除草活性菌株HL-1产孢发酵条件研究

doi:10.3969/j.issn.1004-1524.2021.06.09

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (6): 1042-1048.DOI: 10.3969/j.issn.1004-1524.2021.06.09

除草活性菌株HL-1产孢发酵条件研究

高汉峰(), 刘雨芹, 程亮, 郭青云^*()

青海大学农林科学院植物保护研究所,青海省农业有害生物综合治理重点实验室,农业农村部西宁作物有害生物科学观测实验站,青海西宁 810016

收稿日期:2020-06-29 出版日期:2021-06-25 发布日期:2021-06-25
通讯作者: 郭青云
作者简介:*郭青云,E-mail: guoqingyunqh@163.com
高汉峰(1995—),男,甘肃永登人,硕士研究生,主要从事杂草生物防治研究。E-mail: gaohanfenglz@163.com
基金资助:
国家自然科学基金(31760539);青海省科技厅项目(2018-ZJ-917);第二次青藏高原综合科学考察研究项目(2019QZKK0303)

Study on fermentation conditions of herbicidal active strain HL-1

GAO Hanfeng(), LIU Yuqin, CHENG Liang, GUO Qingyun^*()

Institute of Plant Protection, Academy of Agriculture and Forestry Science, Qinghai University, Key Laboratory of Agricultural Integrated Pest Management of Qinghai Province, Scientific Observing and Experimental Station of Crop Pest of Ministry of Agriculture and Rural Affairs in Xining, Xining 810016, China

Received:2020-06-29 Online:2021-06-25 Published:2021-06-25
Contact: GUO Qingyun

摘要/Abstract

摘要：

以分离自青海患病刺儿菜中具有除草活性的植物内生真菌HL-1菌株为研究对象,通过单因素试验筛选最适合该菌株生长的碳氮源、固态发酵基质、接种量和固-液发酵条件。结果表明:在供试的材料中最适碳源是葡萄糖,氮源是酵母浸膏;该菌株在PDA培养基中的最大产孢量是5.79×10⁸ CFU·mL^-1;而在最适碳源和最适氮源混合培养基中,它的最大产孢量是 4.81×10⁹ CFU·mL^-1。菜籽饼为HL-1在供试材料中产孢最大的培养基质;在体积分数为40%接种量下,它的产孢量最大。通过正交优化试验发现该菌株在培养温度27.5 ℃,培养基质含水量37%,发酵时间8 d条件下,产孢量最大。

关键词: 植物内生真菌HL-1, 固态发酵基质, 固-液发酵条件

Abstract:

Taking endophytic fungus HL-1 strain with herbicidal activity isolated from Qinghai diseased Cirsium setosum as the material, a single factor test was used to screen the carbon and nitrogen sources, solid fermentation substrate, inoculation amount and solid-liquid fermentation conditions that were most suitable for the growth of the strain. The results showed that the most suitable carbon source of HL-1 in the tested materials was glucose and the nitrogen source was yeast extract; when the strain was cultured in PDA medium, its maximum spore production was 5.79×10⁸ CFU·mL^-1 ; and in the optimal carbon source and optimal nitrogen source mixed medium, its maximum spore production was 4.81×10⁹ CFU·mL^-1. Rapeseed cake was the medium material with the largest spore production of HL-1 in the test materials, which had the largest spore production at a volume fraction of 40% inoculation volume. Through orthogonal optimization experiments, the spore production of the strain was found to be the largest under the conditions of culture temperature 27.5 ℃, culture medium water content 37% and fermentation time 8 d.

Key words: endophytic fungus HL-1, solid fermentation substrate, solid-liquid fermentation conditions

中图分类号:

S476.1

高汉峰, 刘雨芹, 程亮, 郭青云. 除草活性菌株HL-1产孢发酵条件研究[J]. 浙江农业学报, 2021, 33(6): 1042-1048.

GAO Hanfeng, LIU Yuqin, CHENG Liang, GUO Qingyun. Study on fermentation conditions of herbicidal active strain HL-1[J]. Acta Agriculturae Zhejiangensis, 2021, 33(6): 1042-1048.

图/表 7

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水平 Level	培养条件Culture condition
水平 Level	培养基含水量 Medium water content/%	培养温度 Culture temperature/℃	培养时间 Culture time/d
上星号臂(+1.6818) Upper asterisk arm (+1.6818)	65	35	14
上水平(+1) Upper level(+1)	58	32	12
零水平(0) Zero level(0)	47.5	27.5	8
下水平(-1) Lower level(-1)	37	23	5
下星号臂(-1.6818) Lower asterisk arm (-1.6818)	30	20	3

水平 Level	培养条件Culture condition
水平 Level	培养基含水量 Medium water content/%	培养温度 Culture temperature/℃	培养时间 Culture time/d
上星号臂(+1.6818) Upper asterisk arm (+1.6818)	65	35	14
上水平(+1) Upper level(+1)	58	32	12
零水平(0) Zero level(0)	47.5	27.5	8
下水平(-1) Lower level(-1)	37	23	5
下星号臂(-1.6818) Lower asterisk arm (-1.6818)	30	20	3

碳氮源 Carbon and nitrogen sources	HL-1菌株的菌落形态 Colony morphology of HL-1 strain
C1	白色的菌落,没有同心轮纹,发达的气生菌丝 White colony without concentric ring pattern, developed aerial mycelium
C2	乳白色的菌落,具有同心轮纹,较少的气生菌丝 Milky white colony with concentric ring pattern, less aerial mycelium
C3	纯白色的菌落,具有同心状轮纹,发达的气生菌丝 Pure white colony with concentric ring pattern, developed aerial mycelium
C4	纯白色的菌落,具有同心轮纹,发达的气生菌丝且铺满整皿 Pure white colony with concentric ring pattern, developed aerial mycelium and full petrie dish
N1	菌落表面有乳白色的绒毛状菌丝,菌落同心轮纹,边缘呈带丝状,基内菌丝呈褐色 Milky white hairy mycelium on the surface of the colony, concentric ring colony with filamentous edge, brown subtrate mycelium
N2	白色的菌丝,基内菌丝部分呈灰褐色,发达的气生菌丝和基内菌丝 White mycelium, gray-brown and developed substrate mycelium, developed aerial mycelium
N3	较小的菌落形态,灰褐色的气生菌丝且表面有隆起,发达的基内菌丝 Small colony morphology, gray-brown aerial mycelium with raised surface, developed substrate mycelium
N4	较大的菌落形态,乳白色的气生菌丝,具有同心轮纹,菌落边缘呈带丝状,黄褐色的基内菌丝 Large colony morphology, milky white aerial mycelium,concentric ring pattern, filamentous edge, yellow-brown substrate mycelium

碳氮源 Carbon and nitrogen sources	HL-1菌株的菌落形态 Colony morphology of HL-1 strain
C1	白色的菌落,没有同心轮纹,发达的气生菌丝 White colony without concentric ring pattern, developed aerial mycelium
C2	乳白色的菌落,具有同心轮纹,较少的气生菌丝 Milky white colony with concentric ring pattern, less aerial mycelium
C3	纯白色的菌落,具有同心状轮纹,发达的气生菌丝 Pure white colony with concentric ring pattern, developed aerial mycelium
C4	纯白色的菌落,具有同心轮纹,发达的气生菌丝且铺满整皿 Pure white colony with concentric ring pattern, developed aerial mycelium and full petrie dish
N1	菌落表面有乳白色的绒毛状菌丝,菌落同心轮纹,边缘呈带丝状,基内菌丝呈褐色 Milky white hairy mycelium on the surface of the colony, concentric ring colony with filamentous edge, brown subtrate mycelium
N2	白色的菌丝,基内菌丝部分呈灰褐色,发达的气生菌丝和基内菌丝 White mycelium, gray-brown and developed substrate mycelium, developed aerial mycelium
N3	较小的菌落形态,灰褐色的气生菌丝且表面有隆起,发达的基内菌丝 Small colony morphology, gray-brown aerial mycelium with raised surface, developed substrate mycelium
N4	较大的菌落形态,乳白色的气生菌丝,具有同心轮纹,菌落边缘呈带丝状,黄褐色的基内菌丝 Large colony morphology, milky white aerial mycelium,concentric ring pattern, filamentous edge, yellow-brown substrate mycelium

碳氮源处理 Carbon and nitrogen source treatments	孢子平均值 Average value of spores/(CFU·mL^-1)	D值 D value
C1	2.30×10⁶	0.110 dD
C2	1.95×10⁶	0.009 dDE
C3	1.05×10⁷	0.250 cC
C4	1.50×10⁸	0.517 bB
N1	4.60×10⁶	0.069 dDE
N2	3.90×10⁶	0.033 deDE
N3	2.20×10⁹	0.650 aA
N4	2.25×10⁵	0.008 eE
PDA	5.79×10⁸	0.528 bB
C4+N3	4.81×10⁹	0.691 aA

除草活性菌株HL-1产孢发酵条件研究

Study on fermentation conditions of herbicidal active strain HL-1

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参考文献 20

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试验号 No.	含水量 Water content	温度 Temperature	培养时间 Culture time	lgCFU均值 lgCFU average/%
1	1	1	1	12.3
2	1	1	-1	31.3
3	1	-1	1	24.0
4	1	-1	-1	17.4
5	-1	1	1	35.7
6	-1	1	-1	32.5
7	-1	-1	1	40.0
8	-1	-1	-1	45.1
9	-1.6818	0	0	23.0
10	1.6818	0	0	31.1
11	0	-1.6818	0	14.0
12	0	1.6818	0	17.2
13	0	0	-1.6818	24.3
14	0	0	1.6818	31.2
15	0	0	0	21.5
16	0	0	0	24.1
17	0	0	0	28.0
18	0	0	0	29.2
19	0	0	0	25.0
20	0	0	0	23.4
21	0	0	0	25.0
22	0	0	0	28.1
23	0	0	0	22.0

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