Identification of N-ethylmaleimide-sensitive and -insensitive phosphatidate phosphatase activity in microspore-derived cultures of oilseed rape

Furukawa-Stoffer, Tara L.; Byers, Susan D.; Hodges, D. Mark; Laroche, André; Weselake, Randall J.

doi:10.1016/s0168-9452(97)00247-1

Cited by 14 publications

(6 citation statements)

References 30 publications

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“…DGAT assays were performed as previously reported by Middleton et al (3). PAP assays were conducted in the presence and absence of 0.5 mM N-ethylmaleimide (NEM) in order to distinguish between PAP-1 and PAP-2 activities (11,12). LPAAT assays were performed on microsomal fractions from the PCD muscle based on the assay published by Schlossman and Bell (13).…”

Section: Methodsmentioning

confidence: 99%

Fatty acid composition of muscle fat and enzymes of storage lipid synthesis in whole muscle from beef cattle

Kazala

Lozeman

Mir

et al. 2006

Lipids

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Enhanced intramuscular fat content (i.e., marbling) in beef is a desirable trait, which can result in increased product value. This study was undertaken with the aim of revealing biochemical factors associated with the marbling trait in beef cattle. Samples of longissimus lumborum (LL) and pars costalis diaphragmatis (PCD) were taken from a group of intact crossbred males and females at slaughter, lipids extracted, and the resulting FAME examined for relationships with marbling fat deposition. For LL, significant associations were found between degree of marbling and myristic (14:0, r = 0.55, P < 0.01), palmitic (16:0, r = 0.80, P < 0.001), stearic (18:0, r = -0.58, P < 0.01), and oleic (18:1c-9, r = 0.79, P < 0.001) acids. For PCD, significant relationships were found between marbling and palmitic (r = 0.71, P < 0.001) and oleic (r = 0.74, P < 0.001) acids. Microsomal fractions prepared from PCD muscle were assayed for diacylglycerol acyltransferase (DGAT), lysophosphatidic acid acyltransferase (LPAAT), and phosphatidic acid phosphatase-1 (PAP-1) activity, and the results examined for relationships with degree of intramuscular fat deposition. None of the enzyme activities from PCD displayed an association with marbling fat content, but DGAT specific activity showed significant positive associations with LPAAT (r = 0.54, P < 0.01), total PAP (r = 0.66, P < 0.001), and PAP-1 (r = 0.63, P < 0.01) specific activities. The results on FA compositions of whole muscle tissues provide insight into possible enzyme action associated with the production of specific FA. The increased proportion of oleic acid associated with enhanced lipid content of whole muscle is noteworthy given the known health benefits of this FA.

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Section: Methodsmentioning

confidence: 99%

Fatty acid composition of muscle fat and enzymes of storage lipid synthesis in whole muscle from beef cattle

Kazala

Lozeman

Mir

et al. 2006

Lipids

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“…Previously, we have used these cell suspension cultures in the study of storage lipid biosynthesis in developing oilseeds (Weselake and Taylor 1999), and have characterized several lipid biosynthetic enzymes from this culture (Furukawa-Stoffer et al 1998;Byers et al 1999;Nykiforuk et al 2002;Furukawa-Stoffer et al 2003). In the present study, we extended this model to the examination of gene expression changes in response to abiotic stress and evaluated the ploidy levels and embryogenic potential of these cultures, which have been maintained via continuous biweekly sub-culture for 23 years.…”

Section: Discussionmentioning

confidence: 99%

“…Jet Neuf have been used in studies involving freezing tolerance (Orr et al 1986;Johnson-Flanagan and Singh 1987;Johnson-Flanagan et al 1991), mutant selection (Saxena et al 1990), nutritional physiology (Moraes and Plaxton 2000) and storage lipid synthesis (Furukawa-Stoffer et al 1998;Byers et al 1999;Weselake and Taylor 1999;Furukawa-Stoffer et al 2003). The cells were originally generated in 1983 during an attempt to induce androgenesis in microspores of B. napus (Simmonds et al 1991).…”

Section: Introductionmentioning

confidence: 99%

Microspore-derived cell suspension cultures of oilseed rape as a system for studying gene expression

Shi

Warrington

et al. 2007

Plant Cell Tiss Organ Cult

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Abiotic stress, such as extreme temperature, drought, or excessive salinity, is one of the leading causes of crop loss worldwide. Microspore-derived (MD) cell suspension cultures of Brassica napus L. cv. Jet Neuf have been shown to be a useful system for studying the biochemistry of developing oilseeds. In the present study, we describe the application of MD cell suspension cultures of B. napus as a system for studying gene expression in response to abiotic stress, and demonstrate emybryogenic competence in cultures that have been continuously subcultured for more than 20 years. MD cell suspension cultures of B. napus L. cv Jet Neuf were exposed to low temperature or osmotic stress and the expression profile of known stress responsive genes was evaluated. The gene expression profile of BN115, a known cold-responsive gene in B. napus, was similar to that described for intact cold-acclimated plants. Likewise, two late embryogenesis abundant (Lea) genes were shown to be upregulated in response to low temperature or osmotic stress. The results demonstrate that B. napus MD cell suspension cultures are a useful system for the investigation of changes in gene expression in plants brought about by abiotic stress.

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“…B. napus (L. cv Jet Neuf) microsporederived (BnaMD) cell suspension cultures were developed in 1984 and maintained in various laboratories. These cultures have been used for many scientific studies in the biochemistry and molecular biology of storage lipid biosynthesis, the enzymology of oilseed carbohydrate metabolism and abiotic stress [3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21]. We are particularly interested in examining molecular strategies for increasing the oil content of plant biomass [22].…”

Section: Introductionmentioning

confidence: 99%

A Rapid Nile Red Fluorescence‐Based Method for Triacylglycerol Content in Microspore‐Derived Cell Suspension Cultures of Brassica napus

Gao

Chen

Weselake

2014

Lipids

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Brassica napus (L. cv Jet Neuf) microspore-derived (BnaMD) cell suspension cultures were used to study the biochemistry and molecular biology of plant lipid metabolism. The cell suspension cultures may also be a potentially useful trait-testing tool for further improvement of oilseed rape, the world's third largest vegetable oil crop. Nile red fluorescence methods have been used to assess neutral lipid accumulation in microorganisms and mammalian oocytes but have not been established for plant cell suspension cultures. In this study, a Nile red method was established and optimized for triacylglycerol (TAG) content analysis in BnaMD cell suspension cultures. Nile red fluorescence was easily determined in 96-well plates using a 490 nm excitation wavelength and a 595 nm emission wavelength without any significant interference from other cellular components. The lipid content of the cell suspension cultures at different growth stages was determined using a conventional lipid analysis method and the Nile red fluorescence method. There was a strong positive correlation between the Nile red fluorescence and TAG content in the cell suspension cultures. Therefore, the TAG content of BnaMD cell suspension cultures can be rapidly and robustly determined using a Nile red fluorescence-based method that can potentially be adapted for application to other plant cell suspension cultures.

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Identification of N-ethylmaleimide-sensitive and -insensitive phosphatidate phosphatase activity in microspore-derived cultures of oilseed rape

Cited by 14 publications

References 30 publications

Fatty acid composition of muscle fat and enzymes of storage lipid synthesis in whole muscle from beef cattle

Fatty acid composition of muscle fat and enzymes of storage lipid synthesis in whole muscle from beef cattle

Microspore-derived cell suspension cultures of oilseed rape as a system for studying gene expression

A Rapid Nile Red Fluorescence‐Based Method for Triacylglycerol Content in Microspore‐Derived Cell Suspension Cultures of Brassica napus

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