参与MAP信号级联、磷酸化WRKY转录因子以及参与超敏反应(HR),在党参响应根腐病过程中发挥重要作用。结论党参NBSLRR家族基因的鉴定及表达分析对于探究党参抗根腐病机制、发掘基因功能具有重要意义。
文摘根据抗病基因核苷酸结合位点(Nuc leotide b ind ing site,NBS)设计简并性引物,从陆地棉cDNA中进行RT-PCR扩增。获得含NBS保守域的EST,进一步用RACE技术和TAIL PCR技术获得其中1个EST的全长基因序列,并获得GHNBS基因的5′调控序列。此基因被命名为GHNBS。该基因的编码区长2 583 bp,编码861个氨基酸,GHNBS编码的氨基酸序列与拟南芥R基因具有28%的同源性。GHNBS与拟南芥的其他几个NBS-LRR基因比较发现,它们在保守区外的相似性相当低。Southern杂交和网上数据库搜索分析都表明GHNBS是1个寡拷贝基因。通过半定量RT-PCR分析发现GHNBS在棉花的蕾、花瓣、韧皮部、根及叶中均有表达且在根、叶中表达量比其它部位强,在木质部基本不表达。
文摘NBS-LRR(nucleotide-binding site and leucine-rich-repeat)是植物中最大类抗病基因家族之一。番茄基因组测序完成为全基因组水平上分析NBS-LRR抗病基因家族提供了机遇。利用生物信息学方法对番茄NBS-LRR抗病基因家族成员数目进行鉴定,并对其染色体定位和系统发育关系进行了分析。随后,将番茄和马铃薯NBS-LRR抗病基因进行了比较基因组学分析。结果表明:番茄基因组共包括252个NBS-LRR抗病基因,分布于番茄12条染色体上;63.5%的基因成簇存在,大部分为串联重复;系统发育关系分析表明番茄CC-NBS-LRR(CNL)亚家族较其他亚家族扩展程度大;同线性分析发现番茄中共79个NBS-LRR抗病基因与马铃薯基因具有同源关系。结果将为番茄NBS-LRR抗病基因家族的深入研究提供依据,同时也为利用番茄NBS-LRR基因进行基因定位以及相关抗病基因克隆等奠定基础。
基金This work was supported by China Postdoctoral Science Foundation (Contact No. 2005037628) and Science and Technology Foun-dation of South China Agricultural University (Contact No. Rnd0401)
