Abstract
Maize is one of the more important agricultural crops in the world and, under certain conditions, prone to attack from pathogenic fungi. One of these, Aspergillus flavus, produces toxic and carcinogenic metabolites, called aflatoxins, as byproducts of its infection of maize kernels. The α-amylase of A. flavus is known to promote aflatoxin production in the endosperm of these infected kernels, and a 36-kDa protein from the Lablab purpureus, denoted AILP, has been shown to inhibit α-amylase production and the growth of A. flavus. Here, we report the isolation of six full-length labAI genes encoding AILP and a detailed analysis of the activities of the encoded proteins. Each of the six labAI genes encoded sequences of 274 amino acids, with the deduced amino acid sequences showing approximately 95–99% identity. The sequences are similar to those of lectin members of a legume lectin–arcelin–α-amylase inhibitor family reported to function in plant resistance to insect pests. The labAI genes did not show any of the structures characteristic of conserved structures identified in α-amylase inhibitors to date. The recombinant proteins of labAI-1 and labAI-2 agglutinated human red blood cells and inhibited A. flavus α-amylase in a manner similar to that shown by AILP. These data indicate that labAI genes are a new class of lectin members in legume seeds and that their proteins have both lectin and α-amylase inhibitor activity. These results are a valuable contribution to our knowledge of plant–pathogen interactions and will be applicable for developing protocols aimed at controlling A. flavus infection.
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Abbreviations
- α-AI:
-
α-Amylase inhibitor
- AIL:
-
α-Amylase inhibitor-like protein
- AILP:
-
α-Amylase inhibitor from Lablab purpureus
- AP:
-
Adaptor primer
- AOX:
-
Alcohol oxidase
- BMGY:
-
Buffered glycerol complex medium
- BMMY:
-
Buffered methanol complex medium
- GSP:
-
Gene-specific primer
- PHA:
-
Phytohemagglutinin
- TI:
-
Trypsin inhibitor
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Acknowledgements
This work was supported by Korea Research Foundation Grant (204-2002-F00001) and by a grant from the BioGreen 21 Program funded by the Rural Development Administration, Republic of Korea. Support was also provided by USDA Cooperative Agreement 58-6435-8-118. We thank Burt Bluhm for providing the labAI antibody.
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Communicated by J. R. Liu
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Kim, YH., Woloshuk, C.P., Cho, E.H. et al. Cloning and functional expression of the gene encoding an inhibitor against Aspergillus flavus α-amylase, a novel seed lectin from Lablab purpureus (Dolichos lablab). Plant Cell Rep 26, 395–405 (2007). https://doi.org/10.1007/s00299-006-0250-2
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DOI: https://doi.org/10.1007/s00299-006-0250-2