金针菇L-赖氨酸合成通路基因鉴定及对不同光质的响应表达  

Identification of Genes in Flammulina filiformis L-lysine Biosynthesis Pathway and Their Expression in Response to Light Conditions

作　　者：陶永新[1,2] 段静怡 李依宁 李自燕 宋寒冰 张祺锶 黄嘉华 高玲玲 谢宝贵[2] TAO Yongxin;DUAN Jingyi;LI Yining;LI Ziyan;SONG Hanbing;ZHANG Qisi;HUANG Jiahua;GAO Lingling;XIE Baogui(College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China;Mycological Research Center,College of Life Sciences,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China)

机构地区：[1]福建农林大学园艺学院,福建福州350002 [2]福建农林大学生命科学学院菌物研究中心,福建福州350002

出　　处：《食用菌学报》2018年第4期1-8,共8页Acta Edulis Fungi

基　　金：福建农林大学园艺学院青年学术骨干培养基金(61201400706);福建农林大学科技创新专项基金(CXZX2017184);国家重点基础研究发展计划项目(2014CB138302);木生型食用菌品种选育与产业化工程项目(fjzycxny2017010);福建省食用菌产业体系项目(K83139296).

摘　　要：为了从分子水平探究提高金针菇赖氨酸含量的环境因子(温度、光照、湿度、CO_2)及其作用机理,本研究首先在金针菇(Flammulina filiformis)单核体L11基因组中鉴定L-赖氨酸合成通路基因,并预测其启动子区顺式元件。根据预测结果,采用红、绿、蓝、红绿蓝复合4种光处理金针菇子实体,并分别用HPLC和定量PCR测定赖氨酸含量变化及其合成通路基因的响应表达。结果表明:金针菇赖氨酸由氨基己二酸(α-Aminoadipate,AAA)途径合成,通路中8步催化酶的编码基因及其序列在L11中全部得到鉴定。关键限速酶基因HCS、HAC1和SHD基因的启动子区存在G-box等光照响应元件。定量PCR表明,总体上看赖氨酸合成通路前7步的酶编码基因对4种光质均有响应表达。进一步比较本试验中4种不同光质的处理效果,发现蓝光照射后的金针菇子实体中,共有7个赖氨酸合成通路基因(HCS、HAC1、HAH、HDH、AAT、AAR和SR)表达量上调,同时赖氨酸含量最高。推测蓝光可能通过激活赖氨酸合成通路基因启动子区的G-box等顺式元件,诱导通路基因上调表达,进而提高了金针菇子实体中赖氨酸含量。In order to explore the environmental inducers(temperature,light,humidity,CO2)of increasing the lysine content in Flammulina filiformis as well as the underlying molecular mechanism,genes in the L-lysine synthesis path way were identified from the genome of L11 monokaryotic strain and then cis-acting elements in the promoter regions of these genes were predicted.According to the predicted results,four light conditions,including red,green,blue and red-green-blue mixed light,were used to illuminate on fruiting bodies of F.filiformis with darkness as the control.Lysine content in pileus and expression of the lysine synthesis pathway genes under each treatment were measured afterwards by HPLC and quantitative PCR,respectively.The results showed that L-lysine was biosynthesized by theα-aminoadipate(AAA)pathway inF.filiformis,and the DNA sequences of the enzymes in the eight steps of AAA pathway were all obtained.There were light response cis-elements such as G-box in the promoter regions of genes HCS(rate limiting key enzyme),HACI and SHD.Quantitative PCR showed that genes in the first seven steps of lysine synthesis(HCS,HAC1,HAH,HDH,AAT,AAR and SR)were up-regulated in response to blue light.Blue light irradiation also resulted in the greatest lysine content compared to other light conditions.It was speculated that blue light may up-regulate genes in the lysine synthesis pathway by activating cis-elements such as G-box in their promoter regions,thereby increasing lysine content in the fruiting body of F.filiformis.

关 键 词：食用真菌 赖氨酸生物合成 光照处理 顺式作用元件 

分 类 号：S646.15[农业科学—蔬菜学]