生物技术通报 ›› 2020, Vol. 36 ›› Issue (10): 127-134.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0236
收稿日期:
2020-03-07
出版日期:
2020-10-26
发布日期:
2020-11-02
作者简介:
杨海霞,女,研究方向:生物技术;E-mail: 基金资助:
YANG Hai-xia(), LIU Xi-min, PAN Yi-chen, ZHAO Xiang-lin, HUANG Hai-dong()
Received:
2020-03-07
Published:
2020-10-26
Online:
2020-11-02
摘要:
为筛选适应滨海盐碱地环境条件的溶磷菌,从天津滨海新区盐碱土壤中分离得到1株耐盐碱溶磷菌Y2R2,多相分类鉴定结果表明其为盐单胞菌属(Halomonas)的一个新种。菌株Y2R2的生长温度范围为10-45℃,能够耐受pH 14和NaCl浓度15%的生长条件。在低于pH 13和NaCl浓度12%的条件下,菌株Y2R2具有较强的溶磷能力,在pH 8和NaCl浓度9%的无机磷培养基中,发酵48 h溶磷能力最高,达到247.6 mg/L。筛选到的菌株Y2R2为适应滨海盐碱地土壤和气候环境的土著微生物新种,具有较高的耐盐碱和溶磷能力,有望用于耐盐碱溶磷菌剂的开发。
杨海霞, 刘希旻, 潘奕臣, 赵香林, 黄海东. 耐盐碱溶磷菌Y2R2的分离鉴定及溶磷特性[J]. 生物技术通报, 2020, 36(10): 127-134.
YANG Hai-xia, LIU Xi-min, PAN Yi-chen, ZHAO Xiang-lin, HUANG Hai-dong. Isolation,Identification and Characterization of Salt-alkali-tolerant and Phosphorus-dissolving Bacterium Y2R2[J]. Biotechnology Bulletin, 2020, 36(10): 127-134.
图4 菌株Y2R2的全细胞脂肪酸分析 主要的脂肪酸组分为2.676 min的C16:0,3.581 min的C19:0 cyclo w8c,3.249 min的C18:1 w7c/C18:1 w6c,2.625 min的C16:1w7c/C16:1w6c;其他为含量较低的脂肪酸组分,包括2.960 min的C17:0 cyclo,1.923 min的C12:0 3OH,1.538 min的C12:0,1.147 min的C10:0
鉴定项目 | 菌株 | 鉴定项目 | 菌株 | |||||
---|---|---|---|---|---|---|---|---|
Y2R2 | H. huangheensis | Y2R2 | H. huangheensis | |||||
Catalase | + | + | Valine arylamidase | + | + | |||
Urease | + | - | Acid phosphatase | + | + | |||
H2S production | - | - | β-glucuronidase | - | - | |||
Aesculin hydrolyzation | + | + | a-mannosidase | - | + | |||
Casein hydrolyzation | - | + | Esterase lipase(C8) | + | + | |||
Tyrosine hydrolyzation | + | - | Cystine arylamidase | + | + | |||
Nitrate reduction | + | + | Naphthol-AS-BI-phosphohydrolase | + | + | |||
Alkaline phosphatase | + | + | a-glucosidase | - | + | |||
Leucine arylamidase | + | + | b-fucosidase | - | - | |||
Chymotrypsin | - | + | Esterase(C14) | w | - | |||
b-galactosidase | - | + | Trypsin | w | + | |||
N-acetyl-b-glucosaminidase | - | - | a-galactosidase | - | + | |||
Lipid enzyme(C4) | + | + | b-glucosidase | - | + |
表1 菌株Y2R2与H. huangheensis的生理生化特征比较
鉴定项目 | 菌株 | 鉴定项目 | 菌株 | |||||
---|---|---|---|---|---|---|---|---|
Y2R2 | H. huangheensis | Y2R2 | H. huangheensis | |||||
Catalase | + | + | Valine arylamidase | + | + | |||
Urease | + | - | Acid phosphatase | + | + | |||
H2S production | - | - | β-glucuronidase | - | - | |||
Aesculin hydrolyzation | + | + | a-mannosidase | - | + | |||
Casein hydrolyzation | - | + | Esterase lipase(C8) | + | + | |||
Tyrosine hydrolyzation | + | - | Cystine arylamidase | + | + | |||
Nitrate reduction | + | + | Naphthol-AS-BI-phosphohydrolase | + | + | |||
Alkaline phosphatase | + | + | a-glucosidase | - | + | |||
Leucine arylamidase | + | + | b-fucosidase | - | - | |||
Chymotrypsin | - | + | Esterase(C14) | w | - | |||
b-galactosidase | - | + | Trypsin | w | + | |||
N-acetyl-b-glucosaminidase | - | - | a-galactosidase | - | + | |||
Lipid enzyme(C4) | + | + | b-glucosidase | - | + |
[1] | 林欣, 王立立, 杨平, 等. 一株溶磷菌对盐地碱蓬修复盐渍土Cd污染的促进效应[J]. 农业环境科学学报, 2018,37(5):883-891. |
Lin X, Wang LL, Yang P, et al. The promoting effect of phosphorus-solubilizing bacteria on remediation of cadmiumcontaminated saline soil by Suaeda salsa[J]. Journal of Agro-Environment Science, 2018,37(5):883-891. | |
[2] | 余海英, 孔亚平, 张科利, 等. 融雪剂在路域土壤中的累积、扩散及其对土壤性质的影响[J]. 水土保持学报, 2009,23(6):182-185. |
Yu HY, Kong YP, Zhang KL, et al. Accumulation and translocation of deicing salts in roadside soils and their effects on soil properties[J]. Journal of Soil and Water Conservation, 2009,23(6):182-185. | |
[3] | 王海江. 玛纳斯河流域土壤盐渍化过程和格局特征及盐渍土改良模式探讨[D]. 北京:中国农业大学, 2014. |
Wang HJ. Processes, patterns and reclamation of soil salinization at Manas river basin, Xinjiang, China[D]. Beijing:China Agricultural University, 2014. | |
[4] | 孟祥坤, 于新, 朱超, 等. 解磷微生物研究与应用进展[J]. 华北农学报, 2018,33(S1):208-214. |
Meng XK, Yu X, Zhu C, et al. Research an application advances in phosphate-solubilizing-microorganisms[J]. Acta Agriculturae Boreali-sinica, 2018,33(S1):208-214. | |
[5] | 陈莎莎, 孙敏, 王文超, 等. 溶磷真菌固体发酵菌肥对玉米生长及根际细菌群落结构的影响[J]. 农业环境科学学报, 2018,37(9):1910-1917. |
Chen SS, Sun M, Wang WC, et al. Effects of solid biofertilizers of phosphate-solubilizing fungi on maize growth and the bacterial community structure in rhizospheres[J]. Journal of Agro-Environment Science, 2018,37(9):1910-1917. | |
[6] | 韩蕾, 杨乐, 唐金铭, 等. 解磷菌发酵及溶磷条件的研究[J]. 生物技术通报, 2019,35(1):98-104. |
Han L, Yang L, Tang JM, et al. Fermentation condition and phosphate-dissolving capacity of phosphate-solubilizing bacterium[J]. Biotechnology Bulletin, 2019,35(1):98-104. | |
[7] | 陶光灿, 王素英, 郭兴强, 等. 以固氮菌和溶磷菌筛选拮抗作物病害的细菌组合[J]. 应用生态学报, 2006(3):3462-3467. |
Tao GC, Wang SY, Guo XQ, et al. Screening of mixed crop disease-resistant bacterial inoculants from N2-fixing and psolubilzing bacterial isolates[J]. Chinese Journal of Applied Ecology, 2006,3:3462-3467. | |
[8] | 秦利均, 杨永柱, 杨星勇. 土壤溶磷微生物溶磷、解磷机制研究进展[J]. 生命科学研究, 2019,23(1):59-64. |
Qin LJ, Yang YZ, Yang XY. Advances in mechanisms of soil phosphorus solubilization and dissolution by phosphate solubilizing microorganisms[J]. Life Science Research, 2019,23(1):59-64. | |
[9] | 邢芳芳, 高明夫, 禚优优, 等. 大麦根际高效溶磷菌的筛选、鉴定及促生效果研究[J]. 华北农学报, 2016,31(S1):252-257. |
Xing FF, Gao MF, Zhuo YY, et al. Screening and identification of phosphate solubilizing bacteria in hordeum vulgare rhizosphere and its growth promoting effect[J]. Acta Agriculturae Boreali-sinica, 2016,31(S1):252-257. | |
[10] | 乔策策, 王甜甜, 王若斐, 等. 高效溶磷菌的筛选及其促生效应研究[J]. 南京农业大学学报, 2017,40(4):664-670. |
Qiao CC, Wang TT, Wang RF, et al. Screening phosphate solubilizing bacterial strains from maize rhizosphere and research on their plant growth promotion effect[J]. Journal of Nanjing Agricultural University, 2017,40(4):664-670. | |
[11] | Vreeland R, Litchfield C, Martin E, et al. Halomonas elongata, a new genus and species of extremely salt-tolerant bacteria[J]. Int J Sys Bacteriol, 1980,30:485-495. |
[12] | 杨永利. 滨海重盐渍荒漠地区生态重建技术模式及效果的研究[D]. 北京:中国农业大学, 2004: 14-16. |
Yang YL. Studies on the technology models and effects for ecological reconstruction in the coast are with heavy soil salinitytake Tianjin coast area as an example[D]. Beijing:China Agricultural University, 2004. | |
[13] | 杨晓潇, 王秀兰, 王计平, 等. 天津市滨海新区土壤水盐空间变异分析[J]. 中国水土保持科学, 2019,17(3):39-47. |
Yang XX, Wang XL, Wang JP, et al. Spatial variation analysis of soil moisture and salinity in Tianjin Binhai New Area[J]. Science of Soil and Water Conservation, 2019,17(3):39-47. | |
[14] | 东秀珠, 蔡妙英. 常见细菌系统鉴定手册[M]. 北京: 科学出版社, 2001: 353-390. |
Dong XZ, Cai MY. Handbook for systematic identification of common bacteria[M]. Beijing: China Science Press, 2001: 353-390. | |
[15] |
Thompson J, Gibson T, Plewniak F, et al. The CLUSTAL_X windows interface:flexible strategies for multiple sequence alignment aided by quality analysis tools[J]. Nucleic Acids Research, 1997,25:4876-4882.
doi: 10.1093/nar/25.24.4876 URL pmid: 9396791 |
[16] |
Huang H, Wang W, Ma T, et al. Sphingomonas sanxanigenens sp. nov., isolated from soil[J]. International Journal of Systematic and Evolutionary Microbiology, 2009,59(4):719-723.
doi: 10.1099/ijs.0.000257-0 URL |
[17] | Ren MN, Li XY, Zhang YQ, et al. Massilia armeniaca sp. nov. , isolated from desert soil[J]. International Journal of Systematic and Evolutionary Microbiology, 2018,68(7):2319-2324. |
[18] | Sasser M. Identification of bacteria by gas chromatography of cellular fatty acids. In methods in phytobacteriology[M]. Budapest:Akademiai Kiado, 1990: 199-204. |
[19] | Vassilev N, Medina A, Azcon R, et al. Microbial solubilization of rock phosphate on media containing agro-industrial wastes and effect of the resulting products on plant growth and P uptake[J]. Plant and Soil, 2006,287:77-84. |
[20] | Chun J, Oren A, Ventosa A, et al. Proposed minimal standards for the use of genome data for the taxonomy of prokaryotes[J]. Int J Sys Evol Microbiol, 2018,68(1):461-466. |
[21] | Miao CH, Jia FF, Wan YS, et al. Halomonas huangheensis sp. nov., a moderately halophilic bacterium isolated from a saline-alkali soil[J]. Int J Sys Evol Microbiol, 2014,64(3):915-920. |
[22] |
Dou G, et al. Halomonas heilongjiangensis sp. nov., a novel moderately halophilic bacterium isolated from saline and alkaline soil[J]. Antonie Van Leeuwenhoek, 2015,108(2):403-413.
URL pmid: 26036672 |
[23] | Xu L, Xu X, Meng F, et al. Halomonas zincidurans sp. nov. , a heavy-metal-tolerant bacterium isolated from the deep-sea environment[J]. Int J Sys Evol Microbiol, 2013,63(11):4230-4236. |
[24] | 王永妍, 赵炳赫, 梁广钰, 等. 不同季节使用微生态制剂后养殖海水细菌群落特征[J]. 生物技术通报, 2020,36(2):126-133. |
Wang YY, Zhao BH, Liang GY, et al. Bacterial community characteristics of cultured seawater with microecological preparations in different seasons[J]. Biotechnology Bulletin, 2020,36(2):126-133. | |
[25] | 汪辉, 周长林. 盐单胞菌Halomonas-GY1的分离鉴定与活性产物的研究[J]. 药学进展, 2005(6):266-270. |
Wang H, Zhou CL. Isolation and identification of a strain of Halomonas and research on activities of its product[J]. Progress in Pharmaceutical Sciences, 2005(6):266-270. | |
[26] | 任世英, 王子峰, 肖天, 等. 一株海洋聚磷菌YSR-3的分离与鉴定[J]. 海洋与湖沼, 2006(5):437-443. |
Ren SY, Wang ZF, Xiao T, et al. Isolation and identification of a marine polyphosphate-accumulating bacterium YSR-3[J]. Oceanologia Et Limnologia Sinica, 2006(5):437-443. | |
[27] | 郑晓梅, 丁旭扬, 黄婷, 等. 一株产酸黑曲霉的分离鉴定及其在盐碱土改良中的应用[J]. 山东农业科学, 2019,51(4):69-73. |
Zheng XM, Ding XY, Huang T, et al. Isolation and identification of an acid-producing Aspergillus niger and its application in improvement of saline-alkali soil[J]. Shandong Agricultural Sciences, 2019,51(4):69-73. | |
[28] | 杨美英, 张婷婷, 武志海, 等. 盐碱土添加外源溶磷菌液对水稻渗透调节能力及光合指标的影响[J]. 西北农林科技大学学报:自然科学版, 2016,44(8):66-74. |
Yang MY, Zhang TT, Wu ZH, et al. Effects of adding exogenous phosphate dissolving bacteria on osmotic adjustment ability and photosynthetic indexes of rice[J]. Journal of Northwest A&F University:Nat. Sci. Ed., 2016,44(8):66-74. | |
[29] | 孙成成, 丁伟, 潘兴兵, 等. 影响微生物菌剂应用稳定性的因素分析[J]. 植物医生, 2019,32(4):19-23. |
Sun CC, Ding W, Pan XB, et al. Inflencing factors for the stability of biocontrol agents[J]. Plant Doctor, 2019,32(4):19-23. | |
[30] | 廉晓娟, 李明悦, 王艳, 等. 滨海盐渍土改良剂的筛选及应用效果研究[J]. 中国农学通报, 2013,29(14):150-154. |
Lian XJ, Li MY, Wang Y, et al. Selection and application effect of soil amendments on coastal saline soil[J]. Chinese Agricultural Science Bulletin, 2013,29(14):150-154. | |
[31] | 韩丽珍, 周静, 王欢. 两株对花生促生的芽孢杆菌的鉴定及溶磷特性研究[J]. 基因组学与应用生物学, 2019,38(9):4066-4076. |
Han LZ, Zhou J, Wang H. Identification and phosphorusdissolving characteristics of two bacillus strains promoted on peanut growth[J]. Genomics and Applied Biology, 2019,38(9):4066-4076. |
[1] | 曾还雄, 方霞, 刘凌燕, 金小宝. 蜚蠊肠道抗真菌活性放线菌的筛选与鉴定[J]. 生物技术通报, 2018, 34(12): 147-151. |
[2] | 李青, 刘洋, 吴刚, 杜慧竟, 游靖, 李红, 柯从玉, 张欣, 余吉良, 赵婷. 一株华北油田地层水细菌HB-1的分离及多相分类鉴定[J]. 生物技术通报, 2013, 0(6): 110-115. |
[3] | 吴刚, 刘洋, 李青, 杜慧竟, 游靖, 李红, 柯丛玉, 张欣, 余吉良, 赵婷. 一株螯合球菌HB-4的分离及多相分类鉴定[J]. 生物技术通报, 2013, 0(5): 165-170. |
[4] | 石小彤,李彦芹,邢国伟,康现江. 一株耐盐好氧反硝化细菌的分离筛选及鉴定[J]. 生物技术通报, 2013, 0(3): 175-180. |
[5] | 刘洋;赵婷;姚粟;葛媛媛;信春晖;许玲;程池;. 一株芝麻香型白酒高温大曲嗜热放线菌的分离与鉴定[J]. , 2012, 0(10): 210-216. |
[6] | 于晓溪;吴慧玲;刘铭;董丹;张殿朋;刘伟成;. 具有杀镰孢菌素合成功能基因菌株的筛选与鉴定[J]. , 2012, 0(10): 217-222. |
[7] | 任大明;张晓轩;董丹;刘伟成;刘德文;. 拮抗菌株Kc-t99的鉴定及其抑菌活性研究[J]. , 2011, 0(04): 153-157. |
[8] | 陈淑吟;胡传明;陆勤勤;孙中响;许广平;丁亚平;. 江苏海域几种绿潮藻类rDNA ITS序列分析[J]. , 2011, 0(03): 135-140. |
[9] | 曲直;洪葵;. 高效液相色谱法测定红树林放线菌DNA的(G+C)mol%含量[J]. , 2009, 0(S1): 209-214. |
[10] | 金人一. 蒙大拿州立大学和Cytoclonal公司从真菌取得紫杉醇[J]. , 1994, 0(04): 24-25. |
阅读次数 | ||||||
全文 |
|
|||||
摘要 |
|
|||||