耐盐碱溶磷菌Y2R2的分离鉴定及溶磷特性

doi:10.13560/j.cnki.biotech.bull.1985.2020-0236

摘要/Abstract

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。

关键词: 耐盐碱溶磷菌, 分类鉴定, 盐单胞菌, 新种

Abstract:

The objective of this study is to screen phosphorus-dissolving bacteria with fine environment adaptability to coastal salt-alkali land. Strain Y2R2 with the ability of salt-alkali-tolerant and phosphorus-dissolving was isolated from salt-alkali soil samples collected from Tianjin Binhai New Area. Strain Y2R2 was identified as a novel species of Halomonas according to poly-phase taxonomic characteristics. Strain Y2R2 grew at the temperature 10℃ to 45℃,and tolerated the condition of pH 14 and 15% NaCl. Strain Y2R2 showed ability of dissolving phosphorus under pH 7-13 and NaCl content 0-12%. In inorganic phosphorus medium,under the fermentation conditions in which the initial pH was 8,NaCl content was 9% and fermentation time was 48 h,the phosphate-dissolving quantity by strain Y2R2 reached its highest value,247.6 mg/L. Strain Y2R2 screened in this study is a novel species of indigenous microorganisms that adapted to the soil and climate environment of coastal salt-alkali land,and has high salt-alkali tolerance and phosphate-dissolving capacities. The strain is expected to be used as salt-alkali phosphorus-dissolving bacterial agent.

Key words: salt-alkali-tolerant phosphorus dissolving bacteria, taxonomic identification, Halomonas, novel species

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/表 8

图1 菌株Y2R2产生的溶磷圈

图2 菌株Y2R2的细胞形态（5000×）

图3 菌株Y2R2的16S rDNA序列系统进化树进化树中的其他菌株是与Y2R2同源关系接近的,Halomonas属其他种的模式菌株,Zymobacter palmae为树根;括号内为菌株16S rDNA序列在GenBank中的登记号;标尺的单位长度代表序列差异度为0.01

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

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
鉴定项目	Y2R2	H. huangheensis	鉴定项目	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	-	+

图5 培养时间对菌株Y2R2溶磷量的影响

图6 NaCl对菌株Y2R2溶磷量的影响

图7 初始pH对菌株Y2R2溶磷量的影响

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