​SSR Based Genetic Diversity Analysis in Fenugreek (Trigonella foenum-graecum L.) Genotypes

Background: Fenugreek (Trigonella foenum-graecum L.) is an important seed spice crop widely grown all over the world. In India, the state of Rajasthan is known for fenugreek production and productivity in the world. A concerted assessment of genetic variability among the germplasm accession is essential for breeding new superior varieties. Molecular markers such as AFLP, RAPD, ISSR, SSR, SCAR, SCoT, SRAP have become for the characterization of the germplasm rapidly and accurately. The present study aimed to characterize 20 elite fenugreek genotypes using simple sequence repeat (SSR) markers to assess the existing genetic diversity of this medicinal crop. Methods: The present study was carried out at the Rajasthan College of Agriculture, Maharana Pratap University of Agriculture and Technology, Udaipur, Rajasthan, India. Total genomic DNA was isolated from old leaves using the CTAB method (Doyle and Doyle, 1990). Further, PCR based genetic diversity was analyzed with using 50 SSR primer pairs. Dendrogram was constructed using NTSYSpc version 2.2 and clustering of the genotypes was done. Result: Twenty genotypes of fenugreek were assessed for genetic diversity analysis using SSR markers. Out of 50 markers 43 primer pairs produced 130 alleles with an average of 84.60% polymorphism. Jaccard’s similarity coefficient lied between and 0.39 to 0.82. Based on UPGMA clustering, a dendrogram consisting of five main clusters was generated with wide variability among the studied genotypes. These diverse genotypes so identified could be gainfully utilized in the fenugreek breeding programme.

Introduction study of Gynostemma pentaphyllum (Thunb.) Makino in the conditions of the Western Caucasus

Relevance. Due to the fact that gynostemma is an endemic plant, providing the pharmaceutical industry with domestic raw materials is not possible, this problem can be solved only through the introduction of this species into the culture.Material and methods. Studies on the introduction of Gynostemma pentaphyllum (Thunb.) Makino in the experimental fields of medicinal crop rotation of the North Caucasus branch of FGBNU VILAR, located in the zone of Western Ciscaucasia, were carried out to study the biomorphological and environmental features of growth and development of the plant, the most optimal conditions for growing and determination of economically valuable indicators. Propagation was carried out through seedlings from seeds received under the contract from South Korea, which were planted in the open ground in early May in areas with direct sunlight and under the canopy of trees (shade). Gynostemma pentaphyllum is a climbing liana with a creeping stem, so it was grown as a trellised crop.Results. It has been established that gynostemma under Krasnodar Territory conditions passes all stages of phenological development, starting from the first year of vegetation, grows and bears fruit every year. The duration of the growing season from 229 to 238 days was determined. Harvesting was carried out in the phase of budding – beginning of flowering. The most optimal condition for growing this crop is shading compared to the option of direct sunlight, where there was a significant increase in growth and development of gynostemma, which contributed to a significant increase in yield. On the rhizome of gynostemma II and III years of vegetation new additional shoots are formed, the number of which is significantly higher in the variant with shading. The conducted introduction studies indicate the possibility of cultivation of Gynostemma pentaphyllum (Thunb.) Makino in the zone of the Western Caucasus.

5-Azacytidine Increases Tanshinone Production in Salvia miltiorrhiza Hairy Roots Through Epigenetic Modulation

Recent studies have indicated strong connections between epigenetic modulation and secondary metabolites in plants. It is vital to understand the roles of epigenetics in the production of secondary metabolites. In this study, the DNA demethylation agent 5-azacytidine (5-Az) was used on the hairy roots of the medicinal crop Salvia miltiorrhiza to investigate its effect on secondary metabolite production, gene expression, and methylation levels in gDNA and promoter regions. Our results showed that the contents of tanshinones in S. miltiorrhiza hairy roots increased by 1.5 to 5 times, and most genes in the MEP pathway showed an upward trend. According to our NGS analysis, the methylation pattern in the copalyl diphosphate synthase (CPS) promoter was altered, and 51 out of 145 cytosines were demethylated during 5-Az treatment. A total of 36 different transcription factors (TFs) were identified in these demethylation sites. Among these TFs, NF-Y and MYB were frequently found in our results. This is the first report to demonstrate a possible mechanism of DNA methylation participating in tanshinone biosynthesis in S. miltiorrhiza hairy roots by modulating the CPS promoter and TFs.

Analysis of Genetic Variability and Agronomic Performance of Indian Lettuce (Lactuca Indica Linn.)

Lactuca indica is a undomesticated medicinal crop in the Asteraceae family. Present study was carried out to identify elite genotype for cultivation and breeding improvement of lettuce. Data was recorded for 19 morphological and developmental traits across 38 accessions (Acc). In addition to this, the morphological cluster was compared with the simple sequence repeat (SSR) marker. The genotypic mean square variance was significant for all characters. The higher extent of genotypic and phenotypic coefficient of variation obtained for basal branch, leaf blade width, and node number. The broad-sense heritability (H2B) ranged from 45.85% (seed length) to 98.59% (node number), whereas genetic advance as a percentage of the mean (GAM%) ranged from 9.33 to 191. Vegetative characters such as node number, plant hight, basal branch were conjugated with high H2B and high GAM% indicating additive gene effect and selection of these traits based on phenotypic observation is effective for better gain. Reproductive traits, including bolting duration, flowering duration, and seed weight were linked with high H2B, and moderate GAM% trevealing that these traits are amenable to genetic improvement, these traits also showed a significant positive correlation. Acc 55 and 8 showed the best performance for the majority of the attributes could be good material for further research and breeding. In the Wards’ phylogenetic tree of morphological traits, accessions were clustered based on their phenotypic characters rather than the geographic origin. Morphological characteristics showed no or very week corelation with the SSR genotyped data.

Tetraploidization Increases the Contents of Functional Metabolites in Cnidium officinale

Cnidium officinale is an important medicinal crop grown in Asia for its pharmacological properties. In this study, tetraploid breeding was conducted to increases the content of medicinal compound and tolerance to the environmental conditions using in vitro shoot culture of C. officinale. For this, we generated tetraploid C. officinale plants using oryzalin, a chromosome doubling agent, and compared the morphological traits, cytological characteristics, and heat stress-responsive gene expression levels between tetraploid and diploid genotypes. Chromosome doubling efficiency was the highest in plantlets treated with 4.0 mg∙L−1 oryzalin for 2 days. Compared with diploids, the plant height of tetraploids was reduced, while the petiole diameter was increased by approximately 39%. The dry matter of tetraploid leaves was significantly higher than that of diploid leaves. Compared with diploids, tetraploids showed higher chloroplast number and stomatal complex size but lower chlorophyll and carotenoid contents. The phenolic content of tetraploid plantlets was significantly higher than that of diploid plantlets. Contents of naringin as well as salicylic acid and gentisic acid, which are strong antioxidant compounds, were dramatically increased upon tetraploidization. Interestingly, liquid chromatography–mass spectrometry (LC–MS) analyses revealed increased levels of senkyunolide F and phthalide in tetraploid roots but not in tetraploid or diploid leaves.

Effect of foliar application of different elicitors on occurrence of pest and diseases during chia (Salvia hispanica L.) cultivation

Chia is an important medicinal crop with lot of nutritional value in seeds and there is lot of demand to chia seeds grown without any chemical application. In this context, present study was concentrated on foliar application of different elicitors at 25 and 50 days after sowing as effective plant protectants by inducing plant defense response. Both black and white chia plants sprayed with chitosan at 200 ppm were un-affected by any pest and diseases. The plants sprayed with other elicitors like 100 ppm of salicylic acid, methyl jasmonic acid, potassium silicate, 200 ppm of boric acid, humic acid and 5000 ppm of dry yeast and PGPR are less prone to pest and disease attack. Whereas, the maximum pest infestation and disease incidence were noticed in plants sprayed with gibberellic acid (pest - 2.38 and 1.93%; disease - 0.89% in both) and in control (pest - 1.34 and 1.04%; disease - 1.34 and 0.74%) in black and white chia respectively.

Planting Density Affects Panax notoginseng Growth and Ginsenoside Accumulation by Balancing Primary and Secondary Metabolism

Adjusting planting density is a common agricultural practice used to achieve maximum yields. However, whether the quality of medicinal herbs can be improved by implementing appropriate planting densities is still uncertain. The medicinal crop Panax notoginseng was used to analyze the effects of planting density on growth and ginsenoside accumulation, and the possible mechanisms of these effects were revealed through metabonomics. The results showed that P. notoginseng achieved high ginsenoside accumulation at high planting densities (8 × 8 and 10 × 10 cm), while simultaneously achieved high biomass and ginsenoside accumulation at moderate planting density of 15 × 15 cm. At the moderate planting density, the primary metabolism (starch and sucrose metabolism) and secondary metabolism (the biosynthesis of phytohormone IAA and ginsenoside) of the plants were significantly enhanced. However, the strong intraspecific competition at the high planting densities resulted in stress as well as the accumulation of phytohormones (SA and JA), antioxidants (gentiobiose, oxalic acid, dehydroascorbic acid) and other stress resistance-related metabolites. Interestingly, the dry biomass and ginsenoside content were significantly lower at low densities (20 × 20 and 30 × 30 cm) with low intraspecific competition, which disturbed normal carbohydrate metabolism by upregulating galactose metabolism. In summary, an appropriate planting density was benefit for the growth and accumulation of ginsenosides in P. notoginseng by balancing primary metabolism and secondary metabolism.