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  • Author or Editor: J. Cai x
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Based on the known GA20-oxidase (GA20ox) cDNAs of barley and wheat, oligonucleotide primers were designed to isolate GA20ox genes from genomic DNA of Dasypyrum villosum. A total of 19 clones were obtained. Each of them contained an open reading frame encoding a putative 40-KDa protein of 359 amino acid residues. Twenty-one SNPs and 4 InDels were found and could divide the 19 sequences into 2 classes, designated as DvGA20ox-1 and DvGA20ox-2, respectively. Q-PCR analyses showed that both DvGA20ox-1 and DvGA20ox-2 were in leaf blade, leaf sheath, stem, eustipes, root and developing spike. Similar expression levels were found between DvGA20ox-1 and DvGA20ox-2 in three stages. The total expression levels of DvGA20ox-1 and DvGA20ox-2 presented downtrend in leaf blade and ascend in stem, eustipes and developing spike along with the development of plants, respectively. However, they were firstly increased and then decreased in root from seeding stage to heading stage. These results revealed that the gene expression profile of DvGA20ox-1 and DvGA20ox-2 closely related to the growth and development of D. villosum.

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Summary

Silica-gel column chromatography and preparative reversed-phase high-performance liquid chromatography (RP-HPLC) were sequentially employed for the isolation of two antioxidants including gallic acid and methyl gallate from Folium Toonea Sinensis. An RP-HPLC-UV method was then developed and validated to rapidly determine their content in this herb with ethyl gallate as internal standard. The quantitation was performed on an XBridge Shield RP18 column (150 mm × 4.6 mm, 5 μm) under 40°C. The mobile phase consisting of acetonitrile and 0.1% formic acid aqueous solution was driven at 1.0 mL min−1 under gradient elution, and 270 nm was selected to monitor the separation. To evaluate the fitness for purpose of the method and to investigate the difference in the content of analytes among different samples, the leaves collected from five production sites were analyzed. The newly established method is suitable for routine analysis of gallic acid and methyl gallate in the herb and, hence, can assist in its quality assessment. It was also found that not only the content of two antioxidants but also the ratio varied significantly among different geographical origins. In addition, three samples from Yantai, Zumadian, and Zhenjiang were distinguished as they have a much higher content ratio than the other two.

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