Diversity and Characteristics of Endophytic Bacteria Isolated from Peperomia dindygulensis Miq
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摘要: 采用菌落表征性状统计和16S rRNA基因序列分析法,对石蝉草根、茎、叶内生细菌的数量分布、群落组成和遗传多样性进行初步研究。从各组织器官中分离出内生细菌383株,通过菌落表征性状,以类平均连锁聚类法进行聚类分析,在欧氏距离为0的水平上可聚为19个亚群。多样性分析结果表明,19个亚群隶属于8属13种;其数量表现为叶>根>茎,多样性表现为茎>叶>根,其中鞘氨醇单胞菌属Sphingomonas和寡养单胞菌属Stenotrophomonas为石蝉草内生细菌的优势属。Abstract: Statistical classification of colony characteristics and 16S rRNA gene sequencing were performed to obtain the information on the quantitative distribution, community composition, and diversity of endophytic bacteria in the roots, stems and leaves of Peperomia dindygulensis Miq. A total of 383 endophytic bacteria were isolated and clustered according to their colony characteristics. After the average linkage clustering, the groups were divided into 19 subgroups at the Euclidean distance of 0 level. The subgroups were phylogenetically classified into 8 genera and 13 species based on their partial 16S rDNA sequences. The quantities of endophytic bacteria in the tissues were in order of leaves > roots > stems; and, the diversity of the bacteria, stems > leaves > roots. The predominant endophytic bacteria in P. dindygulensis Miq were found to belong to the genera of Sphingomonas and Stenotrophomonas.
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Key words:
- Peperomia dindygulensis Miq /
- endophytic bacteria /
- diversity /
- 16S rRNA
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表 1 亚群菌落形态特征
Table 1. Colonial morphology of sub-clusters
亚群 菌株 菌落形态 分离株数 所占比
例/%根 茎 叶 SA-1 Y1 点状、湿、黄色、有光泽、表面凸起、边缘整齐、不透明 2 20 244 69.45 SA-2 Y3 圆形、直径2~3 mm、湿、乳白、有光泽、表面凸起、边缘整齐、半透明 0 6 4 2.61 SA-3 G4 圆形、直径2~3 mm、湿、乳白、有光泽、表面凸起、边缘整齐、不透明 3 0 0 0.78 SA-4 J8 圆形、直径1~2 mm、湿、白色、有光泽、表面凸起、边缘整齐、不透明 0 4 0 1.04 SA-5 G7 圆形、直径1~2 mm、湿、乳黄、有光泽、表面凸起、边缘整齐、透明 51 0 0 13.32 SB-1 Y4 形状不规则、长7~8 mm、黏稠、乳白、无光泽、中间凸起、边缘不整齐、不透明 0 0 3 0.78 SB-2 J5 形状不规则、长6~8 mm、湿、乳白、无光泽、中间凸起、边缘不整齐、不透明 0 7 0 1.83 SB-3 G3 形状不规则、长7~8 mm、黏稠、白色、无光泽、中间凸起、边缘不整齐、不透明 10 0 0 2.61 SB-4 Y5 形状不规则、长6~7 mm、干、白色、无光泽、表面凸起褶皱、边缘不整齐、不透明 0 0 1 0.26 SB-5 J7 形状不规则、长5~7 mm、干、乳白、无光泽、表面凸起褶皱、边缘不整齐、不透明 0 1 0 0.26 SB-6 G6 形状不规则、长5~6 mm、干、乳白、无光泽、中间凸起、边缘不整齐、不透明 1 0 0 0.26 SC-1 Y8 形状不规则、长3~4 mm、湿、乳白、有光泽、表面凸起褶皱、边缘不整齐、半透明 0 0 1 0.26 SD-1 Y6 圆形、直径4~5 mm、黏稠、乳白、无光泽、表面内凹、边缘整齐、不透明 0 0 1 0.26 SD-2 Y9 圆形、直径4~5 mm、湿、乳白、有光泽、中间凸起、边缘整齐、半透明 0 0 4 1.04 SD-3 J4 圆形、直径5~6 mm、黏稠、乳白、有光泽、中间凸起、边缘整齐、半透明 0 6 0 1.57 SD-4 G5 圆形、直径4~5 mm、湿、白色、有光泽、中间凸起、边缘整齐、不透明 1 0 0 0.26 SD-5 J9 圆形、直径3~4 mm、湿、乳白、有光泽、表面内凹、边缘整齐、半透明 0 1 0 0.26 SD-6 Y7 圆形、直径5~6 mm、干、乳白、无光泽、表面扁皱、边缘整齐、半透明 0 1 1 0.52 SE-1 G1 水渍状、长25 mm、湿、乳白、无光泽、表面凹凸不平、边缘不整齐、半透明 8 1 1 2.61 表 2 石蝉草内生细菌的种类组成
Table 2. Species of endophytic bacteria isolated from P. dindygulensis Miq
菌株 相近的属、种 登录号 序列相
似性/%G6 J7 Y5 Bacillus subtilis KT957306.1 100 Y8 Bacillus licheniformis KM289130.1 100 Y7 Bacillus altitudinis KT758448.1 100 Y6 Bacillus pumilus KT003271.1 100 J5 Bacillus anthracis AB190217.1 98 G1 G3 Y4 Bacillus cereus KC692201.1 99~100 J8 Staphylococcus haemolyticus JX293294.1 97 G4 Viridibacillus arenosi JF899298.1 100 G5 J4 Y9 Lysinibacillus fusiformis KP334988.1 99 J9 Paenibacillus sp KF011612.1 99 G7 Stenotrophomonas rhizophila KM096602.1 100 Y3 Rhizobium sp KR067598.1 100 Y1 Sphingomonas aeria KJ572399.1 99 -
[1] 刘学周, 赵智灵, 李绍宾, 等.西洋参内生菌群落结构与多样性[J].微生物学报, 2015, 55(3):330-340. http://www.cnki.com.cn/Article/CJFDTOTAL-WSXB201503013.htm [2] 周佳宇, 贾永, 王宏伟, 等.茅苍术叶片可培养内生细菌多样性及其促生潜力[J].生态学报, 2013, 33(4):1106-1117. http://www.cnki.com.cn/Article/CJFDTOTAL-STXB201304011.htm [3] 杜晓宁, 徐惠娟, 黄盼盼, 等.宁夏枸杞内生细菌的多样性及其抑菌活性研究[J].微生物学通报, 2015, 42(9):1779-1787. http://www.cnki.com.cn/Article/CJFDTOTAL-WSWT201509021.htm [4] 朱妍妍, 艾嫦, 张嘉, 等.具有生物活性的植物内生真菌次生代谢产物[J].化学进展, 2011, 23(4):704-730. http://www.cnki.com.cn/Article/CJFDTOTAL-HXJZ201104012.htm [5] 王娜.乌头内生细菌群落结构及其与宿主生物碱合成关系研究[D].杨凌:陕西师范大学, 2014. http://cdmd.cnki.com.cn/Article/CDMD-10718-1014398431.htm [6] 梁子宁, 赖开平, 朱华, 等.药用植物鸦胆子内生细菌分离及其抑菌活性研究[J].南方农业学报, 2014, 45(3):389-394. http://www.cnki.com.cn/Article/CJFDTOTAL-GXNY201403008.htm [7] 陈立.石蝉草化学成分及抗肿瘤活性的研究[D].北京:中国人民解放军军事医学科学院, 2007. http://cdmd.cnki.com.cn/Article/CDMD-90106-2007121376.htm [8] 朱文君, 林梦感, 杨国红, 等.石蝉草中两个新的聚酮类化合物[J].中草药, 2011, 42(3):420-423. http://www.cnki.com.cn/Article/CJFDTOTAL-ZCYO201103003.htm [9] 方中达.植病研究方法[M].北京:中国农业出版社, 1998:47. [10] 陈泽斌, 夏振远, 雷丽萍, 等.烟草可培养内生细菌的分离及多样性分析[J].微生物学通报, 2011, 38(9):1347-1354. http://www.cnki.com.cn/Article/CJFDTOTAL-WSWT201109006.htm [11] DEVEREUX R, HINES M E, STAHL D A. S cycling:characterization of natural communities of sulfate-reducing bacteria by 16S rRNA sequence comparisons[J]. Microb Ecol, 1996, 32(3):283-292. https://www.ncbi.nlm.nih.gov/pubmed/8849423 [12] FELSENSTEIN J. Confidence limits on phylogenies:An approach using the bootstrap[J]. Evolution, 1985, 39:783-791. doi: 10.2307/2408678 [13] 王志刚, 胡云龙, 徐伟慧, 等.鞘氨醇单胞菌菌株CL01的分离鉴定及其对连作西瓜的促生效应[J].农业生物技术学报, 2015, 23(10):1360-1367. http://www.cnki.com.cn/Article/CJFDTOTAL-NYSB201510012.htm [14] 傅晓方, 韩红江, 郝勇锋, 等.玉米内生固氮菌的分离鉴定及对小麦幼苗的促生效应[J].西北农业学报, 2012, 21(1):66-71. http://www.cnki.com.cn/Article/CJFDTOTAL-XBNX201201017.htm [15] 台喜生, 冯佳丽, 李梅, 等.鞘氨醇单胞菌在生物降解方面的研究进展[J].湖南农业科学, 2011, (7):21-25. http://www.cnki.com.cn/Article/CJFDTOTAL-HNNK201107008.htm [16] 欧阳主才, 王奕恒, 李小妮, 等.鞘氨醇单胞菌株XJ对农药的降解效能[J].华南农业大学学报, 2008, 29(2):47-49. http://www.cnki.com.cn/Article/CJFDTOTAL-HNNB200802012.htm [17] ANDREOTE F D, ROCHA U N, ARAVJO W L. Effect of bacterial inoculation plant genotype and develop mental stage on root-associated and endophytic bacterial communities in potato (Solanum tuberosum L.)[J]. Antonie Van Leeuwenhoek, 2010, 97(4):389-399. doi: 10.1007/s10482-010-9421-9 [18] LEO V O, JAN D E. Effects of plant genotype and growth stage on the structure of bacterial communities associated with potato (Solanum tuberosum L.)[J]. FEMS Microbiology Ecology, 2008, 64(2):183-296. https://www.researchgate.net/publication/5497803_Effects_of_plant_genotype_and_growth_stage_on_the_structure_of_bacterial_communities_associated_with_potato_Solanum_tuberosum_L [19] ROSENBLUETH M, MARTÍNEZ R E. Bacterial endophytes and their interactions with hosts[J]. Molecular Plant-Microbel Interactions, 2006, 19(8):827-837. doi: 10.1094/MPMI-19-0827 [20] HOLLIS J P. Bacteria in the healthy potato tissue[J]. Phytopathology, 1951, (41):197-209. http://digitalcommons.unl.edu/dissertations/AAIDP13795/ [21] AMMANN R R, LUDWIG W, SCHLEIFER K H.Phylogenetic identification and in situ detection of individual microbial cells without cultivation[J]. Microbiol Rev. 1995, 59(1):143-169. https://www.ncbi.nlm.nih.gov/pubmed/7535888