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作 者:叶小真[1] 杨婕 冯丽贞[3] 江仲鹏 陆芝 刘雨菁 李丽红 陈全助[1] YE Xiao-zhen;YANG Jie;FENG Li-zhen;JIANG Zhong-peng;LU Zhi;LIU Yu-jing;LI Li-hong;CHEN Quan-zhu(Jinshan College,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China;Administration Bureau of Fujian Huboliao National Nature Reserve,Zhangzhou 363600,Fujian,China;Forestry College,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China)
机构地区:[1]福建农林大学金山学院,福建福州350002 [2]福建虎伯寮国家级自然保护区管理局,福建漳州363600 [3]福建农林大学林学院,福建福州350002
出 处:《林业科学研究》2019年第6期99-105,共7页Forest Research
基 金:福建农林大学科研启动费(Z150702);福建农林大学科技创新专项基金项目(KFA17278A);福建省财政厅项目(K81150002)
摘 要:[目的]克隆桉树苯丙氨酸解氨酶基因(PAL),分析其序列特征及在焦枯病菌诱导下的表达特征,为探究桉树抗焦枯病机理提供理论支撑。[方法]通过SOE-PCR技术从尾细桉M1中克隆PAL基因,利用real-time PCR分析其在焦枯病菌诱导下的表达特性。[结果]克隆得到尾细桉M1 PAL基因,其ORF序列长2142 bp,编码713个氨基酸,与其他已发表的桉树PAL相似性均在99%以上。序列分析发现,其编码蛋白是一类稳定的亲水性蛋白,含有丝氨酸、苏氨酸和酪氨酸3个磷酸化位点,二级结构以α-螺旋和β-折叠为主,具有典型的苯丙氨酸和组氨酸解氨酶保守结构域。进一步分析桉树不同品系PAL基因的表达趋势,结果发现,不同品系PAL基因在焦枯病菌诱导下均表现为明显的上调表达,且抗性越强,PAL基因越早上调,上调表达量越大。[结论]从尾细桉M1中克隆得到的PAL基因具有典型的PAL家族成员特征,推测该基因可能通过参与酚类防御性物质或者信号分子SA等物质的合成,进而在桉树抵抗焦枯病菌侵染的过程中发挥重要作用。[Objective]This study aimed at cloning the cDNA sequence of Eucalyptus phenylalanin ammonia-lyase(PAL)gene,analyzing its characteristics,and investigating its expressions in Eucalyptus under the infection of Calonectria pseudoreteaudii.[Method]SOE-PCR technology was used to clone PAL gene from E.urophylla×E.tereticornis M1.The expression level of PAL gene in different Eucalyptus cultivars infected with C.pseudoreteaudii were analyzed by RT-PCR.[Result]One PAL gene was cloned.Its ORF sequence was 2142 bp,and encoded 713 amino acids,which showed high similarity to the PAL from other Eucalyptus,with 99%identity.Amino acids sequence analysis indicated that the predicted PAL protein was a stable hydrophilic protein,including three phosphorylation sites(serine,threonine and tyrosine).Its secondary structure was rich inα-helix and random coil.The predicted protein contained phenylalanine and histidine ammonia-lyase conserved structure.The results showed that the PAL gene was significantly up-regulated in all tested cultivars.Moreover,the PAL gene was earlier and more significantly up-regulated in the stronger resistant cultivars.[Conclusion]The PAL gene which was cloned from E.urophylla×E.tereticornis M1 has typical characteristics of PAL family.It is speculated that PAL gene may participant in the synthesis of phenolic compounds or signal molecule salicylic acid,then improves the resistance of Eucalyptus to C.pseudoreteaudii.
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