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.

Grantor dc:publisher

University of North Texas

Year dc:date

2002

Author dc:creator

• Hodson, Jane E.

Contributors dc:contributor

• Chapman, Kent D.
• Pirtle, Robert M.
• Knesek, John

• Tobacco.
• Phospholipases.
• Molecular cloning.
• PLDβ1
• Phospholipase D β1
• phospholipid metabolism
• tobacco
• lipid signaling

Statement dc:rights

• Public
• Copyright
• Hodson, Jane E.
• Copyright is held by the author, unless otherwise noted. All rights reserved.

Language dc:language

English

Identifier

oclc: 52162126
https://digital.library.unt.edu/ark:/67531/metadc3341/
ark: ark:/67531/metadc3341

OAI identifier oai:identifier

info:ark/67531/metadc3341