Diversity and Growth Promotion of Endophytic Bacteria Isolated from Passiflora edulia Sims

doi:10.13560/j.cnki.biotech.bull.1985.2021-0144

Abstract

Abstract:

Endophytes can provide nutrients for plants and also promote plant growth through metabolites. Currently,there are few studies on endophytes in Passiflora edulia Sims. P. edulia Sims is a herbaceous vine of the Passiflora genus of the Passiflora family and mainly distributed in tropical and subtropical regions. By isolating and purifying the endophytes of P. edulia Sims,the obtained strains were clustered according to the IS-PCR results,the 16S rRNA genes of the representative strains were sequenced and phylogenetic analysis was performed,and their nitrogenase activity,auxin-producing capacity,phosphorus-dissolving ability,potassium-dissolving ability,protease production ability and physiological and biochemical characteristics were tested. The isolated and purified 51 strains of P. edulia Sims endophytic bacteria,clustered into 12 groups,belonging to Bacillus altitudinis,Bacillus circulans,Lysinibacillus macrolides,Brevibacillus antibioticus,Paenibacillus illinoisensis,Microbacterium zeae,Rhizobium pusense,Beijerinckia fluminensis,Achromobacter mucicolens,Stenotrophomonas maltophilia,Klebsiella michiganensis and Klebsiella pneumonia,the growth-promoting characteristics of the isolated endophytes were determined,and the results showed that the isolated representative endophytes had abundant functions,10 of them had the ability of secreting auxin,5 of them had nitrogen-fixing function,6 of them had phosphorus-dissolving function,7 of them had potassium-dissolving ability,3 of them had the ability of secreting siderophores,and 5 of them had the ability of producing protease. This experiment shows that the endophytes isolated from P. edulia Sims have both population diversity and functional diversity. These strains with different characteristics are expected to be utilized in agricultural production to exert their values.

Key words: Passiflora edulia Sims, endophytic bacteria, nitrogen fixation, auxin production, growth-promoting effect

TANG Jia-cheng, LIANG Yi-min, MA Jia-si, PENG Gui-xiang, TAN Zhi-yuan. Diversity and Growth Promotion of Endophytic Bacteria Isolated from Passiflora edulia Sims[J]. Biotechnology Bulletin, 2022, 38(1): 86-97.

Figures/Tables 9

References 37

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类群 Group	代表菌株（GenBank登录号） Representative strain（GenBank accession number）	最相似菌株名称（GenBank登录号） Closely related strain（GenBank accession number）	相似性 Similarity/%
Ⅰ	BXG95（MW714902）	Bacillus altitudinis 41KF2b^T（NR_042337）	98.20
Ⅱ	BXG129（MW714912）	Bacillus circulans strain NBRC 13626^T（NR_112632）	97.99
Ⅲ	BXG111（MW714903）	Lysinibacillus macroides strain LMG 18474^T（NR_114920）	98.42
Ⅳ	BXG92（MW714901）	Brevibacillus antibioticus strain TGS2-1^T（NR_165725）	98.70
Ⅴ	BXY92（MW714907）	Paenibacillus illinoisensis strain NBRC 15959^T（NR_113828）	97.49
Ⅵ	BXJ201（MW714905）	Microbacterium zeae strain 1204^T（NR_149816）	97.61
Ⅶ	BXG81（MW714904）	Rhizobium pusense strain NRCPB10^T（NR_116874）	99.00
Ⅷ	BXG212（MW714911）	Beijerinckia fluminensis strain UQM 1685^T（NR_116306）	98.57
Ⅸ	BXG101（MW714910）	Achromobacter mucicolens strain R-46658^T（NR_117613）	98.13
Ⅹ	BXG201（MW714909）	Stenotrophomonas maltophilia strain ATCC 19861^T（NR_040804）	98.29
Ⅺ	BXJ71（MW714906）	Klebsiella michiganensis strain W14^T（NR_118335）	97.13
Ⅻ	BXG53（MW714908）	Klebsiella pneumoniae strain DSM 30104^T（NR_117683）	98.62

类群 Group	代表菌株（GenBank登录号） Representative strain（GenBank accession number）	最相似菌株名称（GenBank登录号） Closely related strain（GenBank accession number）	相似性 Similarity/%
Ⅰ	BXG95（MW714902）	Bacillus altitudinis 41KF2b^T（NR_042337）	98.20
Ⅱ	BXG129（MW714912）	Bacillus circulans strain NBRC 13626^T（NR_112632）	97.99
Ⅲ	BXG111（MW714903）	Lysinibacillus macroides strain LMG 18474^T（NR_114920）	98.42
Ⅳ	BXG92（MW714901）	Brevibacillus antibioticus strain TGS2-1^T（NR_165725）	98.70
Ⅴ	BXY92（MW714907）	Paenibacillus illinoisensis strain NBRC 15959^T（NR_113828）	97.49
Ⅵ	BXJ201（MW714905）	Microbacterium zeae strain 1204^T（NR_149816）	97.61
Ⅶ	BXG81（MW714904）	Rhizobium pusense strain NRCPB10^T（NR_116874）	99.00
Ⅷ	BXG212（MW714911）	Beijerinckia fluminensis strain UQM 1685^T（NR_116306）	98.57
Ⅸ	BXG101（MW714910）	Achromobacter mucicolens strain R-46658^T（NR_117613）	98.13
Ⅹ	BXG201（MW714909）	Stenotrophomonas maltophilia strain ATCC 19861^T（NR_040804）	98.29
Ⅺ	BXJ71（MW714906）	Klebsiella michiganensis strain W14^T（NR_118335）	97.13
Ⅻ	BXG53（MW714908）	Klebsiella pneumoniae strain DSM 30104^T（NR_117683）	98.62

特性 Property	BXG95	BXG129	BXG111	BXG92	BXY92	BXJ201	BXG81	BXG212	BXG101	BXG201	BXJ71	BXG53
革兰氏染色 Gram stain	+	+	-	+	+	+	-	-	-	-	-	-
过氧化氢酶 Catalase	+	+	+	+	+	+	+	+	+	+	+	+
甲基红 Methyl red	+	+	-	+	+	+	+	+	-	-	-	+
产氨 Ammonia production	-	+	-	+	-	+	+	+	+	+	+	+
脲酶 Urease	-	+	-	-	-	+	+	+	+	+	-	+
乙酰甲基甲醇实验 V.P. test	-	-	+	-	-	-	-	-	+	+	+	-
NO₃^-还原测定 Nitrate reduction	-	+	-	+	-	-	+	-	+	+	+	+
明胶液化 Gelatin hydrolysis	+	+	-	-	+	-	-	-	-	+	+	-
固氮酶活性 Nitrogenase activity /（nmol C₂H₄/（mL·h））	0	0	0	0	0	0	23.97±0.43	100.94±0.89	0	21.11±0.66	477.93±3.85	191.33±1.27

特性 Property	BXG95	BXG129	BXG111	BXG92	BXY92	BXJ201	BXG81	BXG212	BXG101	BXG201	BXJ71	BXG53
革兰氏染色 Gram stain	+	+	-	+	+	+	-	-	-	-	-	-
过氧化氢酶 Catalase	+	+	+	+	+	+	+	+	+	+	+	+
甲基红 Methyl red	+	+	-	+	+	+	+	+	-	-	-	+
产氨 Ammonia production	-	+	-	+	-	+	+	+	+	+	+	+
脲酶 Urease	-	+	-	-	-	+	+	+	+	+	-	+
乙酰甲基甲醇实验 V.P. test	-	-	+	-	-	-	-	-	+	+	+	-
NO₃^-还原测定 Nitrate reduction	-	+	-	+	-	-	+	-	+	+	+	+
明胶液化 Gelatin hydrolysis	+	+	-	-	+	-	-	-	-	+	+	-
固氮酶活性 Nitrogenase activity /（nmol C₂H₄/（mL·h））	0	0	0	0	0	0	23.97±0.43	100.94±0.89	0	21.11±0.66	477.93±3.85	191.33±1.27

特性Characteristics	BXG95	BXG129	BXG111	BXG92	BXY92	BXJ201	BXG81	BXG212	BXG101	BXG201	BXJ71	BXG53
可溶性磷含量 Soluble P content/（mg·L^-1）	-	-	9.75	-	12.46	-	-	12.93	13.64	-	12.38	23.47
可溶性钾含量 Soluble K content/（mg·L^-1）	-	22.4	-	-	45.2	42.7	29	23.6	-	-	41	65.6
生长素 IAA/（mg·L^-1）	24.32	28.71	-	3.36	12.38	-	7.55	8.91	20.97	11.46	14.85	17.13
产铁载体 Siderophore production	-	-	-	-	-	-	+	-	-	-	+	+
产蛋白酶 Proteinase production	3.61	1.97	-	-	2.68	-	-	-	-	3.60	2.36	-