A novel Coptis japonica multidrug‐resistant protein preferentially expressed in the alkaloid‐accumulating rhizome1

Yazaki, Kazufumi; Shitan, Nobukazu; Takamatsu, Hideki; Ueda, Kazumitsu; Sato, Fumihiko

doi:10.1093/jexbot/52.357.877

Cited by 40 publications

(31 citation statements)

References 7 publications

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“…In tobacco, the plasma membrane-localized NUP1 moved apoplastic nicotine into the cytoplasm. Similarly, the C. japonica CjMDR1 mobilizes the alkaloid berberine from the apoplast into the cytoplasm (28,41). However, NUP1 and CjMDR1 differ in how the alkaloid uptake is energized.…”

Section: Discussionmentioning

confidence: 99%

Tobacco nicotine uptake permease (NUP1) affects alkaloid metabolism

Hildreth

Gehman

Yang

et al. 2011

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

106

110

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An effective plant alkaloid chemical defense requires a variety of transport processes, but few alkaloid transporters have been characterized at the molecular level. Previously, a gene fragment encoding a putative plasma membrane proton symporter was isolated, because it was coordinately regulated with several nicotine biosynthetic genes. Here, we show that this gene fragment corresponds to a Nicotiana tabacum gene encoding a nicotine uptake permease (NUP1). NUP1 belongs to a plant-specific class of purine uptake permease-like transporters that originated after the bryophytes but before or within the lycophytes. NUP1 expressed in yeast cells preferentially transported nicotine relative to other pyridine alkaloids, tropane alkaloids, kinetin, and adenine. NUP1-GFP primarily localized to the plasma membrane of tobacco Bright Yellow-2 protoplasts. WT NUP1 transcripts accumulated to high levels in the roots, particularly in root tips. NUP1-RNAi hairy roots had reduced NUP1 mRNA accumulation levels, reduced total nicotine levels, and increased nicotine accumulation in the hairy root culture media. Regenerated NUP1-RNAi plants showed reduced foliar and root nicotine levels as well as increased seedling root elongation rates. Thus, NUP1 affected nicotine metabolism, localization, and root growth.

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Section: Discussionmentioning

confidence: 99%

Tobacco nicotine uptake permease (NUP1) affects alkaloid metabolism

Hildreth

Gehman

Yang

et al. 2011

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

106

110

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“…The spatial separation of product formation and storage implicates the intercellular transport of protoberberine alkaloids from the protoderm to cortical and pith cells. A molecular clone for a multidrug-resistance protein (MDR)-type ATP binding cassette (ABC) transporter (Cjmdr1), identified as a putative berberine transporter, has been isolated (Yazaki et al, 2001) and characterized (Shitan et al, 2003) from C. japonica. RNA gel blot and in situ hybridization analyses showed the abundant and preferential expression of Cjmdr1 in the rhizome (Yazaki et al, 2001) and suggested the localization of gene transcripts to xylem tissues (Shitan et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

“…A molecular clone for a multidrug-resistance protein (MDR)-type ATP binding cassette (ABC) transporter (Cjmdr1), identified as a putative berberine transporter, has been isolated (Yazaki et al, 2001) and characterized (Shitan et al, 2003) from C. japonica. RNA gel blot and in situ hybridization analyses showed the abundant and preferential expression of Cjmdr1 in the rhizome (Yazaki et al, 2001) and suggested the localization of gene transcripts to xylem tissues (Shitan et al, 2003). In C. japonica, berberine accumulates mainly in the rhizome, whereas alkaloid biosynthetic enzymes have been associated with roots (Fujiwara et al, 1993).…”

Section: Discussionmentioning

confidence: 99%

Cell Type–Specific Localization of Transcripts Encoding Nine Consecutive Enzymes Involved in Protoberberine Alkaloid Biosynthesis

2005

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Molecular clones encoding nine consecutive biosynthetic enzymes that catalyze the conversion of L-dopa to the protoberberine alkaloid (S)-canadine were isolated from meadow rue (Thalictrum flavum ssp glaucum). The predicted proteins showed extensive sequence identity with corresponding enzymes involved in the biosynthesis of related benzylisoquinoline alkaloids in other species, such as opium poppy (Papaver somniferum). RNA gel blot hybridization analysis showed that gene transcripts for each enzyme were most abundant in rhizomes but were also detected at lower levels in roots and other organs. In situ RNA hybridization analysis revealed the cell type-specific expression of protoberberine alkaloid biosynthetic genes in roots and rhizomes. In roots, gene transcripts for all nine enzymes were localized to immature endodermis, pericycle, and, in some cases, adjacent cortical cells. In rhizomes, gene transcripts encoding all nine enzymes were restricted to the protoderm of leaf primordia. The localization of biosynthetic gene transcripts was in contrast with the tissue-specific accumulation of protoberberine alkaloids. In roots, protoberberine alkaloids were restricted to mature endodermal cells upon the initiation of secondary growth and were distributed throughout the pith and cortex in rhizomes. Thus, the cell type-specific localization of protoberberine alkaloid biosynthesis and accumulation are temporally and spatially separated in T. flavum roots and rhizomes, respectively. Despite the close phylogeny between corresponding biosynthetic enzymes, distinct and different cell types are involved in the biosynthesis and accumulation of benzylisoquinoline alkaloids in T. flavum and P. somniferum. Our results suggest that the evolution of alkaloid metabolism involves not only the recruitment of new biosynthetic enzymes, but also the migration of established pathways between cell types.

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“…Previously, we identified a PM PGP transporter, MDR1, in the model isoquinoline alkaloid producer, Coptis japonica, and demonstrated that the protein mediated ATP-dependent berberine uptake (Yazaki et al, 2001;Sakai et al, 2002;Shitan et al, 2003;Yazaki, 2005). A survey of the Arabidopsis genome for a homolog of CjMDR1 indicated that a predicted protein of 1286 amino acids, AtPGP4 (At2g47000; hereafter referred to as PGP4), was the most similar Arabidopsis homolog (71% amino acid identity, 68% nucleotide identity).…”

Section: Introductionmentioning

confidence: 99%

PGP4, an ATP Binding Cassette P-Glycoprotein, Catalyzes Auxin Transport in Arabidopsis thaliana Roots

et al. 2005

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Members of the ABC (for ATP binding cassette) superfamily of integral membrane transporters function in cellular detoxification, cell-to-cell signaling, and channel regulation. More recently, members of the multidrug resistance P-glycoprotein (MDR/PGP) subfamily of ABC transporters have been shown to function in the transport of the phytohormone auxin in both monocots and dicots. Here, we report that the Arabidopsis thaliana MDR/PGP PGP4 functions in the basipetal redirection of auxin from the root tip. Reporter gene studies showed that PGP4 was strongly expressed in root cap and epidermal cells. PGP4 exhibits apolar plasma membrane localization in the root cap and polar localization in tissues above. Root gravitropic bending and elongation as well as lateral root formation were reduced in pgp4 mutants compared with the wild type. pgp4 exhibited reduced basipetal auxin transport in roots and a small decrease in shoot-to-root transport consistent with a partial loss of the redirective auxin sink in the root. Seedlings overexpressing PGP4 exhibited increased shoot-to-root auxin transport. Heterologous expression of PGP4 in mammalian cells resulted in 1-N-naphthylthalamic acid-reversible net uptake of [ 3 H]indole-3-acetic acid. These results indicate that PGP4 functions primarily in the uptake of redirected or newly synthesized auxin in epidermal root cells.

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A novel Coptis japonica multidrug‐resistant protein preferentially expressed in the alkaloid‐accumulating rhizome1

Cited by 40 publications

References 7 publications

Tobacco nicotine uptake permease (NUP1) affects alkaloid metabolism

Tobacco nicotine uptake permease (NUP1) affects alkaloid metabolism

Cell Type–Specific Localization of Transcripts Encoding Nine Consecutive Enzymes Involved in Protoberberine Alkaloid Biosynthesis

PGP4, an ATP Binding Cassette P-Glycoprotein, Catalyzes Auxin Transport in Arabidopsis thaliana Roots

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