深绿木霉T2菌株发酵液蛋白提取物TraT2A抑菌条件优化及其对百合叶斑病的田间防治效果

陈应娥; 梁巧兰; 魏列新; 同发宇; 王冬

doi:10.16801/j.issn.1008-7303.2022.0031

深绿木霉T2菌株发酵液蛋白提取物TraT2A抑菌条件优化及其对百合叶斑病的田间防治效果

doi: 10.16801/j.issn.1008-7303.2022.0031

1.
甘肃农业大学植物保护学院，甘肃省农作物病虫害生物防治工程实验室，兰州 730070

基金项目: 甘肃省农牧厅项目 (GNSW-2016-25)；兰州市科技局项目 (2018-4-78)；甘肃省科技重大专项 (18ZD2NA010)；甘肃农业大学学科建设基金 (GAU-XKJS-2018-155).

详细信息

作者简介:
陈应娥， 1470857734@qq.com

通讯作者:
梁巧兰，liangql@gsau.edu.cn

中图分类号: S482.2；TQ450.2
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- HTML全文浏览量: 25
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- 被引次数: 0
出版历程
- 收稿日期: 2021-09-26
- 录用日期: 2022-01-13
- 网络出版日期: 2022-04-14
- 刊出日期: 2022-08-03

Optimization of inhibition conditions of Trichoderma atroviride T2 fermentation liquid protein extract TraT2A and its field control efficacy for lily leaf spot disease

1.
College of Plant Protection, Gansu Agricultural University, Biocontrol Engineering Laboratory of Crop Diseases and Pests of Gansu Province, Lanzhou 730070, China

Funds: Agriculture and Animal Husbandry Program of Gansu Province (GNSW-2016-25); Science and Technology Bureau Program of Lanzhou (2018-4-78); The Major Science and Technology Program of Gansu Province (18ZD2NA010); Discipline Construction Fund of Gansu Agricultural University (GAU-XKJS-2018-155).

摘要

摘要: 为了提高深绿木霉Trichoderma atroviride T2菌株发酵液蛋白提取物TraT2A的抑菌活性和光稳定性，采用菌丝生长速率法，通过对紫外保护剂 (抗坏血酸、腐殖酸) 和辅助剂 (IE-08、SP-4821A和SP-4821B) 的筛选，优化了TraT2A对百合叶斑病菌Alternaria alternata的抑菌条件，并测定了其对百合叶斑病的田间防治效果。结果表明：TraT2A在200.00 mg/mL下对百合叶斑病菌A.alternata的抑菌作用较好，抑制率为72.12%；随着紫外光照时间的延长，TraT2A的抑菌活性逐渐降低，添加5.00 mg/mL的腐殖酸对TraT2A有较好的紫外保护作用，抑制率 (69.01%) 比不加紫外保护剂对照 (29.37%) 提高39.64%；添加0.17 mg/mL 的辅助剂SP-4821A对TraT2A活性的提高效果最好，抑制率 (95.13%) 比TraT2A对照 (69.16%) 提高25.97%。按照优化后的配方：200 g TraT2A (质量浓度为200.00 mg/mL) + 5 g腐殖酸 (质量浓度为5.00 mg/mL) + 0.17 g SP-4821A (质量浓度为0.17 mg/mL) + 794.83 mL灭菌水，进行田间药效试验，结果发现其对百合叶斑病的防治效果为80.37%。该研究结果为进一步开发防治百合病害的TraT2A制剂提供了理论依据。
- 深绿木霉T2菌株 /
- 发酵液蛋白提取物 /
- TraT2A /
- 百合叶斑病菌 /
- 抑菌活性 /
- 田间防效
Abstract: In order to improve the inhibition activity and light stability of the protein extract TraT2A from the fermentation broth of Trichoderma atroviride T2 strain, the ultraviolet protective agents (ascorbic acid, humic acid) and the auxiliary agent (IE-08, SP-4821A and SP-4821B) were screened, the inhibition condition of TraT2A for Alternaria alternata were optimized through the colony growth rate test, and the field control efficacy on lily leaf spot disease was determined. The results showed that TraT2A had a good inhibitory effect on A.alternata at 200.00 mg/mL and the inhibitory rate was 72.12%. With the prolongation of the UV illumination time, the inhibition activity of TraT2A gradually decreases. The addition of 5.00 mg/mL humic acid had an excellent UV protection effect on TraT2A and its inhibition rate (69.01%) increased by 39.64% compared with that without a UV protection agent (29.37%). The addition of 0.17 mg/mL adjuvant of SP-4821A increased the inhibition activity of TraT2A most significantly and the inhibition rate (95.13%) increased by 25.97% compared with the TraT2A control (69.16%). According to the optimized formula, 200 g TraT2A (the mass concentration was 200.00 mg/mL), 5 g humic acid (the mass concentration was 5.00 mg/mL), 0.17 g SP-4821A (the mass concentration was 0.17 mg/mL) and 794.83 mL sterilized water was mixed and the field control efficacy of the mixture was tested on lily leaf spot disease, the result showed that its field control efficacy was 80.37%. The research results provide a theoretical basis for the further development of TraT2A preparations for the control of lily diseases.
- Trichoderma atroviride T2 strain /
- fermentation protein extract /
- TraT2A /
- Alternaria alternata /
- inhibition activity /
- field control efficacy

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表 1 TraT2A对百合叶斑病菌的抑菌作用

Table 1. The inhibition effect of TraT2A on A. alternata

TraT2A 质量浓度 TraT2A concentration/(mg/mL)	抑制率 Inhibition rate/%
200.00	72.12 ± 0.25 aA
100.00	67.31 ± 0.68 bB
66.67	26.92 ± 0.57 cC
注：表中数据为平均值 ± 标准误差。同列数据后不同小写和大写字母分别表示经Duncan氏新复极差法检验在0.05和0.01水平上差异显著。Note: Data in the table were shown as mean ± standard error. In the same column of data, different lowercase or capital letters indicated that there were significant differences at 0.05 and 0.01 levels, respectively.

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表 2 紫外光照射时间对TraT2A抑菌活性的影响 (200.00 mg/mL)

Table 2. The influence of UV irradiation time on the inhibition activity of TraT2A (200.00 mg/mL)

处理 Treatment	紫外光照射时间 UV irradiation time/min	抑制率 Inhibition rate/%
TraT2 A	10	59.68 ± 0.23 bB
	20	45.70 ± 0.47 cC
	30	29.57 ± 0.42 dD
	40	20.61 ± 0.59 eE
TraT2A 对照 TraT2A control	0	72.04 ± 0.21 aA
CK	0	—
注：表中数据为平均值 ± 标准误差。同列数据后不同小写和大写字母分别表示经Duncan氏新复极差法检验在0.05和0.01水平上差异显著。Note: Data in the table were shown as mean ± standard error. In the same column of data, different lowercase or capital letters indicated that there were significant differences at 0.05 and 0.01 levels, respectively.

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表 3 紫外保护剂对TraT2A抑菌活性的影响 (200.00 mg/mL)

Table 3. The influence of UV protective agents on the inhibition activity of TraT2A (200.00 mg/mL)

处理 Treatment	紫外光照射时间 UV irradiation time/min	抑制率 Inhibition rate/%
TraT2A + 抗坏血酸 TraT2A plus ascorbic acid	30	54.41 ± 0.17 cC
TraT2A + 腐殖酸 TraT2A plus humic acid	30	69.01 ± 0.67 bB
TraT2 A	30	29.37 ± 0.41 dD
TraT2 A	0	70.45 ± 0.57 aA
抗坏血酸 Ascorbic acid	0	23.24 ± 0.48 eE
腐殖酸 Humic acid	0	3.18 ± 0.43 fF
CK	0	—
注：表中数据为平均值 ± 标准误差。同列数据后不同小写和大写字母分别表示经Duncan氏新复极差法检验在0.05和0.01水平上差异显著。Note: Data in the table were shown as mean ± standard error. In the same column of data, different lowercase or capital letters indicated that there were significant differences at 0.05 and 0.01 levels, respectively.

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表 4 辅助剂对TraT2A抑菌活性的影响 (200.00 mg/mL)

Table 4. The influence of adjuvants on the inhibition activity of TraT2A (200.00 mg/mL)

处理 Treatment	浓度 Concentration/(mg/mL)	抑制率 Inhibition rate/%
处理 Treatment	浓度 Concentration/(mg/mL)	辅助剂 Adjuvant	TraT2A + 辅助剂 TraT2A + adjuvant	与 TraT2A 对照相比抑菌作用提高百分点 Increase percentage points of inhibition effect compared with TraT2A control
IE-08	0.50	15.56 ± 0.89 aA	87.55 ± 0.53 aA	18.36 aA
	0.25	12.81 ± 0.61 bB	81.46 ± 0.99 bB	12.30 bB
	0.13	9.88 ± 0.36 cC	76.74 ± 0.73 cC	7.58 cC
	0.07	5.81 ± 0.82 dD	74.44 ± 0.95 dD	5.28 dD
	0.04	1.62 ± 0.94 eE	70.24 ± 0.63 eE	1.08 eE
SP-4821 B	0.17	13.29 ± 0.60 aA	88.61 ± 0.98 aA	19.45 aA
	0.09	10.96 ± 0.67 bB	80.50 ± 0.61 bB	11.34 bB
	0.05	9.28 ± 0.32 cC	73.35 ± 0.44 cC	4.19 cC
	0.03	6.77 ± 0.79 dD	71.55 ± 0.51 cdCD	2.39 cdCD
	0.02	3.41 ± 0.93 eE	71.03 ± 0.46 deDE	1.87 dD
SP-4821 A	0.17	18.74 ± 0.18 aA	95.13 ± 0.61 aA	25.97 aA
	0.09	14.84 ± 0.07 bB	82.70 ± 0.69 bB	13.54 bB
	0.05	11.49 ± 0.89 cC	75.10 ± 0.58 cC	5.94 cC
	0.03	8.38 ± 1.01 dD	73.33 ± 0.59 dD	4.17 dD
	0.02	5.99 ± 0.99 eE	70.59 ± 0.48 eE	1.43 eE
TraT2A 对照 TraT2A control	200.00	—	69.16 ± 0.32 eE	—
注：表中数据为平均值 ± 标准误差。同列数据后不同小写和大写字母分别表示经Duncan氏新复极差法检验在0.05和0.01水平上差异显著。
Note: Data in the table were shown as mean ± standard error. In the same column of data, different lowercase or capital letters indicated that there were significant differences at 0.05 and 0.01 levels, respectively.

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表 5 TraT2A与紫外保护剂、辅助剂混合液对百合叶斑病的田间防效

Table 5. The field control efficacy of TraT2A + UV protective agents + adjuvants mixture for lily leaf spot

处理 Treatment	施药前病情指数 Disease index before treatment	第 1 次施药后 10 d Ten days after the first treatment		第 2 次施药后 10 d Ten days after the second treatment		第 3 次施药后 10 d Ten days after the third treatment
处理 Treatment	施药前病情指数 Disease index before treatment	病情指数 Disease index	防治效果 Control efficacy/ %	病情指数 Disease index	防治效果 Control efficacy/ %	病情指数 Disease index	防治效果 Control efficacy/ %
200.00 mg/mL TraT2A + 5.00 mg/mL 腐殖酸 + 0.17 mg/mL SP-4821A 200.00 mg/mLTraT2A + 5.00 mg/mL humic acid + 0.17 mg/mL SP-4821 A	3.04	5.17	68.19 aA	5.81	75.41 aA	6.13	80.37 aA
200.00 mg/mL TraT2 A	3.32	8.23	53.64 bB	10.04	61.08 bB	12.46	63.47 bB
5.00 mg/mL腐殖酸 5.00 mg/mL Humic acid	3.09	15.33	7.22 dD	22.03	8.25 dD	28.97	8.75 dD
0.17 mg/mL SP-4821 A	3.25	15.27	12.13 cC	21.76	13.84 cC	28.53	14.55 cC
CK	3.14	16.79	—	24.40	—	32.26	—
注：同列数据后不同小写和大写字母分别表示经Duncan氏新复极差法检验在0.05和0.01水平上差异显著。Note: In the same column of data, different lowercase or capital letters indicated that there were significant differences at 0.05 and 0.01 levels, respectively.

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