Sativa Cultivars Research Articles

Development and validation of a minimal SNP genotyping panel for the differentiation of Cannabis sativa cultivars

BackgroundDue to its previously illicit nature, Cannabis sativa had not fully reaped the benefits of recent innovations in genomics and plant sciences. However, Canada’s legalization of C. sativa and products derived from its flower in 2018 triggered significant new demand for robust genotyping tools to assist breeders in meeting consumer demands. Early molecular marker-based research on C. sativa focused on screening for plant sex and chemotype, and more recent research has sought to use molecular markers to target traits of agronomic interest, to study populations and to differentiate between C. sativa cultivars.ResultsIn this study, we have conducted whole genome sequencing of 32 cultivars, mined the sequencing data for SNPs, developed a reduced SNP genotyping panel to discriminate between sequenced cultivars, then validated the 20-SNP panel using DNA from the sequenced cultivars and tested the assays on commercially available dried flower. The assay conversion rate was higher in DNA extracted from fresh plant material than in DNA extracted from dried flower samples. However, called genotypes were internally consistent, highlighting discrepancies between genotypes detected using sequencing data and observed using genotyping assays. The primary contributions of this work are to clearly document the process used to develop minimal SNP genotyping panels, the feasibility of using such panels to differentiate between C. sativa cultivars, and outline improvements and goals for future iterations of PCR-based, minimal SNP panels to enable efficient development genotyping tools to identify and screen C. sativa cultivars.ConclusionsOur key recommendations are to increase sampling density to account for intra-cultivar variability; leverage higher read length paired-end short-read technology; conduct in-depth pre- and post-processing of reads, mapping, and variant calling data; integrate trait-associated loci to develop multi-purpose panels; and use iterative approaches for in vitro validation to ensure that only the most discriminant and performant SNPs are retained.

Comparison of the rhizospheric soil bacteriomes of Oryza sativa and Solanum melongena crop cultivars reveals key genes and pathways involved in biosynthesis of ectoine, lysine, and catechol meta-cleavage.

Rice (Oryza sativa L.), Poaceae family, forms staple diet of half of world's population, and brinjal (Solanum melongena L.), an important solanaceous crop, are consumed worldwide. Rhizosphere research is gaining importance towards application of knowledge for improving productivity, sustainable agricultural practice, and rhizoremediation for nature restoration. While there are reports on rhizobacteriome of rice, studies comparing structural, functional and metabolomic traits of microbial communities in rhizospheres of rice and brinjal are not yet available. We demonstrated, in Oryza sativa (1144-Hybrid, Dhiren, Local Saran cultivars) and Solanum melongena (Jhiloria, Chandtara, Jotshna cultivars) rhizospheres from Malda, India, using integrated approach of 16S ribosomal sequencing, shotgun metagenomics, and microbial metabolomics to decipher microbial diversity, association with soil physicochemical characteristics, key genes and pathways. Ectoine biosynthesis was significantly expressed in brinjal (Jhiloria), but not in rice rhizosphere. The dominant brinjal rhizobacteriome-specific bacteria comprised Thermus sp., Petrobacter succinatimandens, Thermoanaerobacter sp., and Diaphorobacter sp., that were involved in house-keeping functions including pentose phosphate pathway, biosynthesis of amino acids, lipopolysaccharide, and photosynthesis. The dominant bacteria unique to rice rhizobacteriome (Local Saran) consisted of Aeromonas sp., associated with catechol meta-cleavage, while Clostridium sp., Faecalibacterium prausnitzii, and Roseburia sp. were involved with lysine biosysnthesis in rice (1144-Hybrid). Our results imply novel information for improved breeding of brinjal specific cultivar with enhanced ectoine production associated with osmotic stress tolerance, rice specific cultivars with enhanced lysine production significant to human nutrition and catechol removal for the maintenance of environmental quality.

Intraspecies variation in mycorrhizal response of Medicago sativa to Rhizophagus irregularis under abiotic stress.

Plant partnerships with arbuscular mycorrhizal fungi (AMF) improve plant resilience to stress by increasing the plant's access to and uptake of essential nutrients and water, as well as regulating the plant's stress response. The magnitude and direction of AMF effects during the relationship depend on multiple factors including plant identity and environmental context. To investigate how AMF influence plant responses to environmental stresses, we assessed the effects of drought and salinity on growth, final biomass, and reproduction of nine alfalfa (Medicago sativa) cultivars inoculated with Rhizophagus irregularis or grown alone. In absence of stress, the fungus increased nutrient content, but caused declines in biomass through a reduction in initial growth that was not overcome by a later growth spurt. Mycorrhizal fungus inoculation also magnified stress effects on growth in most scenarios, but this depended on the stress type and cultivar. For salinity, this stress increase in inoculated plants was mediated by increased salt accumulation. Flowering of each cultivar was affected by both inoculation and stress type, albeit erratically, whereas seed production was only affected by inoculation when drought stressed. We found no clear pattern distinguishing differences in mycorrhizal fungus effects on stress among cultivars; however, our results show that mycorrhizal fungus effects on plant stress responses are contingent on the plant performance metric and stress type, highlighting the complexity of responses to mycorrhizas.

A Multidisciplinary Approach for the Assessment of the Last Surviving ‘Marrone di Chiusa Pesio’ Chestnut Trees in the Piemonte Region (Italy)

Chestnut orchards are a multifunctional resource, providing not only fruit or wood but also playing a role in the conservation of mountain and hillside landscapes. In the Piemonte Region, Italy, a rich genetic heritage of chestnut genotypes has contributed to considerable biodiversity and environmental value. The study aimed to valorize an important example of the chestnut agrobiodiversity in the Piemonte Region by focusing on the ‘Marrone di Chiusa Pesio’ (MCP) cultivar (cv). A multidisciplinary approach was applied, involving genetic and morphological analyses, Visual Tree Assessment (VTA), and phytochemical and nutritional profiling. The plant census provided identification and geolocation of 187 MCP specimens; the 20 most representative trees were genetically analyzed, and then, through the VTA, their morpho-functional status was evaluated. The nutraceutical properties and phytochemical composition were assessed by measuring the total polyphenol content (TPC), antioxidant capacity (AOC), and other phytochemical classes through spectrophotometric and chromatographic methods. The results showed significantly higher TPC values (ranged from 36.51 ± 1.60 mgGAE/100 g of dried weight—DW to 103.14 ± 1.24 mgGAE/100 g DW) compared to other ‘Marrone-type’ cultivars, along with high levels of key phenolic markers, bioactive compounds, and nutritional substances. These included tannins (about 22–28 mg/100 g DW) and cinnamic acids (about 23–25 mg/100 g DW), followed by flavonols, benzoic acids, organic acids, monoterpenes, vitamin C, and catechins, listed in order of predominance. A Principal Component Analysis (PCA) was performed to observe the distribution of the samples and their correlations based on the chemical composition. The results confirmed the interesting phytochemical properties of the ‘Marrone di Chiusa Pesio’ nuts, together with their good morphological and functional properties. Given the ongoing genetic erosion of Castanea sativa cultivars, due to cultivation abandonment and climate change, the main factors contributing to the progressive loss of biodiversity worldwide, the presented approach aimed to provide an overview of the conservation status of the local agrobiodiversity. This study highlighted the value of a local chestnut cultivar, presenting the low conservation status of the few remaining specimens. The goal was to define the significant phenotypic variation regarding MCP in the considered area due to environmental variations, which may be of interest in its genetic adaptation to climate change. The study may potentially encourage the development of strategies for actively conserving the forest agrobiodiversity and hillside ecosystem services in the highly diverse landscapes of the Alpine valleys.

De-Methyl Esterification Modification of Root Pectin Mediates Cd Accumulation of Lactuca sativa.

Cadmium (Cd) contamination in agricultural soil brings severe health risks through the dietary intake of Cd-polluted crops. The comprehensive role of pectin in lowering Cd accumulation is investigated through low Cd accumulated (L) and high Cd accumulated (H) cultivars of L. sativa. The significantly different Cd contents in the edible parts of two L. sativa cultivars are accomplished by different Cd transportations. The pectin is the dominant responsive cell wall component according to significantly increased uronic acid contents and the differential Cd absorption between unmodified and modified cell wall. The chemical structure characterization revealed the decreased methyl esterification in pectin under Cd treatment compared with control. Significantly brighter LM19 relative fluorescence density and 40.82% decreased methanol in the root pectin of L cultivar under Cd treatment (p < 0.05) supported that the de-methyl esterification of root pectin is more significant in L cultivar than in H cultivar. The pectin de-methyl esterification of L cultivar is achieved by the upregulation of pectin esterases and the downregulation of pectin esterase inhibitors under Cd treatments, which has facilitated the higher Cd-binding of pectin. Our findings provide deep insight into the differential Cd accumulation of L. sativa cultivars and contribute to the understanding the pollutant behaviors in plants.

Major components of RNAi gene families in Oryza sativa cultivar Kitaake: In-silico discovery and characterization

BackgroundThe Oryza sativa cultivar Kitaake is a promising new model rice for research due to its short life cycle (9 weeks), adaptability to greenhouse conditions, readily accepts foreign genes, and its complete genome sequence is accessible, providing a valuable blueprint for researchers. However, its major RNA interference (RNAi) gene families (DCLs, AGOs, RDRs) have not yet been studied. These gene families influence target-specific protein-coding gene expression and biotic and abiotic stresses, regulating plant growth and development. ObjectivesThis study aims to identify and characterize RNAi gene families from the Kitaake rice. MethodsThis study has been designed by in-silico analysis to explore major RNAi genes highlighting their molecular functions, phylogenetic groups, regulatory factors, and other vital characteristics of Kitaake rice corresponding to OsRNAi genes. ResultsThis study has identified 10 DCLs, 21 AGOs, and 7 RDRs as major RNAi proteins of Kitaake rice corresponding to OsRNAi by BLASTP search. Domain analysis has been revealed the RNase III, PAZ, and Piwi domains are related to gene silencing processes. According to synteny study, Kitaake and rice have the most homology in 33 RNAi gene pairs, suggesting they share a chromosomal order and similar functions. The majority of OsKRNAi proteins have been located in the nucleus and chloroplast, which are related to gene silencing. Gene silencing and ribonuclease III activity are key terms from gene ontology (GO) analysis, which is part of the gene silencing process. Gene regulatory analysis has identified some important transcription factors including ERE, which participates in DNA binding and microRNAs (miRNAs) including 'Osa-MIR168' improves rice resistance to blast disease. The investigation of cis-acting regulatory elements in OsKRNAi genes has shown various crucial components, including MBS, W-box, LTR, ABRE, ARE that are linked with the different stresses. Genes (such as OsKAGO1d) were found to be overexpressed in target of rapamycin (TOR), increasing susceptibility to pathogens. ConclusionThe findings of this study may be useful resources for further experimental investigation on the improvement of Kitaake rice crop against different stresses.

Optimized guidelines for feminized seed production in high-THC Cannabis cultivars.

With the partial legalization of high-THC Cannabis sativa across 23 states for recreational use and 38 states for medical purposes in the United States, the Cannabis industry is poised for significant growth. Projected to reach a sales volume of $50.7 billion by 2028, this growth is driven by the trend of lifting Cannabis prohibition. High-THC C. sativa cultivars, containing more than 0.3% delta-9 tetrahydrocannabinol (Δ9-THC) as defined by the 2018 US Farm Bill, are used for both medicinal and recreational purposes. Cannabis sativa is a short day, dioecious, annual plant, where female plants are favored for THC production, which requires seed feminization techniques to ensure an accurate female plant population. This involves using an ethylene inhibitor to induce sex reversal, leading to male flower development on female plants, allowing for self-pollination and the production of feminized seeds. However, challenges such as seed viability and the occurrence of male flowers in progeny have been noted. This review provides guidelines to enhance the production of viable feminized seeds in high-THC Cannabis cultivars. Literature findings indicate that Silver Thiosulfate (STS) is the most effective ethylene inhibitor for sex reversal and seed feminization in high-THC Cannabis cultivars. Specifically, a single dose of 3 mM STS should be applied during the vegetative stage via foliar spraying until runoff, followed by exposure to a short photoperiod of up to 12 hours to induce flowering and seed production. Progeny plants should be assessed for seed germination rate and compared for growth performance with the original parent plant to assess the declining effects of inbreeding. Adhering to these guidelines can improve the quality and viability of feminized seeds, meeting commercial market standards and industry demands for high-THC Cannabis cultivars.

Physicochemical Properties, Organic Acid, and Sugar Profiles in Edible and Inedible Parts of Parsnip (Pastinaca sativa) Cultivars Harvested in Korea

Parsnip, a root vegetable from the Apiaceae family, is rich in dietary fiber, pectin, and starch but remains relatively unfamiliar in South Korea. This study investigated the physicochemical properties of two Korean-grown parsnip cultivars, ‘Warrior’ and ‘Albion’, focusing on their organic acid and sugar compositions. The ‘Warrior’ cultivar has higher firmness and water content but lower SSC compared to ‘Albion’. In ‘Warrior’, malic and lactic acids were the main organic acids, while ‘Albion’ had predominant oxalic and malic acids. Malic acid was also the primary organic acid in the inedible parts of ‘Warrior’, and oxalic acid in ‘Albion’. In the edible parts of both cultivars, sucrose was identified as the main sugar. In ‘Warrior’, the levels were 88.59%, 90.35%, and 79.13% in the cortex, pith, and skin, respectively, while in ‘Albion’, the levels were 88.56%, 64.40%, and 67.39%. ‘Warrior’ showed higher total sugar content in its cortex (6.66%) compared to ‘Albion’ (3.67%). These results highlight the beneficial compounds in parsnips and suggest their potential for improving dietary strategies and health.

Optimization of transformation conditions of the yeast &lt;i&gt;Saccharomyces cerevisiae&lt;/i&gt; to determine coregulators of the transcription factor NIN in a yeast two-hybrid system

BACKGROUND: The method of protein-protein interaction analysis using the yeast two-hybrid system in Saccharomyces cerevisiae cells is used to search for protein coregulators. This method is also used for mass screening of libraries of cloned fragments of complementary DNA (cDNA) that are translated in the cell. The key factor in the success of such screening is the level of efficiency of yeast cell transformation, since the resolution of the analysis is based on this. AIM: The aim of this study is to search for optimal parameters for chemical transformation of yeast cells to increase the resolution of cDNA library screening. MATERIALS AND METHODS: Plasmids pDEST22 and pDEST32 were used for chemical transformation of the yeast strain S. cerevisiae pJ69-4A. For screening, a cDNA library was used, obtained on the basis of mRNA isolated from the roots of pea Pisum sativum cultivar Finale, inoculated with rhizobia. RESULTS: Factors influencing the efficiency of transformation were identified. Among them are the molecular weight of polyethyleneglycol used for transformation, as well as the number of cell division cycles that the culture undergoes. The effect of the number and size of plasmids used in transformation was also shown. Using the optimized protocol, a cDNA library was successfully screened to find coregulators of the NIN transcription factor. CONCLUSIONS: Based on the data obtained, optimal parameters were determined that allow achieving a high level of competence in yeast cells. The use of the described protocol allowed for successful screening of the library to identify coregulators of the NIN transcription factor.

Quantitative eco-anatomical analysis reveals distinctive stem traits of six Cannabis sativa cultivars in Rif Mountains (Northern Morocco)

Cannabis sativa stands out as an important crop with significant socioeconomic importance in the Rif Mountains of North Morocco. Recently the number of new cultivars developed in this region has gradually increased for different purposes. In this current study, we delve into the eco-anatomical characteristics of stems across six of the most prevalent Cannabis sativa cultivars in Morocco: ‘Avocat’, ‘Beldiya’, ‘Critical Plus’, ‘Industriel’, ‘Khardala’, and ‘Mexicana’. The investigation encompasses the measurement and analysis of six key eco-anatomical traits: plant height, stem diameter, vessel size, vessel density, number of vessels joined in radial files, and coefficient of hydraulic conductivity. Our aim is to elucidate the variations among these cultivars and underscore their agricultural and ecological relevance. Through anatomical examinations of stem cross-sections, our findings reveal notable disparities and distinctive features among the cultivars. The ‘Khardala’ cultivar exhibits the highest values across most measured eco-anatomical traits, while ‘Beldiya’ and ‘Mexicana’ show similar trait values, particularly with low stem diameter and vessel density. In contrast, ‘Industriel’ stands out with the lowest vessel size and highest vessel density. ‘Critical Plus’ and ‘Avocat’ closely resemble ‘Khardala’ in vessel size and stem diameter. Regarding hydraulic conductivity, ‘Khardala’ ranks highest, followed by ‘Avocat’ and ‘Critical Plus’, while ‘Industriel’ registers the lowest. Our findings underscore increased water demand and uptake potential in ‘Khardala’, ‘Critical Plus’, and ‘Avocat’, contrasting with ‘Industriel’, ‘Beldiya’, and ‘Mexicana’. Anatomical diversity among Cannabis cultivars in our study likely stems from genetic lineage and ecological adaptation.

Metabolomics combined with proteomics reveals phytotoxic effects of norfloxacin under drought stress on Oryza sativa

In recent decades, plants enduring abiotic stresses such as drought and chemical stresses. Currently, the mechanism of combined antibiotic and drought stress response and its impact on crop growth and food security remains poorly understood. Here, the mechanism of stress responses under the exposure of norfloxacin (NF) and drought (D) individually and in combination (DNF) were explored on rice (Oryza sativa) cultivar Hanyou73 through proteomics and metabolomic analysis. All treatments adversely affected chlorophyll fluorescence kinetics, antioxidant enzyme activities, rice grain composition and yield. The results showed that in DNF the antibiotic was accumulated 627% more than NF treatment in rice grains while in leaves there was no significant difference under both treatments. The proteomic revealed that differentially expressed identified proteins were involved in carbohydrate metabolism, amino acid metabolism, photosynthesis and mRNA binding. However, the metabolomics results showed that the abundance of metabolites related to RNA biosynthesis and amino acid metabolism were more affected. The disruptions caused in rice plant under DNF treatment become more severe, this makes it more susceptible than individual D and NF treatment. These findings improve our knowledge about the response of rice plant to cope with antibiotic contamination alone and in combination with drought.

Genome-Wide Identification of NAC Family Genes and Their Expression Analyses in Response to Osmotic Stress in Cannabis sativa L.

NAC (NAM, ATAF1/2, and CUC2) transcription factors are unique and essential for plant growth and development. Although the NAC gene family has been identified in a wide variety of plants, its chromosomal location and function in Cannabis sativa are still unknown. In this study, a total of 69 putative CsNACs were obtained, and chromosomal location analysis indicated that the CsNAC genes mapped unevenly to 10 chromosomes. Phylogenetic analyses showed that the 69 CsNACs could be divided into six subfamilies. Additionally, the CsNAC genes in group IV-a are specific to Cannabis sativa and contain a relatively large number of exons. Promoter analysis revealed that most CsNAC promoters contained cis-elements related to plant hormones, the light response, and abiotic stress. Furthermore, transcriptome expression profiling revealed that 24 CsNAC genes in two Cannabis sativa cultivars (YM1 and YM7) were significantly differentially expressed under osmotic stress, and these 12 genes presented differential expression patterns across different cultivars according to quantitative real-time PCR (RT-qPCR) analysis. Among these, the genes homologous to the CsNAC18, CsNAC24, and CsNAC61 genes have been proven to be involved in the response to abiotic stress and might be candidate genes for further exploration to determine their functions. The present study provides a comprehensive insight into the sequence characteristics, structural properties, evolutionary relationships, and expression patterns of NAC family genes under osmotic stress in Cannabis sativa and provides a basis for further functional characterization of CsNAC genes under osmotic stress to improve agricultural traits in Cannabis sativa.

Understanding mechanistic variability in physiological and biochemical responses of pea cultivars (Pisum sativum L.) to ozone exposure

Increasing concentration of ground level O3 and its negative impacts on agricultural output is well documented, however, the response of leguminous crop plants is still sparsely cited. Given their nutritional richness, legume seeds are widely esteemed as a crucial dietary staple worldwide, prized for their abundance of oil, protein, dietary fiber, and low-fat characteristics. Termed as the “poor man's meat” due to their high-quality protein, they hold immense economic value. Acknowledging the significance of legumes, a field experiment was conducted to understand the physiological and antioxidant responses, stomatal characteristics, and yield response in three cultivars of Pisum sativum L. (K Agaiti, K Uday and K Damini), exposed to elevated ozone (O3). In the present study, Pisum sativum cultivars were subjected to ambient (control) and elevated (+15 ppb) concentrations of O3, using separate sets of OTCs. Elevated O3 stimulated the activity of the enzymes of Halliwell Asada pathway, which were responsible for the differential response of the three experimental cultivars. While K Agaiti and K Uday focused on upregulating their antioxidant defense, K Damini followed the strategy of biomass allocation. Test weight showed that K Damini was most efficient in succoring the yield losses under elevated O3. Under elevated O3, test weight reduced by 8.91%, 7.52%, and 5.1%, respectively, in K Agaiti, followed by K Uday and K Damini, rendering K Agaiti most sensitive to O3 stress. The present study not only helps us to elucidate the O3 sensitivity of the selected experimental cultivars, it also helps us in screening O3 tolerant cultivars for future agricultural practices.

Understanding the role of magnetic (Fe3O4) nanoparticle to mitigate cadmium stress in radish (Raphanus sativus L.)

Heavy metals stress particularly cadmium contamination is hotspot among researchers and considered highly destructive for both plants and human health. Iron is examined as most crucial element for plant development, but it is available in inadequate amount because they are present in insoluble Fe3+ form in soil. Fe3O4 have been recently found as growth promoting factor in plants. To understand, a sand pot experiment was conducted in completely randomized design (control, cadmium, 20 mg/L Fe3O4 nanoparticles,40 mg/L Fe3O4 nanoparticles, 20 mg/L Fe3O4 nanoparticles + cadmium, 40 mg/L Fe3O4 nanoparticles + cadmium) to study the mitigating role of Fe3O4 nanoparticles on cadmium stress in three Raphanus sativus cultivars namely i.e., MOL SANO, MOL HOL PARI, MOL DAQ WAL. The plant growth, physiological and biochemical parameters i.e.,shoot length, shoot fresh weight, shoot dry weight, root length, root fresh and dry weight, MDA content, soluble protein contents, APX, CAT, POD activities and ion concentrations, membrane permeability, chlorophyll a, chlorophyll b and anthocyanin content, respectively were studied. The results displayed that cadmium stress remarkably reduces all growth, physiological and biochemical parameters for allcultivars under investigation. However, Fe3O4 nanoparticles mitigated the adverse effect of cadmium by improving growth, biochemical and physiological attributes in all radish cultivars. While, 20 mg/L Fe3O4 nanoparticles have been proved to be more useful against cadmium stress. The outcome of present investigation displayed that Fe3O4 nanoparticles can be utilized for mitigating heavy metal stress.

Low-Cost Imaging to Quantify Germination Rate and Seedling Vigor across Lettuce Cultivars.

The survival and growth of young plants hinge on various factors, such as seed quality and environmental conditions. Assessing seedling potential/vigor for a robust crop yield is crucial but often resource-intensive. This study explores cost-effective imaging techniques for rapid evaluation of seedling vigor, offering a practical solution to a common problem in agricultural research. In the first phase, nine lettuce (Lactuca sativa) cultivars were sown in trays and monitored using chlorophyll fluorescence imaging thrice weekly for two weeks. The second phase involved integrating embedded computers equipped with cameras for phenotyping. These systems captured and analyzed images four times daily, covering the entire growth cycle from seeding to harvest for four specific cultivars. All resulting data were promptly uploaded to the cloud, allowing for remote access and providing real-time information on plant performance. Results consistently showed the 'Muir' cultivar to have a larger canopy size and better germination, though 'Sparx' and 'Crispino' surpassed it in final dry weight. A non-linear model accurately predicted lettuce plant weight using seedling canopy size in the first study. The second study improved prediction accuracy with a sigmoidal growth curve from multiple harvests (R2 = 0.88, RMSE = 0.27, p < 0.001). Utilizing embedded computers in controlled environments offers efficient plant monitoring, provided there is a uniform canopy structure and minimal plant overlap.

Tracing metabolic route of thymoquinone biosynthesis in diverse Nigella sativa L. cultivars using RP-HPLC technique

Although the therapeutic role of Nigella sativa has been known for a long time, for optimal utilization of plant parts, the developmental profiling of bioactive constituents and their dependence on germplasm diversity need to be assessed.Different cultivars of Nigella sativa(covering 3 samples from northern states of India and 3 reference samples) were taken to explore the relative accumulation of thymol (Thy) and thymoquinone (TQ) in different plant parts, i.e., stem, leaf, bud, flower, fruit, and seeds. Amongst all N. sativa cultivars, the maximum TQ content was found in the seed (45.3885 ± 0.3616 mg/g fresh weight) of the RS cultivar. Seeds of the AN1 cultivar showed the maximum Thy content (2.7853± 0.3535 mg/g fresh weight). The minimum Thy content was found in leaves of Haryana (0.0009 ± 0 mg/g fresh weight) and leaves from RS showed the minimum TQ content (0.0029 ± 0.0001 mg/g fresh weight). There are indications that mature seeds are the best source for both Thy and TQ, followed by fresh seeds collected from the intact fruit of N. sativa. In addition, Thy (precursor) and TQ (product)contentin many plant parts are inversely related to each other, especially in the buds. This indicatedthe interdependence of these two bioactive constituents, but not necessarily in all plant parts. It appears that there might be another functional precursor (in addition to Thy)leading to TQ biosynthesis (speculated to be Carvacrol) in Nigella. Such relative dominance of bioactive compounds and their precursors may be used for designing formulations and setting biotransformation experiments for further value addition.

Enhanced arsenic stress tolerance in landrace and improved rice cultivars through modulation of gibberellic acid (GA3) synthesis and antioxidant metabolism via phosphorus and silicon supplementation

Arsenic (As), an environmental pollutant, imposes toxic impacts on plant health. In recent years, researchers have focused on comprehending the As-induced intrusion in growth and metabolic components. However, the interactions between phosphorus (P) and silicon (Si) remain elusive, specifically in rice (Oryza sativa) cultivars. Therefore, the present study investigated the effects of P and Si on the defense mechanisms, nutrients accumulation, carbohydrate metabolism, gibberellic acid (GA3) synthesis, and growth responses in Indica rice cultivars (landrace and improved) under As stress. With this focus, our study potentially indicated that As stress (150 µM) negatively impacted the underlying physiological and growth traits, with more pronounced effects on the improved cultivar compared to the landrace. However, P (30 mg kg−1 soil) and/ or Si (1 mM) treatments have prominently reduced oxidative stress with improved defense mechanisms and GA synthesis. Modulation of enzymatic defense system increased in both cultivars but more pronounced in landrace, including superoxide dismutase (+36.47%, +24.43%), ascorbate peroxidase (+95.11%, +78.26%), along with non-enzymatic antioxidants such as ascorbate (+29.43%, +21.48%), and reduced glutathione (+75.45%, +62.34%) concentrations, respectively. Notably, GA concentration (+23.78%, +16.48%) was substantially increased by the cumulative application of P and Si under As stress in both cultivars but more conspicuous in the landrace, respectively. Further, employing GA biosynthesis inhibitor, paclobutrazol (PBZ; 10 µM), substantiated the input of GA-mediated As tolerance, wherein, PBZ reversed the impacts of P and Si on the endogenous GA concentration and defense metabolism. In summation, landrace outperformed improved cultivar leading to As tolerance. This implies that the landrace could be used for future breeding programs permitting in-depth considerations of P and Si-mediated GA3 synthesis under As stress conditions.

Development of a severity scale and an RT‐qPCR assay for screening resistance levels in rice genotypes against rice stripe necrosis virus and its vector

AbstractRice stripe necrosis virus (RSNV) is the causal agent of the disease ‘rice crinkling’ and is transmitted by the protozoan Polymyxa graminis. Although genetic resistance has been explored, no resistant commercial cultivars are currently available. Oryza glaberrima has been identified as a promising source of resistance. However, it remains unclear whether this resistance is effective against the virus, the vector, or both, as well as whether it can be transferred to Oryza sativa cultivars. Disease‐resistant genotypes are primarily selected through visual observations of symptom expression. The absence of a severity scale for RSNV makes this process difficult, and relying solely on visual assessments can introduce subjectivity. We developed a severity scale and a reverse transcripiton‐quantitative PCR (RT‐qPCR) assay for screening resistance levels in rice genotypes against RSNV and its vector and to analyse the genetic variability of RSNV isolates. To achieve absolute quantification, experiments were conducted using O. glaberrima and three O. sativa cultivars. Inoculation occurred naturally using soil from an area with a history of the disease. Visual symptoms were recorded and disease intensity was evaluated. Subsequently, total nucleic acid extraction was performed on the samples and viral and vector loads were quantified through RT‐qPCR and qPCR, respectively. To characterize the virus variability, symptomatic rice samples were collected in the 2021/2022 crop season. RT‐PCR was conducted to amplify the coat protein gene of RSNV, and molecular variability descriptors were analysed.

Influence of Fungicide Application on Rhizosphere Microbiota Structure and Microbial Secreted Enzymes in Diverse Cannabinoid-Rich Hemp Cultivars.

Microbes and enzymes play essential roles in soil and plant rhizosphere ecosystem functioning. However, fungicides and plant root secretions may impact the diversity and abundance of microbiota structure and enzymatic activities in the plant rhizosphere. In this study, we analyzed soil samples from the rhizosphere of four cannabinoid-rich hemp (Cannabis sativa) cultivars (Otto II, BaOx, Cherry Citrus, and Wife) subjected to three different treatments (natural infection, fungal inoculation, and fungicide treatment). DNA was extracted from the soil samples, 16S rDNA was sequenced, and data were analyzed for diversity and abundance among different fungicide treatments and hemp cultivars. Fungicide treatment significantly impacted the diversity and abundance of the hemp rhizosphere microbiota structure, and it substantially increased the abundance of the phyla Archaea and Rokubacteria. However, the abundances of the phyla Pseudomonadota and Gemmatimonadetes were substantially decreased in treatments with fungicides compared to those without fungicides in the four hemp cultivars. In addition, the diversity and abundance of the rhizosphere microbiota structure were influenced by hemp cultivars. The influence of Cherry Citrus on the diversity and abundance of the hemp rhizosphere microbiota structure was less compared to the other three hemp cultivars (Otto II, BaOx, and Wife). Moreover, fungicide treatment affected enzymatic activities in the hemp rhizosphere. The application of fungicides significantly decreased enzyme abundance in the rhizosphere of all four hemp cultivars. Enzymes such as dehydrogenase, dioxygenase, hydrolase, transferase, oxidase, carboxylase, and peptidase significantly decreased in all the four hemp rhizosphere treated with fungicides compared to those not treated. These enzymes may be involved in the function of metabolizing organic matter and degrading xenobiotics. The ecological significance of these findings lies in the recognition that fungicides impact enzymes, microbiota structure, and the overall ecosystem within the hemp rhizosphere.

Exploring Leaf Anthocyanin Concentrations and Light Effects on Lettuce Growth

Anthocyanins are secondary metabolites classified as water-soluble, non-photosynthetic pigments with the potential ability to shield chloroplasts from excess light energy. This study investigated the morphological and physiological responses of six lettuce (Lactuca sativa) cultivars with different leaf anthocyanin contents grown in a greenhouse under different supplemental photosynthetic photon flux densities (PPFD). Cultivars ‘Cherokee’, ‘Teodore’, ‘Rex’, and ‘Rouxai’ decreased in specific leaf area with increasing PPFD, respectively. We observed that growth in cultivars with higher leaf anthocyanin content (‘Cherokee’ and ‘Rouxai’) increased with increasing PPFD. Light use efficiency (LUE) is an important physiological parameter affecting biomass accumulation, and cultivars ‘Cherokee’, ‘Rex’, ‘Teodore’, and ‘Rouxai’ had the highest LUE and shoot weight. We found that red lettuce varieties, especially ‘Cherokee’ and ‘Rouxai’, showed increased shoot dry weight as light intensity increased. Interestingly, these varieties also had the highest anthocyanin levels in their leaves. This suggests that higher anthocyanin content might contribute to the increased shoot dry weight under higher light intensity, although this was mainly evident in ‘Cherokee’.