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《Journal of Nuclear Agricultural Sciences》 2018-02
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Screening of Enrichment Methods of Endophytic Bacteria From Newhall Navel Orange and Satsuma Mandarin

WU Simeng;LIU Bing;JIANG Junxi;ZHOU Ying;YAN Mingfeng;College of Agriculture,Jiangxi Agriculture University;  
In order to explore the enrichment effect of different methods on citrus endophytic bacteria,the leaves and peels of Newhall Navel orange and Satsuma Mandarin were used as test materials,three methods for enriching the endophytic bacteria were compareed after extracting the macro genomicDNA of plant tissue and detecting the sequence on the Illumina Hi Seq platform. The result showed that compared with the untreated control,three treatments can effectively enrich the endophytic bacteria. When the sample tissues were pre-treated with solution A( 1. 5% Macerozyme R-10 + 1. 5% cellulase R-10 + 8. 55% sucrose),solution B( 1. 5% Macerozyme R-10 + 1. 5% cellulose R-10 + 12. 8% mannitol + 0. 12% MES + 0. 36% CaCl_2·2 H_2O + 0. 011% NaH_2PO_4) and solution C( 8. 55% sucrose) respectively,the proportion of chloroplast DNA to endophytic bacterial genomicDNA decreased,and the lowest chloroplast DNA proportion was obtained in the sample tissues treated with solution A or B. The chloroplast DNA proportion in endophytic bacterial genomicDNA of NNO leave and SM peel treated with solution A were 7. 34% and 74. 89% respectively,while that of NNO peel and SM leaves treated with Solution B were 36. 07% and 9. 65% respectively. Compared to the control,the number of OTU taxa and the amount of Proteobacteria and Firmicutes from the samples treated with three solutions increased,and more types of endogenous bacteria were identified. Three methods are effective at a certain level for the endogenous bacterial enrichment among which solution A or B treatment was the best,the endophytic bacteria were able to be enriched effectively and the interference of plant chloroplast decreased when the genomicDNA of bacteria from citrus was extracted. The results can provide reference for the research of the endophytic bacteria of citrus and other plant.
【Fund】: 国家自然科学基金项目(31460139)
【CateGory Index】: S436.66
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