Rashid Al-Yahyai

The Role of Na+ and K+ Transporters in Salt Stress Adaptation in Glycophytes

Ionic stress is one of the most important components of salinity and is brought about by excess Na+ accumulation, especially in the aerial parts of plants. Since Na+ interferes with K+ homeostasis, and especially given its involvement in numerous metabolic processes, maintaining a balanced cytosolic Na+/K+ ratio has become a key salinity tolerance mechanism. Achieving this homeostatic balance requires the activity of Na+ and K+ transporters and/or channels. The mechanism of Na+ and K+ uptake and translocation in glycophytes and halophytes is essentially the same, but glycophytes are more susceptible to ionic stress than halophytes. The transport mechanisms involve Na+ and/or K+ transporters and channels as well as non-selective cation channels. Thus, the question arises of whether the difference in salt tolerance between glycophytes and halophytes could be the result of differences in the proteins or in the expression of genes coding the transporters. The aim of this review is to seek answers to this question by examining the role of major Na+ and K+ transporters and channels in Na+ and K+ uptake, translocation and intracellular homeostasis in glycophytes. It turns out that these transporters and channels are equally important for the adaptation of glycophytes as they are for halophytes, but differential gene expression, structural differences in the proteins (single nucleotide substitutions, impacting affinity) and post-translational modifications (phosphorylation) account for the differences in their activity and hence the differences in tolerance between the two groups. Furthermore, lack of the ability to maintain stable plasma membrane (PM) potentials following Na+-induced depolarization is also crucial for salt stress tolerance. This stable membrane potential is sustained by the activity of Na+/H+ antiporters such as SOS1 at the PM. Moreover, novel regulators of Na+ and K+ transport pathways including the Nax1 and Nax2 loci regulation of SOS1 expression and activity in the stele, and haem oxygenase involvement in stabilizing membrane potential by activating H+-ATPase activity, favorable for K+ uptake through HAK/AKT1, have been shown and are discussed.

A genome-wide identification of the miRNAome in response to salinity stress in date palm (Phoenix dactylifera L.)

Although date palm is relatively salt-tolerant, little is known about the underlying molecular mechanisms that contribute to its salt tolerance. Only recently, investigators have uncovered microRNA-mediated post-transcriptional gene regulation, which is critical for typical plant development and adaptation to stress conditions such as salinity. To identify conserved and novel miRNAs in date palm and to characterize miRNAs that could play a role in salt tolerance, we have generated sRNA libraries from the leaves and roots of NaCl-treated and untreated seedlings of date palm. Deep sequencing of these four sRNA libraries yielded approximately 251 million reads. The bioinformatics analysis has identified 153 homologs of conserved miRNAs, 89 miRNA variants, and 180 putative novel miRNAs in date palm. Expression profiles under salinity revealed differential regulation of some miRNAs in date palm. In leaves, 54 of the identified miRNAs were significantly affected and the majority (70%) of them were upregulated, whereas in roots, 25 of the identified miRNAs were significantly affected and 76% of them were upregulated by the salinity stress. The salt-responsiveness of some of these miRNAs was further validated using semi-quantitative PCR (qPCR). Some of the predicted targets for the identified miRNA include genes with known functions in plant salt tolerance, such as potassium channel AKT2-like proteins, vacuolar protein sorting-associated protein, calcium-dependent and mitogen-activated proteins. As one of the first cultivated trees in the world that can tolerate a wide range of abiotic stresses, date palm contains a large population of conserved and non-conserved miRNAs that function at the post-transcriptional level. This study provided insights into miRNA-mediated gene expression that are important for adaptation to salinity in date palms.

Phytochemical content, antioxidant capacity and physicochemical properties of pomegranate grown in different microclimates in South Africa

We investigated the antioxidant and physicochemical properties of pomegranate (Punica granatum cv. Bhagwa) fruit grown in three microclimates in South Africa. The physicochemical properties of fruit peel and arils differed among the growing locations, including weight and redness color of both peel and arils. Contents of vitamin C, anthocyanin and total phenolics were 0.67–1.41 mg ml−1, 0.07–0.16 mg ml−1 and 8.54–13.91 mg ml−1, respectively. Fruit grown in locations with high light intensity had higher vitamin C content and lower phenolic content than those of fruit from low-altitude locations and high light intensity. The pH of juice did not significantly differ among locations, whereas significant variation was observed in total soluble solids (TSS), titratible acidity (TA) and TSS:TA ratio. Fruit from locations with low rainfall had high TSS and low TA. Principal component analysis was used successfully to classify fruit by growing location on the basis of antioxidant and phytochemical contents.

Sub-optimal irrigation affects chemical quality attributes of dates during fruit development

Date palm ( Phoenix dactylifera L.) is primarily cultivated in the regions of the Middle East and North Africa that are characterized by high evapotranspiration, low and sporadic rainfall and dependency on irrigation for crop cultivation. The effects of sub-optimal irrigation on the chemical properties of dates were investigated. Chemical quality attributes of date palm (cv. Khalas) grown in northern Oman showed variable responses to reduced frequency of irrigation water applied during fruit development. Trees irrigated on a daily basis showed a significantly higher fruit water content, juice volume, and titratable acidity (TA). Deficit irrigation treatments had the highest total-and reducing sugar content. Trees under water stress that were irrigated once per week showed significantly high dry matter, total soluble solids (TSS) and pectin. Sucrose, tannin and pH were not significantly different among irrigation treatments. Manipulation of irrigation frequency can be used to enhance certain chemical fruit quality attributes of dates.

Date Palm Status and Perspective in Oman

Date palm is the primary agricultural crop in Oman, and it constitutes 80 % of all fruit crops produced and 50 % of the total agricultural area in the country. Oman is the eighth largest producer of dates in the world with an average annual production of 260,000 mt per annum. There are approximately more than over seven million date palms and 250 cultivars in cultivation, primarily in the northern governorates of the sultanate. However, around 70 % of the total date production is harvested from only 10 cultivars, and a small fraction (2.6 %) of the total date production is exported. Only half of the dates produced are used for human consumption, with the other half being utilized primarily for animal feed or considered surplus and wasted. Dates are mainly harvested for fresh fruit consumption; however, alternative uses such as date syrup, date sugar, and other by-products can also be found in the local market. Dubas bug and red palm weevil are the dominant biotic factors that affect date quality and yield in Oman. Traditional methods of cultivation, small farm size, enough labor available, and poor postharvest handling and marketing are the main issues that face date palm production in Oman. New plantlets are produced from tissue culture with a primary focus on superior cultivars that are kept, among other cultivars, in the only date palm ex situ gene bank in the interior of Oman. Enhancing fruit quality by optimizing fruit size and nutritional content and rapid cultivar selectivity based on molecular techniques for better or improved commercial cultivars will increase the marketability of Omani dates. Furthermore, employing modern orchard layouts and mechanization of the labor-dependent cultural practices such as irrigation, pruning, pollination, and harvesting is vital for the sustainable and profitable production of dates in Oman.

Differential DNA methylation and transcription profiles in date palm roots exposed to salinity

As a salt-adaptive plant, the date palm (Phoenix dactylifera L.) requires a suitable mechanism to adapt to the stress of saline soils. There is growing evidence that DNA methylation plays an important role in regulating gene expression in response to abiotic stresses, including salinity. Thus, the present study sought to examine the differential methylation status that occurs in the date palm genome when plants are exposed to salinity, and to identify salinity responsive genes that are regulated by DNA methylation. To achieve these, whole-genome bisulfite sequencing (WGBS) was employed and mRNA was sequenced from salinity-treated and untreated roots. The WGBS analysis included 324,987,795 and 317,056,091 total reads of the control and the salinity-treated samples, respectively. The analysis covered about 81% of the total genomic DNA with about 40% of mapping efficiency of the sequenced reads and an average read depth of 17-fold coverage per DNA strand, and with a bisulfite conversion rate of around 99%. The level of methylation within the differentially methylated regions (DMRs) was significantly (p < 0.05, FDR ≤ 0.05) increased in response to salinity specifically at the mCHG and mCHH sequence contexts. Consistently, the mass spectrometry and the enzyme-linked immunosorbent assay (ELISA) showed that there was a significant (p < 0.05) increase in the global DNA methylation in response to salinity. mRNA sequencing revealed the presence of 6,405 differentially regulated genes with a significant value (p < 0.001, FDR ≤ 0.05) in response to salinity. Integration of high-resolution methylome and transcriptome analyses revealed a negative correlation between mCG methylation located within the promoters and the gene expression, while a positive correlation was noticed between mCHG/mCHH methylation rations and gene expression specifically when plants grew under control conditions. Therefore, the methylome and transcriptome relationships vary based on the methylated sequence context, the methylated region within the gene, the protein-coding ability of the gene, and the salinity treatment. These results provide insights into interplay among DNA methylation and gene expression, and highlight the effect of salinity on the nature of this relationship, which may involve other genetic and epigenetic players under salt stress conditions. The results obtained from this project provide the first draft map of the differential methylome and transcriptome of date palm when exposed to an abiotic stress.

Classification of dates varieties and effect of motion blurring on standardized moment features

Computer vision technology has been used as a successful non-destructive quality assessment tool for various food products. In general, several features are extracted from the images of interest, and used for the classification models. Furthermore, in most of the studies, static images have been used in the calibration and evaluation models. Classification models with a reduced number of features, and a mechanism to test the capability of the algorithm for moving objects by means of simulating the blurring effect on the static images would be beneficial to determine the performance of the system in real-time quality monitoring in industries. Using three date varieties as model food, motion was simulated for the date’s images and a successful neural network classifier was designed with only three statistical features (mean, standard deviation, and skewness). The reduced number of features and simplicity of the classifier yielded a solution that can be potentially implemented in hardware fast enough so that to consider the case of classification of the dates in a conveyor belt. To test the solution under such conditions, a blurring degradation function was used to verify that the classifier would work. The effects that motion blurring causes to these statistical moments in a general sense were examined using random numbers drawn from the distribution in the Pearson system. Because motion blurring showed a tendency to change the distribution to a Gaussian density, the same features and classifier yielded similar results despite of motion.

Aspergillus terreus Inhibits Growth and Induces Morphological Abnormalities in Pythium aphanidermatum and Suppresses Pythium-Induced Damping-Off of Cucumber

The study investigated the efficacy of two isolates of Aspergillus terreus (65P and 9F) on the growth, morphology and pathogenicity of Pythium aphanidermatum on cucumber. In vitro tests showed that the two isolates inhibited the growth of P. aphanidermatum in culture. Investigating P. aphanidermatum hyphae close to the inhibition zone showed that the hyphae showed abnormal growth and loss of internal content. Treating P. aphanidermatum with the culture filtrate (CF) of A. terreus resulted in significant rise in cellular leakage of P. aphanidermatum mycelium. Testing glucanase enzyme activity by both A. terreus isolates showed a significant increase in glucanase activity. This suggests that the cell walls of Pythium, which consist of glucan, are affected by the glucanase enzyme produced by A. terreus. In addition, Aspergillus isolates produced siderephore, which is suggested to be involved in inhibition of Pythium growth. Also, the CFs of 65P and 9F isolates significantly reduced spore production by P. aphanidermatum compared to the control (P < 0.05). In bioassay tests, the two isolates of A. terreus increased the survival rate of cucumber seedlings from 10 to 20% in the control seedlings treated with P. aphanidermatum to 38–39% when the biocontrol agents were used. No disease symptoms were observed on cucumber seedlings only treated with the isolates 65P and 9F of A. terreus. In addition, the A. terreus isolates did not have any negative effects on the growth of cucumber seedlings. This study shows that isolates of A. terreus can help suppress Pythium-induced damping-off of cucumber, which is suggested to be through the effect of A. terreus and its glucanase enzyme on P. aphanidermatum mycelium.

Classification of a new phytoplasmas subgroup 16SrII-W associated with Crotalaria witches’ broom diseases in Oman based on multigene sequence analysis

BackgroundCrotalaria aegyptiaca, a low shrub is commonly observed in the sandy soils of wadis desert and is found throughout all regions in Oman. A survey for phytoplasma diseases was conducted. During a survey in a wild area in the northern regions of Oman in 2015, typical symptoms of phytoplasma infection were observed on C. aegyptiaca plants. The infected plants showed an excessive proliferation of their shoots and small leaves.ResultsThe presence of phytoplasma in the phloem tissue of symptomatic C. aegyptiaca leaf samples was confirmed by using Transmission Electron Microscopy (TEM). In addition the extracted DNA from symptomatic C. aegyptiaca leaf samples and Orosius sp. leafhoppers were tested by PCR using phytoplasma specific primers for the 16S rDNA, secA, tuf and imp, and SAP11 genes. The PCR amplifications from all samples yielded the expected products, but not from asymptomatic plant samples. Sequence similarity and phylogenetic tree analyses of four genes (16S rDNA, secA, tuf and imp) showed that Crotalaria witches’ broom phytoplasmas from Oman is placed with the clade of Peanut WB (16SrII) close to Fava bean phyllody (16SrII-C), Cotton phyllody and phytoplasmas (16SrII-F), and Candidatus Phytoplasma aurantifolia’ (16SrII-B). However, the Crotalaria’s phytoplasma was in a separate sub-clade from all the other phytoplasmas belonging to Peanut WB group. The combination of specific primers for the SAP11 gene of 16SrII-A, −B, and -D subgroup pytoplasmas were tested against Crotalaria witches’ broom phytoplasmas and no PCR product was amplified, which suggests that the SAP11 of Crotalaria phytoplasma is different from the SAP11 of the other phytoplasmas.ConclusionWe propose to assign the Crotalaria witches’ broom from Oman in a new lineage 16SrII-W subgroup depending on the sequences analysis of 16S rRNA, secA, imp, tuf, and SAP11 genes. To our knowledge, this is the first report of phytoplasmas of the 16SrII group infecting C. aegyptiaca worldwide.

Identification of Reference Genes for Quantitative Real-Time PCR in Date Palm (Phoenix dactylifera L.) Subjected to Drought and Salinity

Date palm is an important crop plant in the arid and semi-arid regions supporting human population in the Middle East and North Africa. These areas have been largely affected by drought and salinity due to insufficient rainfall and improper irrigation practices. Date palm is a relatively salt- and drought-tolerant plant and more recently efforts have been directed to identifying genes and pathways that confer stress tolerance in this species. Quantitative real-time PCR (qPCR) is a promising technique for the analysis of stress-induced differential gene expression, which involves the use of stable reference genes for normalizing gene expression. In an attempt to find the best reference genes for date palm’s drought and salinity research, we evaluated the stability of 12 most commonly used reference genes using the geNorm, NormFinder, BestKeeper statistical algorithms and the comparative ΔCT method. The comprehensive results revealed that HEAT SHOCK PROTEIN (HSP), UBIQUITIN (UBQ) and YTH domain-containing family protein (YT521) were stable in drought-stressed leaves whereas GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE (GAPDH), ACTIN and TUBULIN were stable in drought-stressed roots. On the other hand, SMALL SUBUNIT RIBOSOMAL RNA (25S), YT521 and 18S ribosomal RNA (18S); and UBQ, ACTIN and ELONGATION FACTOR 1-ALPHA (eEF1a) were stable in leaves and roots, respectively, under salt stress. The stability of these reference genes was verified by using the abiotic stress-responsive CYTOSOLIC Cu/Zn SUPEROXIDE DISMUTASE (Cyt-Cu/Zn SOD), an ABA RECEPTOR, and a PROLINE TRANSPORTER 2 (PRO) genes. A combination of top 2 or 3 stable reference genes were found to be suitable for normalization of the target gene expression and will facilitate gene expression analysis studies aimed at identifying functional genes associated with drought and salinity tolerance in date palm.