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ARS Home » Research » Publications at this Location » Publication #67441
Title: GENETIC TRANSFORMATION OF CULTIVATED PEANUTS WITH AN ANTIFUNGAL HYDROLASE GENE FROM RICE

Author
item Burns, J
item Melouk, Hassan

Submitted to: Congress on In Vitro Biology
Publication Type: Abstract Only
Publication Acceptance Date: 6/22/1996
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: A chitinase previously shown to reduce fungal disease in transgenic plants was incorporated into the peanut genome as an initial step to enhance disease resistance. Embryogenic tissue cultures of Arachis hypogaea L. cvs Okrun and Tamspan 90 were generated from explants of mature, dry, stored seed for biolistics-mediated (PDS-1000/He) transient expression and transformation with pBI221 and/or pNUT2.5. pNUT2.5 is a pTRA141 derivative containing 35S-hph and 35S*2-rch10 expression cassettes. Rch10 encodes a defense-related class I chitinase from rice. pNUT2.5-bombarded tissue was exposed to 20-27 mg/L hygromycin-B in liquid medium for up to 3 months to isolate resistant embryos. Maximum GUS expression occurred with 1.6 micrometers microcarriers, 1800 psi, and bombarding tissue on empty petri plates. Pre-bombardment treatments of 0.5 M mannitol for 8 or 24 hr followed by a 24-hr post-treatment had little effect on transient GUS expression and was detrimental to the bombarded tissue during hygromycin selection. Maximum stable transformation occurred with 0.7 micrometer W or 1.0 micrometer Au microcarriers at 1800 psi. Over 30 hygromycin-resistant lines of either Okrun or Tamspan 90 transformed with pNUT2.5 were generated. Hygromycin-resistant embryos were used to generate shoot- cultures that were analyzed by PCR and then rooted into plants.