Conventional Breeding Research Articles

Applications of Molecular Marker Implementation for Enhanced Oilseed Breeding through Marker-Assisted Selection (Mas)

Oilseed crops are fundamental for various industrial and food purposes, necessitating advancements in breeding strategies to meet growing demands. Molecular markers and Marker-Assisted Selection (MAS) have emerged as powerful tools in oilseed crop improvement. This review explores the diverse applications of molecular markers and MAS in oilseed crops, highlighting their significance in enhancing breeding efficiency and genetic gains. The primary aim of this review is to provide a comprehensive overview of the application of molecular markers and MAS in oilseed crop breeding programs. Specifically, it aims to synthesize existing literature and case studies to elucidate the methodologies, advancements, and outcomes associated with the integration of molecular markers and MAS in oilseed crop improvement. The review highlights significant advancements in the application of molecular markers and MAS in oilseed crop breeding. Molecular markers have been extensively utilized for trait mapping, genetic diversity analysis, and selection of desirable genotypes with traits such as high oil content, disease resistance, and abiotic stress tolerance. Integration with conventional breeding methods has facilitated the development of elite oilseed crop varieties with improved agronomic performance and stress resilience. In conclusion, molecular markers and MAS represent valuable tools for accelerating oilseed crop breeding programs. Their integration has led to increased breeding efficiency, precision, and genetic gains. However, challenges such as marker validation, cost-effectiveness, and trait complexity remain to be addressed. Future research efforts should focus on refining marker technologies, enhancing trait mapping studies, and promoting the adoption of MAS in oilseed crop breeding to ensure sustainable production and global food security.

Cold‐pressing, ethanol defatting, and saline extraction enhances properties of protein products from new pennycress varieties (covercress™)

AbstractPennycress (Thlaspi arvense L.), a winter oilseed and cover crop, is a source of novel proteins and oil for biofuel. New yellow‐seeded pennycress specialty varieties, named covercress™, were developed via conventional breeding and gene editing, but protein characteristics are still unknown. This work evaluated two covercress lines, TT8 and Y1126, for protein extraction and functionality, and the combination of cold‐pressing, alcohol‐defatting, and saline extraction to produce a protein that could align better with clean labeling. Seeds were first cold‐pressed in a tubular radial expeller then defatted at 60°C with anhydrous ethanol (‐Eth) or hexane (‐Hex) until ≤0.8% residual oil. Protein was extracted from defatted press cake (PC) using the saline method (0.1 M NaCl, 1: 10 w/v, 50°C, 2 h) for hexane‐defatted wild‐type pennycress (WTP) seed meal and PC. TT8 PC had 33%–67% more globulin and glutelin than WTP PC, but ethanol defatting reduced albumin + globulin amounts by 14%–32%. TT8 PC protein recoveries (63% and 40%) were 2‐ to 3‐fold greater than that of Y1126 PC‐Eth and their extracts had greater purity (85%–91% vs. 70% protein). Y1126 PC‐Eth protein showed a nearly constant 68%–70% solubility from pH 2 to 10, but TT8 PC‐Eth protein was more soluble in acidic pH (87%–90% solubility). Y1126 PC‐Eth protein showed greater foaming capacity (146–150 mL), while TT8 PC‐Eth protein had superior emulsifying properties (EAI 141–236 m2/g protein and ESI 13–26 min). This work demonstrated the enhanced functionality of TT8 and Y1126 proteins produced from an environmentally friendly and industrially relevant method.

QTLs and Genes for Salt Stress Tolerance: A Journey from Seed to Seed Continued

Rice (Oryza sativa L.) is a crucial crop contributing to global food security; however, its production is susceptible to salinity, a significant abiotic stressor that negatively impacts plant germination, vigour, and yield, degrading crop production. Due to the presence of exchangeable sodium ions (Na+), the affected plants sustain two-way damage resulting in initial osmotic stress and subsequent ion toxicity in the plants, which alters the cell’s ionic homeostasis and physiological status. To adapt to salt stress, plants sense and transfer osmotic and ionic signals into their respective cells, which results in alterations of their cellular properties. No specific Na+ sensor or receptor has been identified in plants for salt stress other than the SOS pathway. Increasing productivity under salt-affected soils necessitates conventional breeding supplemented with biotechnological interventions. However, knowledge of the genetic basis of salinity stress tolerance in the breeding pool is somewhat limited because of the complicated architecture of salinity stress tolerance, which needs to be expanded to create salt-tolerant variants with better adaptability. A comprehensive study that emphasizes the QTLs, genes and governing mechanisms for salt stress tolerance is discussed in the present study for future research in crop improvement.

An Integrated and Multi-Stakeholder Approach for Sustainable Phosphorus Management in Agriculture

Conventional agriculture relies on non-renewable rock phosphate as a source of phosphorus. The demand for food has led to increased phosphorus inputs, with a negative impact on freshwater biodiversity and food security. The importation of phosphorus fertilizers makes most food systems vulnerable to phosphorus supply risks. The geopolitical instability generated by the pandemic and the current Russia–Ukraine conflict, which has led to a 400% increase in phosphorus commodity prices, offers the international community and institutions an opportunity to embrace the global phosphorus challenge and move towards a more circular system. Here, we discuss an integrated and multi-stakeholder approach to improve phosphorus management in agriculture and increase the efficiency of the whole chain, highlighting the contribution of conventional breeding and genetic engineering, with a particular focus on low-phytic-acid (lpa) crops, whose grains may help in reducing phosphorus-management-related problems. In recent decades, the choice of short-term strategies—such as the use of phytase as a feed additive—rather than lpa mutants, has been carried out without considering the long-term money saving to be derived from lpa crops. Overall, lpa crops have the potential to increase the nutritional quality of foods and feeds, but more research is needed to optimize their performance.

Marker-Assisted Breeding Techniques for the Development of Gluten-Free Wheat Varieties: A Comprehensive Review

Ingestion of gluten proteins from wheat, barley and rye poses a significant health risk. The sole treatment for various disease is a lifelong adherence to a strict gluten-free diet. Conventional breeding methods have limitations in producing wheat varieties that maintain baking quality due to the complexity of the wheat genome and the multitude of gluten genes. This review explores the use of RNA interference (RNAi) silencing and CRISPR/Cas9 gene editing techniques. Efficient screening and selection processes for identifying lines with reduced celiac disease epitopes at both the DNA and protein levels, as well as maintaining baking quality, are discussed. The integration of gene editing for the production of wheat products holds significant potential in meeting the growing demand for safer dietary options while ensuring compliance with regulatory standards and addressing the complex challenges associated with disease management. Marker-assisted selection (MAS) offers several advantages over conventional selection methods in plant breeding, including timesaving, cost-effectiveness and goal-oriented outcomes. This review aims to delineate various molecular markers, encompassing sequence-tagged sites (STS), simple sequence repeats (SSR), genotyping by sequencing (GBS), single nucleotide polymorphism (SNP) arrays, exome capture, Competitive Allele Specific PCR (KASP), cleaved amplified polymorphic sequences (CAPS), semi-thermal asymmetric reverse PCR (STARP), and genotyping by target sequencing (GBTS). Additionally, we compile quantitative trait loci (QTL)/genes and their associated markers, which hold potential utility in MAS applications. The rapid advancement of wheat genomics is poised to expedite marker development, QTL mapping, gene cloning, and wheat breeding processes.

A Sustainable Approach to Combat Micronutrient Deficiencies and Ensure Global Food Security through Biofortification

Micronutrient deficiencies, particularly in essential vitamins and minerals, pose a significant public health challenge, affecting over two billion people worldwide. These deficiencies contribute to various health issues, impaired cognitive development, and reduced productivity, ultimately hindering social and economic progress. Biofortification, a process of enhancing the nutritional content of staple crops through conventional breeding or genetic engineering, has emerged as a promising and sustainable approach to combat micronutrient deficiencies and ensure global food security. This review explores the potential of Biofortification as a cost-effective and sustainable solution to address hidden hunger and improve the nutritional status of vulnerable populations. Biofortification offers several advantages over traditional interventions, such as supplementation and food fortification. By targeting staple crops consumed by the majority of the population, Biofortification ensures a wide reach and sustained nutrient intake without requiring significant changes in dietary habits. Moreover, biofortified crops can be grown locally, reducing the reliance on external interventions and empowering farmers to improve their nutritional status and livelihoods. Numerous studies have demonstrated the efficacy of bio fortified crops in increasing micronutrient intake and improving health outcomes. For instance, iron-biofortified pearl millet has been shown to increase iron absorption and reduce anemia prevalence in children, while zinc-biofortified wheat has improved zinc status and reduced stunting. Additionally, vitamin A-biofortified sweet potato and cassava have significantly increased vitamin A intake and reduced vitamin A deficiency in various populations.
 Despite the promising results, the success of Biofortification relies on several factors, including the development of nutrient-dense varieties, consumer acceptance, and effective dissemination strategies. Collaboration among researchers, policymakers, and stakeholders is essential to scale up Biofortification efforts and ensure their long-term sustainability. By prioritizing Biofortification as a key strategy in combating micronutrient deficiencies, we can work towards a more nourished and food-secure world.

Establishment of an Efficient Sugarcane Transformation System via Herbicide-Resistant CP4-EPSPS Gene Selection

Sugarcane (Saccharum spp.), a major cash crop that is an important source of sugar and bioethanol, is strongly influenced by the impacts of biotic and abiotic stresses. The intricate polyploid and aneuploid genome of sugarcane has shown various limits for conventional breeding strategies. Nonetheless, biotechnological engineering currently offers the best chance of introducing commercially significant agronomic features. In this study, an efficient Agrobacterium-mediated transformation system that uses the herbicide-resistant CP4-EPSPS gene as a selection marker was developed. Notably, all of the plants that were identified by PCR as transformants showed significant herbicide resistance. Additionally, this transformation protocol also highlighted: (i) the high yield of transgenic lines from calli (each gram of calli generated six transgenic lines); (ii) improved selection; and (iii) a higher transformation efficiency. This protocol provides a reliable tool for a routine procedure for the generation of resilient sugarcane plants.

Direct Organogenesis of Citrus Cultivars from Shoot Tip Nodal Segments.

Citrus cultivar improvement via conventional breeding strategies is impeded by factors related to its reproductive biology. The orange is a hybrid between pomelo (Citrus maxima) and mandarin (Citrus reticulata). Among various orange cultivars, Valencia oranges have a bit of bitter tang mixed in with their sweetness, as Navel oranges are, the most widely cultivated citrus species, quite sweeter, and also don't contain any seeds. Tangelo mandarin orange cultivar is a hybrid of C. reticulata × C. maxima or × C. paradisi. The present study was undertaken to optimize the hormonal composition of the media with regard to plant growth regulators for in vitro propagation of sweet orange cultivars from nodal segment explants. The nodal segment explants were collected from three citrus cultivars, Washington Navel, Valencia and Tangelo. Murashige and Skoog (MS) medium supplemented with sucrose and different concentrations of growth regulators was used for shoot proliferation and root induction, and the optimum medium composition was assessed. The patent for Citrus Tissue Culture was obtained from the Office of Research Affairs, Haramaya University. The results indicate that the highest shoot response was recorded for Washington's navel with maximum shoot proliferation rate (99.75%), shoot number per explant (1.76), shoot length (10.70 cm), leaf number per explants (3.54) after three weeks of culture. In all experiments, no growth was observed for the basal MS medium. Phytohormone combinations of IAA (1.2 mg/L) and kinetin (2.0 mg/L) were found to be the best for shoot proliferation. Among the cultivars, there were significant differences for the highest rooting rate (81.255), root number (2.22), and root length (2.95 cm) variables for Washington Navel. The lowest rooting rate (48.45%), root number (1.47) and root length (2.26 cm) were observed for Valencia. The highest rooting rate (84.90%), root number per microshoot (2.22) and root length (3.05 cm) was on MS medium supplemented with 1.5 mg/L NAA. A comparison of different concentrations of IAA and NAA on root induction of microshoots from nodal segments of citrus cultivars demonstrated NAA was a more effective hormone than IAA.

Chromosomal characterization mediated by karyomorphological analysis and differential banding pattern in fenugreek (Trigonella foenum-graecum L.): a neglected legume

Fenugreek or Trigonella foenum-graecum L. is a commercially important yet neglected crop of the family Fabaceae, with potent medicinal applications, and can treat several diseases as well. Conventional breeding studies for higher yields of commercial crops largely depend on chromosomal information of the particular species. Despite a number of cytological research being conducted on T. foenum-graecum, a complete characterization of its chromosomes has not been achieved due to the limitations of traditional karyotype analysis methods. A range of chromosomal markers are advantageous to characterize at full extent and identify individual chromosomes rather than relying on only physical metrics. Thus, in this study, in addition to giemsa staining, other approaches like fluorochrome and silver staining were used for the precise karyomorphological analysis of this species. Enzyme maceration and air drying (EMA) based fluorochrome banding with GC-specific stain Chromomycin A3 (CMA), and AT-specific stain 4’,6-diamidino-2-phenylindole (DAPI) applied for the first time for chromosome characterization. The results showed 2n = 16 chromosomes in metaphase cells, with karyotype formula of 2m+6sm. The unique banding pattern observed in the CMA/DAPI and AgNOR staining highlights the AT and GC-rich regions as well as the nucleolar organizer regions (NORs). All this crucial information can further assist in conducting breeding studies of more precision with simultaneously encouraging similar studies that need to be done in other unexploited species of importance.

Optimization and extraction of DNA from medicinally important plant species Phyllanthus emblica

Indian gooseberry has two scientific names Phyllanthus emblica and Emblica officinalis. Gooseberry has cookery and herbal medicine uses. It grows abundantly in the forests of India and many South Asian countries. Indian gooseberry supplements are used in anti-aging, cancer, heart health, blood sugar levels, liver damage and immune health. Fruit is reputed to probably have the highest content of vitamin C compared with any other naturally occurring substances in nature. Active extracts of P. emblica have been shown to possess several pharmacological properties, e.g., analgesic, anti-inflammatory, antioxidant and chemo protective activities P. emblica being a perennial crop, molecular tools can be of great utility to support the conventional breeding. Therefore, isolation of intact high-quality DNA is essential for carrying out molecular studies. DNA extraction is the isolation of DNA from a sample. The reason includes its role in understanding the plant's genetic makeup, identifying specific genes and developing improved varieties with desirable traits. They contribute to molecular biology research and to the broader understanding of plant genetics and evolution. Different methods include Lysis, Precipitation, Purification and Concentration. DNA is extracted from Indian gooseberry for various purposes such as genetic studies, plant breeding and research. The reagents used in this process are Tris EDTA buffer, cetyltrimethylammonium bromide buffer and some chemicals.

Engineering Brassica Crops to Optimize Delivery of Bioactive Products Postcooking.

Glucosinolates are plant-specialized metabolites that can be hydrolyzed by glycosyl hydrolases, called myrosinases, creating a variety of hydrolysis products that benefit human health. While cruciferous vegetables are a rich source of glucosinolates, they are often cooked before consumption, limiting the conversion of glucosinolates to hydrolysis products due to the denaturation of myrosinases. Here we screen a panel of glycosyl hydrolases for high thermostability and engineer the Brassica crop, broccoli (Brassica oleracea L.), for the improved conversion of glucosinolates to chemopreventive hydrolysis products. Our transgenic broccoli lines enabled glucosinolate hydrolysis to occur at higher cooking temperatures, 20 °C higher than in wild-type broccoli. The process of cooking fundamentally transforms the bioavailability of many health-relevant bioactive compounds in our diet. Our findings demonstrate the promise of leveraging genetic engineering to tailor crops with novel traits that cannot be achieved through conventional breeding and improve the nutritional properties of the plants we consume.

Evaluation of breeding cost in the French maritime pine breeding program and perspectives for alternative strategies using molecular markers

The economic efficiency of conventional breeding strategies for forest trees based on biparental crosses is compared with that of alternative strategies based on pedigree reconstruction using molecular markers. Analyses of economic efficiency is based on comparisons of breeding scenarios corresponding to the same total investment. The first step is the description and cost evaluation of each basic operation, from crossing to genetic selection and clonal archive establishment. Breeding scenarios are then compared by stochastic sampling with a parametric genetic model (POPSIM), the comparison criteria in this case being genetic gain in the seed orchard for a given level of genetic diversity. Additionally, the economic gain resulting from the use of improved material is estimated for different levels of breeding investment. Our analysis shows that genotyping costs account for a much smaller proportion of total investment than phenotyping costs. We also show that, in comparisons of breeding scenarios corresponding to the same total investment, the three main breeding strategies (biparental crosses, polymix crosses, and open pollination) achieve similar genetic gains provided that sufficiently large numbers of parents are considered. These results open up promising perspectives for the wider integration of molecular markers into forest tree breeding strategies.

Grain and shoot iron (Fe) content and root phytosiderophore release are the major determinants of Fe-deficiency tolerance index (FeDTI) and the reliable screening markers to breed Fe-efficient rice (Oryza sativa L.)

There is a need to exploit the genetic variability in rice to improve the Fe deficiency tolerance, particularly under aerobic conditions, and to improve the Fe uptake and accumulation in the grain to facilitate biofortification. The present research was conducted to ascertain the regulators and determinants of Fe-deficiency tolerance in rice. Fifty-seven diverse genotypes were investigated under Fe-deficient (1 µM, FDS, -Fe) and Fe-sufficient (100 µM, FSS, +Fe) nutrient solution cultures for growth and physiological attributes, Fe uptake and accumulation and root release of phytosiderophores (PS), etc., and Fe-deficiency tolerance index (FeDTI). A strong positive correlation was observed between root traits and the PS release. Further, a higher release of PS was evidenced as a major determinant of shoot Fe and grain Fe in rice particularly under Fe deficiency. Based on FeDTI, a total of ten rice genotypes were identified, and their Fe deficiency response was further validated on Fe deficient (∼2.1 ppm Fe, FeDS) and Fe-sufficient soils (∼10 ppm Fe, FeSS) in pot culture. Maintenance of high active Fe content and consequently a higher leaf chlorophyll content throughout the plant growth were extremely important determinants of grain yield and grain Fe fortification in rice under Fe deficiency than under the Fe sufficient condition. Based on the FeDTI, this study further identifies the Fe-deficiency tolerant rice genotypes which can be exploited through conventional and molecular breeding approaches to improve the Fe uptake efficiency and grain Fe content of the high-yielding rice genotypes.

THE CONTRIBUTION OF TRANSGENIC RICE TO ENHANCE GRAIN YIELD

Breeders' main goal is to increase the proportion of high-quality rice produced overall. To create effective rice breeding strategies, possible yield-related loci have been mined. many researchers are using transgenic strategies as cutting-edge methods to increase rice productivity. Quantitative trait loci (QTLs) play a pivotal role in governing grain yield in Oryza sativa L., commonly known as rice. The genes contributing to QTLs that determine grain size, length, and weight have been successfully identified. Numerous genes are upregulated to enhance the overall yield of rice. Recent advancements have led to the discovery of genes and QTLs specifically associated with rice yield. Through an in-depth analysis of various yield characteristics, including grain weight, thousand grain weight, grain length, grain width, grain yield per plant, grain number per panicle, and panicles per plant, we conducted a comprehensive review using extensive literature research and public domain databases. Additionally, we explored the progress made in transgenic technology and advanced genomic techniques. The compiled information on genes and QTLs related to yield enhancement aims to provide a valuable resource. The integrated analysis of existing data on genes and/or QTLs provide evidence on potential combinations for creating superior genotypes that combine high yield across multiple traits. Integration of molecular markers, transgenic techniques and conventional breeding as discussed in this extensive review opens up the prospect of developing high yielding rice varieties

Optimizing speed breeding and seed/pod chip based genotyping techniques in pigeonpea: A way forward for high throughput line development

BackgroundThe challenge of pigeonpea breeding lies in its photosensitivity and seasonal specificity. This poses a problem to the breeder, as it restricts to single generation advancement in a year. Currently, the cross to cultivar gap is twelve to thirteen years resulting in a limited number of varietal releases over the past six decades. Shortening the breeding cycle was need of the hour, unlikely achieved by conventional breeding. To overcome these hindrances speed breeding was a necessary leap. An experiment was planned to optimize the speed breeding coupled with single seed descent and seed or pod chip-based genotyping to shorten the breeding cycle in pigeonpea at ICRISAT, Hyderabad. Monitored photoperiod, light wavelength, temperature and crop management regime were the indicators attributing to the success of speed breeding.ResultA photoperiod of 13 h: 8 h: 13 h at vegetative: flowering and pod filling stages is ideal for shortening the breeding cycle. Broad spectrum light (5700 K LED) hastened early vegetative growth and pod formation. Whereas far-red (735 nm) light favoured early flowering. A significant difference between the photoperiods, genotypes as well as photoperiod x genotype interaction for both days to flowering and plant height was noted.ConclusionThe optimized protocol serves as a road map for rapid generation advancement in pigeonpea. Deploying this protocol, it is possible to advance 2–4 generations per year. The breeding cycle can be reduced to 2–4 years which otherwise takes 7 years under conventional breeding. Single Seed Descent and seed or pod chip-based genotyping for early generation marker assisted selection, strengthened the precision of this technique aiding in high throughput line development.

Recent Advancements in Mitigating Abiotic Stresses in Crops

In recent years, the progressive escalation of climate change scenarios has emerged as a significant global concern. The threat to global food security posed by abiotic stresses such as drought, salinity, waterlogging, temperature stress (heat stress, freezing, and chilling), and high heavy metal accumulation is substantial. The implementation of any of these stresses on agricultural land induces modifications in the morphological, biochemical, and physiological processes of plants, leading to diminished rates of germination, growth, photosynthesis, respiration, hormone and enzyme activity disruption, heightened oxidative stress, and ultimately, a reduction in crop productivity. It is anticipated that the frequency of these stresses will progressively escalate in the future as a result of a rise in climate change events. Therefore, it is crucial to develop productive strategies to mitigate the adverse effects of these challenges on the agriculture industry and improve crop resilience and yield. Diverse strategies have been implemented, including the development of cultivars that are resistant to climate change through the application of both conventional and modern breeding techniques. An additional application of the prospective and emerging technology of speed breeding is the acceleration of tolerance cultivar development. Additionally, plant growth regulators, osmoprotectants, nutrient and water management, planting time, seed priming, microbial seed treatment, and arbuscular mycorrhiza are regarded as effective methods for mitigating abiotic stresses. The application of biochar, kaolin, chitosan, superabsorbent, yeast extract, and seaweed extract are examples of promising and environmentally benign agronomic techniques that have been shown to mitigate the effects of abiotic stresses on crops; however, their exact mechanisms are still not yet fully understood. Hence, collaboration among researchers should be intensified to fully elucidate the mechanisms involved in the action of the emerging technologies. This review provides a comprehensive and current compilation of scientific information on emerging and current trends, along with innovative strategies to enhance agricultural productivity under abiotic stress conditions.

Enhancing Nutrition, Crop Resilience, and Food Security through Biofortification

Biofortification is a process of enhancing the nutritional quality of food crops through conventional plant breeding, genetic engineering, or agronomic practices. It has emerged as an important agricultural strategy to improve public health by increasing the micronutrient density in staple crops and vegetables. Biofortification provides a cost-effective and sustainable approach to combat micronutrient deficiencies, also known as hidden hunger, which affects over 2 billion people worldwide. This review provides an overview of biofortification efforts targeting major micronutrients such as iron, zinc, vitamin A, and folate. The genetic and molecular mechanisms underlying elevated micronutrient accumulation are discussed. The review also summarizes the impacts of biofortification in enhancing micronutrient intake, nutritional status, and health outcomes based on results from efficacy and effectiveness studies. The role of biofortification in building climate resilience and food security is also examined. Overall, biofortification has shown considerable promise in tackling malnutrition sustainably in developing countries. However, continued research and policy support are needed to maximize its impact on nutrition security worldwide.

Development of early maturing salt-tolerant rice variety KKL(R) 3 using a combination of conventional and molecular breeding approaches.

Introduction: Soil salinity poses a severe threat to rice production, resulting in stunted growth, leaf damage, and substantial yield losses. This study focuses on developing an early maturing seedling stage salinity tolerant rice variety by integrating conventional breeding methods with marker assisted breeding (MAB) approaches. Methods: Seedling-stage salinity tolerance Quantitative Trait Locus (QTL) "Saltol" from the salt-tolerant parent FL478 was introduced into the high-yielding but salt-sensitive rice variety ADT 45. This was achieved through a combination of conventional breeding and MAB. The breeding process involved rigorous selection, screening, and physiological parameter assessments. Results: KKL(R) 3 (KR 15066) identified as the top performing Recombinant Inbred Line (RIL), consistently demonstrating maximum mean grain yields under both salinity (3435.6kg/ha) and normal (6421.8kg/ha) conditions. In comparison to the early maturing, salt-tolerant national check variety CSR 10, KKL(R) 3 exhibited a substantial yield increase over 50%. Discussion: The notable improvement observed in KKL(R) 3 positions it as a promising variety for release, offering a reliable solution to maximize yields, ensure food security, and promote agricultural sustainability in both saline and non-saline environments. The study highlights the effectiveness of MAB in developing salt-tolerant rice varieties and emphasizes the significance of the Saltol QTL in enhancing seedling stage salinity tolerance. The potential release of KKL(R) 3 has the capacity to revolutionize rice production in salt affected regions, providing farmers with a reliable solution to maximize yields and contribute to food security while ensuring agricultural sustainability.

CRISPR/Cas9-based genome editing and functional analysis of SlHyPRP1 and SlDEA1 genes of Solanum lycopersicum L. in imparting genetic tolerance to multiple stress factors.

CRISPR/Cas is a breakthrough genome editing system because of its precision, target specificity, and efficiency. As a speed breeding system, it is more robust than the conventional breeding and biotechnological approaches for qualitative and quantitative trait improvement. Tomato (Solanum lycopersicum L.) is an economically important crop, but its yield and productivity have been severely impacted due to different abiotic and biotic stresses. The recently identified SlHyPRP1 and SlDEA1 are two potential negative regulatory genes in response to different abiotic (drought and salinity) and biotic stress (bacterial leaf spot and bacterial wilt) conditions in S. lycopersicum L. The present study aimed to evaluate the drought, salinity, bacterial leaf spot, and bacterial wilt tolerance response in S. lycopersicum L. crop through CRISPR/Cas9 genome editing of SlHyPRP1 and SlDEA1 and their functional analysis. The transient single- and dual-gene SlHyPRP1 and SlDEA1 CRISPR-edited plants were phenotypically better responsive to multiple stress factors taken under the study. The CRISPR-edited SlHyPRP1 and SlDEA1 plants showed a higher level of chlorophyll and proline content compared to wild-type (WT) plants under abiotic stress conditions. Reactive oxygen species accumulation and the cell death count per total area of leaves and roots under biotic stress were less in CRISPR-edited SlHyPRP1 and SlDEA1 plants compared to WT plants. The study reveals that the combined loss-of-function of SlHyPRP1 along with SlDEA1 is essential for imparting significant multi-stress tolerance (drought, salinity, bacterial leaf spot, and bacterial wilt) in S. lycopersicum L. The main feature of the study is the detailed genetic characterization of SlDEA1, a poorly studied 8CM family gene in multi-stress tolerance, through the CRISPR/Cas9 gene editing system. The study revealed the key negative regulatory role of SlDEA1 that function together as an anchor gene with SlHyPRP1 in imparting multi-stress tolerance in S. lycopersicum L. It was interesting that the present study also showed that transient CRISPR/Cas9 editing events of SlHyPRP1 and SlDEA1 genes were successfully replicated in stably generated parent-genome-edited line (GEd0) and genome-edited first-generation lines (GEd1) of S. lycopersicum L. With these upshots, the study's key findings demonstrate outstanding value in developing sustainable multi-stress tolerance in S. lycopersicum L. and other crops to cope with climate change.

Breeding of ornamental orchids with focus on Phalaenopsis: current approaches, tools, and challenges for this century.

Ornamental orchid breeding programs have been conducted to develop commercially valuable cultivars with improved characteristics of commercial interest, such as size, flower color, pattern, shape, and resistance to pathogens. Conventional breeding, including sexual hybridization followed by selection of desirable characteristics in plants, has so far been the main method for ornamental breeding, but other techniques, including mutation induction by polyploidization and gamma irradiation, and biotechnological techniques, such as genetic transformation, have also been studied and used in ornamental breeding programs. Orchids are one of the most commercially important families in floriculture industry, having very particular reproductive biology characteristics and being a well-studied group of ornamentals in terms of genetic improvement. The present review focuses on the conventional and biotechnological techniques and approaches specially employed in breeding Phalaenopsis orchids, the genus with highest worldwide importance as an ornamental orchid, highlighting the main limitations and strengths of the approaches. Furthermore, new opportunities and future prospects for ornamental breeding in the CRISPR/Cas9 genome editing era are also discussed. We conclude that conventional hybridization remains the most used method to obtain new cultivars in orchids. However, the emergence of the first biotechnology-derived cultivars, as well as the new biotechnological tools available, such as CRISPR-Cas9, rekindled the full potential of biotechnology approaches and their importance for improve ornamental orchid breeding programs.