Cloning and characterization of a novel chalcone synthase gene from Phalaenopsis hybrida orchid flowers

Han, Ying; Ming, Feng; Wang, J. W.; Wen, Jianguo; Ye, M. M.; Shen, Daleng

doi:10.1134/s1021443706020129

Cited by 18 publications

(13 citation statements)

References 35 publications

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“…To date, we have cloned three chs genes together with the novel phchs5 gene identified in this study (GeneBank accession number of the other two chs genes are AY954515 (Han et al 2005) and AY825502 (Han et al 2006), respectively). Sequence comparison revealed that each gene contained one intron, which interrupted the codon for cysteine at the same location as reported in all chalcone synthase and chalcone synthase like genes (Feinbaum and Ausubel 1988;Sommer and Saedler 1986).…”

Section: Intron Sequence Comparison Among Phalaenopsis Chs Genesmentioning

confidence: 98%

Molecular evolution and functional specialization of chalcone synthase superfamily from Phalaenopsis Orchid

et al. 2006

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Plant genomes appear to exploit the process of gene duplication as a primary means of acquiring biochemical and developmental flexibility. The best example is the gene encoding chalcone synthase (CHS, EC2.3.1.74), the first committed step in flavonoid biosynthesis. In this study, we examined the molecular evolution of three CHS family members of Phalaenopsis including a novel chs gene (phchs5), which is slowly evolved. The inferred phylogeny of the chs genes of Phalaenopsis with other two orchid plants, Bromoheadia finlaysoniana and Dendrobium hybrid, suggested that gene duplication and divergence have occurred before divergence of these three genera. Relatively quantitative RT-PCR analysis identified expression patterns of these three chs genes in different floral tissues at different developmental stages. Phchs5 was the most abundantly expressed chs gene in floral organs and it was specifically transcribed in petal and lip at the stages when anthocyanin accumulated (stage1-4). Phchs3 and phchs4 were expressed at much lower levels than phchs5. Phchs3 was expressed in pigmented tissue (including lip, petal and sepal) at middle stages (stages 2-4) and in colorless reproductive tissue at late stage (stage 5). Phchs4 was only expressed in petal at earlier stages (stage 1-3) and in lip at middle stage (stage 4). These results present new data on differentiation of gene expression among duplicate copies of chs genes in Phalaenopsis.

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Section: Intron Sequence Comparison Among Phalaenopsis Chs Genesmentioning

confidence: 98%

Molecular evolution and functional specialization of chalcone synthase superfamily from Phalaenopsis Orchid

et al. 2006

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“…Several of these biosynthetic genes in orchids have been identified. Genes involved in the synthesis of anthocyanidin and carotenoid pigments were cloned from the orchids Bromheadia (81), Dendrobium (87), Phalaenopsis (51,52,130), and Oncidium (54). Scent genes in the monoterpene synthesis pathway have been identified from expressed sequence tag (EST) libraries in the orchid Phalaenopsis (59).…”

Section: Candidate Genes Underlying Floral Isolationmentioning

confidence: 99%

Floral Isolation, Specialized Pollination, and Pollinator Behavior in Orchids

Schiestl

Schlüter

2009

Annu. Rev. Entomol.

227

217

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Floral isolation is a form of prepollination reproductive isolation mediated by floral morphology (morphological isolation) and pollinator behavior (ethological isolation). Here we review mechanisms and evolutionary consequences of floral isolation in various pollination systems. Furthermore, we compare key features of floral isolation, i.e., pollinator sharing and specialization in pollination, in different orchid pollination systems. In orchid pollination, pollinator sharing is generally low, indicating strong floral isolation. The pollinators' motivation to visit flowers (specifically) can be due to both foraging or reproductive behavior. In both types of behavior, innate preferences for floral signals can be quickly overruled by learning. In pollination systems in which reproductive behavior of pollinators triggers flower visits, lower pollinator sharing was evident compared with systems with foraging behavior, probably because pollinators displaying reproductive behavior show higher fidelity in their visitation patterns. Orchids pollinated through reproductive behavior also use fewer pollinators than orchids pollinated through foraging behavior. No association between specialization and pollinator sharing was found. Thus, generalized pollination does not impede floral isolation, as orchids with many pollinators may nonetheless have low pollinator sharing. Specialization in pollination was, however, linked to orchid species richness in our analysis. Flower size, spur, and column morphology are most important for morphological isolation, and floral scent is most important for ethological isolation. These traits may be based on few genes, implying that floral isolation can be brought about by few genes of large effect.

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“…Genomic DNA was extracted from leaves by the method of Dellaporta et al (1983) (Han et al 2006). The DNA pellet was washed with 70% ethanol, vacuum dried, and resuspended in TE buffer (10 mM Tris-HCl, 8.0, 1 mM ethylenediaminetetraacetic acid).…”

Section: Dna Extractionmentioning

confidence: 99%

Nested Inverse Polymerase Chain Reactions: An Effective Method for Cloning of Full-Length Sequences of Chalcone Synthase

et al. 2007

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Chalcone synthase is the key enzyme in biosynthesis of flavonoids, which play roles in pigmentation of flowers and protection against ultraviolet and pathogens. Inverse polymerase chain reaction (IPCR) is a method for the rapid in vitro amplification of DNA sequences that flank a region of known sequence. In this study, IPCR united with nested PCR was successfully applied in cloning full-length sequences of three Phalaenopsis chalcone synthase genes (phchs3, phchs4, and phchs5, respectively). Firstly, routine PCR with homologous primers were performed, and gene fragments of phchs3 (1 kb), phchs4 (1.2 kb), and phchs5 (800 bp) were obtained and then sequenced. Then, inverse PCR were carried out for cloning full-length sequence of each gene. Because products were not unique in single round inverse PCR, nested PCR were performed, and the specificity was much enhanced. At last, full-length sequences of 2,499 bp for phchs3, 2,502 bp for phchs4, and 1,855 bp for phchs5 were obtained. This study proved that IPCR could be more efficient if being united with nested PCR.

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Cloning and characterization of a novel chalcone synthase gene from Phalaenopsis hybrida orchid flowers

Cited by 18 publications

References 35 publications

Molecular evolution and functional specialization of chalcone synthase superfamily from Phalaenopsis Orchid

Molecular evolution and functional specialization of chalcone synthase superfamily from Phalaenopsis Orchid

Floral Isolation, Specialized Pollination, and Pollinator Behavior in Orchids

Nested Inverse Polymerase Chain Reactions: An Effective Method for Cloning of Full-Length Sequences of Chalcone Synthase

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