樟子松根际土壤解磷细菌的筛选、鉴定及解磷能力

doi:10.11924/j.issn.1000-6850.casb20191200913

中国农学通报 ›› 2020, Vol. 36 ›› Issue (32): 76-81.doi: 10.11924/j.issn.1000-6850.casb20191200913

• 资源·环境·生态·土壤·气象 • 上一篇下一篇

樟子松根际土壤解磷细菌的筛选、鉴定及解磷能力

宋小双¹^,²(), 遇文婧¹^,², 周琦¹^,², 闵凯³, 邓勋¹^,²()

¹黑龙江省林业科学院森林保护研究所,哈尔滨 150040
²黑龙江省森林草原火灾及病虫害防控重点实验室,哈尔滨 150040
³大庆油田矿区服务事业部,黑龙江大庆 163000

收稿日期:2019-12-05 修回日期:2020-02-15 出版日期:2020-11-15 发布日期:2020-11-19
通讯作者: 邓勋
作者简介:宋小双,女,1976年出生,河北唐山人,副研究员,博士,研究方向:土传病害生物防治和森林土壤微生物。通信地址:150040 黑龙江省哈尔滨市哈平路134号黑龙江省林科院森林有害生物防治研究中心,Tel:0451-86630340,E-mail: sxshappy@126.com。
基金资助:
国家自然科学基金项目“褐环乳牛肝菌与菌根辅助细菌互作对樟子松抗病作用机制”(31700564);国家自然科学基金项目“深色有隔内生真菌提高樟子松抗病性的生理及分子机制”(31670649);黑龙江省林科院人才基金项目“外生菌根菌褐环乳牛肝菌(Suillus luteus)快速分子检测技术”(2015R01)

Isolation, Identification and Phosphate Solubilization Analysis of Phosphate-solubilizing Bacteria Derived from Pinus sylvestris var. mongolica Rhizosphere Soil

Song Xiaoshuang¹^,²(), Yu Wenjing¹^,², Zhou Qi¹^,², Min Kai³, Deng Xun¹^,²()

¹Forest Protection Institute of Heilongjiang Academy of Forestry Sciences, Harbin 150040
²Heilongjiang Key Laboratory of Forest Grassland Fire and Pest Control, Harbin 150040
³Daqing Oilfield Logistic Administration, Daqing Heilongjiang 163000

Received:2019-12-05 Revised:2020-02-15 Online:2020-11-15 Published:2020-11-19
Contact: Deng Xun

摘要/Abstract

摘要：

以磷酸钙为难溶态磷,从樟子松根际土壤分离筛选高效解磷细菌,采用透明圈法分离解磷细菌,以解磷效率为指标定量筛选高效菌株,通过生理生化指标测定结合16S rRNA序列系统发育树构建鉴定菌株,并利用单因素方法分析研究不同碳源、氮源及 C/N比值对菌株解磷能力的影响。结果表明:分离筛选得到11株解磷细菌,筛选得到2株高效解磷细菌A43和A54,初步鉴定菌株A43和A54均为Pseudomonas koreensis,A43在碳源为葡萄糖,氮源为硫酸铵,A54在碳源为蔗糖,氮源为硝酸钠,C/N比均为20:1条件下,菌株解磷效果最佳。试验筛选的高效解磷细菌为进一步制备樟子松促生微生物菌肥打下良好基础。

关键词: 樟子松, 根际解磷细菌, 解磷能力, 菌株鉴定, 微生物菌肥

Abstract:

Using calcium phosphate as insoluble phosphorus, high efficient phosphate solubilizing bacteria were isolated from rhizosphere soil of Pinus sylvestris var. mongolica. The halo ring method was used to isolate the phosphate-solubilizing bacteria, and the phosphorus-solubilizing efficiency was used to quantitatively screen the high efficiency strains. Physiological and biochemical test and 16S rRNA gene sequences analysis and phylogenic tree construction were employed to identify the genus of the phosphate-solubilizing bacteria. The effect of different carbon sources, nitrogen sources and C/N ratio on the phosphorus-solubilizing ability was studied by single factor method. The results showed that eleven strains of phosphate-solubilizing bacteria were isolated from the rhizosphere soil, strains A43 and A54 were quantitatively screened as the high efficiency strain. A43 and A54 were identified to be Pseudomonas koreensis. For strain A43, the phosphate-solubilizing ability was relatively good when the carbon source was glucose, nitrogen source was (NH₄)₂SO₄ and C/N was 20:1. For strain A54 the hosphate-solubilizing ability was relatively good when the carbon source was sucrose, nitrogen source was NaNO₃ and C/N was 20:1. The high efficiency phosphorus-solubilizing bacteria screened from the rhizosphere soil lay a good foundation for further preparation of growth-promoting microbial fertilizer of Pinus sylvestris var. mongolica.

Key words: Pinus sylvestris var. Mongolica, rhizosphere phosphorus-solubilizing bacteria, phosphorus-solubilizing ability, strain identification, microbial fertilizer

中图分类号:

S763.1

宋小双, 遇文婧, 周琦, 闵凯, 邓勋. 樟子松根际土壤解磷细菌的筛选、鉴定及解磷能力[J]. 中国农学通报, 2020, 36(32): 76-81.

Song Xiaoshuang, Yu Wenjing, Zhou Qi, Min Kai, Deng Xun. Isolation, Identification and Phosphate Solubilization Analysis of Phosphate-solubilizing Bacteria Derived from Pinus sylvestris var. mongolica Rhizosphere Soil[J]. Chinese Agricultural Science Bulletin, 2020, 36(32): 76-81.

图/表 9

参考文献 22

[1]	Lobo C, Juárez T, María V, et al. Development of low-cost formulations of plant growth-promoting bacteria to be used as inoculants in beneficial agricultural technologies[J]. Microbiological Research, 2019: 12-25.
[2]	Bushra T, Anwar K, Muhammad T, et al. Review bottlenecks in commercialisation and future prospects of PGPR[J]. Applied Soil Ecology, 2017(10):102-117
[3]	Dissanayaka D, William C, Lambers H, et al. Molecular mechanisms underpinning phosphorus use efficiency in rice[J]. Plant Cell Environment, 2018,41(7):1483-1496.
[4]	杜雷, 王素萍, 陈钢, 等. 一株高效解磷细菌的筛选、鉴定及其溶磷能力的研究[J]. 中国土壤与肥料, 2017(3):136-141.
[5]	Rodriguez H, Fraga R. Phosphate solubilizing bacteria and their role in plant growth promotion[J]. Biotechnology Advance, 1999,17(4):319-39.
[6]	黄敏, 吴金水, 黄巧云, 等. 土壤磷素微生物作用的研究进展[J]. 生态环境, 2003(3):366-370.
[7]	邓勋, 宋瑞清, 宋小双, 等. 高效木霉菌株对樟子松枯梢病的抑菌机理[J]. 中南林业科技大学学报, 2012,32(11):21-27.
[8]	Halifu S, Xun D, Xiaoshuang S, et al. Effects of Sphaeropsis blight on rhizosphere soil bacterial community structure and soil physicochemical properties of Pinus sylvestris var. mongolica in Zhanggutai, China[J]. Forests, 2019,10(11):954.
[9]	柯春亮, 陈宇丰, 周登博, 等. 香蕉根际土壤解磷细菌的筛选、鉴定及解磷能力[J]. 微生物学通报, 2015,42(6):1032-1042.
[10]	徐欢, 俞新玲, 林勇明, 等. 桉树根际土壤解磷细菌的分离、筛选及其解磷效果[J]. 福建农林大学学报:自然科学版, 2016,45(5):529-535.
[11]	王舒, 张林平, 张扬, 等. 红壤区油茶根际解磷细菌的筛选、鉴定及其解磷能力[J]. 林业科学研究, 2015,28(3):409-416.
[12]	屈青松, 彭万里, 翟立明, 等. 马铃薯内生细菌的分离鉴定及种群多样性分析[J]. 中国蔬菜, 2016(6):41-46.
[13]	陈佳兴, 秦琴, 邱树毅, 等. 磷尾矿土壤中解磷细菌的筛选及解磷能力的测定[J]. 生物技术通报, 2018,34(6):183-189.
[14]	陈定安, 魏小武, 张敏, 等. 油茶树根际土壤解有机磷细菌的分离、鉴定及解磷能力分析[J]. 湖南农业科学, 2019(10):8-11.
[15]	沈震. 油松根际土壤解磷细菌及其群落结构特征研究[D]. 陕西杨凌:西北农林科技大学, 2018: 1-79.
[16]	杨倩倩, 姚庆智, 员子晶, 等. 油松菌根际高效解磷钾细菌筛选与鉴定[J]. 基因组学与应用生物学, 2017,36(4):1526-1531.
[17]	肖坤, 崔延, 高丹阳, 等. 核桃根际解磷细菌的筛选及对核桃促生作用研究[J]. 河北农业大学学报, 2018,41(5):49-54.
[18]	吴伟, 张鹏飞, 张桂萍, 等. 连翘根际高效解有机磷细菌的筛选鉴定及促生长特性研究[J]. 西南林业大学学报:自然科学, 2018,38(3):93-100.
[19]	刘春菊, 杜传印, 梁子敬, 等. 高效解磷细菌菌株CT45-1的鉴定及其对烟草的促生作用[J]. 山东农业科学, 2019,51(4):74-78.
[20]	常慧萍, 夏铁骑, 付瑞敏, 等. 小麦根际解磷细菌的筛选鉴定及其促生效果[J]. 江苏农业科学, 2018,46(14):270-273.
[21]	马晓丽, 张婷婷, 郭欢, 等. 红花根际磷细菌筛选、鉴定及其促生效果[J]. 江苏农业科学, 2014,42(9):318-322.
[22]	员子晶. 油松幼苗菌根围优良磷钾细菌的筛选及对油松的促生作用[D]. 呼和浩特:内蒙古农业大学, 2011: 1-82.

菌株	透明圈直径D/mm	菌落直径d/mm	D/d	菌株	透明圈直径D/mm	菌落直径d/mm	D/d
A43	14.333±0.666a	7.227±0.498cd	1.986±0.085b	A37	10.267±0.473d	6.900±0.200cd	1.488±0.030d
A26	12.200±0.700bc	7.700±0.200c	1.587±0.129c	A25	12.467±0.493bc	10.533±0.651b	1.188±0.113e
A22	12.067±1.387bc	6.433±0.503d	1.893±0.357b	52	14.267±1.662a	13.467±0.351a	1.061±0.138e
A20	9.700±0.200d	4.467±0.351e	2.178±0.129a	54	13.167±0.306ab	6.200±0.300d	2.129±0.152a
A58	9.133±0.551d	4.900±0.529e	1.880±0.254b	A17	11.833±0.306c	10.300±0.200b	1.149±0.023e
A12	10.033±0.603d	7.690±0.182c	1.304±0.054d

菌株	透明圈直径D/mm	菌落直径d/mm	D/d	菌株	透明圈直径D/mm	菌落直径d/mm	D/d
A43	14.333±0.666a	7.227±0.498cd	1.986±0.085b	A37	10.267±0.473d	6.900±0.200cd	1.488±0.030d
A26	12.200±0.700bc	7.700±0.200c	1.587±0.129c	A25	12.467±0.493bc	10.533±0.651b	1.188±0.113e
A22	12.067±1.387bc	6.433±0.503d	1.893±0.357b	52	14.267±1.662a	13.467±0.351a	1.061±0.138e
A20	9.700±0.200d	4.467±0.351e	2.178±0.129a	54	13.167±0.306ab	6.200±0.300d	2.129±0.152a
A58	9.133±0.551d	4.900±0.529e	1.880±0.254b	A17	11.833±0.306c	10.300±0.200b	1.149±0.023e
A12	10.033±0.603d	7.690±0.182c	1.304±0.054d

菌株	解磷值/(mg/L)	较CK增幅倍数	pH值
A43	225.65±16.65a	14.57b	4.18±0.35e
A26	80.40±4.98e	6.56g	5.88±0.16ab
A22	88.44±5.48e	7.21f	5.07±0.17bc
A20	178.66±11.07b	18.41a	4.60±0.33d
A58	90.26±6.66e	7.36f	4.84±0.13cd
A37	112.83±8.32d	9.20d	5.30±0.17b
A25	101.54±7.49d	8.28e	4.07±0.15e
A52	125.06±7.75c	10.20c	4.93±0.24c
A54	180.52±13.32b	9.10d	4.44±0.39d
A17	122.79±11.12c	10.02c	4.89±0.42c
A12	111.62±10.11d	14.72b	4.68±0.27d
CK	12.36±1.56f		6.57±0.12a

菌株	解磷值/(mg/L)	较CK增幅倍数	pH值
A43	225.65±16.65a	14.57b	4.18±0.35e
A26	80.40±4.98e	6.56g	5.88±0.16ab
A22	88.44±5.48e	7.21f	5.07±0.17bc
A20	178.66±11.07b	18.41a	4.60±0.33d
A58	90.26±6.66e	7.36f	4.84±0.13cd
A37	112.83±8.32d	9.20d	5.30±0.17b
A25	101.54±7.49d	8.28e	4.07±0.15e
A52	125.06±7.75c	10.20c	4.93±0.24c
A54	180.52±13.32b	9.10d	4.44±0.39d
A17	122.79±11.12c	10.02c	4.89±0.42c
A12	111.62±10.11d	14.72b	4.68±0.27d
CK	12.36±1.56f		6.57±0.12a

樟子松根际土壤解磷细菌的筛选、鉴定及解磷能力

Isolation, Identification and Phosphate Solubilization Analysis of Phosphate-solubilizing Bacteria Derived from Pinus sylvestris var. mongolica Rhizosphere Soil

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