Co-hydrothermal carbonization with process water recirculation as a valuable strategy to enhance hydrochar recovery with high energy efficiency.

This study aims at valorizing the residual aqueous phase from hydrothermal carbonization (HTC) of Sicilian agro-wastes in order to enhance the hydrochar recovery, positively affecting the process energy balance. Process waters (PW) obtained from HTC and co-HTC using orange peel waste and fennel plant residues were used as recycled solvent in experiments carried out at the temperatures of 180 and 230 °C. The results showed that an additional hydrochar formation was promoted during recirculation of solvent, leading to average increments of solid mass yield of 10.5 wt% for tests conducted at 180 °C and 3.9 wt% for 230 °C. After five consecutive recirculation phases in co-HTC runs, the hydrochar yield increased up to 18.2 wt%. The low H/C and O/C atomic ratios values, found after recirculation, indicate that organic acids, accumulated in the PW, may catalyze the process and promote the biomass deoxygenation by boosting dehydration and decarboxylation. The recovered PWs from conversion steps with deionized water were also carbonized in absence of the solid feedstock in order to quantify their contribution in hydrochar formation during recirculation and thus the synergistic interactions. After recirculation, energy recovery averagely augmented by more than threefold, showing that the proposed strategy could significantly improve the sustainability of HTC.

Sustaining rice productivity through weather-resilient agricultural practices.

BACKGROUND: Enhancing productivity and profitability and reducing climatic risk are the major challenges for sustaining rice production. Extreme weather can have significant and varied effects on crops, influencing agricultural productivity, crop yields and food security. RESULTS: In this study, a comparative evaluation of two crop management systems was performed involving farmers adopting a weather forecast-based advisory service (WFBAS) and usual farmers' practice (FP). WFBAS crop management followed the generated weather forecast-based advice whereas the control farmers (FP) did not receive any weather forecast-based advice, rather following their usual rice cultivation practices. The results of the experiments revealed that WFBAS farmers had a significant yield advantage over FP farmers. With the WFBAS technology, the farmers used inputs judiciously, utilized the benefit of favorable weather and minimized the risk resulting from extreme weather events. As a result, besides the yield enhancement, WFBAS provided a scope to protect the environment with the minimum residual effect of fertilizer and pesticides. It also reduced the pressure on groundwater by ensuring efficient water management. Finally, the farmers benefited from higher income through yield enhancement, reduction of the costs of production and reduction of risk. CONCLUSION: A successful and extensive implementation of WFBAS in the rice production system would assist Bangladesh in achieving Sustainable Development Goal 2.4, which focuses on rice productivity and profitability of farmers as well as long-term food security of the country. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

A REML method for the evidence-splitting model in network meta-analysis.

Checking for possible inconsistency between direct and indirect evidence is an important task in network meta-analysis. Recently, an evidence-splitting (ES) model has been proposed, that allows separating direct and indirect evidence in a network and hence assessing inconsistency. A salient feature of this model is that the variance for heterogeneity appears in both the mean and the variance structure. Thus, full maximum likelihood (ML) has been proposed for estimating the parameters of this model. Maximum likelihood is known to yield biased variance component estimates in linear mixed models, and this problem is expected to also affect the ES model. The purpose of the present paper, therefore, is to propose a method based on residual (or restricted) maximum likelihood (REML). Our simulation shows that this new method is quite competitive to methods based on full ML in terms of bias and mean squared error. In addition, some limitations of the ES model are discussed. While this model splits direct and indirect evidence, it is not a plausible model for the cause of inconsistency.

Screening of hepatitis B and C viral infection, recognition of risk factors, and immunization of patients against hepatitis B virus: a module developed for effective hepatitis control.

Introduction: The continually increasing incidence of hepatitis, a worldwide health issue, in Pakistan, has highlighted the need to investigate the epidemiology factors and implement preventive measures accordingly. The purpose of this study was to scrutinize the prevalent and significantly associated risk factors of hepatitis in students and employees, screening them for hepatitis B and C virus and vaccinating them against HBV to make IUB hepatitis free. Methodology: A total of 12,912 participants including students (n = 10,948) and employees (n = 1964) were screened for HBV and HCV via immunochromatographic test. Hepatitis- positive participants' blood samples were further tested and viral load was estimated by quantitative PCR. All the hepatitis-negative participants were vaccinated against HBV. The demographic and risk factors-related data were collected using the questionnaire. Statistical analysis (Chi-square test and bivariate regression analysis) was performed using SPSS software to explore any association between risk factors and hepatitis. Results: Results indicated that 662/12912 participants (students = 478/10,948, employees = 184/1,964) tested positive for hepatitis. Among them, HCV was observed to be more prevalent than HBV among the study participants, employees, and students, and viral count was low in both HBV and HCV-infected participants. However, men were more affected than women. The studied risk factors represented higher frequency among hepatitis-positive participants relative to the hepatitis-negative participants. The Chi-square test revealed that students' gender, history of hepatitis in the family and relatives, dental treatment, sharing cosmetics and shaving blades were significant (p > 0.005) risk factors of hepatitis while in the employees group surgery and age were significant. Moreover, the reused of syringes was found to be associated with hepatitis in both groups. The bivariate analysis helped to identify various new risk factors which were independently, either positively or negatively, associated with hepatitis. Discussion: Our study enabled us to recognize different risk factors of hepatitis among the target population. The information thus generated can be usefully applied in planning hepatitis awareness, targeted screening, and effective control programs for other target populations. In general, this module can be further utilized for any other disease.

Synthesis of a Graphene-Encapsulated Fe3C/Fe Catalyst Supported on Sporopollenin Exine Capsules and Its Use for the Reverse Water-Gas Shift Reaction.

Bioderived materials have emerged as sustainable catalyst supports for several heterogeneous reactions owing to their naturally occurring hierarchal pore size distribution, high surface area, and thermal and chemical stability. We utilize sporopollenin exine capsules (SpECs), a carbon-rich byproduct of pollen grains, composed primarily of polymerized and cross-linked lipids, to synthesize carbon-encapsulated iron nanoparticles via evaporative precipitation and pyrolytic treatments. The composition and morphology of the macroparticles were influenced by the precursor iron acetate concentration. Most significantly, the formation of crystalline phases (Fe3C, α-Fe, and graphite) detected via X-ray diffraction spectroscopy showed a critical dependence on iron loading. Significantly, the characteristic morphology and structure of the SpECs were largely preserved after high-temperature pyrolysis. Analysis of Brunauer-Emmett-Teller surface area, the D and G bands from Raman spectroscopy, and the relative ratio of the C=C to C-C bonding from high-resolution X-ray photoelectron spectroscopy suggests that porosity, surface area, and degree of graphitization were easily tuned by varying the Fe loading. A mechanism for the formation of crystalline phases and meso-porosity during the pyrolysis process is also proposed. SpEC-Fe10% proved to be highly active and selective for the reverse water-gas shift reaction at high temperatures (>600 °C).

50 years of rice breeding in Bangladesh: genetic yield trends.

To assess the efficiency of genetic improvement programs, it is essential to assess the genetic trend in long-term data. The present study estimates the genetic trends for grain yield of rice varieties released between 1970 and 2020 by the Bangladesh Rice Research Institute. The yield of the varieties was assessed from 2001-2002 to 2020-2021 in multi-locations trials. In such a series of trials, yield may increase over time due to (i) genetic improvement (genetic trend) and (ii) improved management or favorable climate change (agronomic/non-genetic trend). In both the winter and monsoon seasons, we observed positive genetic and non-genetic trends. The annual genetic trend for grain yield in both winter and monsoon rice varieties was 0.01 t ha-1, while the non-genetic trend for both seasons was 0.02 t ha-1, corresponding to yearly genetic gains of 0.28% and 0.18% in winter and monsoon seasons, respectively. The overall percentage yield change from 1970 until 2020 for winter rice was 40.96%, of which 13.91% was genetic trend and 27.05% was non-genetic. For the monsoon season, the overall percentage change from 1973 until 2020 was 38.39%, of which genetic and non-genetic increases were 8.36% and 30.03%, respectively. Overall, the contribution of non-genetic trend is larger than genetic trend both for winter and monsoon seasons. These results suggest that limited progress has been made in improving yield in Bangladeshi rice breeding programs over the last 50 years. Breeding programs need to be modernized to deliver sufficient genetic gains in the future to sustain Bangladeshi food security.

Hypothyroidism correlates with dyslipidemia and protein contents in patients with various metabolic disorders.

OBJECTIVES: The present study aimed to investigate the status of abnormalities in thyroid-related hormones, lipid profile parameters and total proteins in the sera of patients suffering from various metabolic disorders. METHODS: To analyze the study parameters, enzyme-linked immunosorbent assays, Bradford assays and standard clinical kits and methods were applied. Data were analyzed through the appropriate statistical tests. RESULTS: In all subjects except those with hypotension, thyroid-stimulating hormone, total cholesterol and low-density lipoprotein were elevated, whereas triiodothyronine levels were downregulated. Thyroxin was downregulated in subjects with diabetes and symptomatic thyroiditis but upregulated in patients with hypertension. High-density lipoprotein was upregulated in men who were diabetic only, and total protein was downregulated in those with hypotension only. Hypothyroidism in patients with diabetes, symptomatic thyroiditis and hypertension was correlated with dyslipidemia. In subjects with hypertension, it was correlated with total protein. CONCLUSION: This study revealed a link between hypothyroidism, dyslipidemia and total protein in patients with various metabolic disorders.

Assessing the between-country genetic correlation in maize yield using German and Polish official variety trials.

KEY MESSAGE: We assess the genetic gain and genetic correlation in maize yield using German and Polish official variety trials. The random coefficient models were fitted to assess the genetic correlation. Official variety testing is performed in many countries by statutory agencies in order to identify the best candidates and make decisions on the addition to the national list. Neighbouring countries can have similarities in agroecological conditions, so it is worthwhile to consider a joint analysis of data from national list trials to assess the similarity in performance of those varieties tested in both countries. Here, maize yield data from official German and Poland variety trials for cultivation and use (VCU) were analysed for the period from 1987 to 2017. Several statistical models that incorporate environmental covariates were fitted. The best fitting model was used to compute estimates of genotype main effects for each country. It is demonstrated that a model with random genotype-by-country effects can be used to borrow strength across countries. The genetic correlation between cultivars from the two countries equalled 0.89. The analysis based on agroecological zones showed high correlation between zones in the two countries. The results also showed that 22 agroecological zones in Germany can be merged into five zones, whereas the six zones in Poland had very high correlation and can be considered as a single zone for maize. The 43 common varieties which were tested in both countries performed equally in both countries. The mean performances of these common varieties in both countries were highly correlated.

Comparative genetic, biochemical and physiological analysis of sodium and chlorine in wheat.

Plant with a great diversity shows several responses towards the biotic and abiotic stresses. Among these abiotic stresses, salinity is the main damaging factor as it reduces the yield of wheat plant with moderate salt tolerance. For its survival, plant undergoes through some genetic, biochemical and physiological changes to tackle the stress. This review mainly describes the conditions where various ions present in the soil, especially sodium and chlorine, enter into the plant and the genes or proteins involved with survival mechanism against the damage in plants. Salt stress causes alteration in enzymatic activity and Photosynthesis, oxidative stress, damage of cellular structure and components and ionic imbalance. Ion toxicity stress occur due to accumulation of excessive sodium ion and chloride ion. Transcriptional factors TaPIMP, TaSRG and TaMYBsdu 1 play key role in gene expression mechanism to overcome the stress. High affinity potassium transporter gene family is responsible for salt tolerance in wheat plant. HKT1;4 and HKT1;5 genes are responsible for Na exclusion in Triticum monococcum. Forty QTLs were found with the marker assisted selection in bread wheat for salinity tolerance and some morphological traits, 5 QTLs were related to sodium ion exclusion. In bread wheat, salt stress tolerance mechanism is mainly an exclusion of Na+ ions but also include K+ ion concentration. The salinity tolerant germplasm MW#293 provides an opportunity for the development of future salinity tolerant bread wheat.

Increasing floral visitation and hybrid seed production mediated by beauty mark in Gossypium hirsutum.

Hybrid crop varieties have been repeatedly demonstrated to produce significantly higher yields than their parental lines; however, the low efficiency and high cost of hybrid seed production has limited the broad exploitation of heterosis for cotton production. One option for increasing the yield of hybrid seed is to improve pollination efficiency by insect pollinators. Here, we report the molecular cloning and characterization of a semidominant gene, Beauty Mark (BM), which controls purple spot formation at the base of flower petals in the cultivated tetraploid cotton species Gossypium barbadense. BM encodes an R2R3 MYB113 transcription factor, and we demonstrate that GbBM directly targets the promoter of four flavonoid biosynthesis genes to positively regulate petal spot development. Introgression of a GbBM allele into G. hirsutum by marker-assisted selection restored petal spot formation, which significantly increased the frequency of honeybee visits in G. hirsutum. Moreover, field tests confirmed that cotton seed yield was significantly improved in a three-line hybrid production system that incorporated the GbBM allele. Our study thus provides a basis for the potentially broad application of this gene in improving the long-standing problem of low seed production in elite cotton hybrid lines.

Morphological, physiological and molecular assessment of cotton for drought tolerance under field conditions.

Climate change could be an existential threat to many crops. Drought and heat stress are becoming harder for cultivated crops. Cotton in Pakistan is grown under natural high temperature and low moisture, could be used as a source of heat and drought tolerance. Therefore, the study was conducted to morphological, physiological and molecular characterization of cotton genotypes under field conditions. A total of 25 cotton genotypes were selected from the gene pool of Pakistan based on tolerance to heat and drought stress. In field trail, the stress related traits like boll retention percentage, plant height, number of nodes and inter-nodal distance were recorded. In physiological assessment, traits such as photosynthesis rate, stomatal conductance, transpiration rate, leaf temperature, relative water content and excised leaf water loss were observed. At molecular level, a set of 19 important transcription factors, controlling drought/heat stress tolerance (HSPCB, GHSP26, HSFA2, HSP101, HSP3, DREB1A, DREB2A, TPS, GhNAC2, GbMYB5, GhWRKY41, GhMKK3, GhMPK17, GhMKK1, GhMPK2, APX1, HSC70, ANNAT8, and GhPP2A1) were analyzed from all genotypes. Data analyses depicted that boll retention percentage, photosynthesis, stomatal conductance, relative water content under the stress conditions were associated with the presence of important drought & heat TF/genes which depicts high genetic potential of Pakistani cotton varieties against abiotic stress. The variety MNH-886 appeared in medium plant height, high boll retention percentage, high relative water content, photosynthesis rate, stomatal conductance, transpiration rate and with maximum number transcription factors under study. The variety may be used as source material for heat and drought tolerant cotton breeding. The results of this study may be useful for the cotton breeders to develop genotype adoptable to environmental stresses under climate change scenario.

Impacts of Carbon Dioxide Enrichment on Landrace and Released Ethiopian Barley (Hordeum vulgare L.) Cultivars.

Barley (Hordeum vulgare L.) is an important food security crop due to its high-stress tolerance. This study explored the effects of CO2 enrichment (eCO2) on the growth, yield, and water-use efficiency of Ethiopian barley cultivars (15 landraces, 15 released). Cultivars were grown under two levels of CO2 concentration (400 and 550 ppm) in climate chambers, and each level was replicated three times. A significant positive effect of eCO2 enrichment was observed on plant height by 9.5 and 6.7%, vegetative biomass by 7.6 and 9.4%, and grain yield by 34.1 and 40.6% in landraces and released cultivars, respectively. The observed increment of grain yield mainly resulted from the significant positive effect of eCO2 on grain number per plant. The water-use efficiency of vegetative biomass and grain yield significantly increased by 7.9 and 33.3% in landraces, with 9.5 and 42.9% improvement in released cultivars, respectively. Pearson's correlation analysis revealed positive relationships between grain yield and grain number (r = 0.95), harvest index (r = 0.86), and ear biomass (r = 0.85). The response of barley to eCO2 was cultivar dependent, i.e., the highest grain yield response to eCO2 was observed for Lan_15 (122.3%) and Rel_10 (140.2%). However, Lan_13, Land_14, and Rel_3 showed reduced grain yield by 16, 25, and 42%, respectively, in response to eCO2 enrichment. While the released cultivars benefited more from higher levels of CO2 in relative terms, some landraces displayed better actual values. Under future climate conditions, i.e., future CO2 concentrations, grain yield production could benefit from the promotion of landrace and released cultivars with higher grain numbers and higher levels of water-use efficiency of the grain. The superior cultivars that were identified in the present study represent valuable genetic resources for future barley breeding.

Appropriate sampling methods and statistics can tell apart fraud from pesticide drift in organic farming.

Pesticide residues are much lower in organic than in conventional food. The article summarizes the available residue data from the EU and the U.S. organic market. Differences between samples from several sources suggest that organic products are declared conventional, when they have residues-but the origin of the residues is not always investigated. A large number of samples are being tested by organic certifiers, but the sampling methods often do not allow to determine if such residues stem from prohibited pesticide use by organic farmers, from mixing organic with conventional products, from short-range spray-drift from neighbour farms, from the ubiquitous presence of such substances due to long-distance drift, or from other sources of contamination. Eight case studies from different crops and countries are used to demonstrate that sampling at different distances from possible sources of short-distance drift in most cases allows differentiating deliberate pesticide application by the organic farmer from drift. Datasets from 67 banana farms in Ecuador, where aerial fungicide spraying leads to a heavy drift problem, were subjected to statistical analysis. A linear discriminant function including four variables was identified for distinguishing under these conditions application from drift, with an accuracy of 93.3%.

A 13-Lipoxygenase, GhLOX2, positively regulates cotton tolerance against Verticillium dahliae through JA-mediated pathway.

Verticillium wilt is a major limiting factor for sustainable production of cotton but the mechanism of controlling this disease is still poorly understood. Lipoxygenase (LOX)-derived oxylipins have been implicated in defense responses against diverse pathogens; however there is limited information about the functional characterization of LOXs in response to Verticillium dahliae infection. In this study, we report the characterization of a cotton LOX gene, GhLOX2, which phylogenetically clustered into 13-LOX subfamily and is closely related to Arabidopsis LOX2 gene. GhLOX2 was predominantly expressed in leaves and strongly induced following V. dahliae inoculation and treatment of methyl jasmonate (MeJA). RNAi-mediated knock-down of GhLOX2 enhanced cotton susceptibility to V. dahliae and was coupled with suppression of jasmonic acid (JA)-related genes both after inoculation with the cotton defoliating strain V991 or MeJA treatment. Interestingly, lignin contents, transcripts of lignin synthesis genes and H2O2 contents were also decreased in GhLOX2-silenced plants. This study suggests that GhLOX2 is involved in defense responses against infection of V. dahliae in cotton and supports that JA is one of the major defense hormones against this pathogen.

Mitigation of bacterial spot disease induced biotic stress in Capsicum annuum L. cultivars via antioxidant enzymes and isoforms.

Bacterial spot, caused by a group of Xanthomonads (Xanthomonas spp.), is a devastating disease. It can adversely affect the Capsicum annum productivity. Scientists are working on the role of antioxidants to meet this challenge. However, research is lacking on the role of antioxidant enzymes and their isoforms in the non-compatible pathogen and host plant interaction and resistance mechanisms in capsicum varieties. The present study was conducted to ascertain the defensive role of antioxidant enzymes and their isoforms in chilli varieties Hybrid, Desi, Serrano, Padron, and Shehzadi against bacterial spot disease-induced Xanthomonas sp. The seedlings were inoculated with bacterial pathogen @ 107 CFU/mL, and samples were harvested after regular intervals of 24 h for 4 days followed by inoculation. Total plant proteins were extracted in phosphate buffer and quantified through Bradford assay. The crude protein extracts were analyzed through quantitative enzymatic assays in order to document activity levels of various antioxidant enzymes, including peroxidase (POD), Catalase (CAT), Ascorbate peroxidase (APX), and Superoxide dismutase (SOD). Moreover, the profiles appearance of these enzymes and their isoforms were determined using native polyacrylamide gel electrophoresis (PAGE) analysis. These enzymes exhibited maximum activity in Hybrid (HiR) cultivar followed by Desi (R), Serrano (S), Padron, and Shehzadi (HS). Both the number of isoforms and expression levels were higher in highly resistant cultivars compared to susceptible and highly susceptible cultivars. The induction of POD, CAT, and SOD occurs at the early stages of growth in resistant Capsicum cultivars. At the same time, APX seems to make the second line of antioxidant defense mechanisms. We found that modulating antioxidant enzymes and isoforms activity at the seedling stage was an important mechanism for mitigating plant growth inhibition in the resistant ones.

Impact of heat stress responsive factors on growth and physiology of cotton (Gossypium hirsutum L.).

Pakistan ranked highest with reference to average temperatures in cotton growing areas of the world. The heat waves are becoming more intense and unpredictable due to climate change. Identification of heat tolerant genotypes requires comprehensive screening using molecular, physiological and morphological analysis. Heat shock proteins play an important role in tolerance against heat stress. In the current study, eight heat stress responsive factors, proteins and genes (HSFA2, GHSP26, GHPP2A, HSP101, HSC70-1, HSP3, APX1 and ANNAT8) were evaluated morphologically and physiologically for their role in heat stress tolerance. For this purpose, cotton crop was grown at two temperature conditions i.e. normal weather and heat stress at 45 °C. For molecular analysis, genotypes were screened for the presence or absence of heat shock protein genes. Physiological analysis of genotypes was conducted to assess net photosynthesis, stomatal conductance, transpiration rate, leaf-air temperature and cell membrane stability under control as well as high temperature. The traits photosynthesis, cell membrane stability, leaf-air temperature and number of heat stress responsive factors in each genotypes showed a strong correlation with boll retention percentage under heat stress. The genotypes with maximum heat shock protein genes such as Cyto-177, MNH-886, VH-305 and Cyto-515 showed increased photosynthesis, stomatal conductance, negative leaf-air temperature and high boll retention percentage under heat stress condition. These varieties may be used as heat tolerant breeding material.

Construction of Gossypium barbadense Mutant Library Provides Genetic Resources for Cotton Germplasm Improvement.

Allotetraploid cotton (Gossypium hirsutum and Gossypium barbadense) are cultivated worldwide for its white fiber. For centuries, conventional breeding approaches increase cotton yield at the cost of extensive erosion of natural genetic variability. Sea Island cotton (G. barbadense) is known for its superior fiber quality, but show poor adaptability as compared to Upland cotton. Here, in this study, we use ethylmethanesulfonate (EMS) as a mutagenic agent to induce genome-wide point mutations to improve the current germplasm resources of Sea Island cotton and develop diverse breeding lines with improved adaptability and excellent economic traits. We determined the optimal EMS experimental procedure suitable for construction of cotton mutant library. At M6 generation, mutant library comprised of lines with distinguished phenotypes of the plant architecture, leaf, flower, boll, and fiber. Genome-wide analysis of SNP distribution and density in yellow leaf mutant reflected the better quality of mutant library. Reduced photosynthetic efficiency and transmission electron microscopy of yellow leaf mutants revealed the effect of induced mutations at physiological and cellular level. Our mutant collection will serve as the valuable resource for basic research on cotton functional genomics, as well as cotton breeding.

Genome-Wide Characterization and Expression Analysis of NHX Gene Family under Salinity Stress in Gossypium barbadense and Its Comparison with Gossypium hirsutum.

Cotton is an important economic crop affected by different abiotic stresses at different developmental stages. Salinity limits the growth and productivity of crops worldwide. Na+/H+ antiporters play a key role during the plant development and in its tolerance to salt stress. The aim of the present study was a genome-wide characterization and expression pattern analysis under the salinity stress of the sodium-proton antiporter (NHX) of Gossypium barbadense in comparison with Gossypium hirsutum. In G. barbadense, 25 NHX genes were identified on the basis of the Na+_H+ exchanger domain. All except one of the G. barbadenseNHX transporters have an Amiloride motif that is a known inhibitor of Na+ ions in plants. A phylogenetic analysis inferred three classes of GbNHX genes-viz., Vac (GbNHX1, 2 and 4), Endo (GbNHX6), and PM (GbNHX7). A high number of the stress-related cis-acting elements observed in promoters show their role in tolerance against abiotic stresses. The Ka/Ks values show that the majority of GbNHX genes are subjected to strong purifying selection under the course of evolution. To study the functional divergence of G. barbadenseNHX transporters, the real-time gene expression was analyzed under salt stress in the root, stem, and leaf tissues. In G. barbadense, the expression was higher in the stem, while in G. hirsutum the leaf and root showed a high expression. Moreover, our results revealed that NHX2 homologues in both species have a high expression under salinity stress at higher time intervals, followed by NHX7. The protein-protein prediction study revealed that GbNHX7 is involved in the CBL-CIPK protein interaction pathway. Our study also provided valuable information explaining the molecular mechanism of Na+ transport for the further functional study of Gossypium NHX genes.