Isolation and Identification of ACC Deamiase Producing Bacteria in the soil of Halostachys caspica

doi:10.6048/j.issn.1001-4330.2018.11.019

Abstract

Abstract: 【Objective】 Plant rhizosphere-promoting bacteria with ACC deaminase activity can increase plant stress resistance and promote plant growth under adverse conditions such as heavy metal pollution, salt stress or drought. Plant rhizosphere-promoting strains with high ACC deaminase activity will being screened.【Method】The rhizosphere soil of the Halostachys caspica soil in Xinjiang saline-alkali soil was used as the sample source, and ACC was the only nitrogen source. The ACC deaminase-producing strain was screened by the method of directional enrichment. Morphological observation was carried out by scanning electron microscopy; physiological and biochemical tests such as Gram staining, nitrate reduction test and citrate utilization, Biolog Gen III plate microwell Identification and 16S rDNA sequence homology analysis were used to identify the isolated strains.【Result】Three strains with ACC deaminase activity were screened. The scanned electron microscopy showed that all the three strains were Brevibacterium, Gram staining was Negative, all spores, nitrate reduction test were positive, and citrate could be used. The three Strains: 1#, 3# and 5#, the specific activities of the ACC deaminase activities were 0.012, 0.012 and 0.014 U/mg. There was significant difference in enzyme activity between 1# and 5#, 3# and 5# (P<0.05), and there was no significant difference in enzyme activity between 1# and 3# (P<0.05). The isolated strains 1#, 3# and 5# were respectively identified as Klebsiella pneumoniae, Klebsiella pneumoniae, Klebsiella oxytoca, using Biolog Gen III plate microwell and 16S rDNA sequence homology analysis. 【Conclusion】The results of this study laid the foundation for further research on the plant growth-promoting effects of ACC deaminase active strains.

Key words: ACC deamiase producing bacteria; rhizobacteria; identification of mycobacterium; Biolog identification

摘要： 【目的】研究从新疆盐碱地的盐穗木根际土壤中筛选到产ACC脱氨酶活性较高的植物根际促生菌。为ACC脱氨酶活性菌株的植物促生作用的研究奠定基础。【方法】以新疆盐碱地的盐穗木根际土壤为样品源,ACC为唯一氮源,利用筛选培养基定向富集的方法,筛选产ACC脱氨酶活性菌株。通过扫描电镜进行形态学的观察;革兰氏染色、硝酸盐还原试验和柠檬酸盐利用等生理生化试验,用Biolog Gen III板微孔鉴定及16S rDNA序列同源性分析对分离的菌株鉴定。【结果】筛选了三株具有ACC脱氨酶活性的菌株,用扫描电镜可以清晰的观察到三株菌株均为短杆菌,革兰氏染色都为阴性,均有芽孢,硝酸盐还原试验均为阳性,都能利用柠檬酸盐。三株菌株分别为1# 、3# 和5#,三株菌株的ACC脱氨酶活性的比活力分别为0.012,0.012,0.014 U/mg;1# 和5#,3# 与5# 之间酶活力差异显著(P<0.05),1#和3#之间酶活力差异不显著(P<0.05),用Biolog Gen III板微孔和16S rDNA序列同源性分析对分离的菌株1#、3# 和5# 菌株分别鉴定为Klebsiella pneumoniae、Klebsiella pneumoniae、Klebsiella oxytoca。【结论】

关键词: ACC脱氨酶, 根际细菌, 菌种鉴定, Biolog鉴定

CLC Number:

S182

ZHANG Pan, WANG Wei-nan, FAN Yong-hong. Isolation and Identification of ACC Deamiase Producing Bacteria in the soil of Halostachys caspica[J]. Xinjiang Agricultural Sciences, 2018, 55(11): 2112-2121.

张盼, 王伟楠, 樊永红. 盐穗木根际土壤产ACC脱氨酶细菌的筛选与鉴定[J]. 新疆农业科学, 2018, 55(11): 2112-2121.

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