利用cDNA-AFLP分析内生解淀粉芽胞杆菌 Fy11诱导辣椒差异表达基因

doi:10.3864/j.issn.0578-1752.2013.12.006

Abstract

Abstract: 【Objective】The objective of this study is to analyze extensive transcription profiling of pepper seedlings inoculated with an endophytic bacterial strain, Bacillus amyloliquefaciens Fy11, and elucidate the molecular mechanism of endophytic Bacillus and plant interaction.【Method】cDNA-AFLP technique was used to conduct transcription profiling of pepper seedlings across five sampling time points after inoculation with Fy11. The validation of cDNA-AFLP expression patterns was analyzed by qRT-PCR.【Result】 A total of 18 620 transcript derived fragments (TDFs) were obtained using cDNA-AFLP with 256 primer pairs, 353 (1.89%) displayed altered expression patterns after inoculation. Two-hundred and fifty seven differentially expressed TDFs produced reliable sequences after cloning and sequencing. Two-hundred and twenty nine expressed sequence tags (ESTs) of unigenes were obtained after assembling, of which 144 showed up-regulated and 85 down-regulated. Blastx analysis and functional annotations were then performed and the results revealed that the 156 ESTs had predicted gene products mainly implicated in energy (10.92%), metabolism and disease/defense (each accounted for 8.73%), signal transduction (7.42%), transporter (6.99%), cell structure (6.55%), transcription (5.68%), cell growth (3.93%), protein destination and storage (3.49%), protein synthesis (2.18%), secondary metabolism and intracellur traffic (each accounted for 1.75%). The 65 ESTs (28.38% of the sequenced total 229 ESTs) had no match to known genes, and the 8 ESTs (3.49% of the sequenced total 229 ESTs) were highly homologous to unknown functional proteins. Ten differential genes related to disease/defense, transcription, and signal transduction were chosen for further qRT-PCR expression patterns, which confirmed the cDNA-AFLP profiles. 【Conclusion】 Endophytic bacteria and plant interaction involved in multifaceted biochemical and physiological reactions, including concerted regulation of the genes involved in different pathways, like disease/defense, transcription, protein metabolism, signal transduction, as well as abiotic stresses. These results provide information for further elucidation of molecular mechanism of endophytic bacteria and plant interaction.

Key words: Capsicum annuum , Bacillus amyloliquefaciens , interaction , gene expression , cDNA-AFLP , qRT-PCR

YANG Rui-Xian, FAN Xiao-Jing, QIU Si-Xin, CAI Xue-Qing, HU Fang-Ping. cDNA-AFLP Analysis of Differential Gene Expression in Pepper Inoculated with Endophytic Bacillus amyloliquefaciens Fy11[J].Scientia Agricultura Sinica, 2013, 46(12): 2449-2458.

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cDNA-AFLP Analysis of Differential Gene Expression in Pepper Inoculated with Endophytic Bacillus amyloliquefaciens Fy11

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