Crops ›› 2024, Vol. 40 ›› Issue (2): 221-227.doi: 10.16035/j.issn.1001-7283.2024.02.027

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Screening and Feature Analysis of Atoxigenic Aspergillus flavus in Reducing Aflatoxin Prodution in Peanuts Soil in Xiangyang

Wang Xue1,2,3(), Li Li1, Zhang Shujuan1, Zhu Mengjie1, Zhang Qi2,3, Li Peiwu2,3, Dong Jing1, Wang Dun1,2,3(), Wu Yanan4   

  1. 1Xiangyang Academy of Agricultural Sciences, Xiangyang 441057, Hubei, China
    2Oil Crops Research Institute of Chinese Academy of Agriculture Sciences, Wuhan 430062, Hubei, China
    3Hubei Hongshan Laboratory, Wuhan 430062, Hubei, China
    4Xiangyang Public Inspection and Testing Center, Xiangyang 441057, Hubei, China
  • Received:2022-04-25 Revised:2023-11-08 Online:2024-04-15 Published:2024-04-15

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Abstract:

Several Aspergillus flavus strains were isolated from Xiangyang, Hubei peanut soil medium. Through characteristic analysis and sequencing identification, the trains were identified as Aspergillus flavus strains. Using liquid fermentation culture, aflatoxin content in the soil was detected, and the atoxigenic Aspergillus flavus strains were obtained at the same time. Through the flat screening, the several good Aspergillus flavus strains were screened at the beginning. The standard and non-virulent strains were used in symbiotic culture to determine the AFT content. The results showed that there were four strains of enterotoxigenic symbiotic strains of Aspergillus flavus, toxins aflatoxin content lower than control strains over 80%, including XZ38 inhibitory reached 99.49%. This indiated that non-toxigenic Aspergillus flavus strains had a great role in the inhibition of enterotoxigenic aspergillus flavus growth, reduction or elimination aflatoxin, and could be used as alternative strains for biological control for the field aflatoxin contamination offers a good alternative.

Key words: Aspergillus flavus, Atoxigenic Aspergillus flavus, Characteristic analysis

Fig.1

PCR amplification identification electropherogram"

Table 1

Classification comparison results of strains on BLAST software"

样品
Sample		 同源性
Homology		 AFPA平板菌落颜色
The colony colour
of AFPA plate		 参考物种
Reference
species
GC-01 702/720(98%) 亮橘色 曲霉属Aspergillus
ZY-06 618/628(98%) 亮橘色 曲霉属Aspergillus
ZY-07 605/608(99%) 亮橘色 曲霉属Aspergillus
ZY-08 597/599(99%) 亮橘色 曲霉属Aspergillus
GX-01 625/631(99%) 亮橘色 曲霉属Aspergillus
NZ-01 598/601(99%) 亮橘色 曲霉属Aspergillus
XZ-34 599/600(99%) 亮橘色 曲霉属Aspergillus
XZ-40 662/672(99%) 亮橘色 曲霉属Aspergillus
XZ-63 604/608(99%) 亮橘色 曲霉属Aspergillus
XZ-69 659/669(99%) 亮橘色 曲霉属Aspergillus
YC-01 601/602(99%) 亮橘色 曲霉属Aspergillus
YC-03 616/623(99%) 亮橘色 曲霉属Aspergillus
YC-11 627/630(99%) 亮橘色 曲霉属Aspergillus
YC-14 621/629(99%) 亮橘色 曲霉属Aspergillus
......

Fig.2

Toxigenic ability by Aspergillus flavus at different regions"

Fig.3

Proportion of distribution of toxin-producing and non-toxic strains in peanut stubble soil"

Fig.4

Antagonism experiments of toxin-producing and non-toxin-producing strains"

Fig.5

Inhibition rates of toxin-producing strains by non-toxin-producing strains"

Fig.6

Fluorescence observation of Aspergillus flavus (a) Blank control of toxin-producing strain, (b) Toxin-producing and non-toxin-producing strain co-cultured."

Fig.7

Liquid phase standard map of the four AFT mixed standards"

Table 2

Standard curve of AFT"

AFT标准品
AFT standard		 标准曲线
Standard curve		 R2 检出限
Limit of detection (LOD) (μg/kg)		 定量限
Limit of quantification (μg/kg)
AFG2 Y=392 96.0X-17 563.0 0.999 968 5 0.26 1.3
AFG1 Y=12 069.0X-9167.47 0.999 956 5 0.36 1.3
AFB2 Y=88 965.4X-57 284.3 0.999 972 8 0.18 1.0
AFB1 Y=44 595.7X-35 890.3 0.999 968 7 0.24 1.0

Fig.8

Status of symbiotic mycelium of toxin- producing and non-toxin-producing strains"

Table 3

Inhibition effects of non-toxic on toxin-producing Aspergillus flavus"

菌株
Strain		 产毒量
Toxin
production
(μg/kg)		 抑制产毒量
Inhibition of
toxin production
(μg/kg)		 抑制产毒率
Inhibition rate of
toxin production
rate (%)
SZ3.4408(SZ)(105) 1072.95±1.67 － －
NZ03:SZ(105:105) 197.26±3.05 875.69±3.05 81.61±1.73a
NZ06:SZ(105:105) 676.07±4.56 396.88±4.56 36.99±2.02a
ZY10:SZ(105:105) 1050.52±5.69 22.43±5.69 2.09±3.06b
ZY11:SZ(105:105) 1053.58±5.04 19.37±5.04 1.81±2.33a
XZ21:SZ(105:105) 936.82±4.83 136.13±4.83 12.69±1.85a
XZ38:SZ(105:105) 5.49±0.19 1067.46±0.19 99.49±0.75c
XZ52:SZ(105:105) 869.79±3.66 203.16±3.66 18.93±1.32d
XZ56:SZ(105:105) 869.47±2.92 203.48±2.92 18.96±1.25d
XZ59:SZ(105:105) 192.45±2.04 880.50±2.04 82.06±0.80c
XZ64:SZ(105:105) 604.78±3.06 468.17±3.06 43.63±1.58d
YC10:SZ(105:105) 75.77±1.35 997.19±1.35 92.94±0.67b

Fig.9

Spore morphology of YC10 (left) and XZ38 (right) under the microscope"

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