黄壤伯克氏溶磷细菌的筛选鉴定及溶磷特性
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摘要
[目的]提高黄壤区土壤磷素有效性。[方法]通过PVK平板法从贵州黄壤分离出多株溶磷细菌。[结果]多次纯化得到14株溶磷能力大于395 mg·L~(-1)的菌株,其中菌株W4的溶磷能力为564.07 mg·L~(-1),显著高于其它菌株(P<0.05)。因此,选择W4作为试验菌株进行下一步黄壤溶磷细菌溶磷特性的研究。通过16S rRNA序列分析,确定W4为伯克氏菌属(Burkholderia sp);通过测定培养基成分含量对溶磷作用的影响,发现当培养液中葡萄糖浓度低于10 g·L~(-1)时,其含量显著影响W4的溶磷能力(P<0.05),当葡萄糖浓度大于10 g·L~(-1)时,其含量对W4的溶磷能力影响较小,培养液中葡萄糖浓度大于15 g·L~(-1)时,微生物活动产生的葡萄糖酸会增加进而对菌体有抑制作用;随着培养液中硫酸铵浓度的增加,W4的溶磷能力逐渐下降,而菌体生长则显著增加(P<0.05);W4以葡萄糖为碳源和硝酸钾为氮源溶磷能力最强,为640.10 mg·L~(-1);外加磷源对W4溶磷能力有一定的抑制作用。[结论]通过以上研究,有助于了解黄壤溶磷细菌的特性并为进一步应用提供理论基础。
[Objective]Present study was conducted in order to discover new phosphorus solubilizing bacteria strains and to improve the bioavailability of soil phosphorus in yellow soil area. [Methods] Phosphorus solubilizing bacteria(PSB) strains were isolated from yellow soil of Guizhou by PVK plate method. [Results] After a few of isolation and purification steps, fourteen bacteria strains with phosphate solubilizing capacity(PSC) greater than 395 mg·L~(-1) were obtained. Among of them, W4 showed PSC at 564.07 mg·L~(-1), which was significantly higher than other strains(P<0.05). Therefore, W4 was selected for further studies on characteristics of PSB. Based on 16 sRNA gene sequence results, W4 was identified as Burkholderia sp. When the glucose concentration of culture medium was lower than 10 g·L~(-1), the PSC of W4 was significantly affected by the content of the glucose(P<0.05), while when glucose concentration was greater than 10 g·L~(-1), no significant effect of glucose content on PSC of W4 was observed. When glucose concentration in culture medium was greater than 15 g·L~(-1), the gluconic acid produced by bacteria activity was increased which inhibited the bacteria growth rate. With the increase of ammonium sulfate concentration in the culture medium, the PSC of W4 was gradually decreased, while the growth rate of bacteria was increased significantly(P<0.05). Among the different carbon and nitrogen sources, glucose and potassium nitrate produced the highest PSC of W4 at 604.10 mg·L~(-1). The addition of phosphorus inhibited PSC of W4. [Conclusion] The results of present study provided evidence for better understanding the characteristics of phosphorus-soluble bacteria in yellow soil and provided theoretical basis for further bacteria application.
[Objective]Present study was conducted in order to discover new phosphorus solubilizing bacteria strains and to improve the bioavailability of soil phosphorus in yellow soil area. [Methods] Phosphorus solubilizing bacteria(PSB) strains were isolated from yellow soil of Guizhou by PVK plate method. [Results] After a few of isolation and purification steps, fourteen bacteria strains with phosphate solubilizing capacity(PSC) greater than 395 mg·L~(-1) were obtained. Among of them, W4 showed PSC at 564.07 mg·L~(-1), which was significantly higher than other strains(P<0.05). Therefore, W4 was selected for further studies on characteristics of PSB. Based on 16 sRNA gene sequence results, W4 was identified as Burkholderia sp. When the glucose concentration of culture medium was lower than 10 g·L~(-1), the PSC of W4 was significantly affected by the content of the glucose(P<0.05), while when glucose concentration was greater than 10 g·L~(-1), no significant effect of glucose content on PSC of W4 was observed. When glucose concentration in culture medium was greater than 15 g·L~(-1), the gluconic acid produced by bacteria activity was increased which inhibited the bacteria growth rate. With the increase of ammonium sulfate concentration in the culture medium, the PSC of W4 was gradually decreased, while the growth rate of bacteria was increased significantly(P<0.05). Among the different carbon and nitrogen sources, glucose and potassium nitrate produced the highest PSC of W4 at 604.10 mg·L~(-1). The addition of phosphorus inhibited PSC of W4. [Conclusion] The results of present study provided evidence for better understanding the characteristics of phosphorus-soluble bacteria in yellow soil and provided theoretical basis for further bacteria application.
引文
[1]Ringeval B, Nowak B, Nesme T, et al. Contribution of anthropogenic phosphorus to agricultural soil fertility and food production [J]. Global Biogeochemical Cycles, 2014, 28(7): 743-756.
[2]伦飞,刘俊国,张丹. 1961-2011年中国农田磷收支及磷使用效率研究[J]. 资源科学, 2016, 38(9):1681-1691.
[3]Gilbert N. The disappearing nutrient[J]. Nature, 2009,461(7265):716-718.
[4]Sattri S Z, Ittersum M K, Giller K E, et al. Key role of China and its agriculture in global sustainable phosphorus management[J]. Environmental Research Letters, 2014, 9(5):54003.
[5]Liu X, Sheng H, Jiang SY, et al. Intensification of phosphorus cycling in China since the 1600s[J]. Proceedings of the National Academy of Sciences of the United States of America, 2016, 113(10): 2609-2614.
[6]Mohammad S K, Almas Z, Mlunees A, et al. Plant growth promotion by phosphate solubilizing fungi - current perspective[J]. Archives of Agronomy and Soil Science, 2010, 56(1):73-98.
[7]Pradhan M, Sahoo R K, Pradhan C, et al. Contribution of native phosphorous-solubilizing bacteria of acid soils on phosphorous acquisition in peanut[J]. Protoplasma, 2017,254(6):2225-2236.
[8]Waseem H, Hina A, Usman I, et al. Analysis of ecological attributes of bacterial phosphorus solubilizers, native to pine forests of Lower Himalaya[J]. Applied Soil Ecology, 2017, 112:51-59.
[9]李渝, 刘彦伶, 张雅蓉, 等. 长期施肥条件下西南黄壤旱地有效磷对磷盈亏的响应[J]. 应用生态学报, 2016, 27(7):2321-2328.
[10]Nautiyal S C. An efficient microbiological growth medium for screening phosphate solubilizing microoraganisms [J]. FEMS Microbiology Letters, 1999, 170(1):265-270.
[11]冯哲叶, 陈莎莎, 王文超, 等. 几株溶磷细菌的筛选鉴定及其溶磷效果[J].南京农业大学学报, 2017, 40(5):842-849.
[12]马骢毓, 张英, 马文彬, 等. 黄芪根际促生菌(PGPR)筛选与特性研究[J]. 草业学报, 2017, 26(1):149-159.
[13]乔志伟, 洪坚平, 谢英荷, 等. 石灰性土壤拉恩式溶磷细菌的筛选鉴定及溶磷特性[J]. 应用生态学报, 2013, 24(8):2294-2300.
[14]Reyes I, Bernier L, Simard R R, et al. Effect of nitrogen source on the solubilization of different inorganic phosphates by an isolate of Penicillium rugulosum and two UV-induced mutants [J]. FEMS Microbiology Ecology, 1999, 28(3): 281-290.
[15]胡山, 牛世全, 龙洋, 等. 河西走廊盐碱土壤中一株高效溶磷菌的鉴定及条件优化[J]. 微生物学通报, 2017, 44(2):358-365.
[16]卫星, 徐鲁荣, 张丹, 等. 一株耐硝酸盐的巨大芽孢杆菌溶磷特性研究[J]. 环境科学学报, 2015, 35(7):2052-2058.
[17]张瑶, 路国兵, 周波, 等.洋葱伯克霍尔德式菌Lu10-1产脂肪酶发酵条件优化及酶学性质研究[J]. 生物技术通报, 2015, 31(9):190-196.
[18]戴沈艳, 申卫收, 贺云举, 等.一株高效解磷细菌的筛选及其在红壤性水稻土中的施用效果[J]. 应用与环境生物学报, 2011, 17(5): 678-683.
[19]张亚经, 马成仓, 郭军康, 等. 接种伯克氏菌D54对东南景天吸收富集Cd的影响[J]. 农业环境科学学报, 2017, 36(3):449-457.
[2]伦飞,刘俊国,张丹. 1961-2011年中国农田磷收支及磷使用效率研究[J]. 资源科学, 2016, 38(9):1681-1691.
[3]Gilbert N. The disappearing nutrient[J]. Nature, 2009,461(7265):716-718.
[4]Sattri S Z, Ittersum M K, Giller K E, et al. Key role of China and its agriculture in global sustainable phosphorus management[J]. Environmental Research Letters, 2014, 9(5):54003.
[5]Liu X, Sheng H, Jiang SY, et al. Intensification of phosphorus cycling in China since the 1600s[J]. Proceedings of the National Academy of Sciences of the United States of America, 2016, 113(10): 2609-2614.
[6]Mohammad S K, Almas Z, Mlunees A, et al. Plant growth promotion by phosphate solubilizing fungi - current perspective[J]. Archives of Agronomy and Soil Science, 2010, 56(1):73-98.
[7]Pradhan M, Sahoo R K, Pradhan C, et al. Contribution of native phosphorous-solubilizing bacteria of acid soils on phosphorous acquisition in peanut[J]. Protoplasma, 2017,254(6):2225-2236.
[8]Waseem H, Hina A, Usman I, et al. Analysis of ecological attributes of bacterial phosphorus solubilizers, native to pine forests of Lower Himalaya[J]. Applied Soil Ecology, 2017, 112:51-59.
[9]李渝, 刘彦伶, 张雅蓉, 等. 长期施肥条件下西南黄壤旱地有效磷对磷盈亏的响应[J]. 应用生态学报, 2016, 27(7):2321-2328.
[10]Nautiyal S C. An efficient microbiological growth medium for screening phosphate solubilizing microoraganisms [J]. FEMS Microbiology Letters, 1999, 170(1):265-270.
[11]冯哲叶, 陈莎莎, 王文超, 等. 几株溶磷细菌的筛选鉴定及其溶磷效果[J].南京农业大学学报, 2017, 40(5):842-849.
[12]马骢毓, 张英, 马文彬, 等. 黄芪根际促生菌(PGPR)筛选与特性研究[J]. 草业学报, 2017, 26(1):149-159.
[13]乔志伟, 洪坚平, 谢英荷, 等. 石灰性土壤拉恩式溶磷细菌的筛选鉴定及溶磷特性[J]. 应用生态学报, 2013, 24(8):2294-2300.
[14]Reyes I, Bernier L, Simard R R, et al. Effect of nitrogen source on the solubilization of different inorganic phosphates by an isolate of Penicillium rugulosum and two UV-induced mutants [J]. FEMS Microbiology Ecology, 1999, 28(3): 281-290.
[15]胡山, 牛世全, 龙洋, 等. 河西走廊盐碱土壤中一株高效溶磷菌的鉴定及条件优化[J]. 微生物学通报, 2017, 44(2):358-365.
[16]卫星, 徐鲁荣, 张丹, 等. 一株耐硝酸盐的巨大芽孢杆菌溶磷特性研究[J]. 环境科学学报, 2015, 35(7):2052-2058.
[17]张瑶, 路国兵, 周波, 等.洋葱伯克霍尔德式菌Lu10-1产脂肪酶发酵条件优化及酶学性质研究[J]. 生物技术通报, 2015, 31(9):190-196.
[18]戴沈艳, 申卫收, 贺云举, 等.一株高效解磷细菌的筛选及其在红壤性水稻土中的施用效果[J]. 应用与环境生物学报, 2011, 17(5): 678-683.
[19]张亚经, 马成仓, 郭军康, 等. 接种伯克氏菌D54对东南景天吸收富集Cd的影响[J]. 农业环境科学学报, 2017, 36(3):449-457.