Cluster Analysis of Ziziphora bungeana from Different Geographical Populations Based on ITS2 and psbA-trnH Sequences / 中国实验方剂学杂志

Objective:To analyze the internal transcribed spacer（ITS）2 and psbA-trnH sequences of Ziziphora bungeana in 18 different geographic populations,in order to provide reference about evaluation of germplasm resources and analysis of genetic diversity of medicinal plants. Method:Genomic DNAs of the Z. bungeana were extracted by kit method. Polymerase chain reaction（PCR）was used to amplify ITS2 and psbA-trnH interstitial region sequences，bidirectional sequencing，splicing，and constructing Neighbor-joining（NJ） based on Kimura 2-parameter（K2P） model. Result:All of sequences of ITS2 and psbA-trnH of Z. bungeana in different geographic populations showed intraspecific variations. The average ITS2 sequence length of Z. bungeana was 236 bp，9 haplotypes were detected，and the genetic distance was 0-0.022. Z. bungeana of different geographical groups gathered into two branches，10 geographic populations,including XTH3，XTH6 and XTH9,were considered as one branch，while 8 geographic populations, including XTH4，XTH5 and XTH10,were the other branch. In addition to XTH6 that lacked 6 in bp psbA-trnH sequence，all of the other geographic populations had a 355 bp sequence of psbA-trnH，4 haplotypes were detected，and the genetic distance was 0-0.023. 12 geographic populations,such as XTH1，XTH3，XTH4,gathered into one branch，while XTH14，XTH17 and XTH18 gathered into the other branch. NJ tree based on ITS2+psbA-trnH combination sequence showed that Z. bungeana of different geographical populations could be divided into two branches，with 12 geographical populations,like XTH11，XTH12，XTH16 as one branch, and XTH14，XTH17 and XTH18 as the other branch. Conclusion:Near or similar geographical locations of different geographical populations implies relatively short genetic distance and relatively close genetic relationship，which indicates that genetic relationship and genetic diversity of Z. bungeana in different geographical populations are related to geographical locations.

Transcriptome analysis reveals candidate genes involved in flavonoid biosynthesis in Ziziphora bungeana / 中国中药杂志

Ziziphora bungeana is a kind of medicinal plants belongs to Labiatae,and it also a kind of geoherbs in Xinjiang. The main active ingredient linarin has a higher content in inflorescence than in other parts. In this study,high-throughput sequencing technology was used to reveal the transcriptome of the inflorescence of Z. bungeana,77 366 unigenes were acquired,of which 56 375 unigenes were annotated based on search of the database and classification. Through the analysis of metabolic pathways,sixty unigenes were probably encoding some enzymes involved in the flavonoid biosynthesis pathways. The contents of linarin in different parts were determined and the key genes were verified by qRT-PCR. The discovery provides the research basis for further analysis of the enzyme genes involved in the biosynthesis of the major flavonoid components in Z. bungeana.

Determination of 3 components in Xinjiang Ziziphora bungeana by HPLC / 中国中药杂志

This study aimed to develop an HPLC method for simultaneous determination of the 3 components in Ziziphora bungeana. The optimum HPLC condition was as follows：ZORBAX SB-C₁₈ column（4.6 mm×250 mm, 5.0 μm）with gradient elution of methanol (A)-0.2% glacial acetic acid (B), detection wavelength 340 nm,column temperature 30 °C, flow rate 1 mL·min⁻¹. There were good linearity between peak areas and injection quantity of caffeic acid, rosmarinic acid, and linarin in the range of 2-40 mg·L⁻¹（=0.999 9）, 3-60 mg·L⁻¹（=1）, 7-140 mg·L⁻¹（r=0.999 9）, respectively. The average recoveries were 100.9%（RSD 1.3%）,98.25%（RSD 2.0%）,and 98.73%（RSD 1.5%）, respectively. The HPLC method was stable and accurate, which could be used to detect caffeic acid,rosmarinic acid, and linarin in Ziziphora bungeana.