Molecular and agro-morphological characterization of new barley genotypes in arid environments.

BACKGROUND: Genetic diversity, population structure, agro-morphological traits, and molecular characteristics, are crucial for either preserving genetic resources or developing new cultivars. Due to climate change, water availability for agricultural use is progressively diminishing. This study used 100 molecular markers (25 TRAP, 22 SRAP, 23 ISTR, and 30 SSR). Additionally, 15 morphological characteristics were utilized to evaluate the optimal agronomic traits of 12 different barley genotypes under arid conditions. RESULTS: Substantial variations, ranging from significant to highly significant, were observed in the 15 agromorphological parameters evaluated among the 12 genotypes. The KSU-B101 barley genotype demonstrated superior performance in five specific traits: spike number per plant, 100-grain weight, spike number per square meter, harvest index, and grain yield. These results indicate its potential for achieving high yields in arid regions. The Sahrawy barley genotype exhibited the highest values across five parameters, namely leaf area, spike weight per plant, spike length, spike weight per square meter, and biological yield, making it a promising candidate for animal feed. The KSU-B105 genotype exhibited early maturity and a high grain count per spike, which reflects its early maturity and ability to produce a high number of grains per spike. This suggests its suitability for both animal feed and human food in arid areas. Based on marker data, the molecular study found that the similarity coefficients between the barley genotypes ranged from 0.48 to 0.80, with an average of 0.64. The dendrogram constructed from these data revealed three distinct clusters with a similarity coefficient of 0.80. Notably, the correlation between the dendrogram and its similarity matrix was high (0.903), indicating its accuracy in depicting the genetic relationships. The combined analysis revealed a moderate correlation between the morphological and molecular analysis, suggesting alignment between the two characterization methods. CONCLUSIONS: The morphological and molecular analyses of the 12 barley genotypes in this study effectively revealed the varied genetic characteristics of their agro-performance in arid conditions. KSU-B101, Sahrawy, and KSU-B105 have emerged as promising candidates for different agricultural applications in arid regions. Further research on these genotypes could reveal their full potential for breeding programs.

Comparative study among Avicennia marina, Phragmites australis, and Moringa oleifera based ethanolic-extracts for their antimicrobial, antioxidant, and cytotoxic activities.

Microbial resistance and other emerging health risk problems related to the side effects of synthetic drugs are the major factors that result in the research regarding natural products. Fruits, leaves, seeds, and oils-based phyto-constituents are the most important source of pharmaceutical products. Plant extract chemistry depends largely on species, plant components, solvent utilized, and extraction technique. This study was aimed to compare the ethanolic extracts of a mangrove plant, i.e., Avicennia marina (1E: Lower half of A. marina's pneumatophores, 2E: A. marina's leaves, 3E: Upper half of A. marina's pneumatophores, and 4E: A. marina's shoots), with non-mangrove plants, i.e., Phragmites australis (5E: P. australis's shoot), and Moringa oleifera (6E: M. oleifera's leaves) for their antimicrobial activities, total phenolic contents, antioxidant activity, and cytotoxicity potential. The antimicrobial activity assays were performed on gram-positive bacteria (i.e., Bacillus subtilis and Staphylococcus aureus), gram-negative bacteria (i.e., Escherichia coli, and Pseudomonas aeruginosa), and fungi (i.e., Aspergillus niger, Candida albicans, and Rhizopus spp.). We estimated antioxidant activity by TAC, DPPH, and FRAP assays, and the cytotoxicity was evaluated by MTT assay. The results of antimicrobial activities revealed that B. subtilis was the most sensitive to the tested plant extracts compared to S. aureus, while it only showed sensitivity to 6E and Imipenem. 5E and 6E showed statistically similar results against P. aeruginosa as compared to Ceftazidime. E. coli was the most resistant bacteria against tested plant extracts. Among the tested plant extracts, maximum inhibition activity was observed by 6E against A. niger (22 ± 0.57 mm), which was statistically similar to the response of 6E against C. albicans and 3E against Rhizopus spp. 2E did not show any activity against tested fungi. We found that 6E (208.54 ± 1.92 mg g-1) contains maximum phenolic contents followed by 1E (159.42 ± 3.22 mg g-1), 5E (131.08 ± 3.10 mg g-1), 4E (i.e., 72.41 ± 2.96 mg g-1), 3E (67.41 ± 1.68 mg g-1), and 2E (48.72 ± 1.71 mg g-1). The results depict a significant positive correlation between the phenolic contents and the antioxidant activities. As a result, phenolic content may be a natural antioxidant source.

Development of SSR Markers Linked to Stress Responsive Genes along Tomato Chromosome 3 (Solanum lycopersicum L.).

This study aimed to develop novel SSR markers in tomato. Several BAC clones along chromosome 3 in tomato were selected based on their content. The criteria was the availability of genes, either directly or indirectly related to stress response (drought, salinity, and heat) in tomato. A total of 20 novel in silico SSR markers were developed and 96 important nearby genes were identified. The identified nearby genes represent different tomato genes involved in plant growth and development and biotic and abiotic stress tolerance. The developed SSR markers were assessed using tomato landraces. A total of 29 determinate and semi-determinate local tomato landraces collected from diverse environments were utilized. A total of 33 alleles with mean of 1.65 alleles per locus were scored, showing 100% polymorphic patterns, with a mean of 0.18 polymorphism information content (PIC) values. The mean of observed and expected heterozygosity were 0.19 and 0.24, respectively. The mean value of the Jaccard similarity index was used for clustering the landraces. The developed microsatellite markers showed potential to assess genetic variability among tomato landraces. The genetic distance information reported in this study can be used by breeders in future genetic improvement of tomato for tolerance against diverse stresses.

Response of Tomato (Solanum lycopersicum L.) Genotypes to Heat Stress Using Morphological and Expression Study.

Due to unfavorable environmental conditions, heat stress is one of the significant production restrictions for the tomato (Solanum lycopersicum L.) crop. The tomato crop is considered an important vegetable crop globally and represents a model plant for fruit development research. The heat shock factor (HSF) gene family contains plant-specific transcription factors (TFs) that are highly conserved and play a key role in plant high-temperature stress responses. The current study was designed to determine the relative response of heat stress under three different temperatures in the field condition to determine its relative heat tolerance. Furthermore, the study also characterized heat shock genes in eight tomato genotypes under different temperature regimes. The expressions of each gene were quantified using qPCR. The descriptive statistics results suggested a high range of diversity among the studied variables growing under three different temperatures. The qPCR study revealed that the SlyHSF genes play an important role in plant heat tolerance pathways. The expression patterns of HSF genes in tomatoes have been described in various tissues were determined at high temperature stress. The genes, SlyHSFs-1, SlyHSFs-2, SlyHSFs-8, SlyHSFs-9 recorded upregulation expression relative to SlyHSFs-3, SlyHSFs-5, SlyHSFs-10, and SlyHSFs-11. The genotypes, Strain B, Marmande VF, Pearson's early, and Al-Qatif-365 recorded the tolerant tomato genotypes under high-temperature stress conditions relative to other genotypes. The heat map analysis also confirmed the upregulation and downregulation of heat shock factor genes among the tomato genotypes. These genotypes will be introduced in the breeding program to improve tomato responses to heat stress.

Cold case: The disappearance of Egypt bee virus, a fourth distinct master strain of deformed wing virus linked to honeybee mortality in 1970's Egypt.

In 1977, a sample of diseased adult honeybees (Apis mellifera) from Egypt was found to contain large amounts of a previously unknown virus, Egypt bee virus, which was subsequently shown to be serologically related to deformed wing virus (DWV). By sequencing the original isolate, we demonstrate that Egypt bee virus is in fact a fourth unique, major variant of DWV (DWV-D): more closely related to DWV-C than to either DWV-A or DWV-B. DWV-A and DWV-B are the most common DWV variants worldwide due to their close relationship and transmission by Varroa destructor. However, we could not find any trace of DWV-D in several hundred RNA sequencing libraries from a worldwide selection of honeybee, varroa and bumblebee samples. This means that DWV-D has either become extinct, been replaced by other DWV variants better adapted to varroa-mediated transmission, or persists only in a narrow geographic or host range, isolated from common bee and beekeeping trade routes.

Morphological Characterization and DNA Barcoding of Duckweed Species in Saudi Arabia.

Duckweeds, or Lemnaceae, are widespread aquatic plants. Morphology-based identification of duckweed species is difficult because of their structural complexity. Hence, molecular tools provide significant advantages for characterizing and selecting species or clones for sustainable commercial use. In this study, we collected and characterized ten duckweed isolates from nine different regions in Saudi Arabia (SA). Based on the morphological characterization and phylogenetic analysis of intergenic spacer sequences of chloroplast DNA using six barcoding markers, the clones were classified into three genera, represented by seven species: Lemna gibba L., Lemna minor L., Lemna japonica Landolt, Lemna aequinoctialis Welw., Lemna perpusilla Torr., Spirodela polyryiza (L.) Schleid., and Landoltia punctate G. Mey. Lemna gibba was revealed to be a distinct dominant duckweed species in many regions of SA. Five barcoding markers showed that L. gibba, L. minor, and L. punctata were the most widely distributed species in the country. However, L. punctata, L. perpusilla, and S. polyryiza were the dominant species in the Al-Qassim, Madinah-1, and Madinah-2 regions, respectively. Moreover, the morphological traits revealed variations for these clones, relative to other studied duckweed clones. According to the results obtained in this study, three out of six plastid markers (trnH-psbA, matK, and atpF-atpH) helped to identify the dominant duckweed species in Saudi Arabia. Further evaluation based on adaptability, molecular genetic studies, and functional genomics is needed for these species to be used at the commercial level in Saudi Arabia.

Evaluation of Frost Damage and Pod Set in Faba Bean (Vicia faba L.) under Field Conditions.

Frost is one factor that causes extensive yield losses globally. A study was conducted to evaluate frost damage under field conditions and assess the genetic variation of flowers converting into pods. Diverse faba bean genotypes were evaluated under four growing seasons in a randomized complete block design: three at the University of Sydney, Narrabri, Australia (2014-2016) with three sowing dates, and one at the Agricultural Research Station, Dirab, Riyadh, Saudi Arabia (2016/2017) in one sowing. Visual methods were used to estimate frost damage and record the development of pods. Radiation frost in 2014 (Narrabri) damaged lower pods, while advection frost in 2016/2017 (Dirab) damaged upper pods. The radiation frost formed immediately above the ground; therefore, flowers and pods of taller plants minimized the damage because of their long distance from the ground. The earliest (mid-April) and middle sowing (7 May) suffered more by frost, while a delay in sowing (last week in May) led to frost escape or minor damage. The genotypes IX474/4-3 and 11NF010a-2 showed low sensitivity to frost at the vegetative and reproductive stages. Flowers developed at the beginning of flowering had a faster and higher pod formation rate (41-43%) than those formed later and contributed more to yields. Therefore, a severe frost at the beginning of flowering can cause a significant yield loss as these flowers are the most productive. The frost-tolerant genotypes, and faster and higher pod forming rates, identified in this study can be exploited to breed better varieties in the future.

Assessment of morphological and molecular variability of some Solanum melongena L. cultivars and wild Solanum incanum L. in Saudi Arabia.

The genus Solanum exhibits a wide range of variability in morphology, flavor, and tolerance to biotic and abiotic stresses. Phenotypic and genetic variability using ISSR and RAPD markers of Solanum incanum distributed in Al-Baha province of the Kingdom of Saudi Arabia is assessed. Thirty samples are representing three different locations: Baljershy, Aqeeq, and Tohama, besides twenty-five samples representing five different commercial cultivars tested. Growth type, the number of leaves per plant, fruit size (phenotypic traits), crude protein, carbohydrates, digestive organic matter, and Mg, Ca, P were the principal contributors in the PCA. Molecular analysis showed that 114 ISSR and 80 RAPD alleles with a 100% polymorphism were recorded. The polymorphism information content (PIC) values ranged from 0.84 to 0.91 for ISSR and from 0.59 to 0.89 for RAPD data. Similarity values ranged from 0.16 to 1.00, with an average of 0.47 for ISSR and from 0.01 to 0.97, with an average of 0.36 for RAPD. It resulted in a positive and significant correlation between morphological, molecular, nutritional, and chemical analysis of fruits using Mantel analysis. UPGMA and PCA for morphological traits and molecular data discriminated commercial cultivars and wild relatives. Solanum incanum was more diverse than commercial varieties. This study revealed a wide genetic diversity among and within collected eggplant accessions and may use in breeding programs of eggplants. There is a need to increase the present eggplant collection to widen the genetic diversity of cultivated eggplant varieties in Saudi Arabia.

Assessment of Morpho-Physiological and Biochemical Responses of Mercury-Stressed Trigonella foenum-gracum L. to Silver Nanoparticles and Sphingobacterium ginsenosidiumtans Applications.

Heavy metals are primarily generated and deposited in the environment, causing phytotoxicity. This work evaluated fenugreek plants' morpho-physiological and biochemical responses under mercury stress conditions toward Ag nanoparticles and Sphingobacterium ginsenosidiumtans applications. The fabrication of Ag nanoparticles by Thymus vulgaris was monitored and described by UV/Vis analysis, FTIR, and SEM. The effect of mercury on vegetative growth was determined by measuring the root and shoots length, the number and area of leaves, the relative water content, and the weight of the green and dried plants; appraisal of photosynthetic pigments, proline, hydrogen peroxide, and total phenols content were also performed. In addition, the manipulation of Ag nanoparticles, S. ginsenosidiumtans, and their combination were tested for mercury stress. Here, Ag nanoparticles were formed at 420 nm with a uniform cuboid form and size of 85 nm. Interestingly, the gradual suppression of vegetal growth and photosynthetic pigments by mercury, Ag nanoparticles, and S. ginsenosidiumtans were detected; however, carotenoids and anthocyanins were significantly increased. In addition, proline, hydrogen peroxide, and total phenols content were significantly increased because mercury and S. ginsenosidiumtans enhance this increase. Ag nanoparticles achieve higher levels by the combination. Thus, S. ginsenosidiumtans and Ag nanoparticles could have the plausible ability to relieve and combat mercury's dangerous effects in fenugreek.

Assessment of Morpho-Physiological, Biochemical and Antioxidant Responses of Tomato Landraces to Salinity Stress.

Understanding salt tolerance in tomato (Solanum lycopersicum L.) landraces will facilitate their use in genetic improvement. The study assessed the morpho-physiological variability of Hail tomato landraces in response to different salinity levels at seedling stages and recommended a tomato salt-tolerant landrace for future breeding programs. Three tomato landraces, Hail 548, Hail 747, and Hail 1072 were tested under three salinity levels: 75, 150, and 300 mM NaCl. Salinity stress reduced shoots' fresh and dry weight by 71% and 72%, and roots were 86.5% and 78.6%, respectively. There was 22% reduced chlorophyll content, carotene content by 18.6%, and anthocyanin by 41.1%. Proline content increased for stressed treatments. The 300 mM NaCl treatment recorded the most proline content increases (67.37 mg/g fresh weight), with a percent increase in proline reaching 61.67% in Hail 747. Superoxide dismutase (SOD) activity decreased by 65% in Hail 548, while it relatively increased in Hail 747 and Hail 1072 treated with 300 mM NaCl. Catalase (CAT) activity was enhanced by salt stress in Hail 548 and recorded 7.6%, increasing at 75 and 5.1% at 300 mM NaCl. It revealed a reduction in malondialdehyde (MDA) at the 300 mM NaCl concentration in both Hail 548 and Hail 1072 landraces. Increasing salt concentrations showed a reduction in transpiration rate of 70.55%, 7.13% in stomatal conductance, and 72.34% in photosynthetic rate. K+/Na+ ratios decreased from 56% for 75 mM NaCl to 85% for 300 mM NaCl treatments in all genotypes. The response to salt stress in landraces involved some modifications in morphology, physiology, and metabolism. The landrace Hail 548 may have better protection against salt stress and observed protection against reactive oxygen species (ROS) by increasing enzymatic "antioxidants" activity under salt stress.

Antioxidant Enzymatic Activities and Growth Response of Quinoa (Chenopodium quinoa Willd) to Exogenous Selenium Application.

Selenium is a trace element essential to many organisms, including higher plants. At low concentrations, it enhances growth and development; however, it is toxic at high concentrations. The development of crops with proper levels of selenium will be worth for both nutrition and Se-based therapeutics. This study aimed to investigate the morphological, physiological, and biochemical responses of the quinoa plant to 0, 2.5, 5, 10, and 20 mg/L of Na2SeO3·5H2O. Selenium at low concentrations (2.5 and 5 mg/L), quinoa plant showed a significant increase of growth parameters, relative water content, photosynthetic pigments, proline, total soluble sugars, and antioxidant enzymes activities as (superoxide dismutase (SOD), catalase (CAT), peroxidase (POD, ascorbate peroxidase (APX), and glutathione reductase (GR)), and contents of malondialdehyde (MDA) and H2O2 were reduced. However, high concentrations (10 and 20) mg/L caused a decrease in plant growth parameters, relative water content, and photosynthetic pigments. In contrast, excess selenium increased the oxidative stress monitored by hydrogen peroxide and lipid peroxidation levels. The enzymatic antioxidant system responded to the selenium supply significantly increased. Osmolytes compounds, such as total sugars and proline, increased in selenium-treated plants. The increase in these osmolytes compounds may show a defense mechanism for the osmotic readjustment of quinoa plants to mitigate the toxicity caused by selenium. This study shows the morphological and physiological responses that must be considered for success in the sustainable cultivation of quinoa plants in environments containing excess selenium.

Phytostimulatory Influence of Comamonas testosteroni and Silver Nanoparticles on Linum usitatissimum L. under Salinity Stress.

They were shifting in land use increases salinity stress, significant abiotic stress affecting plant growth, limiting crop productivity. This work aimed to improve Linum usitatissimum L. (linseed) growth under salinity using Comamonas testosteroni and silver nanoparticles (AgNPs). AgNPs were fabricated exploiting Rosmarinus officinalis and monitored by U.V./Vis spectrophotometry scanning electron microscopy (SEM) and Fourier transforms infrared spectroscopy (FTIR). Photosynthetic pigments, enzymatic and nonenzymatic antioxidants of linseed were investigated under salt stress in treated and untreated plants with C. testosteroni alongside AgNPs. Our findings recorded the formation of AgNPs at 457 nm, which were globular and with a diameter of 75 nm. Notably, chlorophyll-a, b, and total chlorophyll reduction while enhanced carotenoids and anthocyanin contents were attained under salinity stress. Total dissoluble sugars, proline, and dissoluble proteins, H2O2, malondialdehyde, enzymatic and nonenzymatic antioxidants were significantly elevated in NaCl well. Combined AgNPs and C. testosteroni elevated photosynthetic pigments. Also, they led to the mounting of soluble sugars, proline, and soluble proteins. H2O2 and malondialdehyde decreased while enzymatic and nonenzymatic antioxidants increased in response to AgNPs, C. testosteroni, and their combination. Thus, AgNPs and C. testosteroni might bio-fertilizers to improve linseed crop productivity under salinity stress.

Complete mitochondrial genome sequence of Awassi-Jo sheep breed (Ovis aries) in Jordan.

Using high-throughput sequencing technology, the complete mitochondrial genome of Awassi-Jo breed (Ovis aries) was decoded. Mitochondrial genome was 16,617 bp in length. The genome contained 37 genes (13 protein-coding, 22 tRNA, and 2 rRNA) and a control region (D-loop region). The genes were encoded on the H-strand, except for the ND6 gene and 8 tRNA genes, which were encoded on the L-strand. The GC content is 38.9%. Phylogenetic analysis was performed to compare Awassi-Jo with other sheep breeds. The phylogenetic tree showed that Awassi-Jo diverged earlier than related breeds (Turkey, Italy, Germany, and Netherland) with a common ancestor in haplogroup HB. The results revealed the importance of mitochondrial data in studying sheep evolution and domestication.

Complete chloroplast genome sequence of historical olive (Olea europaea subsp. europaea) cultivar Mehras, in Jordan.

The complete chloroplast genome sequence of Olea europaea subsp. europaea cultivar Mehras was determined using high-throughput sequencing technology. Chloroplast genome was 155,897 bp in length, containing a pair of 25,742 bp inverted repeat (IR) regions, which were separated by large and small single-copy regions (LSC and SSC) of 86,622 and 17,791 bp, respectively. The chloroplast genome contained 130 genes (85 protein-coding, 37 tRNA, and eight rRNA). GC content was 37.8%. We performed phylogenetic analysis with other isolates. The analysis showed that O. e. subsp. europaea cultivar Mehras has an ancient common ancestor with cultivated olives in Italy, Spain, and Cyprus.

Evaluation of the genetic diversity of wild Salvadora persica 'Arak' from Saudi Arabia.

Assessment of genetic diversity is crucial for efficient selection genotypes in plant breeding and improvement programs. Studies of genetic diversity of S. persica are rare relative to the large species diversity in Saudi Arabia, despite its valuable importance as one of the most popular medicinal plants. We investigate the genetic variability and genetic differentiation among and within wild Salvadora persica populations distributed in four regions of Saudi Arabia. Twelve sequence-related amplified polymorphism (SRAP) primers combination generated 326 alleles, with an average of 27.2 alleles per primer. All primers showed 100 polymorphism percentage, and higher PIC values exceeded 0.90. Jaccard similarity values varied between 0.04 to 0.43, with an average of 0.31, which showed a weak relationship among the accessions and their origin. Based on UGPMA and principal coordinate analysis, accessions collected from the same region showed less aggregation. Genetic diversity parameters showed that both Aflaj and Joodah populations recorded the highest mean values for the effective number of alleles (1.26 and 1.24). Shannon index and genetic heterozygosity (0.23 and 0.15 for both populations), and percent of polymorphism 45.45% for Aflaj and 43.87 for Joodah population. Most of the genetic variation was because of differences within populations (77%) and 23% among populations. SRAP markers explored the genetic diversity among and within S. persica populations. In this work, genetic diversity within populations was high, and the population structure was weak. We detected no specific geographic structure, which may reveal an active movement of plants among populations.

Detection of genetically modified maize in Jordan.

This study aimed to detect genetically modified maize (GMM) in seeds of eleven imported maize hybrids grown in Jordan. We used promoter 35 S and T-nos terminator for general screening of transgenic materials. Conventional PCR detected the specific events for the screening of Bt 11, MON810, and Bt176 events. Seeds of eleven maize hybrids samples showed a positive response to the 35 S promoter; nine out of eleven showed a positive response for T-nos terminator. Bt11 event was the most used in GMM seeds, where seven out of eleven samples showed positive results. Two out of eleven hybrids showed the presence of the Bt176 event; however, MON810 not detected in any of the tested hybrids. We studied the Bt11 event in imported GMM seeds in Jordan for the first time, reinforcing the need for a mandatory labeling system and a valid simple qualitative method in routine analysis of GMCs.

Legume genomics and transcriptomics: From classic breeding to modern technologies.

Legumes are essential and play a significant role in maintaining food standards and augmenting physiochemical soil properties through the biological nitrogen fixation process. Biotic and abiotic factors are the main factors limiting legume production. Classical breeding methodologies have been explored extensively about the problem of truncated yield in legumes but have not succeeded at the desired rate. Conventional breeding improved legume genotypes but with more resources and time. Recently, the invention of next-generation sequencing (NGS) and high-throughput methods for genotyping have opened new avenues for research and developments in legume studies. During the last decade, genome sequencing for many legume crops documented. Sequencing and re-sequencing of important legume species have made structural variation and functional genomics conceivable. NGS and other molecular techniques such as the development of markers; genotyping; high density genetic linkage maps; quantitative trait loci (QTLs) identification, expressed sequence tags (ESTs), single nucleotide polymorphisms (SNPs); and transcription factors incorporated into existing breeding technologies have made possible the accurate and accelerated delivery of information for researchers. The application of genome sequencing, RNA sequencing (transcriptome sequencing), and DNA sequencing (re-sequencing) provide considerable insights for legume development and improvement programs. Moreover, RNA-Seq helps to characterize genes, including differentially expressed genes, and can be applied for functional genomics studies, especially when there is limited information available for the studied genomes. Genome-based crop development studies and the availability of genomics data as well as decision-making gears look be specific for breeding programs. This review mainly presents an overview of the path from classical breeding to new emerging genomics tools, which will trigger and accelerate genomics-assisted breeding for recognition of novel genes for yield and quality characters for sustainable legume crop production.

Novel inflorescence architecture in gamma radiation-induced faba bean mutant populations.

Purpose: Inflorescence architecture is an important trait in the seed production of grain legumes. As several genes are responsible for this trait, any mutation, on these genes, may cause change in the inflorescence architecture. This study was conducted to evaluate inflorescence architecture in faba bean exposed to gamma radiation and to characterize the inflorescence architecture mutants phenotypically.Materials and methods: Faba bean M2 seeds (4898) generated from M1 generation of cultivars Hassawi 2 and ILB4347 were used in this study. M1 seeds were produced by irradiation treatments at two doses of gamma radiations (25 and 50 Gy). Faba bean M2 seeds were planted under field conditions. A total of 4032 mutant plants out of 4898 M2 seeds were evaluated for their inflorescence architecture.Results: A total of 20 determinate mutants were found and classified into four different types. Determinate type 1 was characterized by the formation of single terminal inflorescence on shoot apical meristem (SAM), type 2 by the formation of multiple inflorescences on SEM and generated upper branches that act as indeterminate type. Type 3 was characterized by the formation of a panicle-like inflorescence. While type 4 was characterized by the formation of primary and secondary panicle-like inflorescence. All of the determinate mutant types had shorter plant height and earlier maturity than control indeterminate type but had lower biological yield and seed yield. Among the determinate mutant types, determinate type 1 was only mutant that had a higher harvest index than the control indeterminate type. This promising mutant can be used to further breeding program to increase biological yield and seed yield.Conclusions: This study indicated potential of gamma radiation in inducing novel inflorescence architecture in faba bean. The mutants developed are valuable resources to study genes related to inflorescence architecture through forward genetics approach.

Identification and Characterization of Salt-Responsive MicroRNAs in Vicia faba by High-Throughput Sequencing.

Salt stress has detrimental effects on plant growth and development. MicroRNAs (miRNAs) are a class of noncoding RNAs that are involved in post-transcriptional gene expression regulation. In this study, small RNA sequencing was employed to identify the salt stress-responsive miRNAs of the salt-sensitive Hassawi-3 and the salt-tolerant ILB4347 genotypes of faba bean, growing under salt stress. A total of 527 miRNAs in Hassawi-3 plants, and 693 miRNAs in ILB4347 plants, were found to be differentially expressed. Additionally, 284 upregulated and 243 downregulated miRNAs in Hassawi-3, and 298 upregulated and 395 downregulated miRNAs in ILB4347 plants growing in control and stress conditions were recorded. Target prediction and annotation revealed that these miRNAs regulate specific salt-responsive genes, which primarily included genes encoding transcription factors and laccases, superoxide dismutase, plantacyanin, and F-box proteins. The salt-responsive miRNAs and their targets were functionally enriched by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, which showed that the miRNAs were involved in salt stress-related biological pathways, including the ABC transporter pathway, MAPK signaling pathway, plant hormone signal transduction, and the phosphatidylinositol signaling system, among others, suggesting that the miRNAs play an important role in the salt stress tolerance of the ILB4347 genotype. These results offer a novel understanding of the regulatory role of miRNAs in the salt response of the salt-tolerant ILB4347 and the salt-sensitive Hassawi-3 faba bean genotypes.

Biochemical and molecular characterization of cowpea landraces using seed storage proteins and SRAP marker patterns.

Seven landraces of cowpea [Vigna unguiculata (L.) Walp.] were assessed for genetic variability in total proteins, protein fractions viz. albumins, globulins, prolamins, and glutelins by SDS-polyacrylamide gel electrophoresis and DNA polymorphism using sequence-related amplified polymorphisms (SRAP) markers. The solubility-based protein fractionation data indicated that the salt soluble fraction (globulin) and water-soluble fraction (albumin) proteins were the predominant fractions in cowpea seeds comprising 45-50.3% and 31.2-35.5% of total soluble proteins, respectively. The electrophoretic pattern revealed the molecular heterogeneity among total proteins as well as different protein fractions. The molecular weights of protein bands obtained by SDS-PAGE varied between 10 to 250, 15 to 110, 15 to 150, and 15 to 130 kDa for total proteins, albumins, globulins, and glutelins, respectively. A large number of bands were found common to the various landraces, indicative of their close relationship with one another. However, a few bands distinctive to some specific landraces were also detected, indicating varietal differences. A 34 SRAP primer pair combination generated a total of 1003 amplicons (loci) showed 100% polymorphism with an average of 0.93 polymorphism information content (PIC) value. Landraces displayed an average 0.50 similarity coefficient which clustered the landraces corresponding to their growth habit in main clusters and to their geographical origin in subcultures. Molecular and biochemical analysis were correlated with a medium level (Mantel test, r = 0.56, P < 0.02). These findings revealed that seed proteins and DNA polymorphism provide valuable information regarding the variability among landraces and this information could be utilized for breeding purposes in the enhancement of protein quality and quantity in grain legumes.