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Peng X.,China Institute of Technology | Wu X.,China Institute of Technology | Ji Q.,Lishui Academy of Agricultural Science | Yang R.,China Institute of Technology | Li Y.,China Institute of Technology
Molecular Biology Reports | Year: 2016

Tetrastigma hemsleyanum is a rare and endangered herb, which is commercialized as the resource of anti-cancer drugs. Wild T. hemsleyanum plants are on the verge of extinction recently, there are increasing numbers of counterfeits on the market. In the present study, inter-simple sequence repeat (ISSR), Cleaved amplified polymorphic sequence (CAPS), and the internal transcribed spacer region II (ITS2) barcode were used for the first time for the authentication of T. hemsleyanum from its commonly counterfeits. ISSR analysis suggested that it was a useful method for distinguishing T. hemsleyanum from its adulterants of different genus. However, it was insufficient to distinguish T. hemsleyanum from those adulterants of the same genus. ITS2 of T. hemsleyanum and the commonly counterfeits were amplified and sequenced. The Neighbor-Joining tree constructed from the ITS2 sequences showed that T. hemsleyanum was clearly differentiated from all counterfeits samples. A mutation site in the ITS2 region of T. hemsleyanum had been found which could be recognized by the restriction endonuclease NcoI. T. hemsleyanum could be readily distinguished from counterfeits as the PCR products from T. hemsleyanum could be digested sufficiently by NcoI, while the PCR products from counterfeits could not be digested. The results indicated that CAPS and ITS2 barcode methods provided effective and accurate identification of T. hemsleyanum from all its adulterants, while ISSR could only distinguish T. hemsleyanum from its adulterants of different genus. The CAPS method developed in the present study will serve as a reliable tool for safe and effective use of T. hemsleyanum in the clinic application. It will also play an important role for the identification, management and conservation of this endangered species. © 2016 Springer Science+Business Media Dordrecht

Peng X.,Zhejiang Pharmaceutical College | Ji Q.,Lishui Academy of Agricultural Science | Wu H.,Zhejiang Pharmaceutical College | Li Y.,Zhejiang Pharmaceutical College
In Vitro Cellular and Developmental Biology - Plant | Year: 2015

Storage under slow growth is one of the most successful techniques for medium-term germplasm conservation and could effectively reduce somaclonal variation during subculture. Tetrastigma hemsleyanum is an endangered medicinal herb with anticancer potential. In the present study, an effective protocol was developed for clonal propagation and medium-term conservation of this endangered medicinal plant. Subculture duration was significantly prolonged to at least 10 mo by using 0.2 mg L−1 of maleic hydrazide (MH) at 8°C. Analyses of inter-simple sequence repeat (ISSR) markers, sequence-related amplified polymorphism (SRAP) markers, total proteins, and peroxidase isozymes together demonstrated the genetic stability of the regenerated germplasm. Total flavonoid content of regenerated microplants also remained stable. These results suggest that MH has potential application in slow-growth conservation of plants. © 2015, The Society for In Vitro Biology.

Peng X.,Zhejiang Pharmaceutical College | Ji Q.,Lishui Academy of Agricultural Science | Fan S.,Zhejiang Pharmaceutical College | Zhang Y.,Zhejiang Pharmaceutical College | Zhang J.,Zhejiang Pharmaceutical College
Genetic Resources and Crop Evolution | Year: 2015

Genetic diversity of 27 Tetrastigma hemsleyanum accessions from wild or cultivated populations was evaluated using SRAP (sequence-related amplified polymorphism) and ISSR (inter-simple sequence repeat) markers. ISSR and SRAP analysis yielded 112 and 66 scorable amplified bands, respectively, of which 85.71 and 77.27 % were polymorphic. ISSR revealed efficiency over SRAP due to high Shannon’s Information Index (I), Nei’s gene diversity (H), and total genetic diversity (Ht). The dendrograms showed clear clustering pattern of geographically distribution. The correlation between the coefficient matrices for the SRAP and ISSR data was high (r = 0.869, P < 0.01). Higher level of genetic similarity was observed in intra-population than in inter-population. The average genetic similarity among cultivated populations were far above that among wild populations, the low level of gene flow, distance isolation, and asexual reproduction were likely related to the low level of differentiation among populations. The promotion of genes exchange will be very beneficial to conserve and utilize the enormous genetic variability contained in the T. hemsleyanum. © 2015, Springer Science+Business Media Dordrecht.

Zhang X.,Zhejiang University | Huang H.,Zhejiang University | Zhang Q.,Zhejiang University | Fan F.,Zhejiang University | And 5 more authors.
International Journal of Molecular Sciences | Year: 2015

In order to fully understand the variations of fruit quality-related phytochemical composition in Chinese bayberry (Myrica rubra Sieb. et Zucc.), mature fruit of 17 cultivars from Zhejiang and Jiangsu provinces was used for the investigation of fruit quality attributes, including fruit color, soluble sugars, organic acids, total phenolics, flavonoids, antioxidant capacity, etc. Sucrose was the main soluble sugar, while citric acid was the main organic acid in bayberry fruit. The content of total phenolics and total flavonoids were positively correlated with 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP) antioxidant activity and 2,2ʹ-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging activity. Five anthocyanidins, i.e., delphinidin–hexoside (Dp–Hex), cyanidin-3–O-galactoside (C-3–Gal), cyanidin-3–O-glucoside (C-3–Glu), pelargonidin-3–O-glucoside (Pg-3–Glu) and peonidin-3-O-glucoside (Pn-3–Glu), and seven flavonols compounds, i.e., myricetin-3-O-rhamnoside (M-3–Rha), myricetin deoxyhexoside–gallate (M-DH–G), quercetin-3-O-galactoside (Q-3–Gal), quercetin-3–O-glucoside (Q-3–Glu), quercetin-3–O-rhamnoside (Q-3–Rha), kaempferol-3–O-galactoside (K-3–Gal) and kaempferol-3–O-glucoside (K-3–Glu), were identified and characterized among the cultivars. The significant differences in phytochemical compositions among cultivars reflect the diversity in bayberry germplasm, and cultivars of good flavor and/or rich in various health-promoting phytochemicals are good candidates for future genetic breeding of bayberry fruit of high quality. In conclusion, our results may provide important information for further breeding or industrial utilization of different bayberry resources. © 2015 by the authors; licensee MDPI, Basel, Switzerland.

He Y.,Zhejiang Academy of Agricultural Sciences | Zhang J.,Zhejiang Academy of Agricultural Sciences | Chen J.,Zhejiang Academy of Agricultural Sciences | Wu Q.,Lishui Academy of Agricultural Science | And 3 more authors.
Journal of Economic Entomology | Year: 2011

Pymetrozine reportedly inhibits feeding of plant sap-sucking insects, such as aphids and brown planthopper (Nilaparvata lugens (Stål)). By using electrical penetration graph (EPG), this study was conducted to investigate any differential effect of pymetrozine on the feeding behaviors of four major rice sap-sucking insect species, 1) N. lugens, 2) white-backed planthopper (Sogatella furcifera (Horváth)), 3) small brown planthopper (Laodelphax striatellus (Falln)), and 4) green rice leafhopper (Nephotettix cincticeps Uhler). On pymetrozine-free TN1 rice plants, white-backed planthopper and small brown planthopper showed a significantly less activity in the phloem phases than brown planthopper or green rice leafhopper while green rice leafhopper engaged in relatively more xylem ingestion than brown planthopper, white-backed planthopper, and small brown planthopper. On the plants treated with 100 mg liter -1 of pymetrozine, all four insect species showed significant increases, in total duration of nonprobing and significant decreases in the activities in phloem tissue, while all species showed similar feeding behavior during the pathway and xylem phases. This study revealed that, regardless of whether the insects on untreated plants spent more time feeding on phloem than xylem (brown planthopper) or more time on xylem than phloem (green rice leafhopper) or similar times on phloem and xylem (white-backed planthopper and small brown planthopper), their feeding behavior was disturbed by pymetrozine and exhibited similar patterns of sharp decline in activity in the phloem tissue and a significant increase the nonprobing. © 2011 Entomological Society of America.

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