Combined Effects of Gold Nanoparticles and Ionizing Radiation on Human Prostate and Lung Cancer Cell Migration.

The effect of 15 nm-sized gold nanoparticles (AuNPs) and/or ionizing radiation (IR) on the migration and adhesion of human prostate (DU145) and lung (A549) cancer cell lines was investigated. Cell migration was measured by observing the closing of a gap created by a pipette tip on cell monolayers grown in 6-well plates. The ratio of the gap areas at 0 h and 24 h were used to calculate the relative migration. The relative migration of cells irradiated with 5 Gy was found to be 89% and 86% for DU145 and A549 cells respectively. When the cells were treated with 1 mM AuNPs this fell to ~75% for both cell lines. However, when the cells were treated with both AuNPs and IR an additive effect was seen, as the relative migration rate fell to ~60%. Of interest was that when the cells were exposed to either 2 or 5 Gy IR, their ability to adhere to the surface of a polystyrene culture plate was significantly enhanced, unlike that seen for AuNPs. The delays in gap filling (cell migration) in cells treated with IR and/or AuNPs can be attributed to cellular changes which also may have altered cell motility. In addition, changes in the cytoskeleton of the cancer cells may have also affected adhesiveness and thus the cancer cell's motility response to IR.

Phenological, nutritional and molecular diversity assessment among 35 introduced lentil (Lens culinaris Medik.) genotypes grown in Saudi Arabia.

Morphological, nutritional and molecular analyses were carried out to assess genetic diversity among 35 introduced lentil genotypes (Lens culinaris Medik.). The genotypes exhibited significant differences for their field parameters and some of them showed noticeable superiority. The nutritional and proximate analysis showed that some genotypes were excellent sources of proteins, essential amino acids, minerals, anti-oxidants, total phenolic contents (TPC) and total flavonoid contents (TFC) and hence, highlights lentil nutritional and medicinal potential. Sequence-related amplified polymorphism (SRAP) and amplified fragments length polymorphism (AFLP) markers were used to estimate the genetic variability at the molecular level. The existence of a considerable amount of genetic diversity among the tested lentil genotypes was also proven at the molecular level. A total of 2894 polymorphic SRAP and 1625 AFLP loci were successfully amplified using six SRAP and four AFLP primer pair combinations. Polymorphism information content (PIC) values for SRAP and AFLP markers were higher than 0.8, indicating the power and higher resolution of those marker systems in detecting molecular diversity. UPGMA (unweighted pair group method with arithmetic average) cluster analysis based on molecular data revealed large number of sub clusters among genotypes, indicating high diversity levels. The data presented here showed that FLIP2009-64L and FLIP2009-69L could be used as a significant source of yield, total protein, essential amino acids, and antioxidant properties. The results suggest potential lentil cultivation in the central region of Saudi Arabia for its nutritional and medicinal properties, as well as sustainable soil fertility crop.

Transcriptomic analysis reveals candidate genes associated with salinity stress tolerance during the early vegetative stage in fababean genotype, Hassawi-2.

Abiotic stresses are a significant constraint to plant production globally. Identifying stress-related genes can aid in the development of stress-tolerant elite genotypes and facilitate trait and crop manipulation. The primary aim of this study was to conduct whole transcriptome analyses of the salt-tolerant faba bean genotype, Hassawi-2, under different durations of salt stress (6 h, 12 h, 24 h, 48 h, and 72 h) at the early vegetative stage, to better understand the molecular basis of salt tolerance. After de novo assembly, a total of 140,308 unigenes were obtained. The up-regulated differentially expressed genes (DEGs) were 2380, 2863, 3057, 3484, and 4820 at 6 h, 12 h, 24 h, 48 h, and 72 h of salt stress, respectively. Meanwhile, 1974, 3436, 2371, 3502, and 5958 genes were downregulated at 6 h, 12 h, 24 h, 48 h, and 72 h of salt stress, respectively. These DEGs encoded various regulatory and functional proteins, including kinases, plant hormone proteins, transcriptional factors (TFs) basic helix-loop-helix (bHLH), Myeloblastosis (MYB), and (WRKY), heat shock proteins (HSPs), late embryogenesis abundant (LEA) proteins, dehydrin, antioxidant enzymes, and aquaporin proteins. This suggests that the faba bean genome possesses an abundance of salinity resistance genes, which trigger different adaptive mechanisms under salt stress. Some selected DEGs validated the RNA sequencing results, thus confirming similar gene expression levels. This study represents the first transcriptome analysis of faba bean leaves subjected to salinity stress offering valuable insights into the mechanisms governing salt tolerance in faba bean during the vegetative stage. This comprehensive investigation enhances our understanding of precise gene regulatory mechanisms and holds promise for the development of novel salt-tolerant faba bean salt-tolerant cultivars.

CRISPR-Cas System, a Possible "Savior" of Rice Threatened by Climate Change: An Updated Review.

Climate change has significantly affected agriculture production, particularly the rice crop that is consumed by almost half of the world's population and contributes significantly to global food security. Rice is vulnerable to several abiotic and biotic stresses such as drought, heat, salinity, heavy metals, rice blast, and bacterial blight that cause huge yield losses in rice, thus threatening food security worldwide. In this regard, several plant breeding and biotechnological techniques have been used to raise such rice varieties that could tackle climate changes. Nowadays, gene editing (GE) technology has revolutionized crop improvement. Among GE technology, CRISPR/Cas (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein) system has emerged as one of the most convenient, robust, cost-effective, and less labor-intensive system due to which it has got more popularity among plant researchers, especially rice breeders and geneticists. Since 2013 (the year of first application of CRISPR/Cas-based GE system in rice), several trait-specific climate-resilient rice lines have been developed using CRISPR/Cas-based GE tools. Earlier, several reports have been published confirming the successful application of GE tools for rice improvement. However, this review particularly aims to provide an updated and well-synthesized brief discussion based on the recent studies (from 2020 to present) on the applications of GE tools, particularly CRISPR-based systems for developing CRISPR rice to tackle the current alarming situation of climate change, worldwide. Moreover, potential limitations and technical bottlenecks in the development of CRISPR rice, and prospects are also discussed.

Molecular diversity assessment using Sequence Related Amplified Polymorphism (SRAP) Markers in Vicia faba L.

Sequence-related amplified polymorphism (SRAP) markers were used to assess the genetic diversity and relationship among 58 faba bean (Vicia faba L.) genotypes. Fourteen SRAP primer combinations amplified a total of 1036 differently sized well-resolved peaks (fragments), of which all were polymorphic with a 0.96 PIC value and discriminated all of the 58 faba bean genotypes. An average pairwise similarity of 21% was revealed among the genotypes ranging from 2% to 65%. At a similarity of 28%, UPGMA clustered the genotypes into three main groups comprising 78% of the genotypes. The local landraces and most of the Egyptian genotypes in addition to the Sudan genotypes were grouped in the first main cluster. The advanced breeding lines were scattered in the second and third main clusters with breeding lines from the ICARDA and genotypes introduced from Egypt. At a similarity of 47%, all the genotypes formed separated clusters with the exceptions of Hassawi 1 and Hassawi 2. Group analysis of the genotypes according to their geographic origin and type showed that the landraces were grouped according to their origin, while others were grouped according to their seed type. To our knowledge, this is the first application of SRAP markers for the assessment of genetic diversity in faba bean. Such information will be useful to determine optimal breeding strategies to allow continued progress in faba bean breeding.

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.

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.

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.

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.

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.

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.

Morphological and chromosomal abnormalities in gamma radiation-induced mutagenized faba bean genotypes.

Purpose: This study was conducted to evaluate and compare the influence of gamma radiations on morphological and chromosomal abnormalities in twenty mutagenized faba bean populations, representing first and second generations (M1 and M2) of five faba bean genotypes.Materials and methods: Five faba bean genotypes were exposed at two doses of gamma radiations (25 and 50 Gy). For determining the types of chromosomal aberrations caused by the gamma radiation, mitotic and meiotic cells were isolated from root tips and pollen mother cells, respectively.Results: The M1 generations of the five genotypes varied for sensitivity to gamma radiations, for seedling emergence. The genotype Skah 2 was more sensitive than other genotypes, the order of sensitivity of other genotypes was Misr 3 > ILB 4347 > Hassawi 2 > Hassawi 3. However, seedling emergence of the M2 generations was not as much reduced as that of the M1 generations. Ten different chlorophyll-deficient mutants were identified among the M2 generations. Gamma radiations also caused the development of abnormal leaflets, flowers and pollen grains. The most common types of chromosome aberrations in the mitotic cells were stickiness, laggard and chromosome breaks, whereas the most common types in the meiotic cells were stickiness and disturbed polarity.Conclusion: The gamma radiation decreased the seedling emergence and induced a wide range of morphological and chromosomal abnormalities in faba bean.

Desi chickpea genotypes tolerate drought stress better than kabuli types by modulating germination metabolism, trehalose accumulation, and carbon assimilation.

Chickpea is mostly grown in rainfed environments and, consequently, its growth is affected by drought stress. This study comprised two independent experiments to investigate the physiological basis of drought tolerance in desi and kabuli chickpea genotypes. In Experiment 1, six genotypes each of desi and kabuli types were planted in soil-filled pots under natural conditions. Ten days after planting, soil moisture was maintained at 75% water holding capacity (well-watered) or 50% water holding capacity (drought stress). Drought stress significantly reduced seedling dry weight, specific leaf area (SLA), and transpiration efficiency (TE) in both chickpea types, relative to the well-watered controls, but their responses varied, with relatively fewer reductions in desi genotypes, Bakhar-2011 and Bitall-2016, and kabuli genotypes, K-70005 and Noor-2013. These four genotypes were used in experiment 2, which was similar to the first but conducted in a climate chamber and the drought was imposed at planting. Drought stress reduced stand establishment, growth, photosynthesis, water relations, α-amylase activity, sugar metabolism, proline, phenolic accumulation, nitrogen and potassium to varying degrees in the four tested genotypes. The reductions were greater in kabuli genotypes than desi genotypes. Under drought stress, desi genotypes germinated better, and had higher trehalose, total and reducing sugars, sucrose, α-amylase activity, photosynthesis, growth, and mineral concentrations than kabuli genotypes. The desi genotype Bakhar-2011 performed better under drought than the desi genotype Bitall-2016 due to better germination metabolism and accumulation of free proline, total phenolics, and trehalose, which maintained carbon assimilation and prevented oxidative damage. In conclusion, desi chickpea types tolerate drought stress better than kabuli types due to better germination metabolism and trehalose accumulation, which prevented oxidative damage, helped with efficient water use, and sustained plant growth.

Impact of Abiotic Stresses on Grain Composition and Quality in Food Legumes.

Grain quality and composition in food legumes are influenced by abiotic stresses. This review discusses the influence of abiotic stresses on grain composition and quality in food grains. Grain protein declines under salt stress due to the restricted absorption of nitrate from the soil solution. Grain phosphorus, nitrogen, and potassium contents declined whereas sodium and chloride increased. However, under drought, grain protein increased whereas the oil contents were decreased. For example, among fatty acids, oleic acid content increased; however, linoleic and/or linolenic acids were decreased under drought. Heat stress increased grain oil content whereas grain protein was decreased. Low temperature during late pod-filling reduced starch, protein, soluble sugar, fat, and fiber contents. However, an elevated CO2 level improved omega-3 fatty acid content at the expense of omega-6 fatty acids. Crop management and improvement strategies, next generation sequencing, and gene manipulation can help improve quality of food legumes under abiotic stresses.

Comparative phytochemical profiling of different soybean (Glycine max (L.) Merr) genotypes using GC-MS.

This study aimed to estimate the proximate, phenolic and flavonoids contents and phytochemicals present in seeds of twenty four soybeans (Glycine max (L.) Merr) genotypes to explore their nutritional and medicinal values. Crude protein composition ranged between 35.63 and 43.13% in Argentinian and USA (Clark) genotypes, respectively. Total phenolic content varied from 1.15 to 1.77 mg GAE/g, whereas flavonoids varied from 0.68 to 2.13 mg QE/g. The GC-MS analysis resulted identification of 88 compounds categorized into aldehydes (5), ketones (13), alcohols (5), carboxylic acids (7), esters (13), alkanes (2), heterocyclic compounds (19), phenolic compound (9), sugar moiety (7) ether (4) and amide (3), one Alkene and one fatty acid ester. Indonesian genotypes (Ijen and Indo-1) had the highest phenolic compounds than others genotype having antioxidant activities, while the Australian genotype contains the maximum in esters compounds. The major phytocompounds identified in majority of genotypes were Phenol, 2,6-dimethoxy-, 2-Methoxy-4-vinylphenol, 3,5-Dimethoxyacetophenone, 1,2-cyclopentanedione and Hexadecanoic acid, methyl ester. The presence of phytochemicals with strong pharmacological actions like antimicrobial and antioxidants activities could be considered as sources of quality raw materials for food and pharmaceutical industries. This study further set a platform for isolating and understanding the characteristics of each compound for it pharmacological properties.

Characterization of drought stress-responsive root transcriptome of faba bean (Vicia faba L.) using RNA sequencing.

Drought and salinity are the major factors that limit the faba bean (Vicia faba L.) production worldwide. The aim of this study is to identify the water stress differentially expressed genes (DEGs) through the root transcriptome analyses of the drought-tolerant Hassawi 2 genotype at vegetative and flowering stages. A total of 624.8 M high-quality Illumina reads were generated and assembled into 198,155 all-unigenes with a mean length of 738 bp and an N50 length of 1347 bp. Among all-unigenes, 78,262 were assigned to non-redundant (Nr), 66,254 to nucleotide (Nt), 54,034 to KEGG, and 43,913 to gene ontology (GO) annotations. A total of 36,834 and 35,510 unigenes were differentially expressed at the vegetative and flowering stages of Hassawi 2 under drought stress, respectively. The majority of unigenes were down-regulated at both developmental stages. However, the number of genes up-regulated (15,366) at the flowering stage exceeded the number of those up-regulated (14,097) at the vegetative stage, and the number of genes down-regulated (20,144) at the flowering stage was smaller than the number of those down-regulated (22,737) at the vegetative stage. The drought stress-responsive differentially expressed unigenes coded for various regulatory proteins, including protein kinases and phosphatases, transcription factors and plant hormones and functional proteins including enzymes for osmoprotectant, detoxification and transporters were differentially expressed, most of which were largely up-regulated. Moreover, a substantial proportion of the DEGs identified in this study were novel, most exhibited a significant change in their expression levels under water stress, making them an unexploited resource that might control specific responses to drought stress in the faba bean. Finally, qRT-PCR results were found almost consistent with the results of next-generation sequencing. Our data will help in understanding the drought tolerance mechanisms in plants and will provide resources for functional genomics.

Seed priming improves chilling tolerance in chickpea by modulating germination metabolism, trehalose accumulation and carbon assimilation.

Chilling stress is one of the major abiotic stresses affecting chickpea productivity worldwide. This study evaluated the potential role of seed priming in improving resistance to chilling stress in chickpea (cv. Punjab, 2008). The priming treatments involved soaking seeds of chickpea cultivar Punjab 2008 in either water for 8 h (on-farm priming), aerated water (hydropriming) for 18 h, or CaCl2 solution (ψs -1.25 MPa; osmopriming) for 18 h. Primed and untreated seeds were grown either at 18/15 °C (control) or 13/10 °C (chilling stress). Chilling stress suppressed the growth of chickpea while seed priming mitigated the adverse effects of chilling stress by improving stand establishment, growth, water relations, photosynthesis, α-amylase activity, sugar metabolism, antioxidant enzyme activity, membrane stability, and leaf accumulation of proline, nitrogen, potassium and soluble phenolics. Seed priming also improved the performance of chickpea under optimal (control) conditions. The overall order of improvement in resistance to chilling by using seed priming was osmopriming > hydropriming > on-farm priming. Osmopriming improved seedling dry weight, specific leaf area, leaf CO2 net assimilation rate, maximal photochemical efficiency of PSII, α-amylase activity, trehalose content and leaf relative water content by 10, 22, 17, 20, 73, 48 and 7%, respectively, relative to the non-primed control under chilling stress. Under optimal temperature conditions, the corresponding values were 30, 32, 16, 10, 83, 75 and 5%, respectively. Sugar metabolism, especially trehalose content, was strongly linked with stand establishment, photosynthesis, antioxidant potential (under chilling stress) and plant biomass. Overall, seed priming improved chickpea performance under both optimal temperature conditions and chilling stress through better germination metabolism and the accumulation of trehalose, which protected from oxidative damage and helped to maintain carbon assimilation and seedling growth.

Faba bean drought responsive gene identification and validation.

This study was carried out to identify drought-responsive genes in a drought tolerant faba bean variety (Hassawi 2) using a suppressive subtraction hybridization approach (SSH). A total of 913 differentially expressed clones were sequenced from a differential cDNA library that resulted in a total of 225 differentially expressed ESTs. The genes of mitochondrial and chloroplast origin were removed, and the remaining 137 EST sequences were submitted to the gene bank EST database (LIBEST_028448). A sequence analysis identified 35 potentially drought stress-related ESTs that regulate ion channels, kinases, and energy production and utilization and transcription factors. Quantitative PCR on Hassawi 2 genotype confirmed that more than 65% of selected drought-responsive genes were drought-related. Among these induced genes, the expression levels of eight highly up-regulated unigenes were further analyzed across 38 selected faba bean genotypes that differ in their drought tolerance levels. These unigenes included ribulose 1,5-bisphosphate carboxylase (rbcL) gene, non-LTR retroelement reverse related, probable cyclic nucleotide-gated ion channel, polyubiquitin, potassium channel, calcium-dependent protein kinase and putative respiratory burst oxidase-like protein C and a novel unigene. The expression patterns of these unigenes were variable across 38 genotypes however, it was found to be very high in tolerant genotype. The up-regulation of these unigenes in majority of tolerant genotypes suggests their possible role in drought tolerance. The identification of possible drought responsive candidate genes in Vicia faba reported here is an important step toward the development of drought-tolerant genotypes that can cope with arid environments.

Assessment of genetic diversity of sesame accessions collected from different ecological regions using sequence-related amplified polymorphism markers.

Sequence-related amplified polymorphism (SRAP) markers were used to assess the genetic diversity among a collection of 52 sesame accessions representing different geographical environments, including eight Saudi landraces. A combination of seventeen primers generated a high number of alleles (365) with 100% polymorphism. The polymorphic information content (PIC) and primer discrimination power (DP) recorded overall means of 0.88 and 5.88, respectively. Genetic similarity values based on Jaccard coefficients ranged from 0.12 to 0.49, with an average similarity value of 0.30, indicating both high genetic distance and a wide genetic basis of the investigated accessions. The unweighted pair group method with arithmetic mean (UPGMA) dendrogram grouped 48 of 52 accessions into seven main clusters, and five accessions failed to form clusters and were separated individually. However, subclusters separated the accessions and, considering the relatedness of accessions and their geographical origin, formed distinct diversity among groups. Saudi landraces showed the widest genetic basis compared with other introduced accessions that were distributed throughout the dendrogram, indicating that agro-ecological zones were indistinguishable by cluster analysis. SRAP analysis revealed a high degree of genetic polymorphism in sesame accessions investigated and showed weak association between geographical origin and SRAP patterns. This wide genetic variability should be considered for sesame breeding programs.