摘要
该研究以泸定百合(Lilium sargentiae Wilson)为材料,构建其组培苗经百合尖孢镰刀菌侵染后的叶片SSH文库,从中筛选镰刀菌枯萎病抗病相关基因。从正向SSH文库中随机挑取300个单克隆测序后得到280条ESTs,进行功能比对分析后,除去未知功能、沉冗蛋白以及无同源序列,得到有功能的ESTs共168条,其功能涉及信号传导、蛋白质合成与代谢、抗病与防御、物质与能量代谢、转录相关等多种途径,其中有31条ESTs与抗病防御相关。从抗病防御相关基因中选取8条ESTs:过氧化氢酶、ATP结合盒转运蛋白(ABC transporter)、Kunitz型胰蛋白酶抑制剂4(Kunitz trypsin inhibitor 4)、丝氨酸乙醛酸氨基转移酶、多聚泛素、脂氧合酶I(Lipoxygenase I)、丝氨酸/苏安酸蛋白激酶(Serine/Threonine-protein kinase)、抗坏血酸过氧化物酶(Arabidopsis thaliana),通过RTPCR对其表达情况进行分析,发现经镰刀菌诱导后均为上调表达,推测它们可能参与了泸定百合镰刀菌枯萎病的抗病反应途径。
In order to screening candidate resistance related genes against Lily Fusarium Wilt,we constructed a suppression subtractive hybridization（SSH）cDNA library from leaves of tissue-culture seedling of Lilium sargentiae Wilson induced by Fusarium oxysporum f.sp.lilii.300 clones of the forward SSH library were picked out randomly and sequenced,and then 280 qualified sequence tags（ESTs）were obtained.After functional annotation,168 ESTs were found putatively encoding functionally known protein.The function-known ESTs related to signal transduction,protein synthesis and transporters,disease resistance and defense,energy metabolism and transcription,etc.Among them,31 ESTs were the defense response genes,and 8defense response ESTs（Catalase,ABC transporter,Kunitz trypsin inhibitor 4,Serine-glyoxy-late aminotransferase,Polyubiquitin,lipoxygenase I,Serine/threonine-protein kinase,Arabidopsis thaliana）were analyzed by the semi-quantitative RT-PCR,the results showed they were upregulated by F.oxysporumf.sp.lilii and supposed to involved in the disease resistance reaction for Lily Fusarium Wilt.
出处
《西北植物学报》
CAS
CSCD
北大核心
2014年第11期2170-2175,共6页
Acta Botanica Boreali-Occidentalia Sinica
基金
863计划(2011AA100208)
云南省应用基础研究计划(2012FB189
2012FD072)
云南省科技计划(2012BB011)
国家观赏园艺工程技术研究中心(2012FU125X10)
云南省农业科学院创新团队(YAAS2014TD002)
关键词
泸定百合
镰刀菌枯萎病
SSH文库
抗病基因
Lilium sargentiae Wilson
Lily Fusarium Wilt
SSH library
resistance relate genes
