Life and death among plant lysophosphatidic acid acyltransferases.

The tetraploid Brassica napus possesses several seed-expressed microsomal lysophosphatidic acid acyltransferases (LPAAT ) including BAT1.5, which has been retained after genome duplication as a consequence of a subfunctionalisation of the gene encoding the ubiquitously expressed Kennedy pathway enzyme BAT1.13. Next, cDNA BAT1.3, encoding a LPAAT was subsequently isolated from an embryo library. The rapeseed LPAAT encoded by BAT1.3 is orthologous to the Arabidopsis thaliana At1g51260 gene product possibly associated with tapetum development and male fertility. However, BAT1.3 expression is predominant during the mid stages of embryo development in seeds of Brassica napus. Functional characterisation of BAT1.3 provides further support for a hypothesis of gene dosage sensitivity of LPAATs as does an analysis of the chromosomal localisation of LPAAT genes in Arabidopsis thaliana. The pattern of retention or loss of LPAAT genes after polyploidisation or segmental duplication is consistent with a model of balanced gene drive.

Sekiguchi lesion gene encodes a cytochrome P450 monooxygenase that catalyzes conversion of tryptamine to serotonin in rice.

Serotonin is a well known neurotransmitter in mammals and plays an important role in various mental functions in humans. In plants, the serotonin biosynthesis pathway and its function are not well understood. The rice sekiguchi lesion (sl) mutants accumulate tryptamine, a candidate substrate for serotonin biosynthesis. We isolated the SL gene by map-based cloning and found that it encodes CYP71P1 in a cytochrome P450 monooxygenase family. A recombinant SL protein exhibited tryptamine 5-hydroxylase enzyme activity and catalyzed the conversion of tryptamine to serotonin. This pathway is novel and has not been reported in mammals. Expression of SL was induced by the N-acetylchitooligosaccharide (chitin) elicitor and by infection with Magnaporthe grisea, a causal agent for rice blast disease. Exogenously applied serotonin induced defense gene expression and cell death in rice suspension cultures and increased resistance to rice blast infection in plants. We also found that serotonin-induced defense gene expression is mediated by the RacGTPase pathway and by the G alpha subunit of the heterotrimeric G protein. These results suggest that serotonin plays an important role in rice innate immunity.

Expression of rapeseed microsomal lysophosphatidic acid acyltransferase isozymes enhances seed oil content in Arabidopsis.

In higher plants, lysophosphatidic acid acyltransferase (LPAAT), located in the cytoplasmic endomembrane compartment, plays an essential role in the synthesis of phosphatidic acid, a key intermediate in the biosynthesis of membrane phospholipids in all tissues and storage lipids in developing seeds. In order to assess the contribution of LPAATs to the synthesis of storage lipids, we have characterized two microsomal LPAAT isozymes, the products of homoeologous genes that are expressed in rapeseed (Brassica napus). DNA sequence homologies, complementation of a bacterial LPAAT-deficient mutant, and enzymatic properties confirmed that each of two cDNAs isolated from a Brassica napus immature embryo library encoded a functional LPAAT possessing the properties of a eukaryotic pathway enzyme. Analyses in planta revealed differences in the expression of the two genes, one of which was detected in all rapeseed tissues and during silique and seed development, whereas the expression of the second gene was restricted predominantly to siliques and developing seeds. Expression of each rapeseed LPAAT isozyme in Arabidopsis (Arabidopsis thaliana) resulted in the production of seeds characterized by a greater lipid content and seed mass. These results support the hypothesis that increasing the expression of glycerolipid acyltransferases in seeds leads to a greater flux of intermediates through the Kennedy pathway and results in enhanced triacylglycerol accumulation.

Interrelationships between dairy product intake, microflora metabolism, faecal properties and plasmid dissemination in gnotobiotic mice.

We previously described the effects of intake of dairy products on plasmid dissemination in the digestive tract of gnotobiotic mice associated with human faecal flora (HFF) and found that yoghurt, heat-treated yoghurt (HTY) and milk reduced population levels of transconjugants compared with findings in mice fed a standard mouse diet. In the case of lactose intake, transconjugants were not detected. The aim of the present study was to assess the possible interrelationships between these observations and other variables (bacterial ecology, pH, moisture, enzyme activities, short-chain fatty acid (SCFA) contents, lactic acid contents). Much of the interest of the present comparison lies in the fact that the animals were homogeneous in terms of age, gender, food and intestinal microflora, owing to the gnotobiotic mouse model maintained in sterile isolators. We observed no variation in SCFA and lactic acid contents or in the population levels of strictly anaerobic strains of Bacteroides and Bifidobacterium, and of the facultative anaerobic recipient Escherichia coli PG1 strain. The main modifications were the reduction of population levels of transconjugants in mice receiving yoghurt, HTY and milk, and concomitantly an increase of beta-galactosidase and a decrease of beta-glucosidase activities, compared with control mice fed a standard diet. Total inhibition of plasmid transfer was observed in HFF mice consuming lactose, and concomitantly the two enzyme activities (beta-glucosidase and beta-galactosidase) were increased, compared with the findings in control mice fed a standard diet. In axenic mice consuming lactose, plasmid transfer occurred, beta-galactosidase was not detected and beta-glucosidase was decreased. It is therefore proposed that these two enzyme activities influence plasmid transfer and persistence of transconjugants in the digestive tract of HFF associated mice. When both activities were increased there was a total inhibition of plasmid transfer (case of lactose intake). When beta-galactosidase increased and beta-glucosidase decreased (case of yoghurt, HTY and milk), plasmid transfer occurred at a lower efficiency than in the control group, resulting in lower population levels of transconjugants.