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High-oleate peanut mutants result from a MITE insertion into the FAD2 gene

Abstract

A high-oleate trait in the cultivated peanut (Arachis hypogaea L.) was reported to rely on the allelic composition of the two homeologous, microsomal oleoyl-PC desaturase genes (ahFAD2A or ahFAD2B). The enzyme activity of either ahFAD2A or ahFAD2B is sufficient for a normal oleate phenotype, and a significant reduction in the levels of ahFAD2B and a mutation in ahFAD2A were reported to be responsible for the high-oleate phenotype in one chemically induced mutant (M2-225) and one derived from a naturally occurring (8-2122) mutant. Here, we report an insertion of the same miniature inverted-repeat transposable element (MITE) in the ahFAD2B gene in another chemically induced mutant (Mycogen-Flavo) and the previously characterized M2-225 mutant. In both cases, this MITE insertion in ahFAD2B causes a frameshift, resulting in a putatively truncated protein sequence in both mutants. The insertion of this MITE in ahFAD2B, in addition to the point mutation in ahFAD2A, appears to be the cause of the high-oleate phenotype in Mycogen-Flavo and M2-225 mutants. Utilizing sequences of the MITE, we developed a DNA marker strategy to differentiate the two insertion-containing mutants from the normal oleate peanut variety (AT-108) and the naturally occurring, high-oleate mutant 8-2122. Reverse transcript-PCR/differential digestion results reveal the expression of both ahFAD2A and ahFAD2B genes in Mycogen-Flavo mutant. This result is in contrast to the observation that ahFAD2B transcripts are greatly reduced in the M2-225 mutant having the MITE insertion further 3′ in ahFAD2B gene.

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Acknowledgements

We would like to thank Dr. Renate Horn for her assistance in preparation of this manuscript. This work was performed in partial fulfillment of M. Patel’s Master of Science degree in genetics. The work was supported by Agra Tech Seeds, Inc.

Author information

Correspondence to A. Abbott.

Additional information

Communicated by C. Möllers

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Patel, M., Jung, S., Moore, K. et al. High-oleate peanut mutants result from a MITE insertion into the FAD2 gene. Theor Appl Genet 108, 1492–1502 (2004). https://doi.org/10.1007/s00122-004-1590-3

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Keywords

  • Kunitz Trypsin Inhibitor
  • Homeologous Gene
  • Diethyl Sulfate
  • Peanut Variety
  • Peanut Genome