RESUMO
Dendrobium nobile Lindl. is an orcid plant with important medicinal values. This is a colourful houseplant, and also a popular herb in traditional Chinese medicine (TCM). The variants of this plant from different geographic regions might be high, and in this study, we aimed to develop specific sequence characterized amplified region (SCAR) markers for the identification of specific variant of this plant. Different cultivars of D. nobile were collected from nine different places of China, and one cultivar from Myanmar. DNA materials were extracted from the plant samples, random amplified polymorphic DNA (RAPD) were developed, cloned and sequenced for the development of SCAR markers. We have developed four SCAR markers, which are specific to the cultivar from Luzhou China, and clearly distinguishable (genetically) from other cultivars. These SCAR markers are deposited in GenBank (accession number MZ417502, MZ484089, MZ417504 and MZ417505). Four SCAR markers for D. nobile are effective molecular technique to genetically identify the different cultivars or species, and this method is applicable for genetic characterization and identification of other plant species too.
Assuntos
Dendrobium , China , DNA , Dendrobium/genética , Marcadores Genéticos/genética , Técnica de Amplificação ao Acaso de DNA PolimórficoRESUMO
Dendrobium nobile Lindl. is an orcid plant with important medicinal values. This is a colourful houseplant, and also a popular herb in traditional Chinese medicine (TCM). The variants of this plant from different geographic regions might be high, and in this study, we aimed to develop specific sequence characterized amplified region (SCAR) markers for the identification of specific variant of this plant. Different cultivars of D. nobile were collected from nine different places of China, and one cultivar from Myanmar. DNA materials were extracted from the plant samples, random amplified polymorphic DNA (RAPD) were developed, cloned and sequenced for the development of SCAR markers. We have developed four SCAR markers, which are specific to the cultivar from Luzhou China, and clearly distinguishable (genetically) from other cultivars. These SCAR markers are deposited in GenBank (accession number MZ417502, MZ484089, MZ417504 and MZ417505). Four SCAR markers for D. nobile are effective molecular technique to genetically identify the different cultivars or species, and this method is applicable for genetic characterization and identification of other plant species too.
Dendrobium nobile Lindl. é uma orquídea com importantes valores medicinais. Esta é uma colorida planta doméstica e também uma erva popular na Medicina Tradicional Chinesa (MTC). As variantes desta planta de diferentes regiões geográficas podem ser altas, e neste estudo, nosso objetivo foi desenvolver marcadores de região amplificada de sequência caracterizada (in English, Sequence Characterized Amplified Region (SCAR)) para a identificação de variante específica desta planta. Diferentes cultivares de D. nobile foram coletadas de nove locais diferentes da China e uma cultivar de Mianmar. Materiais de DNA foram extraídos das amostras de plantas, em que a Amplificação Aleatória de DNA Polimórfico (in English, Random Amplified Polymorphism DNA (RAPD)) foi desenvolvida, clonada e sequenciada para o desenvolvimento de marcadores SCAR. Desenvolvemos quatro marcadores SCAR, que são específicos para a cultivar de Luzhou na China e claramente distinguíveis (geneticamente) de outras cultivares. Esses marcadores SCAR estão depositados no GenBank (números de acesso MZ417502, MZ484089, MZ417504 e MZ417505). Quatro marcadores SCAR para D. nobile compreendem técnicas moleculares eficazes para identificar geneticamente as diferentes cultivares ou espécies, e este método é aplicável para caracterização genética e identificação de outras espécies de plantas também.
Assuntos
Marcadores Genéticos , Orchidaceae , Dendrobium/genéticaRESUMO
Dendrobium officinale is one of the most well-known traditional Chinese medicines, and polysaccharide is its main active ingredient. Many studies have investigated the synthesis and accumulation mechanisms of polysaccharide, but until recently, little was known about the molecular mechanism of how polysaccharide is synthesized because no related genes have been cloned. In this study, we cloned an alkaline/neutral invertase gene from D. officinale (DoNI) by the rapid amplification of cDNA ends (RACE) method. DoNI was 2231 bp long and contained an open reading frame that predicted a 62.8-kDa polypeptide with 554-amino acid residues. An alkaline/neutral invertase conserved domain was predicted from this deduced amino acid sequence, and DoNI had a similar deduced amino acid sequence to Setaria italica and Oryza brachyantha. We also found that DoNI expression in different tissues was closely related to DoNI activity, and more importantly, polysaccharide level. Our results indicate that DoNI is associated with polysaccharide accumulation in D. officinale.
Assuntos
Dendrobium/genética , Genes de Plantas , Proteínas de Plantas/genética , Polissacarídeos/metabolismo , beta-Frutofuranosidase/genética , Sequência Conservada , Dendrobium/enzimologia , Fases de Leitura Aberta , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Polissacarídeos/genética , Domínios Proteicos , beta-Frutofuranosidase/química , beta-Frutofuranosidase/metabolismoRESUMO
In vitro mutagenesis of Dendrobium 'Earsakul' was carried out by incubating the protocorm-like bodies in 0-5 mM sodium azide for 1 h. Twenty-eight putative mutants were evaluated for genetic variability compared to untreated control plants using inter-simple sequence repeat (ISSR) analysis. Polymorphic fragments were produced by 9 of 12 ISSR primers. A total of 173 amplified ISSR fragments varying in size from 140 to 5000 bp were obtained, 39 of which were polymorphic (22.5%). Of the 28 putative mutants, 15 (53.6%) showed altered genetic profiles compared to control and were identified as mutants. These results suggest that sodium azide can be effectively utilized to generate mutants in Dendrobium 'Earsakul', and ISSR provides a powerful tool that allows efficient early detection of these mutants. The identified mutants are currently being multiplied for further evaluation of their horticultural characteristics.
Assuntos
Dendrobium/efeitos dos fármacos , Mutagênicos/farmacologia , Polimorfismo Genético , Azida Sódica/farmacologia , Dendrobium/genética , Marcadores Genéticos , Repetições de Microssatélites , Mutação , Análise de Componente Principal , Seleção GenéticaRESUMO
The Orchidaceae is one of the largest and most diverse families of flowering plants. The Dendrobium genus has high economic potential as ornamental plants and for medicinal purposes. In addition, the species of this genus are able to produce large crops. However, many Dendrobium varieties are very similar in outward appearance, making it difficult to distinguish one species from another. This study demonstrated that the 12 Dendrobium species used in this study may be divided into 2 groups by internal transcribed spacer (ITS) sequence analysis. Red and yellow flowers may also be used to separate these species into 2 main groups. In particular, the deciduous characteristic is associated with the ITS genetic diversity of the A group. Of 53 designed simple sequence repeat (SSR) primer pairs, 7 pairs were polymorphic for polymerase chain reaction products that were amplified from a specific band. The results of this study demonstrate that these 7 SSR primer pairs may potentially be used to identify Dendrobium species and their progeny in future studies.
Assuntos
Dendrobium/genética , Variação Genética , Repetições de Microssatélites/genética , Filogenia , DNA de Plantas/genética , Reação em Cadeia da Polimerase , Especificidade da EspécieRESUMO
Expressed sequence tag (EST)-derived simple sequence repeat (SSR) markers could enrich the current resource of molecular markers. In this study, microsatellite markers were developed for Dendrobium nobile Lindl. by mining the ESTs. Twenty-eight EST-SSRs amplified 2 to 6 nucleotide repeats with a mean number of 2.82 alleles per locus. The observed and expected heterozygosities per locus ranged from 0.158 to 0.579 and 0.422 to 0.752, respectively. These novel EST-SSRs enriched the current resource of molecular markers for the Dendrobium genus and would facilitate further applications in germplasm appraisal, evolution and genetic diversity studies, genetic mapping, and plant breeding of D. nobile and other congeneric species.
Assuntos
Dendrobium/genética , Etiquetas de Sequências Expressas , Repetições de Microssatélites , DNA de Plantas/genética , Dendrobium/classificação , Marcadores Genéticos , Genoma de PlantaRESUMO
Dendrobium officinale is an endangered orchid from southeast Asia that is known for its medicinal properties in traditional Chinese medicine. We constructed an integrated genetic linkage map of an F(1) population derived from an interspecific cross between D. officinale and D. aduncum (both, 2n = 38), using expressed sequence tag-simple sequence repeats (EST-SSR) and sequence-related amplified polymorphism (SRAP). A total of 349 polymorphic loci, including 261 SRAP loci and 88 EST-SSR loci, were identified for genetic linkage analysis. The software JoinMap 4.0 was used to construct the genetic maps. A total of 157 loci were arranged into 27 major linkage groups, each containing a minimum of four markers, and a further 23 markers were distributed to five triplets and four doublets, the frame map covered a total distance of 1580.4 cM, with a mean of 11.89 cM between adjacent markers. This primary map of the D. officinale and D. aduncum hybrid provides a basis for genetic studies and should facilitate future studies of medical traits mapping and marker-assisted selection in Dendrobium species breeding programs.