Cloning and Characterization of the 2-<i>C</i>-Methyl-<scp>D</scp>-erythritol 4-Phosphate (MEP) Pathway Genes of a Natural-Rubber Producing Plant,<i>Hevea brasiliensis</i>

Sando, Tomoki; Takeno, Shinya; Watanabe, Nobuyuki; Okumoto, Hiroshi; Kuzuyama, Tomohisa; Yamashita, Atsushi; Hattori, Masahira; Ogasawara, Nagahisa; Fukusaki, Eiichiro; Kobayashi, Akio

doi:10.1271/bbb.80387

Cited by 68 publications

(46 citation statements)

References 40 publications

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“…3a and Supplementary Table 18). In contrast, at least one gene for each MVA pathway enzyme shows latex-biased abundant expression, supporting the proposition that it is the MVA rather than the MEP pathway that is involved in rubber biosynthesis 22,23 . Three gene families, REF/SRPP, CPT (cis-prenyltransferase) and DXS, have more than ten genes.…”

Section: Resultssupporting

confidence: 59%

The rubber tree genome reveals new insights into rubber production and species adaptation

et al. 2016

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The Para rubber tree (Hevea brasiliensis) is an economically important tropical tree species that produces natural rubber, an essential industrial raw material. Here we present a high-quality genome assembly of this species (1.37 Gb, scaffold N50 = 1.28 Mb) that covers 93.8% of the genome (1.47 Gb) and harbours 43,792 predicted protein-coding genes. A striking expansion of the REF/SRPP (rubber elongation factor/small rubber particle protein) gene family and its divergence into several laticifer-specific isoforms seem crucial for rubber biosynthesis. The REF/SRPP family has isoforms with sizes similar to or larger than SRPP1 (204 amino acids) in 17 other plants examined, but no isoforms with similar sizes to REF1 (138 amino acids), the predominant molecular variant. A pivotal point in Hevea evolution was the emergence of REF1, which is located on the surface of large rubber particles that account for 93% of rubber in the latex (despite constituting only 6% of total rubber particles, large and small). The stringent control of ethylene synthesis under active ethylene signalling and response in laticifers resolves a longstanding mystery of ethylene stimulation in rubber production. Our study, which includes the re-sequencing of five other Hevea cultivars and extensive RNA-seq data, provides a valuable resource for functional genomics and tools for breeding elite Hevea cultivars.

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Section: Resultssupporting

confidence: 59%

The rubber tree genome reveals new insights into rubber production and species adaptation

et al. 2016

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“…Enzymes in the plastid-localized methylerythritol phosphate (MEP) pathway are also expressed in H. brasiliensis latex (Seetang-Nun et al, 2008). However, there was no significant incorporation of [1][2][3][4][5][6][7][8][9][10][11][12][13] C]1-deoxyxylulose into cis-polyisoprene in H. brasiliensis seedlings, even though this compound is closely related to the early MEP pathway intermediate 1-deoxyxylulose 5-phosphate, whereas large amounts of labeled carotenoids were detected in the same experiment, suggesting that the H. brasiliensis MEP pathway contributes to diterpenoid and carotenoid biosynthesis but not rubber biosynthesis (Sando et al, 2008).…”

mentioning

confidence: 84%

“…Therefore, we investigated additional regulatory models. The MVA pathway is thought to be the main source of IPP in rubberproducing plants (Bandurski and Teas, 1957;Hepper and Audley, 1969;Sando et al, 2008), so we studied the expression of hydroxymethylglutaryl-coenzyme A reductase (HMGR), the rate-limiting enzyme of the MVA pathway (Chappell et al, 1995). This enzyme is known to be feedback regulated by isoprenoid compounds (for review, see Stermer et al, 1994).…”

Section: Rubber Biosynthesis Is Linked To Triterpene and Inulin Contentmentioning

confidence: 99%

Laticifer-Specific cis-Prenyltransferase Silencing Affects the Rubber, Triterpene, and Inulin Content of Taraxacum brevicorniculatum

et al. 2012

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Certain Taraxacum species, such as Taraxacum koksaghyz and Taraxacum brevicorniculatum, produce large amounts of high-quality natural rubber in their latex, the milky cytoplasm of specialized cells known as laticifers. This high-molecular mass biopolymer consists mainly of poly(cis-1,4-isoprene) and is deposited in rubber particles by particle-bound enzymes that carry out the stereospecific condensation of isopentenyl diphosphate units. The polymer configuration suggests that the chain-elongating enzyme (rubber transferase; EC 2.5.1.20) is a cis-prenyltransferase (CPT). Here, we present a comprehensive analysis of transgenic T. brevicorniculatum plants in which the expression of three recently isolated CPTs known to be associated with rubber particles (TbCPT1 to -3) was heavily depleted by laticifer-specific RNA interference (RNAi). Analysis of the CPT-RNAi plants by nuclear magnetic resonance, size-exclusion chromatography, and gas chromatography-mass spectrometry indicated a significant reduction in rubber biosynthesis and a corresponding 50% increase in the levels of triterpenes and the main storage carbohydrate, inulin. Transmission electron microscopy revealed that the laticifers in CPT-RNAi plants contained fewer and smaller rubber particles than wild-type laticifers. We also observed lower activity of hydroxymethylglutaryl-coenzyme A reductase, the key enzyme in the mevalonate pathway, reflecting homeostatic control of the isopentenyl diphosphate pool. To our knowledge, this is the first in planta demonstration of latex-specific CPT activity in rubber biosynthesis.

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“…3). Additionally, we compared the recently reported rubberbiosynthesis related genes with the assembled unigenes and found that one or more unigenes matched with all of the genes involved in the six steps of the mevalonate (MVA) pathway (Sando et al 2008a) and seven genes related to the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway (Sando et al 2008b). Six different sucrose transporter genes (HbSUTs), which were recently reported to correlate with rubber yield (Tang et al 2010) and other rubberbiosynthesis related genes were also assembled (Table S6).…”

Section: Annotation and Classification Of H Brasiliensis Unigenesmentioning

confidence: 99%

RNA-Seq analysis and de novo transcriptome assembly of Hevea brasiliensis

et al. 2011

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Hevea brasiliensis, being the only source of commercial natural rubber, is an extremely economically important crop. In an effort to facilitate biological, biochemical and molecular research in rubber biosynthesis, here we report the use of next-generation massively parallel sequencing technologies and de novo transcriptome assembly to gain a comprehensive overview of the H. brasiliensis transcriptome. The sequencing output generated more than 12 million reads with an average length of 90 nt. In total 48,768 unigenes (mean size = 436 bp, median size = 328 bp) were assembled through de novo transcriptome assembly. Out of 13,807 H. brasiliensis cDNA sequences deposited in Genbank of the National Center for Biotechnology Information (NCBI) (as of Feb 2011), 11,746 sequences (84.5%) could be matched with the assembled unigenes through nucleotide BLAST. The assembled sequences were annotated with gene descriptions, Gene Ontology (GO) and Clusters of Orthologous Group (COG) terms. In all, 37,432 unigenes were successfully annotated, of which 24,545 (65.5%) aligned to Ricinus communis proteins. Furthermore, the annotated uingenes were functionally classified according to the GO, COG and Kyoto Encyclopedia of Genes and Genomes databases. Our data provides the most comprehensive sequence resource available for the study of rubber trees as well as demonstrates effective use of Illumina sequencing and de novo transcriptome assembly in a species lacking genomic information.

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Cloning and Characterization of the 2-C-Methyl-D-erythritol 4-Phosphate (MEP) Pathway Genes of a Natural-Rubber Producing Plant,Hevea brasiliensis

Cited by 68 publications

References 40 publications

The rubber tree genome reveals new insights into rubber production and species adaptation

The rubber tree genome reveals new insights into rubber production and species adaptation

Laticifer-Specific cis-Prenyltransferase Silencing Affects the Rubber, Triterpene, and Inulin Content of Taraxacum brevicorniculatum

RNA-Seq analysis and de novo transcriptome assembly of Hevea brasiliensis

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