HighlightArabidopsis glycerol-3-phosphate acyltransferase 9 (GPAT9) is an sn-1 specific acyl-CoA:GPAT that contributes to intracellular glycerolipid biosynthesis in seeds, developing leaves and pollen grains, but not to extracellular glycerolipid biosynthesis.
We report n-6 monounsaturated primary alcohols (C 26:1 , C 28:1 , and C 30:1 homologs) in the cuticular waxes of Arabidopsis (Arabidopsis thaliana) inflorescence stem, a class of wax not previously reported in Arabidopsis. The Arabidopsis cer17 mutant was completely deficient in these monounsaturated alcohols, and CER17 was found to encode a predicted ACYL-COENZYME A DESATURASE LIKE4 (ADS4). Studies of the Arabidopsis cer4 mutant and yeast variously expressing CER4 (a predicted fatty acyl-CoA reductase) with CER17/ADS4, demonstrated CER4's principal role in synthesis of these monounsaturated alcohols. Besides unsaturated alcohol deficiency, cer17 mutants exhibited a thickened and irregular cuticle ultrastructure and increased amounts of cutin monomers. Although unsaturated alcohols were absent throughout the cer17 stem, the mutation's effects on cutin monomers and cuticle ultrastructure were much more severe in distal than basal stems, consistent with observations that the CER17/ADS4 transcript was much more abundant in distal than basal stems. Furthermore, distal but not basal stems of a double mutant deficient for both CER17/ADS4 and LONG-CHAIN ACYL-COA SYNTHETASE1 produced even more cutin monomers and a thicker and more disorganized cuticle ultrastructure and higher cuticle permeability than observed for wild type or either mutant parent, indicating a dramatic genetic interaction on conversion of very long chain acyl-CoA precursors. These results provide evidence that CER17/ADS4 performs n-6 desaturation of very long chain acyl-CoAs in both distal and basal stems and has a major function associated with governing cutin monomer amounts primarily in the distal segments of the inflorescence stem. Plant cuticle coats most aerial surfaces of vascular plants and plays a major role in coordinating interactions between the plant and its environment (Rensing et al., 2008; Yeats and Rose, 2013). The cuticle is primarily composed of two lipid classes, the non-polymerized (free) cuticular waxes and the cutin polyester, both of which are synthesized by epidermal cells. Common plant wax compounds are the very long chain fatty acids (VLCFAs) and their derivatives including aldehydes, primary alcohols, alkanes, secondary alcohols, ketones, and esters (Samuels et al., 2008). As much as 4.0% of total waxes on Arabidopsis (Ara-bidopsis thaliana) inflorescence stems are composed of numerous yet unidentified wax compounds (Jenks et al., 1995). Cutin consists primarily of C 16 and C 18 fatty acid derivatives (e.g. hydroxy fatty acids and dicar-boxylic acids), which are linked by ester bonds; however , glycerol and small amounts of longer chain cutin monomers have also been reported (Pollard et al., 2008). In the past decade, there has been significant progress toward understanding the molecular mechanisms controlling the cuticular wax biosynthetic pathway based primarily on studies of wax-deficient mutants (Bernard and Joubès, 2013; Yeats and Rose, 2013). Synthesis of wax occurs in epidermal cells and begins with C 16 and C 18 lon...
Background It is important to explore renewable alternatives (e.g. biofuels) that can produce energy sources to help reduce reliance on fossil oils, and reduce greenhouse gases and waste solids resulted from fossil oils consumption. Camelina sativa is an oilseed crop which has received increasing attention due to its short life cycle, broader adaptation regions, high oil content, high level of omega-3 unsaturated fatty acids, and low-input requirements in agriculture practices. To expand its Camelina production areas into arid regions, there is a need to breed for new drought-tolerant cultivars. Leaf cuticular wax is known to facilitate plant development and growth under water-limited conditions. Dissecting the genetic loci underlying leaf cuticular waxes is important to breed for cultivars with improved drought tolerance. Results Here we combined phenotypic data and single nucleotide polymorphism (SNP) data from a spring C. sativa diversity panel using genotyping-by-sequencing (GBS) technology, to perform a large-scale genome-wide association study (GWAS) on leaf wax compositions. A total of 42 SNP markers were significantly associated with 15 leaf wax traits including major wax components such as total primary alcohols, total alkanes, and total wax esters as well as their constituents. The vast majority of significant SNPs were associated with long-chain carbon monomers (carbon chain length longer than C 28 ), indicating the important effects of long-chain carbon monomers on leaf total wax biosynthesis. These SNP markers are located on genes directly or indirectly related to wax biosynthesis such as maintaining endoplasmic reticulum (ER) morphology and enabling normal wax secretion from ER to plasma membrane or Golgi network-mediated transport. Conclusions These loci could potentially serve as candidates for the genetic control involved in intracellular wax transport that might directly or indirectly facilitate leaf wax accumulation in C. sativa and can be used in future marker-assisted selection (MAS) to breed for the cultivars with high wax content to improve drought tolerance.
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