林木根际细菌JYZ-SD5的促生抗逆性能及种类鉴定
|
摘要
旨在探究林木根际细菌JYZ-SD5促生抗逆性能,以期揭示该菌在野外应用的价值。采用特定培养基定性检测该菌的固氮解磷解钾特性,测定该菌产IAA能力;平板对峙法检测菌株拮抗病原真菌活性;检测该菌重金属耐受性;将根际细菌JYZSD5施用于水杉盆栽幼苗,观测对水杉生长的影响;采用形态、生理生化测试及16S rRNA基因序列分析,对菌株JYZ-SD5进行种类鉴定。结果显示,菌株JYZ-SD5具有固氮、解磷、解钾的能力;不加色氨酸前提下产IAA量可达6.818 6μg/mL;对链格孢(Alternaria sp.)、异色状拟盘多毛孢(Pestalotiopsis vesicolor)有一定拮抗活性,可产生蛋白酶和纤维素酶;对重金属Mn7+、Cr6+耐受性强,最高均可达400 mg/L;温室盆栽试验表明,根际细菌JYZ-SD5可提高水杉苗木叶绿素相对含量,对水杉株高和地径增长具有促进作用。结合形态学、生理生化和分子鉴定的方法初步确定该菌为拟蕈状芽孢杆菌(Bacillus paramycoides)。根际细菌JYZ-SD5具有良好的促生抗逆特性,具有成为新型生物菌肥的开发潜力和野外应用价值。
This work aims to explore the performance of promoting growth and stress resistance of JYZ-SD5,a tree rhizobacterium,and to reveal the value of this bacterium in the field. Specific culture medium was used to detect its ability of nitrogen fixation and decomposition of phosphorus and potassium,also to measure its capacity of producing IAA. Then plate confrontation was employed to check the antagonistic activity to pathogenic fungi,and growth was observed to determine its tolerance to heavy metals. Further,the tree rhizobacterium JYZ-SD5 was applied to the seedlings of Metasequoia glyptostroboides Hu et Cheng in plots to test its effects on the growth of the seedlings. Morphology observation,physiological and biochemical tests,and 16 S rRNA gene sequence analysis were combined to identify the strain JYZ-SD5.Results showed that JYZ-SD5 had the ability of fixing nitrogen and decomposing phosphorus and potassium. The production of IAA without adding tryptophan reached 6.818 6 μg/mL. JYZ-SD5 presented the antagonistic activity to Alternaria sp and Pestalotiopsis vesicolor,and protease and cellulose were also detected. This strain also demonstrated strong tolerance to heavy metals Mn7+ and Cr6+,to the highest content of400 mg/L. The greenhouse plot experiment revealed that JYZ-SD5 resulted in the increase of relative content of chlorophyll in the seedlings of M.glyptostroboides Hu et Cheng,and it promoted the growth and ground diameter of M.glyptostroboides Hu et Cheng. Finally,the rhizobacterium was identified as Bacillus paramycoides by morphological,physiological,biochemical,and molecular approaches. In sum,the rhizobacterium JYZ-SD5 shows favorable performance on promoting growth and stress resistance,and thus it has the potential to be a novel biological fertilizer and meets the requirement of applying it in field.
This work aims to explore the performance of promoting growth and stress resistance of JYZ-SD5,a tree rhizobacterium,and to reveal the value of this bacterium in the field. Specific culture medium was used to detect its ability of nitrogen fixation and decomposition of phosphorus and potassium,also to measure its capacity of producing IAA. Then plate confrontation was employed to check the antagonistic activity to pathogenic fungi,and growth was observed to determine its tolerance to heavy metals. Further,the tree rhizobacterium JYZ-SD5 was applied to the seedlings of Metasequoia glyptostroboides Hu et Cheng in plots to test its effects on the growth of the seedlings. Morphology observation,physiological and biochemical tests,and 16 S rRNA gene sequence analysis were combined to identify the strain JYZ-SD5.Results showed that JYZ-SD5 had the ability of fixing nitrogen and decomposing phosphorus and potassium. The production of IAA without adding tryptophan reached 6.818 6 μg/mL. JYZ-SD5 presented the antagonistic activity to Alternaria sp and Pestalotiopsis vesicolor,and protease and cellulose were also detected. This strain also demonstrated strong tolerance to heavy metals Mn7+ and Cr6+,to the highest content of400 mg/L. The greenhouse plot experiment revealed that JYZ-SD5 resulted in the increase of relative content of chlorophyll in the seedlings of M.glyptostroboides Hu et Cheng,and it promoted the growth and ground diameter of M.glyptostroboides Hu et Cheng. Finally,the rhizobacterium was identified as Bacillus paramycoides by morphological,physiological,biochemical,and molecular approaches. In sum,the rhizobacterium JYZ-SD5 shows favorable performance on promoting growth and stress resistance,and thus it has the potential to be a novel biological fertilizer and meets the requirement of applying it in field.
引文
[1]Ambardar S, Vakhlu J. Plant growth promoting bacteria from Crocus sativus rhizosphere[J]. World Journal of Microbiology&Biotechnology, 2013, 29(12):2271-2279.
[2]SabatéDC, Brandan CP, Petroselli G, et al. Decrease in the incidence of charcoal root rot in common bean(Phaseolus vulgaris L.)by Bacillus amyloliquefaciens, B14, a strain with PGPR properties[J]. Biological Control, 2017, 113:1-8.
[3]Mayak S, Tirosh T, Glick BR. Plant growth-promoting bacteria confer resistance in tomato plants to salt stress[J]. Plant Physiology&Biochemistry, 2004, 42(6):565-572.
[4]Dinesh R, Srinivasan V, Hamza S, et al. Isolation and characterization of potential Zn solubilizing bacteria from soil and its effects on soil Zn release rates, soil available Zn and plant Zn content[J].Geoderma, 2018, 321:173-186.
[5]Islam MR, Sultana T, Joe MM, et al. Nitrogen-fixing bacteria with multiple plant growth-promoting activities enhance growth of tomato and red pepper[J]. Journal of Basic Microbiology, 2013, 53(12):1004-1015.
[6]Li Y, Liu X, Hao T, et al. Colonization and maize growth promotion induced by phosphate solubilizingbacterialisolates[J].International Journal of Molecular Sciences, 2017, 18(7):1253.
[7]Liu H, Wu X, Ren J, et al. Phosphate-dissolving characteristics and growth promoting effect of Pseudomonads fluorescent JW-JSI on poplar seedlings[J]. Scientia Silvae Sinicae, 2013, 49(9):112-118.
[8]宋芳旭,吴小芹,赵群.水拉恩氏菌JZ-GX1对杨树溃疡病菌的拮抗作用[J].南京林业大学学报:自然科学版, 2017, 41(4):42-48.
[9]Sengupta K, Maiti TK, Saha P. Degradation of 4-nitrophenol in presence of heavy metals by a halotolerant Bacillus sp. strain BUPNP2, having plant growth promoting traits[J]. Symbiosis,2015, 65(3):157-163.
[10]Kuramshina ZM, Smirnova YV, Khairullin RM. Cadmium and nickel toxicity for Sinapis alba, plants inoculated with endophytic strains of Bacillus subtilis[J]. Russian Journal of Plant Physiology, 2018, 65(2):269-277.
[11]宋金秋,刘淑娇,崔丽红,等.根际细菌溶磷、产IAA及其抑菌作用的研究[J].基因组学与应用生物学, 2017(11):4722-4728.
[12]Xu J, Kloepper JW, Huang P, et al. Isolation and characterization of N2-fixing bacteria from giant reed and switchgrass for plant growth promotion and nutrient uptake[J]. Journal of Basic Microbiology,2018, 58:459-471.
[13]韩晓阳,周波,董玉惠,等.山东茶园土壤高活性解钾细菌的筛选鉴定及肥效研究[J].茶叶科学, 2018, 38(1):78-86.
[14]李进,钟志军,苏怀益,等.大熊猫肠道芽孢杆菌的分离鉴定及部分生物学特性[J].微生物学通报, 2016, 43(2):351-359.
[15]李伟,龚劲松,李恒,等.一株产蛋白酶不动杆菌的分离与酶学性质[J].食品与生物技术学报, 2017, 36(1):8-14.
[16]MeloAL,SoccolVT,SoccolCR.Bacillusthuringiensis:mechanism of action, resistance, and new applications:a review[J]. Critical Reviews in Biotechnology, 2016, 36(2):317.
[17]Pérezflores P, Valenciacantero E, Altamiranohernández J, et al.Bacillus methylotrophicus M4-96 isolated from maize(Zea mays)rhizoplane increases growth and auxin content in Arabidopsis thaliana via emission of volatiles[J]. Protoplasma, 2017:1-13.
[18]Yasmi A, Radziah O, Hawa ZE, et al. Characterisation of plant growth-promoting bacteria from Kacip Fatimah(Labisia pumila)under natural tropical forest[J]. Universiti Putra Malaysia Press,2016, 39(4):557-575.
[19]Kuffner M, Puschenreiter M, Wieshammer G, et al. Rhizosphere bacteriaaffectgrowthandmetaluptakeofheavymetal accumulating willows[J]. Plant&Soil, 2008, 304(1/2):35-44.
[20]Krishnendu P, Soumik M, Anumita S, Tushar KM. Alleviation of phytotoxic effects of cadmium on rice seedlings by cadmium resistant PGPR strain Enterobacter aerogenes MCC 3092[J].Journal of Hazardous Materials, 2018, 351:317-329.
[21]Elshafie HS, Ippolito C, Rocco R, et al. In vitro antifungal activity of Burkholderia gladiolipv. agaricicola against some phytopathogenic fungi[J]. International Journal of Molecular Sciences, 2012, 13(12):16291-16302.
[22]Liu Y, Du J, Lai Q, et al. Proposal of nine novel species of the Bacillus cereus group[J]. International Journal of Systematic&Evolutionary Microbiology, 2017, 67(8):2499.
[2]SabatéDC, Brandan CP, Petroselli G, et al. Decrease in the incidence of charcoal root rot in common bean(Phaseolus vulgaris L.)by Bacillus amyloliquefaciens, B14, a strain with PGPR properties[J]. Biological Control, 2017, 113:1-8.
[3]Mayak S, Tirosh T, Glick BR. Plant growth-promoting bacteria confer resistance in tomato plants to salt stress[J]. Plant Physiology&Biochemistry, 2004, 42(6):565-572.
[4]Dinesh R, Srinivasan V, Hamza S, et al. Isolation and characterization of potential Zn solubilizing bacteria from soil and its effects on soil Zn release rates, soil available Zn and plant Zn content[J].Geoderma, 2018, 321:173-186.
[5]Islam MR, Sultana T, Joe MM, et al. Nitrogen-fixing bacteria with multiple plant growth-promoting activities enhance growth of tomato and red pepper[J]. Journal of Basic Microbiology, 2013, 53(12):1004-1015.
[6]Li Y, Liu X, Hao T, et al. Colonization and maize growth promotion induced by phosphate solubilizingbacterialisolates[J].International Journal of Molecular Sciences, 2017, 18(7):1253.
[7]Liu H, Wu X, Ren J, et al. Phosphate-dissolving characteristics and growth promoting effect of Pseudomonads fluorescent JW-JSI on poplar seedlings[J]. Scientia Silvae Sinicae, 2013, 49(9):112-118.
[8]宋芳旭,吴小芹,赵群.水拉恩氏菌JZ-GX1对杨树溃疡病菌的拮抗作用[J].南京林业大学学报:自然科学版, 2017, 41(4):42-48.
[9]Sengupta K, Maiti TK, Saha P. Degradation of 4-nitrophenol in presence of heavy metals by a halotolerant Bacillus sp. strain BUPNP2, having plant growth promoting traits[J]. Symbiosis,2015, 65(3):157-163.
[10]Kuramshina ZM, Smirnova YV, Khairullin RM. Cadmium and nickel toxicity for Sinapis alba, plants inoculated with endophytic strains of Bacillus subtilis[J]. Russian Journal of Plant Physiology, 2018, 65(2):269-277.
[11]宋金秋,刘淑娇,崔丽红,等.根际细菌溶磷、产IAA及其抑菌作用的研究[J].基因组学与应用生物学, 2017(11):4722-4728.
[12]Xu J, Kloepper JW, Huang P, et al. Isolation and characterization of N2-fixing bacteria from giant reed and switchgrass for plant growth promotion and nutrient uptake[J]. Journal of Basic Microbiology,2018, 58:459-471.
[13]韩晓阳,周波,董玉惠,等.山东茶园土壤高活性解钾细菌的筛选鉴定及肥效研究[J].茶叶科学, 2018, 38(1):78-86.
[14]李进,钟志军,苏怀益,等.大熊猫肠道芽孢杆菌的分离鉴定及部分生物学特性[J].微生物学通报, 2016, 43(2):351-359.
[15]李伟,龚劲松,李恒,等.一株产蛋白酶不动杆菌的分离与酶学性质[J].食品与生物技术学报, 2017, 36(1):8-14.
[16]MeloAL,SoccolVT,SoccolCR.Bacillusthuringiensis:mechanism of action, resistance, and new applications:a review[J]. Critical Reviews in Biotechnology, 2016, 36(2):317.
[17]Pérezflores P, Valenciacantero E, Altamiranohernández J, et al.Bacillus methylotrophicus M4-96 isolated from maize(Zea mays)rhizoplane increases growth and auxin content in Arabidopsis thaliana via emission of volatiles[J]. Protoplasma, 2017:1-13.
[18]Yasmi A, Radziah O, Hawa ZE, et al. Characterisation of plant growth-promoting bacteria from Kacip Fatimah(Labisia pumila)under natural tropical forest[J]. Universiti Putra Malaysia Press,2016, 39(4):557-575.
[19]Kuffner M, Puschenreiter M, Wieshammer G, et al. Rhizosphere bacteriaaffectgrowthandmetaluptakeofheavymetal accumulating willows[J]. Plant&Soil, 2008, 304(1/2):35-44.
[20]Krishnendu P, Soumik M, Anumita S, Tushar KM. Alleviation of phytotoxic effects of cadmium on rice seedlings by cadmium resistant PGPR strain Enterobacter aerogenes MCC 3092[J].Journal of Hazardous Materials, 2018, 351:317-329.
[21]Elshafie HS, Ippolito C, Rocco R, et al. In vitro antifungal activity of Burkholderia gladiolipv. agaricicola against some phytopathogenic fungi[J]. International Journal of Molecular Sciences, 2012, 13(12):16291-16302.
[22]Liu Y, Du J, Lai Q, et al. Proposal of nine novel species of the Bacillus cereus group[J]. International Journal of Systematic&Evolutionary Microbiology, 2017, 67(8):2499.