Special High-efficient Microorganisms for Dissolving Phosphorus, Potassium and Growth-promoting of Walnut: Screening and Compounding

doi:10.11924/j.issn.1000-6850.casb2020-0252

Abstract

Abstract:

To solve the problem that phosphorus and potassium in soil are difficult to be absorbed and utilized by plants, the functional microorganisms for dissolving phosphorus, potassium and growth promotion were screened out by the method of coating dilution. The results showed that 1326 strains of microorganisms were screened out from soil, 536 strains remained by removing the strains with the same morphology and gram staining results. The ability to dissolve phosphorus and potassium and secrete hormones was detected, and 4 functional strains were identified to study the effects of their combinations on walnut seedlings. Through the observation of the morphological characteristics and the physiological and biochemical identification of the 4 strains, and with the combination of 16S rDNA sequence analysis, ht1 and ht4 were confirmed as Bacillus amyloliquefaciens and ht2 and ht3 as Bacillus subtilis. Plate confrontation test was carried out on the 4 functional strains, and it was found that the 4 strains had no inhibitory effect. After the combination of different strains, the IAA secretion of the 4 functional strains was significantly increased. The compounding effect of the 4 strains with different proportions was explored, and the strong colonization ability of the strains was detected in the soil. The measurement of agronomic traits of walnut seedlings showed that the compounding effect of the 4 strains with 1:1:1:1 ratio was the best, and the indexes of tree height, sub-branch height and crown width were significantly higher than those of other treatments.

Key words: walnut, screening of functional microorganisms, identification, functional verification, agronomic traits

CLC Number:

S144.1

Zhang Qi, Xu Siqi, Zhao Xuetong, Pan Wenzheng, Shang Haili, Wang Jingxian, Zhao Rongbiao. Special High-efficient Microorganisms for Dissolving Phosphorus, Potassium and Growth-promoting of Walnut: Screening and Compounding[J]. Chinese Agricultural Science Bulletin, 2020, 36(34): 64-70.

Figures/Tables 8

References 25

[1]	郗荣庭, 张毅萍. 中国果树志·核桃卷[M]. 北京: 中国林业出版社,1996:24- 25, 45-46.
[2]	崔军, 李桂凤, 李坚. 曲靖市核桃生产现状及发展对策[J].防护林科技,2019(10):57-59,61.
[3]	有祥亮, 邢世岩, 张友朋, 等. 核桃抗病育种研究进展[J].山东林业科技,2001(6):35-36.
[4]	代佳和, 田洋, 杨舒雯, 等. 云南省核桃产业发展现状及对策[J]. 农产品加工, 2019,472(1):79-82.
[5]	耿树香, 宁德鲁, 韩明珠, 等. 云南核桃主要栽培品种蛋白质及脂肪酸综合评价分析[J]. 中国油脂, 2019,44(10):116-120,141.
[6]	陈少瑜, 吴涛, 肖良俊. 三江并流区金沙江流域云南核桃种质资源遗传多样性及分布特征[J]. 南方农业学报, 2019,50(12):2656-2664.
[7]	陈伟, 李江, 裴艳辉, 等. 云南省核桃栽培的技术问题及对策[J]. 林业调查规划, 2014,39(5):123-128.
[8]	刘云高. 云南省核桃产业发展现状及对策分析[J]. 林业调查规划, 2019,44(3):143-147,151.
	铁学江, 唐永奉, 刘世平, 等. 一种核桃肥的生产以应用试验[J]. 中国南方果树, 2012,14(6):58-60.
[10]	刘文干. 三株红壤高效溶磷菌的分离、鉴定、溶磷特性及其对花生促生效应的研究[D]. 江苏:南京农业大学, 2012.
[11]	王孝峰, 蔡恩明. 我国钾肥产业发展思路探讨-由必和必拓收购加拿大钾肥公司引发的思考[J].中国石油和化工经济分析,2010(11):32-35.
[12]	金月波. 水稻根际促生菌筛选及其对水稻促生效果研究[J]. 辽宁农业职业技术学院学报, 2020,22(2):4-7.
[13]	吴日章, 杨琼, 代燕丽, 等. 蚕沙分离的溶磷解钾菌株OP6发酵配方[J]. 农业工程, 2019,9(3):45-49.
[14]	张雨. 生姜内生细菌分离鉴定及促生效果研究[D]. 雅安:四川农业大学, 2015.
[15]	Thilagar G, Bagyaraj D J, Rao M S. Selected microbial consortia developed for chilly reduces application of chemical fertilizers by 50% under field conditions[J]. Scientia Horticulturae, 2016,198:27-35. doi: 10.1016/j.scienta.2015.11.021 URL
[16]	Mc Steen P. Auxin and monocot development[J]. Cold Spring Harbor Perspectives in Biology, 2010,2(3):a001479. doi: 10.1101/cshperspect.a001479 URL pmid: 20300208
[17]	Dodd I C, Zinovkina N Y, Safronova V I, et al. Rhizobacterial mediation of plant hormone status[J]. Annals of Applied Biology, 2010,157(3):361-379. doi: 10.1111/aab.2010.157.issue-3 URL
[18]	Madhaiyan M, Poonguzhali S, Ryu J, et al. Regulation of ethylene levels in canola (Brassica campestris) by 1-aminocyclopropane-1-carboxylate deaminase-containing Methylobacterium fujisawaense[J]. Planta, 2006,224(2):268-278. doi: 10.1007/s00425-005-0211-y URL pmid: 16416316
[19]	Lorteau M A, Ferguson B J, Frédérique Catherine Guinel. Effects of cytokinin on ethylene production and nodulation in pea (Pisum sativum) cv. Sparkle[J]. Physiologia Plantarum, 2010,112(3):421-428. URL pmid: 11473700
[20]	高亚敏, 姚拓, 李海云, 等. 高寒草甸蒿草、珠芽蓼根际优良植物根际促生菌的分离筛选及促生特性研究[J]. 草业学报, 2019,28(11):114-123.
[21]	江红梅, 殷忠伟, 史发超, 等. 一株耐盐溶磷真菌的筛选、鉴定及其生物肥料的应用效果[J]. 植物营养与肥料学报, 2018,24(3):728-742.
[22]	Chinheya Cleopas C, Yobo Kwasi S, Laing Mark D. Biological control of the rootknot nematode, Meloidogyne javanica (Chitwood) using Bacillus isolates, on soybean[J]. Biological Control, 2017: 109.
[23]	汪筱雪, 韦继光, 杨振德, 等. 广西西北部核桃真菌性病害调查及核桃炭疽病防治试验[J]. 南方农业学报, 2018,49(8):1531-1540.
[24]	赵玉美, 赵宁, 冯小飞, 等. 云南核桃感病枝干真菌多样性初步研究[J]. 中国森林病虫, 2017,36(4):11-16.
[25]	杨越. 新疆核桃复合微生物菌种的筛选[D]. 乌鲁木齐:新疆农业大学, 2016.

序号	菌种编号	溶磷圈直径/cm	解钾圈直径/cm	IAA显色程度
1	Y1	0.5	0.3	++
2	Y7	1.2	1.3	+++++
3	Y8	0.2	0.4	+
4	5-7	0.5	0.2	+
5	33-4	0.4	0.2	++
6	YC-2	0.9	1.0	++++
7	YC-5	0.8	0.7	++++
8	KC-7	0.2	0.6	+
9	ML-5	0.1	0.5	++
10	ML-23	0.3	0.4	+
11	YN9	0.3	0.4	++
12	YN13	1.1	1.0	+++
13	BS-2	0.4	0.3	++
14	BS-6	0.2	0.6	+

序号	菌种编号	溶磷圈直径/cm	解钾圈直径/cm	IAA显色程度
1	Y1	0.5	0.3	++
2	Y7	1.2	1.3	+++++
3	Y8	0.2	0.4	+
4	5-7	0.5	0.2	+
5	33-4	0.4	0.2	++
6	YC-2	0.9	1.0	++++
7	YC-5	0.8	0.7	++++
8	KC-7	0.2	0.6	+
9	ML-5	0.1	0.5	++
10	ML-23	0.3	0.4	+
11	YN9	0.3	0.4	++
12	YN13	1.1	1.0	+++
13	BS-2	0.4	0.3	++
14	BS-6	0.2	0.6	+

特征	ht1	ht2	ht3	ht4
革兰氏染色	+	+	+	+
KOH反应	-	-	-	-
芽孢染色	+	+	+	+
淀粉水解	+	+	+	+
甲基红	-	-	-	-
V-P试验	+	+	+	+
吲哚产生	-	-	-	-
明胶液化	+	+	+	+
柠檬酸盐利用	+	+	+	+
硝酸盐还原	+	+	+	+
葡萄糖利用	+	+	+	+
丙二酸盐利用	-	-	-	-

特征	ht1	ht2	ht3	ht4
革兰氏染色	+	+	+	+
KOH反应	-	-	-	-
芽孢染色	+	+	+	+
淀粉水解	+	+	+	+
甲基红	-	-	-	-
V-P试验	+	+	+	+
吲哚产生	-	-	-	-
明胶液化	+	+	+	+
柠檬酸盐利用	+	+	+	+
硝酸盐还原	+	+	+	+
葡萄糖利用	+	+	+	+
丙二酸盐利用	-	-	-	-

序号	处理	可溶性磷	速效钾	IAA
1	ht1+ht2	718.31	12.41	88.25
2	ht1+ht3	751.24	10.94	91.21
3	ht1+ht4	775.55	14.75	68.32
4	ht2+ht3	674.33	9.92	85.98
5	ht2+ht4	712.67	12.19	64.41
6	ht3+ht4	687.99	11.47	47.38
7	ht1+ht2+ht3	701.12	12.58	67.35
8	ht1+ht2+ht4	721.66	14.84	65.32
9	ht1+ht3+ht4	695.85	13.52	127.32
10	ht2+ht3+ht4	678.74	10.29	87.21
11	ht1+ht2+ht3+ht4	776.13	15.88	187.46