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Plants use several pathways to synthesize phosphatidylcholine (PC), the major phospholipid of eukaryotic cells. PC has important structural and signaling roles. One pathway plants use for synthesis is the phospho-base methylation pathway, which forms the head-group phosphocholine through the triple methylation of phosphoethanolamine (PEA) catalyzed by phosphoethanolamine N-methyltransferases (PEAMTs). Our understanding of that pathway and its physiological importance remains limited. We recently reported that disruption of Arabidopsis thaliana PEAMT1/NMT1 and PEAMT3/NMT3 induces severe PC deficiency leading to dwarfism and impaired development. However, the double nmt1 nmt3 knock-out mutant is viable. Here, we show that this is enabled by residual PEAMT activity through a third family member, NMT2. The triple nmt1 nmt2 nmt3 knock-out mutant cannot synthesize PC from PEA and is lethal. This shows that, unlike mammals and yeast, Arabidopsis cannot form PC from phosphatidyl ethanolamine (PE), and demonstrates that methylation of PEA is the sole, and vital, entry point to PC synthesis. We further show that Arabidopsis has evolved an expanded family of four nonredundant PEAMTs through gene duplication and alternate use of the NMT2 promoter. NMT2 encodes two PEAMT variants, which greatly differ in their ability to perform the initial phospho-base methylation of PEA. Five amino acids at the N terminus of PEAMTs are shown to each be critical for the catalysis of that step committing to PC synthesis. As a whole, these findings open new avenues for enzymatic engineering and the exploration of ways to better tune phosphocholine and PC synthesis to environmental conditions for improved plant performance. [ABSTRACT FROM AUTHOR]

Phosphatidylcholine (PC) is a major membrane phospholipid and a precursor for major signaling molecules. Understanding its synthesis is important for improving plant growth, nutritional value, and resistance to stress. PC synthesis is complex, involving several interconnected pathways, one of which proceeds from serine-derived phosphoethanolamine to form phosphocholine through three sequential phospho-base methylations catalyzed by phosphoethanolamine N-methyltransferases (PEAMTs). The contribution of this pathway to the production of PC and plant growth has been a matter of some debate. Although a handful of individual PEAMTs have been described, there has not been any in planta investigation of a PEAMT family. Here, we provide a comparative functional analysis of two Arabidopsis (Arabidopsis thaliana) PEAMTs, NMT1 and the little known NMT3. Analysis of loss-of-function mutants demonstrates that NMT1 and NMT3 synergistically regulate PC homeostasis, phase transition at the shoot apex, coordinated organ development, and fertility through overlapping but also specific functions. The nmt1 nmt3 double mutant shows extensive sterility, drastically reduced PC concentrations, and altered lipid profiles. These findings demonstrate that the phospho-base methylation pathway makes a major contribution to PC synthesis in Arabidopsis and that NMT1 and NMT3 play major roles in its catalysis and the regulation of PC homeostasis as well as in plant growth and reproduction. [ABSTRACT FROM AUTHOR]

Arsenic is a ubiquitous environmental poison that inhibits root elongation and seed germination to a variable extent depending on the plant species. To understand the molecular mechanisms of arsenic resistance, a genetic screen was developed to isolate arsenate overly sensitive (aos) mutants from an activation-tagged Arabidopsis (Arabidopsis thaliana) population. Three aos mutants were isolated, and the phenotype of each was demonstrated to be due to an identical disruption of plastidial LIPOAMIDE DEHYDROGENASE1 (ptLPD1), a gene that encodes one of the two E3 isoforms found in the plastidial pyruvate dehydrogenase complex. In the presence of arsenate, ptlpd1-1 plants exhibited reduced root and shoot growth and enhanced anthocyanin accumulation compared with wild-type plants. The ptlpd1-1 plants accumulated the same amount of arsenic as wild-type plants, indicating that the aos phenotype was not due to increased arsenate in the tissues but to an increase in time innate sensitivity to the poison. Interestingly, a ptlpd1-4 knockdown allele produced a partial aos phenotype. Two loss-of-function alleles of ptLPD2 in Arabidopsis also caused elevated arsenate sensitivity, but the sensitivity was less pronounced than for the ptlpd1 mutants. Moreover, both the ptlpd1 and ptlpd2 mutants were more sensitive to arsenite than wild-type plants, and time LPD activity in isolated chloroplasts from wild-type plants was sensitive to arsenite but not arsenate. These findings show that the ptLPD isoforms are critical in vivo determinants of arsenite-mediated arsenic sensitivity in Arabidopsis and possible strategic targets for increasing arsenic tolerance. [ABSTRACT FROM AUTHOR]

Alterations of mitochondrial-encoded subunits of the FoF1-ATP synthase are frequently associated with cytoplasmic male sterility (CMS) in plants; however, little is known about the relationship of the nuclear encoded subunits of this enzyme with CMS. In the present study, the full cDNA of the gene TaFAd that encodes the putative FAd subunit of the FoF1-ATP synthase was isolated from the wheat (Triticum aestivum) fertility restorer ‘2114’ for timopheevii cytoplasm-based CMS. The deduced 238 amino acid polypeptide is highly similar to its counterparts in dicots and other monocots but has low homology to its mammalian equivalents. TaFAd is a single copy gene in wheat and maps to the short arm of the group 6 chromosomes. Transient expression of the TaFAd–GFP fusion in onion epidermal cells demonstrated TaFAd's mitochondrial location. TaFAd was expressed abundantly in stem, leaf, anther, and ovary tissues of 2114. Nevertheless, its expression was repressed in anthers of CMS plants with timopheevii cytoplasm. Genic male sterility did not affect its expression in anthers. The expression of the nuclear gene encoding the 20 kDa subunit of Fo was down-regulated in a manner similar to TaFAd in the T-CMS anthers while that of genes encoding the 6 kDa subunit of Fo and the γ subunit of F1 was unaffected. These observations implied that TaFAd is under mitochondrial retrograde regulation in the anthers of CMS plants with timopheevii cytoplasm. [ABSTRACT FROM PUBLISHER]