白木香信号肽肽酶AsSPP1基因的克隆及表达分析

Cloning and expression analysis of signal peptide peptidase (AsSPP1) from Aquilaria sinensis

信号肽肽酶在生物的生命过程中起到重要作用。本研究根据白木香转录组高通量测序结果设计引物,结合RACE及RT-PCR技术从白木香中克隆得到信号肽肽酶AsSPP1基因全长,并对其进行生物信息学分析;利用实时荧光定量PCR检测AsSPP1基因在不同组织以及在NaCl、重金属、低温、干旱胁迫和ABA、GA3、MeJA、SA处理下的表达差异。本研究中克隆获得的AsSPP1基因全长1 063 bp,编码353个氨基酸。AsSPP1基因在茎中的表达量最高,其次是叶和茎尖,在根中的表达量最低。盐、重金属和低温胁迫能够诱导AsSPP1基因的表达,而干旱胁迫则降低AsSPP1基因的表达。ABA、SA和MeJA能够诱导AsSPP1基因的表达,而GA3对AsSPP1基因表达影响不大。这些研究结果表明AsSPP1基因可能在白木香防御反应中起重要作用。

Signal peptide peptidase(SPP)plays an important role in biological processes of organisms.In present study,the full length of AsSPP1 gene was cloned by RACE and reverse transcription PCR from Aquilaria sinensis calli based on the high-throughput sequencing results of the transcriptome.The SPP transmembrane domain and phylogenetic analysis were predicted by different software to analysize the bioinformatics of SPP proteins.Transcript level of AsSPP1 was performed with real-time quantitative RT-PCR in different tissues and in responds to abiotic stresses including salt,heavy metal,cold and drought stress,as well as plant growth regulators including abscisic acid(ABA),gibberellins(GA3),methyl jasmonate(MeJA)and salicylic acid(SA)treatments.Our results demonstrated that the AsSPP1 cDNA sequence had an ORF of 1 063 bp,encoding a protein of 353 amino acids.Tissue expression analysis indicated that AsSPP1 was primarily expressed in stems,followed by leaves and shoot tips,and then roots.The expression of AsSPP1 was induced bp salt,heavy metal and cold stresses,while decreased under drought stress.Furthermore,the transcript level of AsSPP1 was increased treated by ABA,MeJA and SA,while GA3 had little influence on the expression of AsSPP1.These results suggested the AsSPP1 may play an important role in plant defense and hormone signal transduction in A.sinensis.