Functional Analysis of a β-Ketoacyl-CoA Synthase Gene,<i>MpFAE2</i>, by Gene Silencing in the Liverwort<i>Marchantia polymorpha</i>L.

Kajikawa, Masataka; Yamaoka, Shohei; Yamato, Katsuyuki T.; Kanamaru, Hiroyuki; Sakuradani, Eiji; Shimizu, Sakayu; Fukuzawa, Hideya; Ohyama, Kanji

doi:10.1271/bbb.67.605

Cited by 26 publications

(24 citation statements)

References 32 publications

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“…Total RNA was isolated from 100 mg of 28-day-old A. thaliana leaves with the RNeasy plant minikit (Qiagen, Valencia, CA) according to manufacturer instructions and used to synthesize first-strand cDNA with NotI-d(T) 18 primers and Moloney murine leukemia virus reverse transcriptase (Amersham Pharmacia); 2 l of this cDNA was used directly for PCR with the appropriate primers. PCR products were cloned blunt into the pCR-Blunt II-TOPO vector (Invitrogen), cleaved with the appropriate restriction enzymes, and ligated in-frame with the C-terminal His-tag to the pYES2 vector (Invitrogen) digested with the same enzymes.…”

Section: Methodsmentioning

confidence: 99%

“…Several plant VLCFAEs have been characterized, including A. thaliana FAE1 (5, 9-11), KCS1 (12), KCS2 (13), CUT1 (14), Brassica napus FAE1 (15), Lesquerella fendleri KCS3 (16), Simmondsia chinensis KCS (17), and FAE2 from the moss Marchantia polymorpha (18). These VLCFAEs differ in terms of their tissue-and ontogeny-specific expression as well as their substrate specificity.…”

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confidence: 99%

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Specific and differential inhibition of very-long-chain fatty acid elongases from Arabidopsis thaliana by different herbicides

Trenkamp

Martin²,

Tietjen³

2004

Proc. Natl. Acad. Sci. U.S.A.

221

220

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In higher plants, very-long-chain fatty acids (VLCFAs) are the main constituents of hydrophobic polymers that prevent dessication at the leaf surface and provide stability to pollen grains. Of the 21 genes encoding VLCFA elongases (VLCFAEs) from Arabidopsis thaliana, 17 were expressed heterologously in Saccharomyces cerevisiae. Six VLCFAEs, including three known elongases (FAE1, KCS1, and KCS2) and three previously uncharacterized gene products (encoded by At5g43760, At1g04220, and At1g25450) were found to be enzymatically active with endogenous yeast fatty acid substrates and to some extent with externally supplied unsaturated substrates. The spectrum of VLCFAs accumulated in expressing yeast strains was determined by gas chromatography͞mass spectrometry. Marked specificity was found among elongases tested with respect to their elongation products, which encompassed saturated and monounsaturated fatty acids 20 -30 carbon atoms in length. The active VLCFAEs revealed highly distinct patterns of differential sensitivity to oxyacetamides, chloroacetanilides, and other compounds tested, whereas yeast endogenous VLCFA production, which involves its unrelated elongase (ELO) in sphingolipid synthesis, was unaffected. Several compounds inhibited more than one VLCFAE, and some inhibited all six active enzymes. These findings pinpoint VLCFAEs as the target of the widely used K3 class herbicides, which have been in commercial use for 50 years, provide important clues as to why spontaneous resistance to this class is rare, and point to complex patterns of substrate specificity and product spectrum among members of the Arabidopsis VLCFAE family.

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Specific and differential inhibition of very-long-chain fatty acid elongases from Arabidopsis thaliana by different herbicides

Trenkamp

Martin²,

Tietjen³

2004

Proc. Natl. Acad. Sci. U.S.A.

221

220

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“…Preparation of nucleic acids from M. polymorpha thalli and P. patens protonemata The procedures used for isolation of genomic DNA, total RNA, and poly(A) + RNA from M. polymorpha have been described previously [23]. Genomic DNA of P. patens protonemata was isolated using a DNeasy Plant Mini kit (Qiagen, Valencia, CA).…”

Section: Sequence and Phylogenetic Analysesmentioning

confidence: 99%

“…Genomic DNA and total RNA were used in Southern and Northern blot analysis as described elsewhere [23]. For detection of MpELO2 transcript and gene, its coding sequence was amplified by PCR with the primers MpELO2-03F and MpELO2-04R, using the pPICZAMpELO2 as a template.…”

Section: Northern and Southern Blot Analysesmentioning

confidence: 99%

Isolation and functional characterization of fatty acid Δ5‐elongase gene from the liverwort Marchantia polymorpha L

et al. 2005

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Bryophyte Marchantia polymorpha L. produces C22 very-long-chain polyunsaturated fatty acid (VLCPUFA). Thus far, no enzyme that mediates elongation of C20 VLCPUFAs has been identified in land plants. Here, we report the isolation and characterization of the gene MpELO2, which encodes an ELO-like fatty acid elongase in M. polymorpha. Heterologous expression in yeast demonstrated that MpELO2 encodes D5-elongase, which mediates elongation of arachidonic (20:4) and eicosapentaenoic acids (20:5). Phylogenetic and gene structural analysis indicated that the MpELO2 gene is closely related to bryophyte D6-elongase genes for C18 fatty acid elongation and diverged from them by local gene duplication.

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“…Moreover, a continuing project for M. polymorpha genome sequencing under the Community Sequencing Program (CSP) at the Joint Genome Institute designates that various biological events found in higher land plants are conserved in a very uncomplicated form. In recent times, the development of transformation techniques for M. polymorpha enables the utilization of other genetic tools such as gene targeting, gene silencing, homologous recombination and gene editing (Kajikawa et al, 2003;Ishizaki et al, 2008;2013a;2013b). The central criteria required for versatile model organisms include forward genetics, reverse genetics, genetic transformation and genome-wide studies.…”

Section: Introductionmentioning

confidence: 99%

Marchantia polymorpha L.: An Emerging Model Plant System to Study Contemporary Plant Biology – A Review

Alam

Pandey

2016

Plant Sci Today

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The liverwort, Marchantia polymorpha L., one of the species of first land plants is a promising model plant system for the analysis of diverse facets of contemporary plant biology. The unique characteristics of the plant such as dominant haploid gametophytic generation enables the isolation and disruption of mutant for genetic analysis, rapid sexual and asexual reproduction can be induced under controlled conditions which leads to the formation of genetically homogenous lines and also the complete organelle genome sequence of chloroplast and mitochondria has been established. In addition, the ongoing whole genome sequencing of M. polymorpha by the community sequencing plan at the Joint Genome Institute specifies the conservation of several mechanisms of biological science that are instituted in other terrestrial plants in a smaller extent of intricacy. Thus, with the development of several feasible and reliable genetic transformation strategies, in vitro cell culture, gene silencing, targeted gene modification and its critical evolutionary position make this plant as a potential model plant to study evolutionary and developmental biology in detail.

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Functional Analysis of a β-Ketoacyl-CoA Synthase Gene,MpFAE2, by Gene Silencing in the LiverwortMarchantia polymorphaL.

Cited by 26 publications

References 32 publications

Specific and differential inhibition of very-long-chain fatty acid elongases from Arabidopsis thaliana by different herbicides

Specific and differential inhibition of very-long-chain fatty acid elongases from Arabidopsis thaliana by different herbicides

Isolation and functional characterization of fatty acid Δ5‐elongase gene from the liverwort Marchantia polymorpha L

Marchantia polymorpha L.: An Emerging Model Plant System to Study Contemporary Plant Biology – A Review

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