不同浓度吐温-80对烟草叶面微生物洗脱效果的影响  

Effects of Tween-80 at different concentrations on elution of microorganisms from tobacco leaf surface

作　　者：张振远 滕环瑜 潘影 王召军 张洪映[1] 闫筱筱 崔红[1] ZHANG Zhenyuan;TENG Huanyu;PAN Ying;WANG Zhaojun;ZHANG Hongying;YAN Xiaoxiao;CUI Hong(College of Tobacco Science,Henan Agricultural University,Zhengzhou 450046,China)

机构地区：[1]河南农业大学烟草学院,郑州市450046

出　　处：《烟草科技》2023年第12期7-13,共7页Tobacco Science & Technology

基　　金：中国烟草总公司烤烟基因组计划和生物育种重大专项项目“烤烟叶面化学成分的分子调控和定向改良”[110202101005(JY-05)]。

摘　　要：为建立高效的烟草叶面微生物收集体系,以高分泌型烟草T.I.1068为材料,分别采用浓度(体积分数)为0、2%、8%、10%、15%、20%、30%吐温-80(Tween-80)溶液进行叶面微生物的洗脱和收集,并通过平板培养和扩增子测序研究了吐温-80对高分泌型烟草的叶面微生物洗脱效果的影响。结果表明:(1)随着吐温-80浓度的提高,收集的可培养微生物数量呈现先增加后降低的趋势,15%吐温-80处理的细菌菌落密度最大,10%吐温-80处理的真菌菌落密度最大。(2)随着吐温-80浓度的增加,叶面微生物总DNA浓度呈现增长态势,吐温-80浓度达到20%后进入平台期。(3)16S rDNA扩增子测序分析发现,与无菌水相比,20%吐温-80洗脱的细菌物种丰富度提高,群落结构发生明显改变,黄单胞菌科(Xanthomonadaceae)、黄杆菌科(Xanthobacteraceae)、鞘脂菌科(Sphingomonadaceae)、根瘤菌科(Rhizobiaceae)、假单胞菌科(Pseudomonadaceae)等的丰度显著增加。(4)ITS rDNA扩增子测序分析发现,与无菌水相比,20%吐温-80洗脱的真菌物种丰富度下降,群落结构明显改变,白粉菌科(Erysiphaceae)和法夫酵母科(Phaffomycetaceae)等的丰度显著增加。表明吐温-80可提高烟草叶面微生物洗脱效率和微生物总DNA浓度,对叶面微生物培养以及群落结构和多样性的分析具有积极作用。To establish an efficient collection system for tobacco leaf surface microorganisms,high-secreting tobacco T.I.1068 was used as the research material.Solutions of Tween-80 at concentrations(volume fraction)of 0,2%,8%,10%,15%,20%,and 30%were used to elute and collect leaf surface microorganisms.The effects of Tween-80 on the elution of leaf surface microorganisms from high-secreting tobacco were studied by plate culture and amplicon sequencing.The results are as follows:1)With the increase of Tween-80 concentration,the number of cultivable microorganisms on the plate first increased and then decreased.The bacteria colony density was the highest under the treatment of 15%Tween-80,and that of fungi was the highest under the treatment of 10%Tween-80.2)With increasing Tween-80 concentration,the total DNA concentration of the foliar microorganisms showed an increasing trend and reached a plateau at 20%or above.3)16S rDNA amplicon sequencing analysis showed that the richness and evenness of bacterial species obtained by 20%Tween-80 elution were higher than those of aqueous solutions,but the structure of the community of bacteria changed,and the abundance of bacterial families such as Xanthomonadaceae,Xanthomonadaceae,Sphingomonadaceae,Rhizobiaceae and Pseudomonadaceae increased significantly.4)Amplicon sequencing and ITS rDNA analysis showed that the richness of fungal species obtained by 20%Tween-80 elution was lower than that of the aqueous solution,but the structure of the fungal community changed,and the abundance of Erysiphaceae and Phaffomycetaceae,the fungal families,increased significantly.In conclusion,Tween-80 could effectively improve the efficiency of microbial elution and the concentration of total microbial DNA,and had a positive effect on microbial culture and community structure and diversity analysis.

关 键 词：烟草 叶面微生物 吐温-80 微生物收集 扩增子测序 

分 类 号：TS414[农业科学—烟草工业]