Phospholipase D (PLD) initiates phospholipid (PL) catabolism in plant cells and is also involved in signal transduction and retailoring of membrane PL. Total PL declines and phosphatidic acid increases in pericarp tissue during tomato fruit ripening, suggesting that increased PLD activity alters membrane structure. To assess the role of PLD in tomato ripening, we have begun a molecular genetic approach. Using a castor bean PLDα cDNA as a probe, a PLDα cDNA (LEPLD2) was isolated from our tomato fruit library. It has an open reading frame of 2421 nucleotides, encoding a polypeptide of 807 amino acids with a molecular mass of 92 kDa. The deduced LEPLD2 PLDα shares > 75% sequence identity with PLDαs from castor bean, tobacco and tomato. LEPLD2 transcript, detected by RNA gel-blot analysis, was very low in roots, low in stems, moderate in leaves, high in flowers, and increased in fruit during development and ripening. Expression of LEPLD2 in Escherichia coli yielded active enzyme, and a FLAG-PLDα fusion protein produced by transformed E. coli migrated close to the calculated 94 kDa on SDS/PAGE.
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Conference Article|
December 01 2000
Characterization of a phospholipase Dα cDNA from tomato fruit
B. D. Whitaker;
B. D. Whitaker
1
1Horticultural Crops Quality Laboratory, Agricultural Research Service, USDA 10300 Baltimore Avenue, Beltsville, MD 20705-2350, U.S.A.
1To whom correspondence should be addressed (e-mail WhitakeB@ba.ars.usda.gov)
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D. L. Smith;
D. L. Smith
1Horticultural Crops Quality Laboratory, Agricultural Research Service, USDA 10300 Baltimore Avenue, Beltsville, MD 20705-2350, U.S.A.
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K. C. Green
K. C. Green
1Horticultural Crops Quality Laboratory, Agricultural Research Service, USDA 10300 Baltimore Avenue, Beltsville, MD 20705-2350, U.S.A.
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Publisher: Portland Press Ltd
Received:
June 16 2000
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© 2000 Biochemical Society
2000
Biochem Soc Trans (2000) 28 (6): 819–821.
Article history
Received:
June 16 2000
Citation
B. D. Whitaker, D. L. Smith, K. C. Green; Characterization of a phospholipase Dα cDNA from tomato fruit. Biochem Soc Trans 1 December 2000; 28 (6): 819–821. doi: https://doi.org/10.1042/bst0280819
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