Conventional Breeding Research Articles

120 Gene editing: From the general public perspective

Abstract The public controversy around genetically engineered (GE) products, aka genetically modified organisms (GMO), has been ongoing for almost 40 yr fueled in part by well-funded misinformation campaigns disseminated by activist organizations. Polls reveal that public perception of GE animals is generally negative, with biomedical applications being more positively perceived than agricultural applications. Opposition to GE animals is often conflated with opposition to use of animals in research in general, as well as opposition to aspects of intensive animal agriculture. In general, concerns about animal biotechnology are influenced by views around the moral status of animals, the boundary between “natural” and “unnatural,” and perceived risks and benefits of GE animals to health and the environment (personal and cultural characteristics); the purpose of the application (e.g., disease resistance > fast growth), the method(s) being used, and the motivation of the research group making the genetic modification (public or private); and the species being modified (animal characteristics). There were three factors that aggravated opposition to GE products including 1) a lengthy and expensive regulatory step uniquely associated with commercializing products developed using that breeding method, 2) competing business interests (e.g., organic, non-GMO project) that were able to monetarize fear around the method to extract value (rent seeking) and selling their value-added (more expensive) product that avoided the use of GE, and 3) there is a mandatory labeling requirement to track/differentiate products produced with (or without) that breeding method to enable value-added marketing. Even so, when GE products that people wanted were allowed to reach the market (e.g., Impossible Burger with GE heme, GloFish, AquAdvantage salmon), they met with commercial acceptance. Now it has been suggested that the public will similarly reject animal products derived from gene edited (GnEd) animals, but that idea has only recently been put to the test as such food products were not previously available. A 2022 Japanese paper noted that with regard to GnEd food (gene knock-out tomatoes and fish) there was a “gap between anticipated and actual public response”. They noted that “even though there were some petitions against the use of genome-editing made by consumer groups; they did not develop into a mass mobilization, and media coverage was mostly positive.” These GnEd food products were developed by startups from Japanese universities, required no unique regulatory step before commercialization or mandatory labeling with Japanese regulatory agencies transparently sharing a summary of the information and notification on their websites, and businesses were encouraged to share information on products to respect The right of consumers to know. Successful commercialization of GnEd animal products has so far occurred in countries (e.g., Argentina, Brazil, Japan) where regulations treat simple GnEd products with no “foreign DNA” to be “non-GMO” and analogous to conventional breeding.

Exploring natural genetic variation in photosynthesis-related traits of barley in the field.

Optimizing photosynthesis is considered an important strategy for improving crop yields to ensure food security. To evaluate the potential of using photosynthesis-related parameters in crop breeding programs, we measured chlorophyll fluorescence along with growth-related and morphological traits of 23 barley inbreds across different developmental stages in field conditions. The photosynthesis-related parameters were highly variable, changing with light intensity and developmental progression of plants. Yet, the variations in photosystem II (PSII) quantum yield observed among the inbreds in the field largely reflected the variations in CO2 assimilation properties in controlled climate chamber conditions, confirming that the chlorophyll fluorescence-based technique can provide proxy parameters of photosynthesis to explore genetic variations under field conditions. Heritability (H2) of the photosynthesis-related parameters in the field ranged from 0.16 for the quantum yield of non-photochemical quenching to 0.78 for the fraction of open PSII center. Two parameters, the maximum PSII efficiency in light-adapted state (H2 0.58) and the total non-photochemical quenching (H2 0.53), showed significant positive and negative correlations, respectively, with yield-related traits (dry weight per plant and net straw weight) in the barley inbreds. These results indicate the possibility of improving crop yield through optimizing photosynthetic light use efficiency by conventional breeding programs.

Adaptive features of winter triticale accessions from the VIR collection

Background. In the context of global climate change, the study and development of genetic material with environmental plasticity and stability in the manifestation of its traits becomes relevant. The scientific and practical results achieved in triticale breeding place this crop among the most popular cereals in terms of economic importance. However, the main problem for the use of triticale in breeding practice is its limited genetic resources.Materials and methods. The research materials were 161 winter triticale (× Triticosecale Wittm. ex A. Camus) accessions from the global collection of VIR. The study and breeding-oriented assessment of the material were based on conventional breeding techniques and carried out according to the guidelines approved by VIR with minor amendments. The obtained research data underwent statistical analysis using the method by A. V. Kilchevsky and L. V. Khotyleva.Results. Environmental and genetic parameters were analyzed under the conditions of the Middle Volga Region of Russia. Genotypes were identified for their high general adaptability, i. e., securing the highest average yield in the entire set of environments. Cultivars were also selected for their least variable high yields under the impact of weather conditions. The analysis of the studied set of accessions showed that both high-yielding and low-yielding genotypes were stable. The character’s mean value and its environmental sensitivity were relatively independent and could be incorporated in one genotype in various combinations. This finding can be used in the process of triticale breeding.

Androgenesis and gynogenesis in tomato (<I>Solanum lycopersicum</I> L.) <I>in vitro</I>

Tomato (Solanum lycopersicum L.) is one of the most consumed vegetable crops worldwide. Tomato fruits are rich in vitamins, minerals, and pigments, including lycopene. The high demand and the need to enhance tomato production call for new improved cultivars and F1 hybrids.Biotechnological methods reduce the time for source material development and the labor intensity of breeding efforts. Obtaining doubled haploid plants makes it possible to fix and analyze new gene combinations faster than with conventional breeding techniques, and produce homozygous genotypes. Tomato is highly unsusceptible to haploid induction, which has been continuously studied for more than 40 years and is still of special interest. The main methods for producing haploids are based on androgenesis and gynogenesis. Androgenesis is the production of haploids from the cells of the male gametophyte, and gynogenesis from the cells of the female gametophyte.The objective was to review the research on the induction of tomato haploids based on androgenesis and gynogenesis. No standardized, efficient or reproducible protocols are currently available to produce doubled haploids of tomato. It is necessary to determine the incubation conditions, physicochemical environments, dependence of the genotype in vitro, physiological state of the donor plant, and development of the anther, which affect the reproducibility of protocols to achieve haploid induction. Anther culture for obtaining haploid tomato plants has not yielded successful results, and the studies on microspore culture were too few, so it is difficult to understand the effectiveness of this technique. The method of gynogenesis is poorly investigated, but the culture of unfertilized ovules can become a successful way to obtain tomato haploids, with more research on this subject.

Genomic Advances in Biofortification of Iron and Zinc in Wheat (Triticum aestivum L.)

Majority of population that depend on wheat are found to be deficient in iron and zinc. About 20% of calories worldwide are derived from wheat. Milling of wheat reduces the bioavaibility of Fe and Zn in wheat grains. Micronutrient deficiency is the main cause of malnutrition worldwide and hidden hunger in underdeveloped countries. To overcome this problem it becomes necessary to develop varieties that are rich in iron and zinc content. Biofortification gets around these issues by enhancing the micronutrient content of wheat. The genetic approaches of biofortification particularly the non-conventional methods have proved to be the most efficient ones. These method includes QTL mapping approach, GWAS and transgenics. Although conventional methods for a long time has been used for developing varieties of wheat rich in Fe and Zn but conventional breeding for certain traits may be inconvenient and time-consuming and are prone to errors due to their reliance on traditional phenotyping or visual assessments. Several research have been undertaken to map QTLs of Fe and Zn concentration in wheat grains and have been used in elite genetic backgrounds to improve micronutrient content. Genomic studies revealed through GWAS exhibit high QTL resolution and allele coverage, which can be used for improvement of Fe and Zn content in wheat. Genetic engineering techniques or transgenics approaches allows genetic alteration of crops to improve the nutritional quality of wheat. Genomic approach of biofortification of wheat has proved to be the most efficient methods for increasing iron and zinc content in the crop.

Multidrug resistance in Salmonella isolates of swine origin: mobile genetic elements and plasmids associated with cephalosporin resistance with potential transmission to humans.

The emergence of foodborne Salmonella strains carrying antimicrobial resistance (AMR) in mobile genetic elements (MGE) is a significant public health threat in a One Health context requiring continuous surveillance. Resistance to ciprofloxacin and cephalosporins is of particular concern. Since pigs are a relevant source of foodborne Salmonella for human beings, we studied transmissible AMR genes and MGE in a collection of 83 strains showing 9 different serovars and 15 patterns of multidrug resistant (MDR) previously isolated from pigs raised in the conventional breeding system of Northern Spain. All isolates were susceptible to ciprofloxacin and three isolates carried blaCMY-2 or blaCTX-M-9 genes responsible for cefotaxime resistance. Filter mating experiments showed that the two plasmids carrying blaCTX-M-9 were conjugative while that carrying blaCMY-2 was self-transmissible by transformation. Whole-genome sequencing and comparative analyses were performed on the isolates and plasmids. The IncC plasmid pSB109, carrying blaCMY-2, was similar to one found in S. Reading from cattle, indicating potential horizontal transfer between serovars and animal sources. The IncHI2 plasmids pSH102 in S. Heidelberg and pSTM45 in S. Typhimurium ST34, carrying blaCTX-M-9, shared similar backbones and two novel "complex class 1 integrons" containing different AMR and heavy metal genes. Our findings emphasize the importance of sequencing techniques to identify emerging AMR regions in conjugative and stable plasmids from livestock production. The presence of MGE carrying clinically relevant AMR genes raises public health concerns, requiring monitoring to mitigate the emergence of bacteria carrying AMR genes and subsequent spread through animals and food.IMPORTANCEThe emergence of foodborne Salmonella strains carrying antimicrobial resistance (AMR) in mobile genetic elements (MGE) is a significant public health threat in a One Health context. Since pigs are a relevant source of foodborne Salmonella for humans, in this study, we investigate different aspects of AMR in a collection of 83 Salmonella showing nine different serovars and 15 patterns of multidrug resistant (MDR) isolated from pigs raised in the conventional breeding system. Our findings emphasize the importance of sequencing techniques to identify emerging AMR regions in conjugative and stable plasmids from livestock production. The presence of MGE carrying clinically relevant AMR genes raises public health concerns, requiring monitoring to mitigate the emergence of bacteria carrying AMR genes and subsequent spread through animals and food.

Finger millet (Eleusine coracana) improvement: Challenges and prospects—A review

AbstractFinger millet is a climate‐resilient and highly nutritious small grain crop widely grown in the semi‐arid tropics. It has multiple uses, including for food, feed and beverage preparations. However, finger millet is an under‐utilized and under‐researched crop with a mean yield of <1.0 t/ha despite a potential productivity of up to 8 t/ha. The yield gap is attributed to several production constraints, such as biotic and abiotic stresses, a lack of access to improved seeds and production inputs and poor agronomic management practices. There are valuable genetic resources and genetic variability of finger millet in its centres of diversity and global gene banks for variety design, product development and commercialization. The genetic variability can be harnessed further to integrate essential traits into candidate varieties through conventional and modern breeding methods. Breeding and genetic innovations such as genomics‐assisted breeding, mutation breeding and genome editing would accelerate finger millet breeding and new variety design and deployment. The objective of this review was to document the opportunities, challenges and prospects of finger millet improvement as a guide for variety development and deployment with enhanced grain yield and nutritional contents. The first section describes global production status and yield gains, major production and productivity challenges in finger millet. This is followed by an in‐depth presentation on breeding and genetic progress on variety development with improved agronomic and nutritional quality traits, drought and salinity tolerance, and fungal diseases, weeds and insect pest resistance. Further, the review summarized finger millet's genetic and genomic resources, reference genomes, whole genome re‐sequencing and transcriptomics of finger millet technologies, genetic engineering and genome editing and their integration with conventional breeding methods for variety design with desired end‐use traits. The review provides foundational information to expedite the development of new‐generation finger millet cultivars with desirable product profiles, including high grain yield potential, early maturity, desirable seed colour, compact head type, food and feed nutrients quality and high marketability through modern breeding approaches.

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.