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Background The mechanisms underlying rice root responses to drought during the early developmental stages are yet unknown. Results This study aimed to determine metabolic differences in IR64, a shallow-rooting, drought-susceptible genotype, and Azucena, a drought-tolerant and deep-rooting genotype under drought stress. The morphological evaluation revealed that Azucena might evade water stress by increasing the lateral root system growth, the root surface area, and length to access water. At the same time, IR64 may rely mainly on cell wall thickening to tolerate stress. Furthermore, significant differences were observed in 49 metabolites in IR64 and 80 metabolites in Azucena, for which most metabolites were implicated in secondary metabolism, amino acid metabolism, nucleotide acid metabolism and sugar and sugar alcohol metabolism. Among these metabolites, a significant positive correlation was found between allantoin, galactaric acid, gluconic acid, glucose, and drought tolerance. These metabolites may serve as markers of drought tolerance in genotype screening programs. Based on corresponding biological pathways analysis of the differentially abundant metabolites (DAMs), biosynthesis of alkaloid-derivatives of the shikimate pathway, fatty acid biosynthesis, purine metabolism, TCA cycle and amino acid biosynthesis were the most statistically enriched biological pathway in Azucena in drought response. However, in IR64, the differentially abundant metabolites of starch and sucrose metabolism were the most statistically enriched biological pathways. Conclusion Metabolic marker candidates for drought tolerance were identified in both genotypes. Thus, these markers that were experimentally determined in distinct metabolic pathways can be used for the development or selection of drought-tolerant rice genotypes.

Pomegranate has been considered a medicinal plant due to its rich nutrients and bioactive compounds. Since environmental conditions affect the amount and composition of metabolites, selecting suitable locations for cultivation would be vital to achieve optimal production. In this study, data on the diversity of targeted metabolites and morphological traits of 152 Iranian pomegranate genotypes were collected and combined in order to establish the first core collection. The multivariate analyses were conducted including principal component analysis (PCA), and cluster analysis. In addition, the current and future geographical distribution of pomegranate in Iran was predicted to identify suitable locations using the MaxEnt model. The results showed high diversity in the studied morphological and metabolic traits. The PCA results indicated that FFS, NFT, JA, and AA are the most important traits in discriminating the studied genotypes. A constructed core collection using maximization strategy consisted of 20 genotypes and accounted for 13.16% of the entire collection. Shannon-Weaver diversity index of a core collection was similar or greater than the entire collection. Evaluation of the core collection using four parameters of MD, VD, CR, and VR also indicated the maintenance of the genetic diversity of the original set. According to the MaxEnt model, altitude, average temperature of coldest quarter, and isothertmality were the key factors for the distribution of pomegranate. The most suitable areas for pomegranate cultivation were also determined which were located in arid and semi-arid regions of Iran. The geographic distribution of pomegranate in the future showed that the main provinces of pomegranate cultivation would be less affected by climatic conditions by the middle of the century. The results of this study provide valuable information for selection of elite genotypes to develop the breeding programs to obtain the cultivars with the highest levels of metabolic compounds for pharmaceutical purposes, as well as identification of the most suitable agro-ecological zones for orchard establishment.

Trigonella foenum-graecum is widely distributed worldwide and grown under a wide range of climatic conditions. The current research was conducted to study the effects of the environmental variables on yield related traits and metabolite contents of 50 different Persian fenugreeks at various geographical locations. Accordingly, multivariate statistical techniques including canonical correspondence analysis (CCA), hierarchical clustering on principal components, and partial least squares regression (PLSR) were applied to determine important proxy variables and establish a relevant model to predict bioactive compounds in fenugreeks. The interrelation of clustered groups emphasized the importance of functional groups of bioactive compounds and several yield related traits. The CCA indicated that two climatic variables of temperature and solar irradiation contributed prominently to 4-hydroxyisoleucine accumulation. The predicted model based on PLSR revealed climatic variables such as temperature, solar, and rain. The precursor of isoleucine was the predictive power for 4-hydroxyisoleucine accumulation while seed weight predicted trigonelline content. The current study's findings may provide helpful information for the breeding strategies of this multipurpose crop.

Background The encapsulation of metagenome-derived multi-enzymes presents a novel approach to improving poultry feed by enhancing nutrient availability and reducing anti-nutritional factors. By integrating and encapsulated enzymes such as carbohydrate-hydrolyzing enzymes, protease, lipase, and laccase into feed formulations, this method not only improves feed digestibility but also potentially contributes to animal health and productivity through antimicrobial properties. Results This study investigates the encapsulation of metagenome-derived enzymes, including carbohydrate-hydrolyzing enzymes, protease, lipase, and laccase, using Arabic and Guar gums as encapsulating agents. The encapsulated multi-enzymes exhibited significant antimicrobial activity, achieving a 92.54% inhibition rate against Escherichia coli at a concentration of 6 U/mL. Fluorescence tracking with FITC-labeled enzymes confirmed efficient encapsulation and distribution, while physical characterization, including moisture content and solubility assessments, along with Atomic Force Microscopy (AFM) imaging, validated successful encapsulation. The encapsulated enzymes also effectively hydrolyzed poultry feed, leading to an increase in phenolic content and antioxidant activity, as confirmed by 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays. Conclusions The encapsulated multi-enzymes improved the overall feed quality by increasing reducing sugars and enhancing physical properties such as solubility and water-holding capacity. The encapsulated multi-enzymes improved the overall feed quality by increasing reducing sugars, antioxidant activity and enhancing physical properties such as solubility and water-holding capacity. Scanning Electron Microscopy (SEM) and Fourier-Transform Infrared Spectroscopy (FTIR) analyses confirmed the enzymatic breakdown of the feed structure. These results suggest that supplementing poultry feed with encapsulated multi-enzymes can enhance its physical, nutritional, and functional properties, leading to improved digestibility and overall feed quality.

This study focuses on the genetic diversity and population structure of Prunus dulcis (almond tree), a crucial agricultural component with widespread cultivation and commercial importance, particularly in Iran, a region with a longstanding tradition of almond cultivation. The diverse almond collection in Iran encompasses many local varieties, breeding selections, rootstocks, and international cultivars. This diversity necessitates advanced genotyping techniques to gain insights into genetic diversity, population structure, and linkage disequilibrium (LD). In this paper, genotyping-by-sequencing (GBS) was employed to analyze 62 almond germplasm samples, identifying approximately 63,537 high-quality single nucleotide polymorphisms (SNPs) distributed across the eight chromosomes of the almond genome. On average, there were 30,225 SNPs per chromosome. The analysis yielded an average polymorphism information content (PIC) of 0.315 and an expected heterozygosity (He) rate of 0.28, indicating a significant level of genetic diversity within the studied almond germplasm. The LD analysis demonstrated a rapid decline, with an average LD decay spanning approximately 300 kb for an r 2 value of 0.2. This suggests substantial hybridization among the sampled almond varieties. Principal Component Analysis (PCA) and structure analysis could not differentiate genotypes based on geographical origin, providing further evidence of genetic mixing among the studied almond populations. An Analysis of Molecular Variance (AMOVA) highlighted significant genetic diversity within populations but revealed minimal differences. This comprehensive study of Iran’s almond genotypes offers valuable insights for future breeding and conservation efforts, emphasizing this agriculturally significant species abundant genetic diversity and intricate population structure.

Viral diseases affect quince plant productivity and fruit quality. This study evaluated the effectiveness of droplet-vitrification cryotherapy and thermotherapy methods in the eradication of apple chlorotic leaf spot virus (ACLSV) and apple stem grooving virus (ASGV) from virus-infected in vitro cultures of quince rootstock ‘QA’ and cultivar ‘Neyshabour’. In vitro cultures infected with ACLSV and ASGV were thermo-treated at 38 °C for 0, 7, 10, and 15 days followed by apical shoot tip isolation. In the cryotherapy procedure, shoot tips were incubated on preculture medium and then exposed to plant vitrification solution 2 for 30 min at 0 °C prior to liquid nitrogen (LN) exposure. Shoot tips were warmed in unloading solution and placed on recovered medium. The frequency of virus eradication was determined using RT-PCR in plantlets recovered from thermo-and cryo-treatments and controls that were grown under in vitro conditions for 4 months. Droplet-vitrification cryotherapy and thermotherapy resulted in high frequencies of ACLSV and ASGV eradication from in vitro cultures of quince. The results showed that increasing the duration of thermotherapy significantly increased virus eradication. In vitro shoots of quince rootstock ‘QA’ and cultivar ‘Neyshabour’ heat-treated for 10 days were 64% and 67% free of ACLSV and 55% and 33% free of ASGV, respectively. All quince rootstock ‘QA’ plantlets regenerated from droplet-vitrification cryotherapy and RT-PCR assessed were free of ACLSV and 67% were free of ASGV. Cryotherapy and thermotherapy may be considered as promising methods for virus eradication programs in quince genotypes facilitating the production of healthy stock plants.

Iran is a center of origin and diversity for walnuts ( Juglans regia L.) with very good potential for breeding purposes. The rich germplasm available, creates an opportunity for study and selection of the diverse walnut genotypes. In this study, the population structure of 104 Persian walnut accessions was assessed using AFLP markers in combination with phenotypic variability of 17 and 18 qualitative and quantitative traits respetively. The primers E-TG/M-CAG, with high values of number of polymorphic bands, polymorphic information content, marker index and Shannon’s diversity index, were the most effective in detecting genetic variation within the walnut germplasm. Multivariate analysis of variance indicated 93.98% of the genetic variability was between individuals, while 6.32% of variation was among populations. A relatively new technique, an advanced maximization strategy with a heuristic approach, was deployed to develop the core collection. Initially, three independent core collections (CC1–CC3) were created using phenotypic data and molecular markers. The three core collections (CC1–CC3) were then merged to generate a composite core collection (CC4). The mean difference percentage, variance difference percentage, variable rate of coefficient of variance percentage, coincidence rate of range percentage, Shannon’s diversity index, and Nei’s gene diversity were employed for comparative analysis. The CC4 with 46 accessions represented the complete range of phenotypic and genetic variability. This study is the first report describing development of a core collection in walnut using molecular marker data in combination with phenotypic values. The construction of core collection could facilitate the work for identification of genetic determinants of trait variability and aid effective utilization of diversity caused by outcrossing, in walnut breeding programs.

Background The cuticular wax serves as a primary barrier that protects plants from environmental stresses. The Eceriferum ( CER ) gene family is associated with wax production and stress resistance. Results In a genome-wide identification study, a total of 52 members of the CER family were discovered in four Gossypium species: G. arboreum , G. barbadense , G. raimondii , and G. hirsutum . There were variations in the physicochemical characteristics of the Gossypium CER ( GCER ) proteins. Evolutionary analysis classified the identified GCERs into five groups, with purifying selection emerging as the primary evolutionary force. Gene structure analysis revealed that the number of conserved motifs ranged from 1 to 15, and the number of exons varied from 3 to 13. Closely related GCERs exhibited similar conserved motifs and gene structures. Analyses of chromosomal positions, selection pressure, and collinearity revealed numerous fragment duplications in the GCER genes. Additionally, nine putative ghr-miRNAs targeting seven G. hirsutum CER (GhCER) genes were identified. Among them, three miRNAs, including ghr-miR394 , ghr-miR414d , and ghr-miR414f , targeted GhCER09A , representing the most targeted gene. The prediction of transcription factors (TFs) and the visualization of the regulatory TF network revealed interactions with GhCER genes involving ERF, MYB, Dof, bHLH, and bZIP. Analysis of cis -regulatory elements suggests potential associations between the CER gene family of cotton and responses to abiotic stress, light, and other biological processes. Enrichment analysis demonstrated a robust correlation between GhCER genes and pathways associated with cutin biosynthesis, fatty acid biosynthesis, wax production, and stress response. Localization analysis showed that most GCER proteins are localized in the plasma membrane. Transcriptome and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) expression assessments demonstrated that several GhCER genes, including GhCER15D , GhCER04A , GhCER06A , and GhCER12D , exhibited elevated expression levels in response to water deficiency stress compared to control conditions. The functional identification through virus-induced gene silencing ( VIGS ) highlighted the pivotal role of the GhCER04A gene in enhancing drought resistance by promoting increased tissue water retention. Conclusions This investigation not only provides valuable evidence but also offers novel insights that contribute to a deeper understanding of the roles of GhCER genes in cotton, their role in adaptation to drought and other abiotic stress and their potential applications for cotton improvement.

Potato is considered a key component of the global food system and plays a vital role in strengthening world food security. A major constraint to potato production worldwide is the Potato Virus Y (PVY), belonging to the genus Potyvirus in the family of Potyviridae . Selective breeding of potato with resistance to PVY pathogens remains the best method to limit the impact of viral infections. Understanding the genetic diversity and population structure of potato germplasm is important for breeders to improve new cultivars for the sustainable use of genetic materials in potato breeding to PVY pathogens. While, genetic diversity improvement in modern potato breeding is facing increasingly narrow genetic basis and the decline of the genetic diversity. In this research, we performed genotyping-by-sequencing (GBS)-based diversity analysis on 10 commercial potato cultivars and weighted gene co-expression network analysis (WGCNA) to identify candidate genes related to PVY-resistance. WGCNA is a system biology technique that uses the WGCNA R software package to describe the correlation patterns between genes in multiple samples. In terms of consumption, these cultivars are a high rate among Iranian people. Using population structure analysis, the 10 cultivars were clustered into three groups based on the 118343 single nucleotide polymorphisms (SNPs) generated by GBS. Read depth ranged between 5 and 18. The average data size and Q30 of the reads were 145.98 Mb and 93.63%, respectively. Based on the WGCNA and gene expression analysis, the StDUF538 , StGTF3C5 , and StTMEM161A genes were associated with PVY resistance in the potato genome. Further, these three hub genes were significantly involved in defense mechanism where the StTMEM161A was involved in the regulation of alkalization apoplast, the StDUF538 was activated in the chloroplast degradation program, and the StGTF3C5 regulated the proteins increase related to defense in the PVY infected cells. In addition, in the genetic improvement programs, these hub genes can be used as genetic markers for screening commercial cultivars for PVY resistance. Our survey demonstrated that the combination of GBS-based genetic diversity germplasm analysis and WGCNA can assist breeders to select cultivars resistant to PVY as well as help design proper crossing schemes in potato breeding.

Crossbreeding with Booroola or Texel sheep harboring major genes for prolificacy and muscularity enhances productivity but it may limit adaptation and survivorship in crossbred lambs. Thus, the trade-offs that impact productivity and adaptability have not been quantified or modeled and remain largely elusive at the genetic level, limiting the development of optimized breeding strategies. This study investigates the genomic architecture of purebred Moghani sheep and the first paternal backcross (PBC1) generation of crossbred lambs, including Booroola Merino × Moghani (BMM), Booroola Romney × Moghani (BRM), Texel Dalzell × Moghani (TDM), and Texel Tamlet × Moghani (TTM). Genotyping-by-sequencing (GBS) was used to assess genetic diversity, admixture patterns, and selection signatures. Structure analysis revealed complex admixture in BMM and BRM, while TDM and TTM were more homogeneous. Purebred Moghani sheep exhibited the highest genetic diversity (H O = 0.521 ± 0.10) and the lowest inbreeding (F IS = − 0.474), serving as a key genetic bridge among the groups. In contrast, BRM and TTM showed lower heterozygosity (H O = 0.410 ± 0.09 and 0.431 ± 0.10) and increased inbreeding (F ROH ), with extended runs of homozygosity (ROH), suggesting recent inbreeding and reduced effective population sizes. The functional annotation of ROH islands connected TDM and TTM to immune response and muscle development pathways like VEGF and insulin signaling, while BMM and BRM were linked to metabolic and reproductive pathways like central carbon metabolism in cancer ( mTOR ) and prolactin signaling ( LHB ). Taken together, these results highlight the need for improved breeding methods that prioritize trade-offs associated with reduced genetic diversity in crossbred populations. Nevertheless, given the limited genotype representation, the results should be taken with caution; so, next research should cover a larger panel of genotypes in order to have a more complete knowledge.