响应面法优化禾谷镰刀菌产脱氧雪腐镰刀菌烯醇及其衍生物的液体培养条件

doi:10.13304/j.nykjdb.2021.1055

摘要/Abstract

摘要：

为研究不同液体培养条件对禾谷镰刀菌脱氧雪腐镰刀菌烯醇（deoxynivalenol，DON）和15-乙酰基脱氧雪腐镰刀菌烯醇（15-acetyldeoxynivalenol，15-ADON）产量的影响，采用单因素试验和响应面法进行研究。结果表明，培养时间、pH及两者间交互作用对DON产量影响极显著；培养时间及培养时间与培养温度的交互作用对15-ADON产量影响极显著。DON的最佳产毒条件为：培养温度26.3 ℃，初始pH 6.35，培养22 d，此条件下DON的产量达5 739.69 μg·L^-1；15-ADON的最佳产毒条件为：培养温度26.3 ℃，初始pH 5.58，培养21 d，此条件下15-ADON的产量达10 453.41 μg·L^-1，为DON及15-ADON的生物合成、日常监管和污染防控等提供理论依据和数据支持。

关键词: 禾谷镰刀菌, 液体培养, 脱氧雪腐镰刀菌烯醇, 乙酰化脱氧雪腐镰刀菌烯醇, 响应面法

Abstract:

To investigate the effects of different liquid culture conditions on the production of deoxynivalenol （DON） and 15-acetyldeoxynivalenol （15-ADON） by Fusarium graminearum， the single factor experiment and response surface methodology were used. The results showed that the production of DON was significantly influenced by incubation time， initial pH and their interactive effect， while the production of 15-ADON was significantly influenced by incubation time， incubation temperature and their interactive effect. The optimal conditions for DON production was followed with incubation time 22 d， temperature 26.3 ℃， initial pH 6.35， which the highest amount of DON was 5 739.69 μg·L^-1. The highest production of 15-ADON with 10 453.41 μg·L^-1 was achieved when the incubation time， the temperature and the initial pH were set as 21 d， 26.3 ℃ and 5.58， respectively. The above results provided theoretical basis and data support for the biosynthesis， routine supervision and control of DON and 15-ADON.

Key words: Fusarium graminearum, liquid incubation, deoxynivalenol, acetylated deoxynivalenol, response surface methodology

中图分类号:

TS201.3

赵秀英, 黄晴雯, 曹浩杰, 王杰, 李瑞姣, 聂冬霞, 韩铮, 赵志辉. 响应面法优化禾谷镰刀菌产脱氧雪腐镰刀菌烯醇及其衍生物的液体培养条件[J]. 中国农业科技导报, 2023, 25(7): 222-233.

Xiuying ZHAO, Qingwen HUANG, Haojie CAO, Jie WANG, Ruijiao LI, Dongxia NIE, Zheng HAN, Zhihui ZHAO. Optimization of Liquid Culture Conditions for the Production of Deoxynivalenol and Its Derivatives by Fusarium graminearum Using Response Surface Methodology[J]. Journal of Agricultural Science and Technology, 2023, 25(7): 222-233.

图/表 12

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名称Name	5 μg·L^-1
名称Name	回收率 Recovery/%	日内精密度 Intra-day precision/%	日间精密度 Inter-day precision/%	回收率 Recovery/%	日内精密度 Intra-day precision/%	日间精密度 Inter-day precision/%	回收率 Recovery/%	日内精密度 Intra-day precision/%	日间精密度 Inter-day precision/%
DON	103.97	7.02	5.67	83.28	7.53	7.36	81.03	11.98	3.55
3-ADON	86.06	3.32	5.26	82.90	4.50	4.56	80.18	3.18	7.69
15-ADON	90.72	5.28	6.78	87.93	4.31	7.36	81.34	2.80	6.33

名称Name	5 μg·L^-1
名称Name	回收率 Recovery/%	日内精密度 Intra-day precision/%	日间精密度 Inter-day precision/%	回收率 Recovery/%	日内精密度 Intra-day precision/%	日间精密度 Inter-day precision/%	回收率 Recovery/%	日内精密度 Intra-day precision/%	日间精密度 Inter-day precision/%
DON	103.97	7.02	5.67	83.28	7.53	7.36	81.03	11.98	3.55
3-ADON	86.06	3.32	5.26	82.90	4.50	4.56	80.18	3.18	7.69
15-ADON	90.72	5.28	6.78	87.93	4.31	7.36	81.34	2.80	6.33

试验号 Test number	培养时间 Incubation time/d	培养温度 Incubation temperature/℃	初始pH Initial pH	DON产量 Production of DON/（μg·L^-1）
1	30.00	28.00	7.50	3 428.56
2	20.00	28.00	6.00	5 508.33
3	20.00	28.00	6.00	5 480.50
4	30.00	20.00	6.00	2 427.21
5	10.00	35.00	6.00	68.60
6	30.00	28.00	4.50	378.45
7	30.00	35.00	6.00	1 006.98
8	10.00	20.00	6.00	1 003.23
9	20.00	20.00	4.50	678.46
10	10.00	28.00	7.50	578.55
11	10.00	28.00	4.50	57.98
12	20.00	28.00	6.00	6 677.33
13	20.00	35.00	7.50	1 689.00
14	20.00	35.00	4.50	500.78
15	20.00	28.00	6.00	5 363.87
16	20.00	20.00	7.50	2 677.33
17	20.00	28.00	6.00	5 143.87

试验号 Test number	培养时间 Incubation time/d	培养温度 Incubation temperature/℃	初始pH Initial pH	DON产量 Production of DON/（μg·L^-1）
1	30.00	28.00	7.50	3 428.56
2	20.00	28.00	6.00	5 508.33
3	20.00	28.00	6.00	5 480.50
4	30.00	20.00	6.00	2 427.21
5	10.00	35.00	6.00	68.60
6	30.00	28.00	4.50	378.45
7	30.00	35.00	6.00	1 006.98
8	10.00	20.00	6.00	1 003.23
9	20.00	20.00	4.50	678.46
10	10.00	28.00	7.50	578.55
11	10.00	28.00	4.50	57.98
12	20.00	28.00	6.00	6 677.33
13	20.00	35.00	7.50	1 689.00
14	20.00	35.00	4.50	500.78
15	20.00	28.00	6.00	5 363.87
16	20.00	20.00	7.50	2 677.33
17	20.00	28.00	6.00	5 143.87

来源 Source	平方和 Sum of squares	自由度 Degree of freedom	均方 Mean squares	F值 F value	P值 P value
模型 Model	82 250 000.00	9	9 139 000.00	36.98^**	< 0.000 1
A	3 860 000.00	1	3 860 000.00	15.62^**	0.005 5
B	1 550 000.00	1	1 550 000.00	6.27^*	0.040 7
C	5 786 000.00	1	5 786 000.00	23.41^**	0.001 9
AB	52 476.25	1	52 476.25	0.21	0.658 9
AC	1 600 000.00	1	1 600 000.00	6.47^*	0.038 4
BC	158 800.00	1	158 800.00	0.64	0.449 1
A²	24 090 000.00	1	24 090 000.00	97.48^**	<0.000 1
B²	19 340 000.00	1	19 340 000.00	78.27^**	<0.000 1
C²	19 140 000.00	1	19 140 000.00	77.45^**	<0.000 1
残差 Residual	1 730 000.00	7	247 100.00
失拟项 Lack of Fit	288 700.00	3	96 221.44	0.27	0.846 6
纯误差 Pure error	1 441 000.00	4	360 300.00
总误差 Total error	83 980 000.00	16