Screening of Enrichment Methods of Endophytic Bacteria From Newhall Navel Orange and Satsuma Mandarin
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 genomic DNA of plant tissue and detecting the sequence on the Illumina HiSeq 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％ CaC12·2H2O + 0.011％ NaH2PO4) and solution C (8.55％ sucrose) respectively,the proportion of chloroplast DNA to endophytic bacterial genomic DNA 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 genomic DNA 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 genomic DNA of bacteria from citrus was extracted.The results can provide reference for the research of the endophytic bacteria of citrus and other plant.
国家科技期刊平台
NSTL
万方数据
维普
