Using ITS2 PCR‐RFLP to generate molecular markers for authentication of <i>Sophora flavescens</i> Ait.

Lin, Tzu Che; Yeh, Mau Shing; Cheng, Ya-Ming; Lin, Li; Sung, Jih Min

doi:10.1002/jsfa.4667

Cited by 10 publications

(6 citation statements)

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“…The regions of ITS 1 and ITS 2 are commonly used as character sources to determine the phylogenetic relationship. Amplification of target band on PCR product the region of ITS-1 and ITS-2 rDNA on more than 500 base pair (Baldwin et al, 1995;Baldwin & Markos, 1998;Lin et al, 2007Lin et al, , 2011.…”

Section: Introductionmentioning

confidence: 99%

Phylogenetic Relationship of Indonesian Water Yam (Dioscorea alata L.) Cultivars Based on DNA Marker Using ITS-rDNA Analysis

Purnomo¹,

Daryono²,

Shiwachi³

2017

JAS

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Research on genetic diversity and intra-species classification of Indonesian Dioscorea alata L. based on morphological characters has been done, and the result shows that there are 4 sub-groups of green cultivars group, and 5 sub-groups of purplish-red cultivars group. The objectives of this study are to determine the phylogenetic relationship cultivars of D. alata Indonesia compared to D. bulbifera as nearest species as well as 3 cultivars from GenBank. The young leaves of 18 water yam cultivar accessions were collected from Java, Madura, South Sumatera, South Kalimantan, Centre Celebes (Sulawesi), Ternate, West Papua, and Nusa Tenggara islands of Indonesia. DNA Isolation was conducted using Phytopure reagent. DNA amplification was conducted using thermocycler, predenaturation at 95 o C 5 minutes, denaturation at 95 o C 1 minutes, annealing at 60 o C 3 minutes, and elongation at 72 o C 2 minutes along with 30 cycles. The PCR products were electrophoresed in 1.5% agarose and visualized under UV transiluminator. Fifty-five µl of PCR products with positive targeted band between 700-800 bp were sent to 1 st Base Singapore for purification and sequencing of 18S, ITS-1, 5.8S, ITS-2, and 28S rDNA. The DNA sequences were compared and aligned by BioEdit program (version 7.0.5.2) and MEGA programs. Comparison of entire sequences of the tested samples were aligned by software ClustalW (version 1.83). Phylogenetic trees were based on hierarchical clustering of the alignments of 18S, ITS-1, 5.8S, ITS-2, and 28S rDNA and produced by Neigbor-Joining using MEGA 5 software of the bootstrap values (1000 replicates). The study result shows that D. alata cultivars have high genetic variability on ITS-1, 5.8S, and ITS-2 rDNA region. Groups of green and purplish red cultivars formed based on morphological characters are not formed based on ITS-rDNA markers. Sub-groups were formed based on ITS-rDNA molecular markers derived from both the green and purplish-red cultivar groups. This result revealed that two cultivar groups are not similar with RAPD and morphological characters.

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Section: Introductionmentioning

confidence: 99%

Phylogenetic Relationship of Indonesian Water Yam (Dioscorea alata L.) Cultivars Based on DNA Marker Using ITS-rDNA Analysis

Purnomo¹,

Daryono²,

Shiwachi³

2017

JAS

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“…These results are not surprising, because the molecular assessments on internal transcribed spacer sequence showed that the tested S. flavescens populations have extremely low genetic variability (Lin et al, 2012). Since the karyological characteristics for the tested populations were similar, therefore, the karyotype data were presented as the means of the tested four populations (Table 1).…”

Section: Resultsmentioning

confidence: 86%

Karyological, morphological and phytochemical characteristics of medicinal plants Sophora flavescens Aiton grown from seeds collected at different localities

2014

Self Cite

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BackgroundDried roots of Sophora flavescens Aiton contain many phytochemicals that exhibit beneficial effects on human health. This study examined and compared the karyological, morphological and phytochemical characteristics of three S. flavescens populations collected from the Danda, Hualien and Yuli of Taiwan and a population collected from Gansu, China.ResultsKaryotypes of the four populations were similar, with a diploid number of 2n = 18. The Hualien population produced more roots but with less matrine and oxymatrine contents in its root tissue than others. However, only the root of Danda population had a measurable level of naringenin. The dried root of Yuli population had greater ferric reducing antioxidant power and trolox equivalent antioxidant capacity than that of the other populations.ConclusionsThus, the collected S. flavescens populations, particularly the population collected from Danda, have the potential to be used in breeding programs.Electronic supplementary materialThe online version of this article (doi:10.1186/1999-3110-55-5) contains supplementary material, which is available to authorized users.

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“…The advantages of the ITS regions made them a preferred choice in the past, as the ITS sequences were used as genetic markers for various species (Gao et al 2010;Sun et al 2011;Lin et al 2012;Ganopoulos et al 2012;Liu et al 2012). Slanc et al (2006) used ITS for the identification of four herbal drugs in tea mixtures.…”

Section: Resultsmentioning

confidence: 99%

Identification of Smallanthus sonchifolius in herbal tea mixtures by PCR and DART/TOF-MS methods

Žiarovská¹,

Rajchl²,

Lojka³

et al. 2016

Czech J. Food Sci.

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AbstractŽiarovská J., Rajchl A., Fernández E., Prchalová J., Milella L. (2016): Identification of Smallanthus sonchifolius in herbal tea mixtures by PCR and DART/TOF-MS methods. Czech J. Food Sci., 34: 495-502.The identification of yacon, a medicinal plant, in tea mixtures by rapid Polymerase Chain Reaction (PCR) and the Direct Analysis in Real Time coupled with Time-of-Flight Mass Spectrometry (DART/TOF-MS) method were evaluated. Three tea products and a pure yacon tea were analysed using the molecular method PCR, concretely the intraspecific variation of the internal transcribed spacer (ITS) regions of rDNA and the DART method coupled with TOF-MS. The results show the reliability of PCR and restriction cleavage of the ITS as a combined approach to confirm the presence of yacon in herbal tea mixtures. Three fragments of approximately 700, 408, and 235 bp in length are present when yacon is a part of the herbal tea mixture. The Principal Component Analysis (PCA) based on the fingerprints of the complete Total Ion Current (TIC) mass spectra shows sufficient separation of herbal teas with and without yacon leaves. The reported methods are technically rapid and can be used as an effective tool for the purposes of yacon identification or authentication.

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Using ITS2 PCR‐RFLP to generate molecular markers for authentication of Sophora flavescens Ait.

Abstract: The present study demonstrates the usefulness of ITS2 PCR-RFLP coupled with pre-selected restriction enzymes for practical and accurate authentication of S. flavescens Ait. The technique is also suitable for analysing S. flavescens Ait. mixed with other adulterants.

Cited by 10 publications

References 14 publications

Phylogenetic Relationship of Indonesian Water Yam (Dioscorea alata L.) Cultivars Based on DNA Marker Using ITS-rDNA Analysis

Phylogenetic Relationship of Indonesian Water Yam (Dioscorea alata L.) Cultivars Based on DNA Marker Using ITS-rDNA Analysis

Karyological, morphological and phytochemical characteristics of medicinal plants Sophora flavescens Aiton grown from seeds collected at different localities

Identification of Smallanthus sonchifolius in herbal tea mixtures by PCR and DART/TOF-MS methods

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