中华补血草内生与根际具ACC脱氨酶活性细菌的筛选及其生物多样性

Screening and biodiversity of endophytic and rhizosphere bacteria containing ACC deaminase from halophyte Limonium sinense (Girard) Kuntze

摘要:

[目的] 获得江苏沿海滩涂盐生药用植物中华补血草内生及根际具有1-氨基环丙烷-1-羧酸(ACC)脱氨酶活性的细菌,研究其遗传多样性和潜在促生活性。[方法] 从中华补血草和根际土壤分离筛选具有ACC脱氨酶活性的菌株,对其ACC脱氨酶活性定量检测,通过16S rRNA基因序列分析确定菌株系统发育地位。同时研究其固氮、溶磷、产植物生长素吲哚乙酸(IAA)及耐盐能力。[结果] 分离筛选获得18株具有ACC脱氨酶活性的内生与根际细菌,定量检测发现其中有13株菌的ACC脱氨酶含量在20 nmol α-KA/(mg Pr·h)以上,有11株菌可以固氮,7株菌能够解磷,9株菌产生IAA。菌株的NaCl盐耐受范围多数在0-13%之间。16S rRNA基因测序表明,活性菌株分属于7个属,多样性丰富,节杆菌属(Arthrobacter)为优势类群。其中菌株KLBMP 5180为节杆菌属的潜在新种。[结论] 江苏沿海滩涂盐生药用植物中华补血草共生环境中具有丰富多样的具ACC脱氨酶活性的菌株,并存在潜在新物种资源,具有进一步研究价值。

Abstract:

[Objective] We isolated and screened endophytic and rhizosphere bacteria with 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase from halophyte Limonium sinense (Girard) Kuntze collected from Jiangsu coastal area and investigated their diversity and plant growth promoting potential. [Methods] Strains were obtained from inner tissues and rhizosphere soils using pure culture cultivation method and identified by 16S rRNA gene sequencing and phylogenetic analysis. Their potential plant growth promoting index of nitrogen fixation, phosphate solubilization, indoleacetic acid (IAA) production and NaCl tolerance ability were evaluated. [Results] Eighteen strains with ACC deaminase were obtained and 13 of them exhibited more than 20 nmol α-KA/(mg Pr·h) ACC deaminase activity. Nine isolates produced IAA, 11 had nitrogen fixation ability and 7 of them had phosphate solubilization ability. Most of the isolates could grow under 0%-13% NaCl. The results of 16S rRNA sequencing showed that these strains belonged to seven genera, with Arthrobacter as the most predominant genus. Among them, strain KLBMP 5180 was found to be a potential novel species of the genus Arthrobacter. [Conclusion] The halophyte plants Limonium sinense (Girard) Kuntze located in the area of coastal shoal contain a variety of symbiotic bacteria with ACC deaminase as well as the source of novel species. Some of them had good research prospect in the future.