scholarly journals Authentication of Shan-Dou-Gen Based on DNA Analysis

2019 ◽  
Vol 14 (6) ◽  
pp. 1934578X1985854
Author(s):  
Masashi Kitamura ◽  
Naoki Muramatsu ◽  
Ryuichiro Suzuki ◽  
Yoshiaki Shirataki
Keyword(s):  
Dna Analysis ◽  
Genetic Approach ◽  
Chain Reaction ◽  
Analgesic Effects ◽  
Specific Polymerase Chain Reaction ◽  
Crude Drug ◽  
Crude Drugs ◽  
Sophora Tonkinensis ◽  
Polymerase Chain ◽  
Species Specific

Shan-Dou-Gen (山豆根) is a crude drug mainly derived from the roots of Leguminosae plants, and it has antipyretic, antidotal, anti-inflammatory, and analgesic effects. In Japan, the root of Euchresta japonica has been used as a material of Shan-Dou-Gen. However, E. japonica is not used for medicinal purposes today, and commercial Shan-Dou-Gen products are imported from China. In China, several plant species have been used as Shan-Dou-Gen materials, but a crude drug derived from the root of Sophora tonkinensis is now officially used as Shan-Dou-Gen. However, it is difficult to morphologically identify the species used in Shan-Dou-Gen. In the present study, we showed that the Shan-Dou-Gen products commercially available in Japan are derived from S. tonkinensis using phylogenetic and sequencing analyses of the chloroplast trnH–psbA region. Furthermore, we performed species-specific polymerase chain reaction using conserved sequences of S. tonkinensis. Amplification was observed with Shan-Dou-Gen, whereas no amplification occurred with other crude drugs derived from the roots of S. flavescens and S. japonica. These results indicated that the genetic approach can be useful to authenticate Shan-Dou-Gen.

Routine identification of four sympatric species of calanoid copepods Pseudocalanus spp. in the Atlantic Arctic using a species-specific polymerase chain reaction

2020 ◽  
Vol 48 (1) ◽  
pp. 62-72
Author(s):  
E. A. Ershova
Keyword(s):  
Polymerase Chain Reaction ◽  
Life Cycles ◽  
The Arctic ◽  
Relative Distribution ◽  
Chain Reaction ◽  
Specific Polymerase Chain Reaction ◽  
Routine Identification ◽  
Polymerase Chain ◽  
Species Specific ◽  
Plankton Communities

Сalanoid copepods of the genus Pseudocalanus play an important role in the plankton communities of the Arctic and boreal seas, often dominating in numbers and constituting a significant proportion of the biomass of zooplankton. Despite their high presence and significance in the shelf plankton communities, species-specific studies of the biology of these are significantly hampered by extremely small morphological differences between them, especially at the juvenile stages, at which they are virtually indistinguishable. In this paper, we describe a new, routine and low-cost molecular method for identifying all Pseudocalanus species found in the Atlantic sector of the Arctic: the Arctic P. acuspes, P. minutus and the boreal P. moultoni and P. elongatus, and apply it to describe the relative distribution of these species in four locations of the Arctic and sub-Arctic. With this method, species-specific polymerase chain reaction (ssPCR), mass identification of individuals of any developmental stage, including nauplii, is possible. This method can serve as an excellent tool for studying the species-specific biology of this group, describing their life cycles, as well as monitoring changes in Arctic marine ecosystems under the influence of changing climate.

scholarly journals A Species-Specific Polymerase Chain Reaction Assay for Rapid Detection of Phytophthora nicotianae in Irrigation Water

2003 ◽  
Vol 93 (7) ◽  
pp. 822-831 ◽  
Author(s):  
Ping Kong ◽  
Chuanxue Hong ◽  
Steven N. Jeffers ◽  
Patricia A. Richardson
Keyword(s):  
Polymerase Chain Reaction ◽  
Irrigation Water ◽  
Phytophthora Nicotianae ◽  
Detection Sensitivity ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Specific Polymerase Chain Reaction ◽  
Wide Range ◽  
Polymerase Chain ◽  
Species Specific

Phytophthora nicotianae is a common and destructive pathogen of numerous ornamental, agronomic, and horticultural crops such as tobacco, tomato, and citrus. We have developed a species-specific polymerase chain reaction (PCR) assay for rapid and accurate detection of this pathogen in irrigation water, a primary source of inoculum and an efficient means of propagule dissemination. This PCR assay consists of a pair of species-specific primers (PN), customization of a commercial soil DNA extraction kit for purification of DNA from propagules in irrigation water, and efficient PCR protocols for primer tests and sample detection. The PN primers proved adequately specific for P. nicotianae in evaluations with 131 isolates of P. nicotianae, 102 isolates from 15 other species of Phytophthora, and 64 isolates from a variety of other oomycetes, true fungi, and bacteria. These isolates originated from a wide range of host plants, three substrates (plant tissue, soil, and irrigation water), and numerous geographic locations. The detection sensitivity is between 80 and 800 fg DNA/μl. The assay detected the pathogen in naturally infested water samples from Virginia and South Carolina nurseries more rapidly and accurately than standard isolation methods. Use of this PCR assay can assist growers in making timely disease management decisions with confidence.

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