Evaluation of genetic diversity using iPBS-SCoT marker methods in native hawthorn genetic resources and species identification by using DNA barcoding method.

Hawthorn is an important medicinal plant that spreads around the world and is used in traditional Chinese medicine. Its flowers and leaves contain flavonoids, vitamins, organic acids and essential oils. Its fruit is consumed as fresh and dried and is an important plant for human health. In this study, iPBS (Inter Primer Binding Site) and SCoT (Start Codon Target Polymorphism) markers were used to analyze genetic variation among 101 hawthorn genotypes collected from Çoruh Valley, Türkiye and ITS markers were used for DNA barcoding.  Ten iPBS primers were used and a total of 400 alleles were identified from ten iPBS primers with an average of 40 alleles. PIC values ranged from 0.239 (iPBS 2387) to 0.272 (iPBS 2244). Twenty SCoT primers were used and have an average of 50.05 alleles. The PIC values of the primers ranged from 0.251 (SCoT 2) to 0.297 (SCoT 34). For the DNA barcoding study, it was confirmed that the correct region was amplified and sequenced. The genotypes we used in the study matched 14 different accession numbers by searching a BLASTN in the NCBI. NCBI similarity rates of hawthorn genotypes are between 90.83% and 100%. The study emphasizes the genetic diversity of hawthorn grown from seed and the importance of preserving plant genetic resources.

Botanical, Phytochemical, Anti-Microbial and Pharmaceutical Characteristics of Hawthorn (Crataegusmonogyna Jacq.), Rosaceae.

Hawthorn (Crataegus monogyna Jacq.) is a wild edible fruit tree of the genus Crataegus, one of the most interesting genera of the Rosaceae family. This review is the first to consider, all together, the pharmaceutical, phytochemical, functional and therapeutic properties of C. monogyna based on numerous valuable secondary metabolites, including flavonoids, vitamin C, glycoside, anthocyanin, saponin, tannin and antioxidants. Previous reviews dealt with the properties of all species of the entire genera. We highlight the multi-therapeutic role that C. monogyna extracts could have in the treatment of different chronic and degenerative diseases, mainly focusing on flavonoids. In the first part of this comprehensive review, we describe the main botanical characteristics and summarize the studies which have been performed on the morphological and genetic characterization of the C. monogyna germplasm. In the second part, the key metabolites and their nutritional and pharmaceutical properties are described. This work could be an essential resource for promoting future therapeutic formulations based on this natural and potent bioactive plant extract.

[Cloning and prokaryotic expression analysis of squalene synthase CpSQS1 and CpSQS2 from Crataegus pinnatifida].

In order to understand the structural characteristics of squalene synthase genes in the triterpenoids biosynthesis pathway of Crataegus pinnatifida, the squalene synthase genes of C. pinnatifida was cloned and analyzed by bioinformatics and prokaryotic expression. Two squalene synthase genes CpSQS1 and CpSQS2 were cloned from C. pinnatifida fruit by RT-PCR. The ORF length of CpSQS1 and CpSQS2 were 1 239 bp and 1 233 bp respectively, encoding 412 aa and 410 aa respectively. CpSQS1 and CpSQS2 were predicted to be stable acidic proteins by online tools. The secondary structure was mainly composed of α-helix structure, and the tertiary structure was predicted by homology modeling. Structural functional domain analysis showed that 35-367 aa of CpSQS1 and CpSQS2 cDNA containing conserved trans-isoprenyl pyrophosphate synthase domains. Transmembrane domain analysis predicted that two transmembrane domains were founded in CpSQS1 and CpSQS2. The squalene synthase amino sequence of C. pinnatifida had higher homology with the known SQS of Salvia miltiorrhiza and Glycyrrhiza glabra. Phylogenetic tree analysis showed that CpSQS1 and CpSQS2 were clustered into one branch of MdSQS1 and MdSQS2, which were consistent with the phylogenetic rule. Prokaryotic expression vector pGEX-4 T-1-CpSQS1 and pGEX-4 T-1-CpSQS2 were transformed into Escherichia coli Transetta(DE3) for induction, and the target protein was successfully expressed at 65 kDa. The expression levels of CpSQS2 were significantly higher than that of CpSQS1 in three different developmental stages of C. pinnatifida. In this study, the full-length cDNA sequences of C. pinnatifida SQS1 and SQS2 were cloned and analyzed for the first time, which provided the foundation for further study on the metabolic pathway of C. pinnatifida triterpenoids.

Designing universal primers for the isolation of DNA sequences encoding Proanthocyanidins biosynthetic enzymes in Crataegus aronia.

BACKGROUND: Hawthorn is the common name of all plant species in the genus Crataegus, which belongs to the Rosaceae family. Crataegus are considered useful medicinal plants because of their high content of proanthocyanidins (PAs) and other related compounds. To improve PAs production in Crataegus tissues, the sequences of genes encoding PAs biosynthetic enzymes are required. FINDINGS: Different bioinformatics tools, including BLAST, multiple sequence alignment and alignment PCR analysis were used to design primers suitable for the amplification of DNA fragments from 10 candidate genes encoding enzymes involved in PAs biosynthesis in C. aronia. DNA sequencing results proved the utility of the designed primers. The primers were used successfully to amplify DNA fragments of different PAs biosynthesis genes in different Rosaceae plants. CONCLUSION: To the best of our knowledge, this is the first use of the alignment PCR approach to isolate DNA sequences encoding PAs biosynthetic enzymes in Rosaceae plants.

Composition and health effects of phenolic compounds in hawthorn (Crataegus spp.) of different origins.

Epicatechin, aglycons and glycosides of B-type oligomeric procyanidins and flavonols, phenolic acids and C-glycosyl flavones are the major groups of phenolic compounds in hawthorn (Crataegus spp). The total content of phenolic compounds is higher in the leaves and flowers than in the fruits. Procyanidins dominate in the fruits, whereas flavonol glycosides and C-glycosyl flavones are most abundant in the leaves. Genotype and developmental/ripening stage have strong impacts. Procyanidin glycosides and C-glycosyl flavones may be chemotaxonomic markers differentiating species and varieties of hawthorn. Future research shall improve the separation, identification and quantification of procyanidins with degree of polymerisation (DP) ≥ 6, procyanidin glycosides, C-glycosyl flavones and some flavonol glycosides. In vitro and animal studies have shown cardioprotective, hypolipidaemic, hypotensive, antioxidant, radical-scavenging and anti-inflammatory potentials of hawthorn extracts, suggesting different phenolic compounds as the major bioactive components. However, the varying and insufficiently defined composition of the extracts investigated, as a result of different raw materials and extraction methods, makes comparison of the studies very difficult. Clinical evidence indicates that some hawthorn extracts may increase the exercise tolerance of patients with congestive heart failure. More clinical studies are needed to establish the effects of hawthorn, especially in healthy humans.

[Phytochemical research of hawthorn species introduced in Lithuania]. / Lietuvoje introdukuotu gudobeliu rusiu ziedu ir lapu fitocheminis tyrimas.

UNLABELLED: The natural vegetable resources of Republic of Lithuania can be supplied by imported hawthorn species, subspecies and varieties, not growing naturally in the territory of country, but well growing and able to produce the crude drug with local conditions of the climate. OBJECTIVE: Determinate amounts of phenolic substances (favonoid glycosides and phenolic acids) in the flowers and leaves of collected species of genus Crataegus and Crataegomespilus. MATERIALS AND METHODS: Different examples of crude drug from some Lithuanian regions were extracted with 96% of ethanol and flavonoids were detected spectroscopically and with HPLC (High performance liquid chromatography). RESULTS: In leaves and flowers were found different quantities of common amounts of flavonoids (1.07-2.53%) and different quantities of individual substances: flavonoids (hyperosid, vitexin-O-rhamnosid, rutin, quercitrin, quercetin, vitexin) and phenolic acids (chlorogenic and caffeic acids). CONCLUSION: Flowers and leaves of hawthorn species, introduced in Lithuania, can be used for preparation of crude drugs procurements; the marks of phytochemical compounds of phenotypes from different genetic lines are sufficient grounds for planning of production of crude drug with special phytotherapeutic influences.