Molecular cloning and characterization of the <i>CER2</i> gene of <i>Arabidopsis thaliana</i>

Negruk, V. I.; Yang, Ping; Subramanian, Mohan Raj; McNevin, John; Lemieux, Bertrand

doi:10.1046/j.1365-313x.1996.09020137.x

Cited by 94 publications

(76 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As a complement to reverse genetic approaches, forward genetic strategies can be a highly effective means of gene discovery. In such cases, phenotypic changes in the fruit cuticle can lead to identification of the underlying gene, as was the case with several of the eceriferum Arabidopsis mutants (Negruk et al, 1996). In tomato, these strategies enabled the discovery of new cuticle-associated genes and functions, including genes involved in cutin synthesis and polymerization (Yeats et al, 2012b;Shi et al, 2013;Petit et al, 2014) and in cuticle regulation (Adato et al, 2009;Isaacson et al, 2009;Ballester et al, 2010;Nadakuduti et al, 2012).…”

Section: Discussionmentioning

confidence: 99%

The glycerol-3-phosphate acyltransferase GPAT6 from tomato plays a central role in fruit cutin biosynthesis

et al. 2016

View full text Add to dashboard Cite

The thick cuticle covering and embedding the epidermal cells of tomato (Solanum lycopersicum) fruit acts not only as a protective barrier against pathogens and water loss but also influences quality traits such as brightness and postharvest shelf-life. In a recent study, we screened a mutant collection of the miniature tomato cultivar Micro-Tom and isolated several glossy fruit mutants in which the abundance of cutin, the polyester component of the cuticle, was strongly reduced. We employed a newly developed mapping-by-sequencing strategy to identify the causal mutation underlying the cutin deficiency in a mutant thereafter named gpat6-a (for glycerol-3-phosphate acyltransferase6). To this end, a backcross population (BC 1 F 2 ) segregating for the glossy trait was phenotyped. Individuals displaying either a wild-type or a glossy fruit trait were then pooled into bulked populations and submitted to whole-genome sequencing prior to mutation frequency analysis. This revealed that the causal point mutation in the gpat6-a mutant introduces a charged amino acid adjacent to the active site of a GPAT6 enzyme. We further showed that this mutation completely abolished the GPAT activity of the recombinant protein. The gpat6-a mutant showed perturbed pollen formation but, unlike a gpat6 mutant of Arabidopsis (Arabidopsis thaliana), was not male sterile. The most striking phenotype was observed in the mutant fruit, where cuticle thickness, composition, and properties were altered. RNA sequencing analysis highlighted the main processes and pathways that were affected by the mutation at the transcriptional level, which included those associated with lipid, secondary metabolite, and cell wall biosynthesis.

show abstract

Section: Discussionmentioning

confidence: 99%

The glycerol-3-phosphate acyltransferase GPAT6 from tomato plays a central role in fruit cutin biosynthesis

et al. 2016

View full text Add to dashboard Cite

show abstract

“…CER1 has been proposed to encode an aldehyde decarbonylase (Aarts et al, 1995) mainly because of the increase in aldehydes and reduced levels of alkanes, secondary alcohols, and alkanes in the cer1 mutant. The CER2 gene might code for a CoAdependent acyltransferase, but its precise function is unknown (Negruk et al, 1996;Xia et al, 1996;Kunst and Samuels, 2003). The cer2 mutant accumulates C26 and C28 acyl groups, primary alcohols, and wax esters and shows reduced levels of the decarbonylation pathway products.…”

Section: Introductionmentioning

confidence: 99%

The SHINE Clade of AP2 Domain Transcription Factors Activates Wax Biosynthesis, Alters Cuticle Properties, and Confers Drought Tolerance when Overexpressed in Arabidopsis[W]

Aharoni¹,

Dixit²,

et al. 2004

View full text Add to dashboard Cite

The interface between plants and the environment plays a dual role as a protective barrier as well as a medium for the exchange of gases, water, and nutrients. The primary aerial plant surfaces are covered by a cuticle, acting as the essential permeability barrier toward the atmosphere. It is a heterogeneous layer composed mainly of lipids, namely cutin and intracuticular wax with epicuticular waxes deposited on the surface. We identified an Arabidopsis thaliana activation tag gain-of-function mutant shine (shn) that displayed a brilliant, shiny green leaf surface with increased cuticular wax compared with the leaves of wild-type plants. The gene responsible for the phenotype encodes one member of a clade of three proteins of undisclosed function, belonging to the plant-specific family of AP2/EREBP transcription factors. Overexpression of all three SHN clade genes conferred a phenotype similar to that of the original shn mutant. Biochemically, such plants were altered in wax composition (very long fatty acid derivatives). Total cuticular wax levels were increased sixfold in shn compared with the wild type, mainly because of a ninefold increase in alkanes that comprised approximately half of the total waxes in the mutant. Chlorophyll leaching assays and fresh weight loss experiments indicated that overexpression of the SHN genes increased cuticle permeability, probably because of changes in its ultrastructure. Likewise, SHN gene overexpression altered leaf and petal epidermal cell structure, trichome number, and branching as well as the stomatal index. Interestingly, SHN overexpressors displayed significant drought tolerance and recovery, probably related to the reduced stomatal density. Expression analysis using promoter-b-glucuronidase fusions of the SHN genes provides evidence for the role of the SHN clade in plant protective layers, such as those formed during abscission, dehiscence, wounding, tissue strengthening, and the cuticle. We propose that these diverse functions are mediated by regulating metabolism of lipid and/or cell wall components.

show abstract

“…Four of these-CER2 , CER3 , GL2 , and GL15 -appear to encode regulatory loci (Tacke et al, 1995;Hannoufa et al, 1996;Moose and Sisco, 1996;Negruk et al, 1996;Xia et al, 1996), three-CER6 , KCS1 , and GL8 -may encode metabolic enzymes (Xu et al, 1997;Millar et al, 1999;Todd et al, 1999;Fiebig et al, 2000), and two-CER1 and GL1 -may be involved in either the transport or the metabolism of wax compounds (Aarts et al, 1995;Hansen et al, 1997). None of these genes has been associated with cuticle membrane synthesis.…”

Section: Introductionmentioning

confidence: 99%

Cloning and Characterization of the WAX2 Gene of Arabidopsis Involved in Cuticle Membrane and Wax Production

et al. 2003

View full text Add to dashboard Cite

Insertional mutagenesis of Arabidopsis ecotype C24 was used to identify a novel mutant, designated wax2 , that had alterations in both cuticle membrane and cuticular waxes. Arabidopsis mutants with altered cuticle membrane have not been reported previously. Compared with the wild type, the cuticle membrane of wax2 stems weighed 20.2% less, and when viewed using electron microscopy, it was 36.4% thicker, less opaque, and structurally disorganized. The total wax amount on wax2 leaves and stems was reduced by Ͼ 78% and showed proportional deficiencies in the aldehydes, alkanes, secondary alcohols, and ketones, with increased acids, primary alcohols, and esters. Besides altered cuticle membranes, wax2 displayed postgenital fusion between aerial organs (especially in flower buds), reduced fertility under low humidity, increased epidermal permeability, and a reduction in stomatal index on adaxial and abaxial leaf surfaces. Thus, wax2 reveals a potential role for the cuticle as a suppressor of postgenital fusion and epidermal diffusion and as a mediator of both fertility and the development of epidermal architecture (via effects on stomatal index). The cloned WAX2 gene (verified by three independent allelic insertion mutants with identical phenotypes) codes for a predicted 632-amino acid integral membrane protein with a molecular mass of 72.3 kD and a theoretical pI of 8.78. WAX2 has six transmembrane domains, a His-rich diiron binding region at the N-terminal region, and a large soluble C-terminal domain. The N-terminal portion of WAX2 is homologous with members of the sterol desaturase family, whereas the C terminus of WAX2 is most similar to members of the short-chain dehydrogenase/reductase family. WAX2 has 32% identity to CER1, a protein required for wax production but not for cuticle membrane production. Based on these analyses, we predict that WAX2 has a metabolic function associated with both cuticle membrane and wax synthesis. These studies provide new insight into the genetics and biochemistry of plant cuticle production and elucidate new associations between the cuticle and diverse aspects of plant development.

show abstract

Molecular cloning and characterization of the CER2 gene of Arabidopsis thaliana

Cited by 94 publications

References 0 publications

The glycerol-3-phosphate acyltransferase GPAT6 from tomato plays a central role in fruit cutin biosynthesis

The glycerol-3-phosphate acyltransferase GPAT6 from tomato plays a central role in fruit cutin biosynthesis

The SHINE Clade of AP2 Domain Transcription Factors Activates Wax Biosynthesis, Alters Cuticle Properties, and Confers Drought Tolerance when Overexpressed in Arabidopsis[W]

Cloning and Characterization of the WAX2 Gene of Arabidopsis Involved in Cuticle Membrane and Wax Production

Contact Info

Product

Resources

About