Analysis on YABBY Transcription Factor Family of Original Plant Cannabis sativa of Traditional Chinese Medicine Cannabis Fructus / 中国实验方剂学杂志

Objective:To analyze the sequence characteristics，chromosomal location，gene structure，conserved motifs，phylogenetic evolution and differential gene expressions of the <italic>Cannabis sativa</italic> YABBY transcription factor family，in order to provide a molecular basis for in-depth study of <italic>YABBY</italic> gene function and theoretical support for the selection and breeding of superior hemp varieties. Method:The bio-informatics method was used to identify and analyze the <italic>CsYABBY </italic>gene family of the original hemp seed plant. PlantTFDB，ExPASy，MEME，CELLO，PLANTCARE and other online websites and TBtools，MEGA，DNAMAN and other software were used for prediction，visualization and analysis. Result:<italic>C. sativa</italic> contains 6 <italic>YABBY</italic> gene members distributed on 5 chromosomes，in which 5 members are localized in the nucleus and 1 in extracellular， they consist of 185-235 amino acids， and the isoelectric point is between 5.05 and 9.34， the molecular weight is between 20 582.45-26 282.7 Da. All of CsYABBY proteins contain two conserved domains， namely Zinc finger domain and YABBY domain. <italic>CsYABBY</italic> genes have multiple cis-acting elements，and their expressions differ in different tissues and cultivars. Conclusion:The expressions of CsYABBY may be affected by hormones and externally environmental factors. <italic>CsYABBY</italic> gene expressions are tissue-specific. In addition，YABBY transcription factor family may play an important role in regulating the development of <italic>C. sativa</italic> female flowers，and subfamilies YAB1 and YAB5 may be involved in the synthesis of cannabinoids.

Identification and expression analysis of the C2H2 gene family in Cannabis sativa L / 药学学报

C2H2 transcription factors play an important role in plant growth, development and the regulation of secondary metabolism. This article identifies members of the C2H2 gene family in Cannabis sativa L. at the genome level. Chromosomal location and linkage, evolutionary relationships, and identification of conserved motifs was determined from the C. sativa genome and transcriptome data using bioinformatics tools and on-line websites such as TBtools, MEGA software, NCBI, PlantTFDB, ExPASy, HMMSCAN, MEME, WoLFPSORT and PlantCARE. The results show that C. sativa contains 30 members of the C2H2 gene family (named CsC2H2-1-CsC2H2-30) distributed on 9 chromosomes. The encoded proteins range in length from 138 to 635 amino acids, and the theoretical isoelectric points range from 5.85 to 9.52. Molecular weights range from 15 909.48 to 68 445.53 Da. Transcriptome analysis showed that CsC2H2 was differentially expressed in the female flowers, bracts, leaves, and stems of the Diku variety and female flowers of nine different varieties of C. sativa. Quantitative real-time PCR verified that CsC2H2-1, CsC2H2-5, and CsC2H2-19 were significantly expressed in the female flowers and bracts of the Diku variety. This provides a theoretical basis for in-depth study of the function of the C2H2 gene family and the breeding of high-quality C. sativa varieties.

Application of MALDI-mass spectrometry imaging in spatial distribution of secondary metabolites in medicinal plants -- a case study of Lepdium meyenii root / 中国中药杂志

Matrix-assisted laser desorption/ionization mass spectrometry imaging(MALDI-MSI) is a novel technique for in-situ distribution of various substances in tissue without labeling. This technique is increasingly applied to the study of medicinal plants owing to its high spatial resolution and its potential of in-situ analysis in small molecules. In this study, the structural information and their fragmentation patterns of the midazole alkaloids(1,3-dibenzyl-4,5-dimethylimidazolium chloride and 1,3-dibenzyl-2,4,5-trimethylimi-dazolium chloride) and benzylglucosinolate in the medicinal plant Maca(Lepdium meyeni) root were analyzed by ultra-high-performance liquid phase combined with LTQ-Orbitrap mass spectrometry(UHPLC-HR-MS). The localization of these active ingredients in the cross-sections of Maca root was performed by MALDI-MSI. These results demonstrated that the two types of imidazole alkaloids had a similar distributed pattern. They were located more in the cortex and the periderm than those in the medulla of a lateral root, while the localization of benzylglucosinolate was concentrated in the center of the root rather than in the cortex and the periderm. The precise spatial distribution of various secondary metabolites in tissue provides an important scientific basis for the accumulation of medicinal plant active ingredients in tissues. In addition, this imaging method is a promising technique for the rapid evaluation and identification of the active ingredients of traditional Chinese medicine in plant tissues, as well as assisting the research on the processing of medicinal plants.

Advances in biosynthesis of indigo in plants / 中国中药杂志

Natural indigo, as one of the oldest dyes, is also a pivotal dye utilized in cotton fabrics today. A diversity of plants rich in indigo compounds belong to traditional Chinese herbal medicines. Indigo compounds have a variety of biological and pharmacological activities, including anticonvulsant, antibacterial, antifungal, antiviral and anticancer activities. A substantial progress in indigo biosynthesis has been made lately. This paper summarizes the value of indigo from the aspects of cultural history, biosynthetic pathways and the medicinal activities of its related derivatives involved in the pathways. In addition, the latest research advancements in indigo biosynthetic pathways is demonstrated in this paper, which would lay the theoretical foundation for the exploration and utilization of natural indigo.

Genome-wide Identification and Characterization of TIFY Gene Family in Medicinal Plant Cannabis sativa / 中国实验方剂学杂志

Objective:The TIFY gene family will be identified and characterized from the whole genome level in Cannabis sativa，which will lay the foundation for gene function study on TIFY family genes and their regulation mechanism on the biosynthesis of cannabinoids and other secondary metabolites. Method:Using the existing genomic data of cannabis，the CsTIFY genes were identified through bioinformatics analysis tools such as NCBI，PlantTFDB，MEME and TBtools etc.，and physicochemical properties，phylogenetic trees，gene structures，chromosome locations and gene expression patterns were analyzed and visualized. Result:Fourteen TIFY family genes（CsTIFY1-CsTIFY14） were identified in Cannabis sativa，which belong to four subfamilies：TIFY，JAZ，ZML，and PPD. The CsTIFYs are composed of 365-1 369 bp nucleotides encoding 118-442 amino acid residues，and their isoelectric points are 4.64-9.96. The 14 CsTIFYs are unevenly distributed on 8 chromosomes，and their proteins are all located in the nucleus. The promoter of CsTIFYs contain multiple abiotic stress responsive cis-acting elements，which indicated that CsTIFYs might involved in the regulation of different abiotic stresses. Transcriptome profiling revealed that CsTIFYs expressed differently in female flowers of 10 differently cannabis varieties，or in flowers，bracts，stems，and leaves of the same variety. Conclusion:Fourteen TIFY family genes were characterized from the whole genome level in C. sativa，and their phylogenetic evolutions and gene expression patterns were analyzed，indicating that CsTIFYs may play important regulatory roles in JA signal transduction，abiotic stress and cannabinoid biosynthesis. This study will provide valuable reference for gene function study of the TIFY family genes in cannabis and cannabis breeding.