杨树PeAFB基因克隆及表达模式初步分析

doi:10.11707/j.1001-7488.20150804

Abstract

Abstract:

[Objective] The TIR1/AFB F-box proteins are auxin receptors, which directly involve in auxin perception by degrading Aux/IAA proteins, thereby regulating biological processes of stress responses and development. In order to reveal the auxin signal transduction and regulation mechanisms of AFB genes in poplar, AFB gene family members of poplar were cloned and their expression patterns under non-host pathogen, plant hormone and drought treatment were analyzed.[Method] Using PCR and RACE methods, AFB gene family members were cloned from a poplar hybrid clone NL-895 (Populus deltoides ×P. euramericana cl. NL-895). Online softwares and databases were used for gene predictions, conserve domain prediction, tertiary structure prediction, multiple-sequence alignments, and phylogenetic analysis. Real-time RT-PCR was conducted for gene expression pattern analysis of poplar AFB genes.[Result] Four AFB gene family members in NL-895 were cloned and named PeAFB2-1,PeAFB2-2,PeAFB3 and PeAFB 5 respectively. The full-length cDNA sequences of the PeAFB genes were 2 461, 3 115, 2 441,and 2 818 bp and the corresponding lengths of ORF (open reading frames) were 1 546, 1 716, 1 716 and 1 914 bp. AFBs are usually leucine-rich proteins, and the 4 PeAFBs also show high leucine content (10%-13.4%). The analysis of deduced protein sequences showed that 4 PeAFBs all contained a typical conserved F-box domain (IPR001810) and five LRR-rich repeats (IPR006553). A phylogenetic analysis using the 47 AFB proteins from land plants revealed that all AFB proteins fell into four clades, and the 4 PeAFBs resided in the first clade. Furthermore, four PeAFB genes' expressions were down-regulated under Pseudomonas syringae DC3000, MeJA(500 μmol·L^-1), and PEG(20%, m/V) treatment.[Conclusion] Four PeAFB gene family members in poplar NL-895 were cloned,and their phylogenetic relationships were revealed. Four PeAFB genes' expressions were down-regulated under biotic and abiotic stresses, which indicate the four genes may play a role in negative regulation of poplar abiotic/biotic stress responses.

Key words: poplar NL-895, TIR1/AFB F-box protein, PeAFB, stress responses

CLC Number:

S718.46

Chen Ying, Shao Zhilong, Wang Haoran, Zhu Yanyu, Zhu Sheng, Huang Minren. Cloning and Expression of PeAFB Genes in Populus[J]. Scientia Silvae Sinicae, 2015, 51(8): 26-32.

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Cloning and Expression of PeAFB Genes in Populus

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