WRKY transcription factors in plant defense.

Environmental stressors caused by climate change are fundamental barriers to agricultural sustainability. Enhancing the stress resilience of crops is a key strategy in achieving global food security. Plants perceive adverse environmental conditions and initiate signaling pathways to activate precise responses that contribute to their survival. WRKY transcription factors (TFs) are essential players in several signaling cascades and regulatory networks that have crucial implications for defense responses in plants. This review summarizes advances in research concerning how WRKY TFs mediate various signaling cascades and metabolic adjustments as well as how epigenetic modifications involved in environmental stress responses in plants can modulate WRKYs and/or their downstream genes. Emerging research shows that clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas)-mediated genome editing of WRKYs could be used to improve crop resilience.

Exogenous application of salicylic acid ameliorates salinity stress in barley (Hordeum vulgare L.).

Barley (Hordeum vulgare L.) is a significant cereal crop belonging to Poaceae that is essential for human food and animal feeding. The production of barley grains was around 142.37 million tons in 2017/2018. However, the growth of barley was influenced by salinity which was enhanced by applying a foliar spray of salicylic acid. The current study investigated to evaluated the potential effect of SA on the barley (Hordeum vulgare L.) plants under salinity stress and its possible effects on physiological, biochemical, and growth responses. The experiment was conducted at Postgraduate Research Station (PARS), University of Agriculture; Faisalabad to assess the influence of salicylic acid on barley (Hordeum vulgare L.) under highly saline conditions. The experiment was conducted in a Completely Randomized Design (CRD) with 3 replicates. In plastic pots containing 8 kg of properly cleaned sand, two different types of barley (Sultan and Jau-17) were planted. The plants were then watered with a half-strength solution of Hoagland's nutritional solution. After the establishment of seedlings, two salt treatments (0 mM and 120 mM NaCl) were applied in combining three levels of exogenously applied salicylic acid (SA) (0, 0.5, and 1 mg L-1). Data about morphological, physiological, and biochemical attributes was recorded using standard procedure after three weeks of treatment. The morpho-physiological fresh weight of the shoot and root (48%), the dry mass of the shoot and root (66%), the plant height (18%), the chlorophyll a (30%), the chlorophyll b (22%), and the carotenoids (22%), all showed significant decreases. Salinity also decreased yield parameters and the chl. ratio (both at 29% and 26% of the total chl. leaf area index). Compared to the control parameters, the following data was recorded under salt stress: spike length, number of spikes, number of spikelets, number of tillers, biological yield, and harvest index. Salicylic acid was used as a foliar spray to lessen the effects of salinity stress, and 1 mg L-1 of salicylic acid proved more effective than 0.5 mg L-1. Both varieties show better growth by applying salicylic acid (0 mg L-1) as a control, showing normal growth. By increasing its level to (0.5 mg L-1), it shows better growth but maximized growth occurred at a higher level (1 mg L-1). Barley sultan (Hordeum vulgare L.) is the best variety as compared to Jau-17 performs more growth to mitigate salt stress (0mM and 120mM NaCl) by improving morpho-physiological parameters by enhancing plan height, Root and shoot fresh and dry weights, as well as root and shoot lengths, photosynthetic pigments, area of the leaves and their index, and yield attributes and reduce sodium ions.

Effect of salinity stress and surfactant treatment with zinc and boron on morpho-physiological and biochemical indices of fenugreek (Trigonella foenum-graecum).

Micronutrient application has a crucial role in mitigating salinity stress in crop plants. This study was carried out to investigate the effect of zinc (Zn) and boron (B) as foliar applications on fenugreek growth and physiology under salt stress (0 and 120 mM). After 35 days of salt treatments, three levels of zinc (0, 50, and 100 ppm) and two levels of boron (0 and 2 ppm) were applied as a foliar application. Salinity significantly reduced root length (72.7%) and shoot length (33.9%), plant height (36%), leaf area (37%), root fresh weight (48%) and shoot fresh weight (75%), root dry weight (80%) and shoot dry weight (67%), photosynthetic pigments (78%), number of branches (50%), and seeds per pod (56%). Fenugreek's growth and physiology were improved by foliar spray of zinc and boron, which increased the length of the shoot (6%) and root length (2%), fresh root weight (18%), and dry root weight (8%), and chlorophyll a (1%), chlorophyll b (25%), total soluble protein content (3%), shoot calcium (9%) and potassium (5%) contents by significantly decreasing sodium ion (11%) content. Moreover, 100 ppm of Zn and 2 ppm of B enhanced the growth and physiology of fenugreek by reducing the effect of salt stress. Overall, boron and zinc foliar spray is suggested for improvement in fenugreek growth under salinity stress.

Pathogenesis related-1 proteins in plant defense: regulation and functional diversity.

Climate change-related environmental stresses can negatively impact crop productivity and pose a threat to sustainable agriculture. Plants have a remarkable innate ability to detect a broad array of environmental cues, including stresses that trigger stress-induced regulatory networks and signaling pathways. Transcriptional activation of plant pathogenesis related-1 (PR-1) proteins was first identified as an integral component of systemic acquired resistance in response to stress. Consistent with their central role in immune defense, overexpression of PR-1s in diverse plant species is frequently used as a marker for salicylic acid (SA)-mediated defense responses. Recent advances demonstrated how virulence effectors, SA signaling cascades, and epigenetic modifications modulate PR-1 expression in response to environmental stresses. We and others showed that transcriptional regulatory networks involving PR-1s could be used to improve plant resilience to stress. Together, the results of these studies have re-energized the field and provided long-awaited insights into a possible function of PR-1s under extreme environmental stress.

Alleviation potential of green-synthesized selenium nanoparticles for cadmium stress in Solanum lycopersicum L: modulation of secondary metabolites and physiochemical attributes.

KEY MESSAGE: Selenium nanoparticles reduce cadmium absorption in tomato roots, mitigating heavy metal effects. SeNPs can efficiently help to enhance growth, yield, and biomolecule markers in cadmium-stressed tomato plants. In the present study, the effects of selenium nanoparticles (SeNPs) were investigated on the tomato plants grown in cadmium-contaminated soil. Nanoparticles were synthesized using water extract of Nigella sativa and were characterized for their size and shape. Two application methods (foliar spray and soil drench) with nanoparticle concentrations of 0, 100, and 300 mg/L were used to observe their effects on cadmium-stressed plants. Growth, yield, biochemical, and stress parameters were studied. Results showed that SeNPs positively affected plant growth, mitigating the negative effects of cadmium stress. Shoot length (SL), root length (RL), number of branches (NB), number of leaves per plant (NL), and leaf area (LA) were significantly reduced by cadmium stress but enhanced by 45, 51, 506, 208, and 82%, respectively, by soil drench treatment of SeNPs. Similarly, SeNPs increased the fruit yield (> 100%) and fruit weight (> 100%), and decreased the days to fruit initiation in tomato plants. Pigments were also positively affected by the SeNPs, particularly in foliar treatment. Lycopene content was also enhanced by the addition of NPs (75%). Furthermore, the addition of SeNPs improved the ascorbic acid, protein, phenolic, flavonoid, and proline contents of the tomato plants under cadmium stress, whereas stress enzymes also showed enhanced activities under cadmium stress. It is concluded from the present study that the addition of selenium nanoparticles enhanced the growth and yield of Cd-stressed plants by reducing the absorption of cadmium and increasing the stress management of plants.

Pathogenesis-Related 1 (PR1) Protein Family Genes Involved in Sugarcane Responses to Ustilago scitaminea Stress.

Plant resistance against biotic stressors is significantly influenced by pathogenesis-related 1 (PR1) proteins. This study examines the systematic identification and characterization of PR1 family genes in sugarcane (Saccharum spontaneum Np-X) and the transcript expression of selected genes in two sugarcane cultivars (ROC22 and Zhongtang3) in response to Ustilago scitaminea pathogen infection. A total of 18 SsnpPR1 genes were identified at the whole-genome level and further categorized into four groups. Notably, tandem and segmental duplication occurrences were detected in one and five SsnpPR1 gene pairs, respectively. The SsnpPR1 genes exhibited diverse physio-chemical attributes and variations in introns/exons and conserved motifs. Notably, four SsnpPR1 (SsnpPR1.02/05/09/19) proteins displayed a strong protein-protein interaction network. The transcript expression of three SsnpPR1 (SsnpPR1.04/06/09) genes was upregulated by 1.2-2.6 folds in the resistant cultivar (Zhongtang3) but downregulated in the susceptible cultivar (ROC22) across different time points as compared to the control in response to pathogen infection. Additionally, SsnpPR1.11 was specifically upregulated by 1.2-3.5 folds at 24-72 h post inoculation (hpi) in ROC22, suggesting that this gene may play an important negative regulatory role in defense responses to pathogen infection. The genetic improvement of sugarcane can be facilitated by our results, which also establish the basis for additional functional characterization of SsnpPR1 genes in response to pathogenic stress.

Maize/peanut rotation intercropping improves ecosystem carbon budget and economic benefits in the dry farming regions of China.

Monoculture is widely practiced to increase crop productivity, but long-term adaptation has drawbacks as it increases the depletion of soil nutrients and reduces soil quality, especially in dryland areas. Conversion from traditional maize monoculture to intercropping improves sustainable production. However, maize/peanut intercropping, especially rotation of planting strips impacts of maize/peanut intercropping in dryland on carbon (C) budgets and economic benefits remain unclear. In this study, a 5-year field experiment was conducted to evaluate the influence of maize/peanut intercropping with rotation of planting strips on soil health, indirect CO2-eq greenhouse gas emissions, and ecosystem C inputs. Four intercropping treatments viz. maize monoculture, peanut monoculture, maize/peanut intercropping, and maize/peanut rotation-intercropping were tested from 2018 to 2022. Maize/peanut rotation intercropping significantly improved the land equivalent ratio followed by intercropping and monoculture. Rotation-intercropping also improved economic benefits over intercropping and monoculture which were mainly associated with increased peanut yield where the border rows contributed the maximum, followed by the middle rows. Moreover, rotation-intercropping significantly increased the soil organic C and nitrogen (N) content. Rotation-intercropping decreased indirect CO2-eq greenhouse gas emissions and ecosystem C inputs by 3.11% and 18.04%, whereas increased ecosystem C outputs and net ecosystem C budget by 10.38% and 29.14%, respectively, over the average of monoculture. On average for intercropping and monoculture, rotation-intercropping increased ecosystem C emission efficiency for economic benefits by 51.94% and 227.27% in 2021 and 2022, respectively, showing the highest C utilization efficiency than other treatments. In the long run, maize/peanut rotation-intercropping can be practiced in dryland agriculture to achieve sustainable agriculture goals.

Shattering and yield expression of sesame (Sesamum indicum L) genotypes influenced by paclobutrazol concentration under rainfed conditions of Pothwar.

Seed shattering is a critical challenge that significantly reduces sesame production by 50%. These shattering losses can be reduced by selecting shattering resistant genotypes or by incorporating modern agronomic management such as paclobutrazol, which can boost productivity and prevent seed shattering in sesame. Two-years of field trials were conducted to examine the effect of sesame genotypes, environment, and paclobutrazol (PBZ) concentrations. Twelve sesame genotypes were used in a four-way factorial RCBD with three replications and five PBZ concentrations (T0 = Control; T1 = 150; T2 = 300; T3 = 450; and T4 = 600 mg L- 1) under rainfed conditions of Pothwar. The findings revealed significant variations in the major effects of all examined variables (genotypes, locations, years, and PBZ levels). Sesame genotypes PI-154304 and PI-175907 had the highest plant height, number of capsule plant- 1, seed capsule- 1, 1000 seed weight, biological yield, and seed yield, while also having the lowest seed losses and shattering percentage. Regarding environments, NARC-Islamabad generated the highest plant height, number of capsule plant- 1, shattering percentage, and biological yield; however, the URF-Koont produced the highest seed yield with the lowest shattering percentage. Additionally, plant height, capsules plant- 1, and biological yield were higher in 2021, while seed capsule- 1, 1000 seed weight, seed losses, shattering percentage, and seed yield were higher in 2020. PBZ concentration affected all measured parameters; plant height and number of seed capsule- 1 decreased with increasing PBZ concentrations. 450 mg L- 1 PBZ concentration generated the highest biomass, number of capsules plant- 1, and seed yield. At the same time, PBZ concentration 600 mg L- 1 generated the smallest plant, the lowest seed capsules- 1, the greatest thousand seed weight, and the lowest shattering percentage. The study concluded that paclobutrazol could dramatically reduce shattering percentage and shattering losses while increasing economic returns through better productivity. Based on the findings, the genotypes PI-154304 and PI-175907 with paclobutrazol level 450 mgL- 1 may be suggested for cultivation in Pothwar farming community under rainfed conditions, as they showed promising shattering resistance as well as enhanced growth and yield.

Enhancing drought stress tolerance in Camelina (Camelina sativa L.) through exogenous application of potassium.

The current study was performed under controlled conditions to study the effects of exogenous potassium application on carotenoid contents and drought tolerance in Camelina. Water deficit levels such as 100% FC (control) and 40% FC (drought stress) were imposed after germination of Camelina plants grown to maturity, and different treatments of exogenous K+ were applied at the vegetative stage. We have reported 17 traits of plant growth, physiology, antioxidant enzyme activity, focusing on carotenoids in Camelina to explore their potential yield and yield components. For this purpose, we used multivariate analysis techniques (descriptive statistics, correlation matrix, analysis of variance [ANOVA] and principal components analysis [PCA] to determine the best relation between potassium and studied traits). The results showed a large number of variations in the studied trait under control and water deficit condition. Plant fresh weight (g) was negatively correlated with shoot length and SOD insignificantly correlated with plant fresh weight (g) under water deficit conditions. Potassium loading predicted that foliar application (3 mM K2 SO4 ), foliar application (6 mM KNO3 ), foliar application (12 mM KNO3 ) and foliar application (12 mM K2 SO4 ) are the important doses that contribute the most to enhance the growth, physiological and biochemical activities and carotenoids to improve the Camelina yield under water deficit condition. These doses should be considered in the future to improve the Camelina yield under semi-arid conditions with increased genetic diversity (varietal selection).

Single nucleotide polymorphism based assessment of genetic diversity in local and exotic onion genotypes.

BACKGROUND: Onion is an economically important vegetable cultivated worldwide on a large scale. Liberal exchange of germplasm and frequent selection caused narrow genetic diversity in most crops, including onion. Thus, it is essential to estimate and understand genetic diversity before launching of any breeding program. The current study was conducted to explore genetic diversity among 39 short-day onion genotypes (indigenous and exotic). METHODS AND RESULTS: All the genotypes were evaluated for various phenotypic traits by using single nucleotide polymorphism (SNP) genotyping based on KASPar assays. Principal component analysis (PCA) was performed to determine the variability among genotypes. The four principal components with eigenvalue greater than 1 accounted for 67.5656% variability for quantitative traits, whereas first five principal components with eigenvalue greater than 0.7 accounted for 86.24% variation among the genotypes for qualitative traits. The principal component analysis identified diverse traits including bulb weight, bulb diameter, plant height, number of survived plants and vitamin C. These traits were further analyzed through ANOVA (Analysis of Variance) following augmented block design to describe genotypic variability for selected traits. Onion genotypes showed significant variation for bulb weight, bulb diameter and Vitamin C. Genotypic clustering based on PCA showed that 15 indigenous genotypes were clustered with exotic genotypes (14) while remaining indigenous genotypes (10) were distant. A total of 30 SNPs were used for assessment of genetic diversity out of these, 24 SNPs were detected with polymorphic loci (0.8%, heterozygosity), while only six markers were with monomorphic sites (0.2% heterozygosity). Subsequently, population structure analysis revealed three different populations indicating significant variability. CONCLUSION: Conclusively, a significant similarity between exotic and a group of indigenous genotypes indicates direct adoption of exotic genotypes or their sister lines. A further broadening of the genetic base is required and could be done by crossing distant genotypes.

Endorsement and phylogenetic analysis of some Fabaceae plants based on DNA barcoding.

BACKGROUND: DNA barcoding have been considered as a tool to facilitate species identification based on its simplicity and high-level accuracy in compression to the complexity and subjective biases linked to morphological identification of taxa. MaturaseK gene (MatK gene) of the chloroplast is very vital in the plant system which is involved in the group II intron splicing. The main objective of this study is to determine the relative utility of the "MatK" chloroplast gene for barcoding in 15 legume as a tool to facilitate species identification based on their simplicity and high-level accuracy linked to morphological identification of taxa. METHODS AND RESULTS: MatK gene sequences were submitted to GenBank and the accession numbers were obtained with sequence length ranging from 730 to 1545 nucleotides. These DNA sequences were aligned with database sequence using PROMALS server, Clustal Omega server and Bioedit program. Maximum likelihood and neighbor-joining algorithms were employed for constructing phylogeny. Overall, these results indicated that the phylogenetic tree analysis and the evolutionary distances of an individual dataset of each species were agreed with a phylogenetic tree of all each other consisting of two clades, the first clade comprising (Enterolobium contortisiliquum, Albizia lebbek), Acacia saligna, Leucaena leucocephala, Dichrostachys Cinerea, (Delonix regia, Parkinsonia aculeata), (Senna surattensis, Cassia fistula, Cassia javanica) and Schotia brachypetala were more closely to each other, respectively. The remaining four species of Erythrina humeana, (Sophora secundiflora, Dalbergia Sissoo, Tipuana Tipu) constituted the second clade. CONCLUSION: Moreover, their sequences could be successfully utilized in single nucleotide polymorphism or as part of the sequence as DNA fragment analysis utilizing polymerase chain reaction in plant systematic. Therefore, MatK gene is considered promising a candidate for DNA barcoding in the plant family Fabaceae and provides a clear relationship between the families.

Genome-wide analysis of mitogen-activated protein (MAP) kinase gene family expression in response to biotic and abiotic stresses in sugarcane.

To systematically analyze mitogen-activated protein (MAP) kinase gene families and their expression profiles in sugarcane (Saccharum spp. hybrids; Sh) under diverse biotic and abiotic stresses, we identified 15 ShMAPKs, 6 ShMAPKKs and 16 ShMAPKKKs genes in the sugarcane cultivar R570 genome. These were also confirmed in one S. spontaneum genome and two transcriptome datasets of sugarcane trigged by Acidovorax avenae subsp. avenae (Aaa) and Xanthomonas albilineans (Xa) infections. Phylogenetic analysis revealed that four subgroups were present in each ShMAPK and ShMAPKK family and three sub-families (RAF, MEKK and ZIK) presented in the ShMAPKKK family. Conserved protein motif and gene structure analyses supported the evolutionary relationships of the three families inferred from the phylogenetic analysis. All of the ShMAPK, ShMAPKK and ShMAPKKK genes identified in Saccharum spp. R570 were distributed on chromosomes 1-7 and 9-10. RNA-seq and qRT-PCR analyses indicated that ShMAPK07 and ShMAPKKK02 were defense-responsive genes in sugarcane challenged by both Aaa and Xa stimuli, while some genes were upregulated specifically by Aaa and Xa infection. Additionally, ShMAPK05 acted as a negative regulator under drought and salinity stress, but served as a positive regulator under salicylic acid (SA) treatment. ShMAPK07 plays a positive role under drought stress, but a negative role under SA treatment. ShMAPKKK01 was negatively modulated by both salinity stress and SA treatment, whereas ShMAPKKK06 was positively regulated by both of the two stress stimuli. Our results suggest that members of MAPK cascade gene families regulate adverse stress responses through multiple signal transduction pathways in sugarcane.

Sustainable Development of Chitosan/Calotropis procera-Based Hydrogels to Stimulate Formation of Granulation Tissue and Angiogenesis in Wound Healing Applications.

The formation of new scaffolds to enhance healing magnitude is necessarily required in biomedical applications. Granulation tissue formation is a crucial stage of wound healing in which granulation tissue grows on the surface of a wound by the formation of connective tissue and blood vessels. In the present study, porous hydrogels were synthesized using chitosan incorporating latex of the Calotropis procera plant by using a freeze-thaw cycle to stimulate the formation of granulation tissue and angiogenesis in wound healing applications. Structural analysis through Fourier transform infrared (FTIR) spectroscopy confirmed the interaction between chitosan and Calotropis procera. Latex extract containing hydrogel showed slightly higher absorption than the control during water absorption analysis. Thermogravimetric analysis showed high thermal stability of the 60:40 combination of chitosan (CS) and Calotropis procera as compared to all other treatments and controls. A fabricated scaffold application on a chick chorioallantoic membrane (CAM) showed that all hydrogels containing latex extract resulted in a significant formation of blood vessels and regeneration of cells. Overall, the formation of connective tissues and blood capillaries and healing magnitude decreased in ascending order of concentration of extract.

Magnetic Field Treatments Improves Sunflower Yield by Inducing Physiological and Biochemical Modulations in Seeds.

Magnetic seed enhancement has been practicing as a promising tool to improve germination and seedling growth of low vigor seeds stored under suboptimal conditions, but there is still ambiguity regarding the prospects for magnetism in oilseeds. Present study elucidates the potential of magnetic seed stimulation to improve sunflower germination, growth and yield. Germination and emergence tests were performed to optimize the strength of the magnetic field to sunflower seed enhancement. The seeds were directly exposed to magnetic field strengths of 50, 100 and 150 millitesla (mT) for 5, 10 and 15 min (min) and then standard germination tests were performed. Secondly, the emergence potential of untreated seeds was compared with seed exposed to hydropriming, priming with 3% moringa leaf extract (MLE), priming with magnetically treated water (MTW) for 10 min and priming with 3% MLE solution prepared in magnetically treated water (MTW + MLE). Germination, emergence, seedling growth and seed biochemical properties were used to select the best treatment for field evaluation. The results of the study revealed that magnetic seed treatment with 100 mT for 10 min and seed priming with 3% MLE solution in magnetically treated water (MTW + MLE) significantly improved emergence, crop growth rate and sunflower yield.

Ornamental Plant Efficiency for Heavy Metals Phytoextraction from Contaminated Soils Amended with Organic Materials.

Accumulation of heavy metals (HMs) by ornamental plants (OPs) from contaminated agriculture soils is a unique technique that can efficiently reduce the metal load in the food chain. Amaranthus tricolor L. has attractive characteristics acquiring a higher growth rate and large biomass when grown at heavy metal contaminated soils. Site-specific detailed information is not available on the use of A. tricolor plant in metal phytoremediation from the polluted sites. The study aimed to enhance the uptake of HMs (Pb, Zn, and Cu) via amending poultry litter extract (PLE), vinasse sugarcane (VSC), and humic acid (HA) as natural mobilized organic materials compared to ethylene diamine tetraacetic acid (EDTA), as a common mobilized chemical agent by A. tricolor plant. The studied soils collected from Helwan, El-Gabal El-Asfar (Cairo Governorate), Arab El-Madabeg (Assiut Governorate), Egypt, and study have been conducted under pot condition. Our results revealed all organic materials in all studied soils, except EDTA in EL-Gabal El-Asfar soil, significantly increased the dry weight of the A. tricolor plant compared to the control treatment. The uptake of Pb and Zn significantly (p > 0.05) increased due to applying all organic materials to the studied soils. HA application caused the highest uptake as shown in Pb concentration by more than 5 times in Helwan soil and EDTA by 65% in El-Gabal El-Asfar soil while VSC increased it by 110% in El-Madabeg soil. Also, an increase in Zn concentration due to EDTA application was 58, 42, and 56% for Helwan, El-Gabal El-Asfar, and El-Madabeg soil, respectively. In all studied soils, the application of organic materials increased the remediation factor (RF) than the control. El-Madabeg soil treated with vinasse sugarcane gave the highest RF values; 6.40, 3.26, and 4.02% for Pb, Zn, and Cu, respectively, than the control. Thus, we identified A. tricolor as a successful ornamental candidate that, along with organic mobilization amendments, most efficiently develop soil health, reduce metal toxicity, and recommend remediation of heavy metal-contaminated soils. Additionally, long-term application of organic mobilization amendments and continued growth of A. tricolor under field conditions could be recommended for future directions to confirm the results.

First Report of Pantoea stewartii subsp. stewartii Causing Bacterial Leaf Wilt of Sugarcane in China.

The pathogen of Pantoea stewartii subsp. stewartii (Pss) that is the causal agent of Stewart's bacterial wilt of corn also infects numerous experimental hosts of graminaceous plants (Pepper et al., 1967; Wang et al., 2012). However, little is known about this pathogen naturally infecting sugarcane. In 2017, we observed some sugarcane cultivars showing leaf blade bleaching at the disease initiation stage, which further resulted in development of blight and necrotic lesions (Figure 1-A and -B) in Zhanjiang, Guangdong province of China. To diagnose this putative disease, five symptomatic leaf samples were collected from different sugarcane cultivars. The Pss was found to infect these samples using the nested PCR with Pss-specific outer primers PS1/PS4 and inner primers Ps2r/Ps3r that targeted at the 16S rRNA gene of this pathogen (Wang et al., 2009). The expected 262-bp fragments from positive samples were amplified, cloned, and sequenced (GenBank accession no. MW015795-MW015799). BLASTN analysis revealed that these isolates had more than 99.5% nucleotide identify (222 bp out of 262 bp) with each other and with Pss strains (ATCC 8199 and DC283) as well as P. stewartii subsp. indologenes strains (SR2-12 and LMG 2632) after sequences were trimmed at the 5'- and 3'-terminal of inner primer sequences. In addition, these leaf samples were surface-sterilized with 75% alcohol followed by macerated and chopped in sterile water. Upon plating on solid NA medium at 28 °C for 24-36 h, the bacterial colonies exhibited yellow color with circular, convex, smooth and translucent edges (Figure 1-C). Straight rods and non-encapsulated cells were detected under transmission electron microscopy (Figure 1-D). Moreover, an identical colony termed as PSCN1 was isolated from sugarcane cultivar YZ08-1095 and was further confirmed by the PCR with a universal primer pair 63F (5'-CAGGCCTAACACATGCAAGTC-3') and 1387R (5'-GGGCGGWGTGTACAAGGC-3') that targeted at bacterial 16S rRNA gene (Marchesi et al., 1998). A 1362-bp DNA fragment sequence was obtained from PSCN1 strain and deposited on GenBank library (accession no. MW015767). Sequence analysis showed that PSCN1 shared 99.9-100% nucleotide identity (1315 bp out of 1362 bp) with the two reference strains of Pss (ATCC 8199 and DC283) after sequences were trimmed at the 5'- and 3'-terminal of primer sequences. According to Koch's postulates, pathogenicity test was carried out on YZ08-1095 plants with 3-5 fully developed leaf inoculated with the suspended cells (108 cells/ml) of PSCN1 strain by cutting the one-third of leaves before spraying with a suspension. Control plants were mock-inoculated with serial liquid nutrition agar medium. Two independent experiments were performed for pathogenicity assay and more than 28 plants of YZ08-1095 were used in each treatment. Plants were cultured in a growth chamber at 28 °C and 60% humidity under a 16 h light/8 h dark photoperiod. Leaves inoculated by the PSCN1 initially showed bleached, blight and wilting symptoms on leaf edges at seven days post-inoculation (dpi) (Figure 1-E and -F), which were similar to those symptoms observed in the fields. Control plants remained asymptomatic (Figure 1-G). The average incidence of diseased plant was 51.9% at 21 dpi. The bacteria were subsequently re-isolated from diseased leaves, and yielded colonies were completely identical to the PSCN1. Taken together, our data provides the valuable information for diagnosis and controlling this disease in sugarcane.

Interaction between Sugarcane and Environmental Stressors: From Identification to Molecular Mechanism.

An increase in the vulnerability of crops to a wide range of biotic and abiotic stresses can have a marked influence on the productivity and quality of major crops, especially sugarcane (Saccharum spp [...].

Establishment of an Efficient Sugarcane Transformation System via Herbicide-Resistant CP4-EPSPS Gene Selection.

Sugarcane (Saccharum spp.), a major cash crop that is an important source of sugar and bioethanol, is strongly influenced by the impacts of biotic and abiotic stresses. The intricate polyploid and aneuploid genome of sugarcane has shown various limits for conventional breeding strategies. Nonetheless, biotechnological engineering currently offers the best chance of introducing commercially significant agronomic features. In this study, an efficient Agrobacterium-mediated transformation system that uses the herbicide-resistant CP4-EPSPS gene as a selection marker was developed. Notably, all of the plants that were identified by PCR as transformants showed significant herbicide resistance. Additionally, this transformation protocol also highlighted: (i) the high yield of transgenic lines from calli (each gram of calli generated six transgenic lines); (ii) improved selection; and (iii) a higher transformation efficiency. This protocol provides a reliable tool for a routine procedure for the generation of resilient sugarcane plants.

Risk Factors Associated With Anastomotic Stricture in Patients Undergoing Minimally Invasive Esophagectomy: Experience From a High-Volume Center.

BACKGROUND: Esophageal cancer is a prevalent cancer, with a high incidence in low socioeconomic category countries. Minimally invasive esophagectomy is increasingly being used to treat this malignant condition. However, anastomotic stricture is a serious complication post esophagectomy. The study aims to enhance diagnostic consistency, improve treatment methods, guide patient management, stratify outcomes, and offer evidence-based preventive interventions. METHODS: A retrospective analysis of 550 patients who had minimally invasive esophagus surgery was carried out at Shaukat Khanum Memorial Cancer Hospital and Research Centre in Lahore between 2015 and 2020. All patients were treated with radical resection. For tumors of the lower esophagus and gastroesophageal junction, transhiatal esophagectomy was used; for tumors of the middle and upper thoracic esophagus, right video-assisted thoracoscopic surgery (VATS) was used in a three-stage procedure. Patients were routinely followed up two weeks after discharge, then every three, six, and 12 months. RESULTS: The mean age and BMI were 46.7 years and 19.4 kg/m2, respectively. Anastomotic leaks were rare, with only 13 patients experiencing them. The grade of tumor differentiation was poor, moderate, and well-differentiated. The study found that older age, high Eastern Cooperative Oncology Group (ECOG) performance status, and malignancies located in the upper and middle one-third of the esophagus had significant associations with anastomotic stricture. However, some variables, like sex, did not show significant associations in either analysis. CONCLUSION: The study reveals that factors such as older patient age, high ECOG performance status, single comorbidity, and malignancies located in the upper and middle one-third of the esophagus significantly influence anastomotic stricture. The study suggests that measures against anastomotic stricture such as endoscopic procedures and minimally invasive esophagectomy should be implemented to minimize the complications.

Insights into Reactive Oxygen Species Production-Scavenging System Involved in Sugarcane Response to Xanthomonas albilineans Infection under Drought Stress.

Plants must adapt to the complex effects of several stressors brought on by global warming, which may result in interaction and superposition effects between diverse stressors. Few reports are available on how drought stress affects Xanthomonas albilineans (Xa) infection in sugarcane (Saccharum spp. hybrids). Drought and leaf scald resistance were identified on 16 sugarcane cultivars using Xa inoculation and soil drought treatments, respectively. Subsequently, four cultivars contrasting to drought and leaf scald resistance were used to explore the mechanisms of drought affecting Xa-sugarcane interaction. Drought stress significantly increased the occurrence of leaf scald and Xa populations in susceptible cultivars but had no obvious effect on resistant cultivars. The ROS bursting and scavenging system was significantly activated in sugarcane in the process of Xa infection, particularly in the resistant cultivars. Compared with Xa infection alone, defense response via the ROS generating and scavenging system was obviously weakened in sugarcane (especially in susceptible cultivars) under Xa infection plus drought stress. Collectively, ROS might play a crucial role involving sugarcane defense against combined effects of Xa infection and drought stress.