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Tobacco Phospholipase D β1: Molecular Cloning and Biochemical Characterization

Transgenic tobacco plants were developed containing a partial PLD clone in antisense orientation. The PLD isoform targeted by the insertion was identified. A PLD clone was isolated from a cDNA library using the partial PLD as a probe: Nt10B1 shares 92% identity with PLDβ1 from tomato but lacks the C2 domain. PCR analysis confirmed insertion of the antisense fragment into the plants: three introns distinguished the endogenous gene from the transgene. PLD activity was assayed in leaf homogenates in PLDβ/g conditions. When phosphatidylcholine was utilized as a substrate, no significant difference in transphosphatidylation activity was observed. However, there was a reduction in NAPE hydrolysis in extracts of two transgenic plants. In one of these, a reduction in elicitor- induced PAL expression was also observed.

Identiferoai:union.ndltd.org:unt.edu/info:ark/67531/metadc3341
Date12 1900
CreatorsHodson, Jane E.
ContributorsChapman, Kent D., Pirtle, Robert M., Knesek, John
PublisherUniversity of North Texas
Source SetsUniversity of North Texas
LanguageEnglish
Detected LanguageEnglish
TypeThesis or Dissertation
FormatText
RightsPublic, Copyright, Hodson, Jane E., Copyright is held by the author, unless otherwise noted. All rights reserved.

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