, Volume 40, Issue 4, pp 305–310 | Cite as

Comparison of different inoculating methods to evaluate the pathogenicity and virulence of Aspergillus niger on two maize hybrids



A two-year field study was conducted to determine the effects of artificial inoculation techniques on the pathogenicity and virulence of Aspergillus niger kernel infection on two maize hybrids. Test plants included in the study were hybrids resistant and susceptible to Aspergillus flavus to determine if the host resistance mechanisms that limited A. flavus infection would also suppress A. niger infection. Ears were inoculated with the silk-channel, side-needle, and spray techniques 7 days after midsilk (50% of the plants in a plot had silk emerging). Ears were also inoculated with a modified-pinbar technique 21 days after midsilk. Kernel infection in 2008 in inoculated plants ranged from 2% to 11% and from 2% to 45% in the resistant and susceptible hybrids, respectively. In 2009, kernel infection in inoculated plants ranged from 13% to 32% and from 10% to 67% in the resistant and susceptible, respectively. The silk-channel, side-needle, and modified-pinbar techniques produced significantly higher levels of kernel infection in the susceptible hybrid in both years than the spray technique. When hybrids were compared, the silk-channel, side-needle, and modified-pinbar techniques induced significantly higher levels of infections in the susceptible hybrid than in the resistant hybrid in 2008 and 2009. The level of A. niger pathogenicity and virulence increased when conidia were placed inside the husks of developing ears by wounding (modified-pinbar and side-needle techniques) or non-wounding (silk-channel technique) inoculation methods. Although A. niger kernel infection was significantly lower in the A. flavus resistant hybrid compared to the A. flavus susceptible hybrid, A. niger infection levels were much higher than A. flavus infection levels typically observed in both of these hybrids in past studies.


Aspergillus flavus Corn Kernel infection Zea mays 



The authors thank Mike Alpe, Greg Flint, Gerald Matthews, and Ladonna Owens for excellent technical assistance. This article reports the results of research only. Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the USDA or Mississippi State University. This article is a contribution of the USDA-ARS in cooperation with the Mississippi Agricultural and Forestry Experiment Station, Mississippi State, MS. It is published with the approval of both agencies.


  1. Abarca, M. L., Accensi, F., Bragulat, M. R., & Cabanes, F. J. (1994). Ochratoxin a production by strains of Aspergillus niger var. niger. Applied and Environmental Microbiology, 60, 2650–2652.PubMedGoogle Scholar
  2. Bacon, C., & Palencia, E. (2010). Production of mycotoxins by members of the Aspergillus section Nigri isolated from peanuts and maize in the United States. Phytopathology, 100, S10.Google Scholar
  3. Fennell, D. I., Bothast, R. J., Lillehoj, E. B., & Peterson, R. E. (1973). Bright greenish-yellow fluorescence and associated fungi in white corn naturally contaminated with aflatoxin. Cereal Chemistry, 50, 404–414.Google Scholar
  4. Frisvad, J. C., Smedsgaard, J., Samson, R. A., Larsen, T. O., & Thrane, U. (2007). Fumonisin B2 production by Aspergillus niger. Journal of Agricultural and Food Chemistry, 55, 9727–9732.PubMedCrossRefGoogle Scholar
  5. Hesseltine, R. F., Rogers, R. F., & Shotwell, O. L. (1981). Aflatoxin and mold flora in North Carolina in 1977 corn crop. Mycologia, 73, 216–228.CrossRefGoogle Scholar
  6. Magnoli, C., Hallak, C., Astoreca, A., Ponsone, L., Chiacchiera, S., & Dalcero, A. M. (2006). Occurrence of ochratoxin a-producing fungi in commercial corn kernels in Argentina. Mycopathologia, 161, 53–58.PubMedCrossRefGoogle Scholar
  7. Mogensen, J. M., Frisvad, J. C., Thrane, U., & Nielsen, K. F. (2010). Production of fumonisin B2 and B4 by Aspergillus niger on grapes and raisins. Journal of Agricultural and Food Chemistry, 58, 954–958.PubMedCrossRefGoogle Scholar
  8. Mukanga, M., Derera, J., Tongoona, P., & Laing, M. D. (2010). A survey of pre-harvest ear rot diseases of maize and associated mycotoxins in south and central Zambia. International Journal of Food Microbiology, 141, 213–221.PubMedCrossRefGoogle Scholar
  9. Palencia, E. R., Glenn, A. E., & Bacon, C. W. (2009). Colonization of maize seedlings under drought conditions by two ochratoxin A producer species within the A. section Nigri. Phytopathology, 99, S99.Google Scholar
  10. Palencia, E. R., Hinton, D. M., & Bacon, C. W. (2010). The black Aspergillus species of maize and peanuts and their potential for mycotoxin production. Toxins, 2, 399–416.PubMedCrossRefGoogle Scholar
  11. Robertson-Hoyt, L. A., Betrán, J., Payne, G. A., White, D. G., Isakeit, T., Maragos, C., et al. (2007). Relationships among resistances to Fusarium and Aspergillus ear rots and contamination by fumonisin and aflatoxin in maize. Phytopathology, 97, 311–317.PubMedCrossRefGoogle Scholar
  12. Scott, G. E., & Zummo, N. (1988). Sources of resistance in maize to kernel infection by Aspergillus flavus in the field. Crop Science, 28, 504–507.CrossRefGoogle Scholar
  13. Scott, G. E., & Zummo, N. (1990). Preharvest kernel infection by Aspergillus flavus for resistant and susceptible maize hybrids. Crop Science, 30, 381–383.CrossRefGoogle Scholar
  14. Scott, G. E., Zummo, N., Lillehoj, E. B., Widstrom, N. W., Kang, M. S., West, D. R., et al. (1991). Aflatoxin in corn hybrids field inoculated with Aspergillus flavus. Agronomy Journal, 83, 595–598.CrossRefGoogle Scholar
  15. Taubenhaus, J. J. (1920). A study of the black and yellow molds of ear corn. Texas Agricultural Experiment Station Bulletin, 270, 3–10.Google Scholar
  16. Varga, J., Kocsube, S., Suri, K., Szigeti, G., Szekeres, A., Varga, M., et al. (2010). Fumonisin contamination and fumonisin producing black Aspergilli in dried vine fruits of different origin. International Journal of Food Microbiology, 143, 143–149.PubMedCrossRefGoogle Scholar
  17. Williams, W. P., Windham, G. L., & Buckley, P. M. (2005). Enhancing maize with resistance to Aspergillus flavus infection and aflatoxin accumulation. In H. K. Abbas (Ed.), Aflatoxin and food safety (pp. 370–394). New York, NY: CRC Press.Google Scholar
  18. Williams, W. P., Windham, G. L., Buckley, P. M., & Daves, C. A. (2002). Aflatoxin contamination in conventional and transgenic corn hybrids infested with southwestern corn borer (Lepidoptera: Crambidae). Journal of Agricultural and Urban Entomology, 19, 227–236.Google Scholar
  19. Windham, G. L., Hawkins, L. K., & Williams, W. P. (2010). Aflatoxin accumulation and kernel infection of maize hybrids inoculated with Aspergillus flavus and Aspergillus parasiticus. World Mycotoxin Journal, 3, 89–93.CrossRefGoogle Scholar
  20. Windham, G. L., & Williams, W. P. (1998). Aspergillus flavus infection and aflatoxin accumulation in resistant and susceptible maize hybrids. Plant Disease, 82, 281–284.CrossRefGoogle Scholar
  21. Windham, G. L., & Williams, W. P. (2002). Evaluation of corn inbreds and advanced breeding lines for resistance to aflatoxin contamination in the field. Plant Disease, 86, 232–234.CrossRefGoogle Scholar
  22. Windham, G. L., & Williams, W. P. (2007). A comparison of inoculation techniques for inducing aflatoxin contamination and Aspergillus flavus kernel infection on corn hybrids in the field. Phytoparasitica, 35, 277–281.CrossRefGoogle Scholar
  23. Windham, G. L., Williams, W. P., Buckley, P. M., Abbas, H. K., & Hawkins, L. K. (2005). Techniques used to identify aflatoxin-resistant corn. In H. K. Abbas (Ed.), Aflatoxin and food safety (pp. 407–421). New York, NY: CRC Press.Google Scholar
  24. Zummo, N., & Scott, G. E. (1989). Evaluation of field inoculation techniques for screening maize genotypes against kernel infection by Aspergillus flavus in Mississippi. Plant Disease, 73, 313–316.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. (outside the USA) 2012

Authors and Affiliations

  1. 1.USDA-ARS Corn Host Plant Resistance Research UnitMississippi StateUSA

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