Banana fruit bruise detection using fractal dimension based image processing.

The study used the fractal dimension (FD), browning incidence, and grayscale values using machine vision to describe the bruise magnitude and quality of mechanically damaged 'Fard' bananas bruised from 20, 40, 60 cm drop heights by 66, 98, and 110 g ball weights conditioned at different storage temperatures (5, 13, 22 °C) after 48 h. Conventional analyses like bruise area (BA), bruise volume (BV), and bruise susceptibility (BS) were also conducted. A correlation was performed to determine the relationship between image processing and conventional assessment of bruise damage in bananas. Weight, firmness, color, sugar content, and acidity were investigated. The results demonstrated that bananas bruised from the highest force and stored at 5 and 22 °C reported the lowest FD with values of 1.7162 and 1.7403, respectively. Increasing the level of damage reduced the fractal dimension and grayscale values and increased browning incidence and bruise susceptibility values after 48 h of storage. The total color change values showed a strong Pearson's correlation coefficient (r≥-0.81) with image analysis fractal dimension and grayscale values. The findings also indicated that higher bruising and temperature can induce weight loss, firmness reduction, lightness, and yellowness increment, and sugar and acidity changes. Overall, the fractal image analysis conducted in this study was highly effective in describing the bruising magnitude of bananas under different conditions.

Effect on physiological properties of banana fruit based on pendulum impact test and storage.

This study investigated the effect of mechanical damage of different impact energies (0.074 J, 0.160 J, 0.273 J) generated by a pendulum impactor and evaluated the bruise intensity and quality changes of bananas after 12 days of storage at 5, 13, and 22 °C. The bruise area (BA) and bruise susceptibility (BS) in the banana fruit were significantly increased as impact damage increased. The results of weight loss % and electrolyte leakage (EL%) were highly dependent on all studied factors; in the order of storage temperature > storage days > impact level. Storage temperature and impact damage accelerated the incidence of transpiration rate (TRm) during storage. Regarding color measurements, the results of image processing emphasized that storage at 13 °C exhibited the ideal color change in bruised fruit, which revealed a slow increment in lightness (L*) and yellowness (b*) until the last day of storage. The ethylene production rate (EPR) was recorded at its maximum peak on days 2 and 12 with values of 3.85 × 10-3 and 3.08 × 10-3 mg kg-1h-1 in banana fruit bruised by high-impact (0.273 J) and stored at 22 °C. Regardless of impact level, all size measurements reduction % including, surface area (As), geometric mean diameter (Dg), and fruit volume (Vf) of bruised bananas were higher at ambient storage conditions than those stored at 5 and 13 °C. The results of regression analysis can confirm the possibility of bruise susceptibility (BS) to estimate the quality changes of bruised fruit during storage.

Genome-Wide Analysis and Expression Profiling of DUF668 Genes in Glycine max under Salt Stress.

The DUF668 gene performs a critical role in mitigating the impact of abiotic stress factors. In this study, we identified 30 DUF668 genes in a soybean genome, distributed across fifteen chromosomes. The phylogenetic analysis classified the DUF668 genes into three groups (group I, group II, and group III). Interestingly, gene structure analysis illustrated that several GmDUF668 genes were without introns. Furthermore, the subcellular localization results suggested that GmDUF668 proteins were present in the nucleus, mitochondria, cytoplasm, and plasma membrane. GmDUF668 promoters were analyzed in silico to gain insight into the presence of regulatory sequences for TFs binding. The expression profiling illustrated that GmDUF668 genes showed expression in leaves, roots, nodules, and flowers. To investigate their response to salt stress, we utilized the RNA sequencing data of GmDUF668 genes. The results unveiled that GmDUF668-8, GmDUF668-20, and GmDUF668-30 genes were upregulated against salt stress treatment. We further validated these findings using qRT-PCR analysis. These findings provide a scientific basis to explore the functions of GmDUF668 genes against different stress conditions.

Physiological Response of Stored Pomegranate Fruit Affected by Simulated Impact.

Mechanical damage resulting from excessive impact force during handling and other postharvest operations from harvesting to consumption is a critical quality problem in fresh produce marketing. The study investigates the impact of bruise damage, storage temperature, and storage period on the physiological responses of Omani pomegranate fruit cultivar 'Helow'. Fruits were subjected to low (45°; 1.18 J) and high (65°; 2.29 J) impact levels using a pendulum test by hitting the fruit on the cheek side. Bruised and non-bruised fruit were stored at 5 and 22 °C for 28 days. Bruise measurements, water loss per unit mass, water loss per surface area, firmness, fruit size measurements, geometric mean diameter, surface area, fruit volume, color parameters, respiration rate, and ethylene production rate were evaluated. Bruise area, bruise volume, and bruise susceptibility of damaged pomegranate fruit were increased as impact level, storage duration, and storage temperature increased. Pomegranates damaged at a high impact level and conditioned at 22 °C showed 20.39% weight loss on the last day of storage compared to the control and low-impact-bruised fruit. Firmness and geometric mean diameter were significantly (p < 0.05) reduced by bruising at a high impact level. Impact bruising level and storage temperature decreased lightness, yellowness, browning index, and increased redness over time. Furthermore, the respiration rate was five times higher in the non-bruised and low- and high-impact-injured fruit stored at 22 °C than that stored at 5 °C. The ethylene production rate recorded its highest value on day 21 in high-level-impact-bruised pomegranate fruit. The bruise susceptibility was strongly correlated with the majority of the studied parameters. This study can confirm that bruising can affect not only the visual quality characteristics but also the physiological attributes of pomegranate fruit; therefore, much care is required to preserve fresh produce and avoid any mechanical damage and losses during postharvest handling.

Gamma rays induced genetic variability in tomato (Solanum lycopersicum L.) germplasm.

The present study assessed the effectiveness of gamma radiation in inducing favorable genetic variability in tomato (Solanum lycopersicum L.). An experiment was conducted in a randomized complete block design to produce M1 generation. Significant differences were observed among the genotypes as well as between the treatments at individual plant level based on observed traits (seed germination percentage, seedling survival, plant height, number of flower clusters plant-1, number of flowers and fruits plant-1). All observed characters in the mutagenized population were adversely affected with increasing radiation dose. Results identified 450 Gy as the most damaging radiation dose followed by 300 Gy and 150 Gy. Moreover, 300 Gy treatment was identified as lethal dose (LD50) as it caused a 50% germination inhibition in almost all the evaluated genotypes. The 150 Gy treatment showed the least damaging impact and induced maximum genetic variability in almost all the genotypes under study. Character association studies were also conducted which could be utilized in the selection of desirable mutants. Correlation studies revealed an altered association among the observed parameters from positive to negative direction in 300 Gy and 450 Gy treatments as compared to control. These deviations in correlation coefficients proved that mutagenesis can break the linkage among specific loci. Furthermore, path coefficient analysis identified the growth attributes with an effective direct and indirect contribution in yield.

Corrigendum: Genome-Wide Identification and Expression Profiling of Germin-Like Proteins Reveal Their Role in Regulating Abiotic Stress Response in Potato.

[This corrects the article DOI: 10.3389/fpls.2021.831140.].

Protective effect of jasmonic acid and potassium against cadmium stress in peas (Pisum sativum L.).

A combination of mineral nutrients and plant growth regulators should be assessed to improve crop performance under various abiotic stresses. There is a need to include plant growth regulators in fertilization regime of various crops along with essential mineral nutrients, especially when they are irrigated with polluted water with higher levels of heavy metals. The performance of pea was evaluated under cadmium (Cd) stress coupled with potassium (K) and jasmonic acid (JA) supplementation. The Cd stress (50 µM) was applied to soil (sandy loam) grown pea plants as basal dose after a month of sowing. The control and stressed plants were then supplemented with K (5 M), JA (0.5 mM) and their collective application along with control as distilled water. Cd stress showed a marked reduction in growth pattern, however, the collective supplementation sufficiently improved the growth pattern of stressed peas plants as evidenced by improvement in shoot length (cm), root length (cm), number of leaves per plant, leaf area (cm2), plant fresh and dry weight (gm). Potassium application under Cd stress significantly enhanced internodal distance (cm) while the number of seeds per pod and relative water contents remained nonsignificant. The applied treatment (JA + K) under Cd stress prominently improved enzymatic activities, which were measured as nitrate reductase activity (NRA), nitrite reductase activity (NiRA), superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT). Cd stress impacted the biochemical profile by enhancing antioxidant capacity (AC), antioxidant activity (AA), total phenols (TP), while reducing total soluble protein (TSP), chlorophyll 'a', chlorophyll 'b' and carotenoids. The combined application of JA and K under Cd stress enhanced AC, AA, TP, Chl a and b, TSP and carotenoids. The results indicate that foliar application of JA and K efficiently negated the harmful effects of Cd stress on peas.

Seed priming with growth regulators modulates production, physiology and antioxidant defense of Indian squash (Praecitrullus fistulosus) under semi-arid conditions.

Indian squash (Praecitrullus fistulosus) crop faces heat and drought during its growth that is considered the most important abiotic stress in semi-arid areas. Seed priming with growth regulators enhances stress tolerance; hence, mitigates the adverse effects of unpredictable stresses due to adverse weather conditions. This two-year (2019 and 2020) study was conducted to infer the role of seed priming in improving heat tolerance of Indian squash (cultivar Sahavi) through improvement in physiological and antioxidant defense systems. Six treatments that included no priming (control), hydropriming, priming with indole acetic acid (IAA) at 100 mg L-1, salicylic acid (SA) at 50 mg L-1, ascorbic acid (AA) at 100 mg L-1 and thiourea at 500 mg L-1 each for 06 hours) were included in the study. Results revealed that priming with AA and SA significantly (P ≤ 0.05) enhanced germination (39 and 47%), germination index (57 and 58%), plant height (23 and 22%), vine length (15 and 14%), number of fruits per plant (62%), fruit weight per plant (66 and 67%), economic yield (32%), photosynthesis rate (18 and 17%), protein content (10%), proline (23%), glycine betaine (3%), malondialdehyde content (11 and 10%) and catalase activity (24%) compared to control treatment. Furthermore, seed priming with AA and SA significantly (P ≤ 0.05) shortened the mean germination time (25 and 28%) compared to the control. The results indicated that AA and SA had significant potential to mitigate adverse effects of heat stress in Indian squash. Findings from this study showed that seed priming with AA and SA promoted heat-stress tolerance and enhanced growth and productivity of Indian squash.

Thin layer drying kinetics and quality dynamics of persimmon (Diospyros kaki) treated with preservatives and solar dried under different temperatures.

Poor postharvest handling, microbial infestation, and high respiration rate are some the factors are responsible for poor storage life of perishable commodities. Therefore, effective preservation of these commodities is needed to lower the damages and extend shelf life. Preservation is regarded as the action taken to maintain desired properties of a perishable commodity as long as possible. Persimmon (Diospyros kaki) is perishable fruit with high nutritive value; however, has very short shelf-life. Therefore, effective preservation and drying is needed to extend its storage life. Drying temperature and preservatives significantly influence the quality of perishable vegetables and fruits during drying. The current study investigated the effect of different temperatures and preservatives on drying kinetics and organoleptic quality attributes of persimmon. Persimmon fruits were treated with preservatives (25% honey, 25% aloe vera, 2% sodium benzoate, 1% potassium metabisulfite, and 2% citric acid solutions) under different drying temperatures (40, 45, and 50°C). All observed parameters were significantly affected by individual effects of temperatures and preservatives, except ash contents. Similarly, interactive effects were significant for all parameters except total soluble sugars, ash contents, and vitamin C. Generally, fruits treated with citric acid and dried under 50°C had 8.2% moisture loss hour-1, 14.9 drying hours, 0.030 g H2O g-1 hr-1, 1.23° Brix of total soluble solids, 6.71 pH, 1.35% acidity, and 6.3 mg vitamin C. These values were better than the rest of the preservatives and drying temperatures used in the study. Therefore, treating fruits with citric acid and drying at 50°C was found a promising technique to extend storage life of persimmon fruits. It is recommended that persimmon fruits dried at 50°C and preserved in citric acid can be used for longer storage period.

Expression profiling of pathogenesis-related Protein-1 (PR-1) genes from Solanum tuberosum reveals its critical role in phytophthora infestans infection.

Pathogen-related (PR) proteins are an integral part of plants' defense mechanisms against various types of biotic and abiotic stresses. A little is known about the importance of these PR proteins in potato defense mechanisms. In the current study, a total of 22 pathogenesis-related 1 genes were identified in the potato genome. All identified proteins possessed the CAP superfamily domain with some other motifs. The cis-acting elements analysis identified several stress-responsive elements, including MYB, ABRE, and MeJRE. The gene duplication events demonstrated purifying and positive selection pressure. Expression profiling showed high transcripts level in root compared to other tissues; however, some genes have tissue-specific expression. Furthermore, the PR-1-5 gene is transcriptionally induced under Phytophthora infestans stress and hormonal (ABA and IAA) treatments. The Real-Time qPCR analysis also validated the RNA-seq data results of genes with maximum expression in roots compared to leaves and stems. The current study results provided basic data for functional characterization and can also use as a reference study for other important crops.

Genome-Wide Identification and Expression Profiling of DUF221 Gene Family Provides New Insights Into Abiotic Stress Responses in Potato.

The domain of the unknown function 221 proteins regulate several processes in plants, including development, growth, hormone transduction mechanism, and abiotic stress response. Therefore, a comprehensive analysis of the potato genome was conducted to identify the deafness-dystonia peptide (DDP) proteins' role in potatoes. In the present study, we performed a genome-wide analysis of the potato domain of the unknown function 221 (DUF221) genes, including phylogenetic inferences, chromosomal locations, gene duplications, gene structures, and expression analysis. In our results, we identified 10 DDP genes in the potato genome. The phylogenetic analysis results indicated that StDDPs genes were distributed in all four clades, and clade IV was the largest clade. The gene duplication under selection pressure analysis indicated various positive and purifying selections in StDDP genes. The putative stu-miRNAs from different families targeting StDDPs were also predicted in the present study. Promoter regions of StDDP genes contain different cis-acting components involved in multiple stress responses, such as phytohormones and abiotic stress-responsive factors. The analysis of the tissue-specific expression profiling indicated the StDDPs gene expression in stem, root, and leaf tissues. We subsequently observed that StDDP4, StDDP5, and StDDP8 showed higher expressions in roots, stems, and leaves. StDDP5 exhibited high expression against heat stress response, and StDDP7 showed high transcript abundance against salt stress in potatoes. Under abscisic acid (ABA) and indole acetic acid (IAA) treatments, seven StDDP genes' expressions indicated that ABA and IAA performed important roles in immunity response. The expression profiling and real-time qPCR of stems, roots, and leaves revealed StDDPs' significant role in growth and development. These expression results of DDPs are primary functional analysis and present basic information for other economically important crops.

Genome-Wide Identification and Expression Profiling of Germin-Like Proteins Reveal Their Role in Regulating Abiotic Stress Response in Potato.

Germin and germin-like proteins (GLPs) perform a significant role in plants against biotic and abiotic stress. To understand the role of GLPs in potato, a comprehensive genome-wide analysis was performed in the potato genome. This study identified a total of 70 StGLPs genes in the potato genome, distributed among 11 chromosomes. Phylogenetic analysis exhibited that StGLPs were categorized into six groups with high bootstrap values. StGLPs gene structure and motifs analysis showed a relatively well-maintained intron-exon and motif formation within the cognate group. Additionally, several cis-elements in the promoter regions of GLPs were hormones, and stress-responsive and different families of miRNAs target StGLPs. Gene duplication under selection pressure also exhibited positive and purifying selections in StGLPs. In our results, the StGLP5 gene showed the highest expression in response to salt stress among all expressed StGLPs. Totally 19 StGLPs genes were expressed in response to heat stress. Moreover, three genes, StGLP30, StGLP17, and StGLP14, exhibited a relatively higher expression level in the potato after heat treatment. In total, 22 genes expressed in response to abscisic acid (ABA) treatment indicated that ABA performed an essential role in the plant defense or tolerance mechanism to environmental stress. RNA-Seq data validated by RT-qPCR also confirm that the StGLP5 gene showed maximum expression among selected genes under salt stress. Concisely, our results provide a platform for further functional exploration of the StGLPs against salt and heat stress conditions.

Association of the 16SrII-D Phytoplasma with African Marigold (Tagetes erecta) Phyllody in Oman.

The African marigold (Tagetes erecta L.) is an ornamental, herbaceous plant commonly found in Oman. In 2019, African marigold plants showing phyllody and virescence symptoms, which are typical symptoms of phytoplasmas disease, were found in at Sultan Qaboos University in Oman. Transmission electron microscopy of marigold leaf midrib from phyllody disease plants showed the presence of numerous phytoplasma bodies in the sieve tube of all of the symptomatic samples. DNA was extracted from asymptomatic and symptomatic marigold plant samples, followed by PCR of the 16S ribosomal RNA (rRNA) and imp genes. The PCR assays showed that the symptomatic plants are positive for phytoplasma. The DNA sequence analysis and phylogenetic trees showed that the 16S rDNA and imp gene sequences from all marigold phyllody strains shared 100% sequence identity to 16SrII-D subgroup sequences in the GenBank. This is the first report of a phytoplasma of the 16SrII-D subgroup associated with the African marigold (T. erecta) worldwide.

Molecular Characterization of a Date Palm Vascular Highway 1-Interacting Kinase (PdVIK) Under Abiotic Stresses.

The date palm (Khalas) is an extremophile plant that can adapt to various abiotic stresses including drought and salinity. Salinity tolerance is a complex trait controlled by numerous genes. Identification and functional characterization of salt-responsive genes from the date palm is fundamental to understand salinity tolerance at the molecular level in this plant species. In this study, a salt-inducible vascular highway 1-interacting kinase (PdVIK) that is a MAP kinase kinase kinase (MAPKKK) gene from the date palm, was functionally characterized using in vitro and in vivo strategies. PdVIK, one of the 597 kinases encoded by the date palm genome possesses an ankyrin repeat domain and a kinase domain. The recombinant PdVIK protein exhibited phosphotyrosine activity against myelin basic protein (MBP) substrate. Overexpression of PdVIK in yeast significantly improved its tolerance to salinity, LiCl, and oxidative stresses. Transgenic Arabidopsis seedlings overexpressing PdVIK displayed improved tolerance to salinity, osmotic, and oxidative stresses as assessed by root growth assay. The transgenic lines grown in the soil also displayed modulated salt response, compared to wild-type controls as evaluated by the overall plant growth and proline levels. Likewise, the transgenic lines exhibited drought tolerance by maintaining better relative water content (RWC) compared to non-transgenic control plants. Collectively, these results implicate the involvement of PdVIK in modulating the abiotic stress response of the date palm.

A novel tonoplast Na+/H+ antiporter gene from date palm (PdNHX6) confers enhanced salt tolerance response in Arabidopsis.

KEY MESSAGE: A sodium hydrogen exchanger (NHX) gene from the date palm enhances tolerance to salinity in Arabidopsis plants. Plant sodium hydrogen exchangers/antiporters (NHXs) are pivotal regulators of intracellular Na+/K+ and pH homeostasis, which is essential for salt stress adaptation. In this study, a novel orthologue of Na+/H+ antiporter was isolated from date palm (PdNHX6) and functionally characterized in mutant yeast cells and Arabidopsis plants to assess the behavior of the transgenic organisms in response to salinity. Genetically transformed yeast cells with PdNHX6 were sensitive to salt stress when compared to the empty vector (EV) yeast cells. Besides, the acidity value of the vacuoles of the transformant yeast cells has significantly (p ≤ 0.05) increased, as indicated by the calibrated fluorescence intensity measurements and the fluorescence imagining analyses. This observation supports the notion that PdNHX6 might regulate proton pumping into the vacuole, a crucial salt tolerance mechanism in the plants. Consistently, the transient overexpression and subcellular localization revealed the accumulation of PdNHX6 in the tonoplast surrounding the central vacuole of Nicotiana benthamiana leaf epidermal cells. Stable overexpression of PdNHX6 in Arabidopsis plants enhanced tolerance to salt stress and retained significantly higher chlorophyll, water contents, and increased seed germination under salinity when compared to the wild-type plants. Despite the significant increase of Na+, transgenic Arabidopsis lines maintained a balanced Na+/K+ ratio under salt stress conditions. Together, the results obtained from this study imply that PdNHX6 is involved in the salt tolerance mechanism in plants by controlling K+ and pH homeostasis of the vacuoles.

An endophytic Talaromyces omanensis enhances reproductive, physiological and anatomical characteristics of drought-stressed tomato.

The effects of a newly discovered endophytic fungus, Talaromyces omanensis, on the drought tolerance of tomato is presented in this study. The fungus was obtained from a desert plant Rhazya stricta in Oman. Drought stress was induced by a 15% solution of Polyethylene glycol-6000 (PEG-6000). Several parameters were measured including pollen sterility, pollen tube length, growth, flowering, and yield characteristics, the biochemical analysis of the leaves and fruits, as well as other physiological and anatomical parameters. The results showed that T. omanensis provided multiple advantages to tomato grown under drought stress, including improved reproductive characteristics, chlorophyll fluorescence, and some anatomical characteristics such as increased phloem and cortex width and a reduction of pith autolysis that leads to hollow stem. In addition, T. omanensis significantly increased drought-stress related characteristics such as shoot dry weight, root length, the number of flowers, and fruit weight. A significantly higher concentration of gibberellic acid (GA3) was found in tomato plants treated by T. omanensis, which may enhance their drought tolerance. These results suggest that T. omanensis is a potential biological anti-stress stimulator for important horticultural crops such as tomatoes. This study is the first to report the beneficial effects of T. omanensis in alleviating drought stress in tomatoes.

Metabolomic analysis of date palm seedlings exposed to salinity and silicon treatments.

Silicon is known to promote plant growth as well as stress tolerance of plants. The current study was undertaken to assess the growth promoting effect of silicon on date palm seedling development as well as its ability to abate some of the negative effects of salinity. In this study, date palm seedlings were treated with silicon and sodium chloride salts, and the effect of these salts on some physiological parameters of the plants was determined. In addition, a global nontargeted metabolomics analysis was performed for the leaf and root tissues using liquid chromatography-mass spectrometry (LC-MS). The results showed that under non-stress conditions, silicon treatment enhanced the growth of the date palm seedlings, however, under salinity, silicon slightly mitigates the negative effects of salt stress on the date palm seedlings although it enhances the potassium accumulation under this condition. The global metabolomics analysis has identified a total of 1,101 significant differentially accumulated (p, q ≤ 0.05) metabolites in leaves and roots under silicon, salinity or their combination. A differential pairwise metabolic profile comparison revealed the accumulation of distinct metabolites in response to silicon and salinity treatments such as antioxidant compounds pyridoxine, cepharanthine, allithiamine, myristic acid and boldine; osmoregulators such as mucic acid; along with the accumulation of detoxification intermediates such as S-D-lactoylglutathione, beta-cyano-L-alanine and gamma-glutamyl-conjugates. In addition, histochemical analyses revealed that application of silicon significantly (p ≤ 0.05) enhanced the formation of the Casparian strip. Identification of the differentially accumulated metabolites could offer an insight into how silicon is able to promote growth and salinity tolerance in date palms.

Talaromyces variabilis interferes with Pythium aphanidermatum growth and suppresses Pythium-induced damping-off of cucumbers and tomatoes.

Pythium-induced damping-off disease is a major disease limiting cucumber and tomato production in different parts of the world. The current study investigated the efficiency of Talaromyces variabilis and its bioactive metabolites in suppressing Pythium-induced damping-off of cucumbers and tomatoes. T. variabilis inhibited the in vitro growth of P. aphanidermatum in solid and liquid media. In addition, abnormalities in P. aphanidermatum hyphae were observed as a result of T. variabilis. Extracts from T. variabilis induced cellular leakage and suppressed oospore production of P. aphanidermatum. Biochemical analyses of T. variabilis metabolites showed that T. variabilis produces glucanase, cellulase and siderophores, suggesting the contribution of these metabolites in the inhibition of P. aphandermatum growth and in hyphal abnormalities. Treating cucumber seeds with spore and mycelial suspension of T. variabilis isolates led to a significant improvement in the seedling survival of P. aphanidermatum-inoculated seedlings from 18 to 52% (improvement by 34%) for isolate 48 P and from 30-66% (improvement by 36%) for isolate 28 R. Similarly, treating tomato seeds with spore and mycelial suspension of T. variabilis isolates led to a significant improvement in the seedling survival of P. aphanidermatum-inoculated seedlings from 7 to 36% (improvement by 29%) for isolate 28 R and from 20 to 64% (improvement by 44%) for isolate 48 P. Differences in the percent improvement in seedling survival between experiments may be related to difference in the efficacy of the two different isolates or their interaction with the hosts and pathogen. The use of T. variabilis in the biocontrol of Pythium-induced diseases may offer alternatives to the currently used chemical control.

Overexpression of a Metallothionein 2A Gene from Date Palm Confers Abiotic Stress Tolerance to Yeast and Arabidopsis thaliana.

Although the date palm tree is an extremophile with tolerance to drought and certain levels of salinity, the damage caused by extreme salt concentrations in the soil, has created a need to explore stress-responsive traits and decode their mechanisms. Metallothioneins (MTs) are low-molecular-weight cysteine-rich proteins that are known to play a role in decreasing oxidative damage during abiotic stress conditions. Our previous study identified date palm metallothionein 2A (PdMT2A) as a salt-responsive gene, which has been functionally characterized in yeast and Arabidopsis in this study. The recombinant PdMT2A protein produced in Escherichia coli showed high reactivity against the substrate 5'-dithiobis-2-nitrobenzoic acid (DTNB), implying that the protein has the property of scavenging reactive oxygen species (ROS). Heterologous overexpression of PdMT2A in yeast (Saccharomyces cerevisiae) conferred tolerance to drought, salinity and oxidative stresses. The PdMT2A gene was also overexpressed in Arabidopsis, to assess its stress protective function in planta. Compared to the wild-type control, the transgenic plants accumulated less Na+ and maintained a high K+/Na+ ratio, which could be attributed to the regulatory role of the transgene on transporters such as HKT, as demonstrated by qPCR assay. In addition, transgenic lines exhibited higher chlorophyll content, higher superoxide dismutase (SOD) activity and improved scavenging ability for reactive oxygen species (ROS), coupled with a better survival rate during salt stress conditions. Similarly, the transgenic plants also displayed better drought and oxidative stress tolerance. Collectively, both in vitro and in planta studies revealed a role for PdMT2A in salt, drought, and oxidative stress tolerance.

Functional Characterization of Date Palm Aquaporin Gene PdPIP1;2 Confers Drought and Salinity Tolerance to Yeast and Arabidopsis.

Recent studies on salinity tolerance in date palm revealed the discovery of salt-responsive genes including PdPIP1;2, a highly conserved aquaporin gene in plants, which was functionally characterized in this study to investigate its precise role in drought and salinity tolerance. Immunoblot assay showed a high level of PIP1 protein accumulation only in the leaves of date palm plants when grown under drought, an observation which may imply the involvement of PIP1;2 in CO2 uptake. Heterologous overexpression of PdPIP1;2 in yeast (Saccharomyces cerevisiae) improved tolerance to salinity and oxidative stress. While, heterologous overexpression of PdPIP1;2 in Arabidopsis had significantly (p < 0.05) increased biomass, chlorophyll content, and root length under drought and salinity. In addition, a significantly (p < 0.05) higher percentage of transgenic plants could be recovered by rewatering after drought stress, indicating the ability of the transgenic plants to maintain water and viability under drought. Transgenic plants under drought and salinity maintained significantly (p < 0.05) higher K+/Na+ ratios than wild type (WT) plants, an observation which may represent an efficient tolerance mechanism controlled by the transgene. Collectively, this study provided an insight on the mechanism by which PdPIP1;2 conferred tolerance to salt and drought stresses in date palm.