The Impact of Various Organic Phosphorus Carriers on the Uptake and Use Efficiency in Barley.

Organic phosphorus (OP) is an essential component of the soil P cycle, which contributes to barley nutrition after its mineralization into inorganic phosphorus (Pi). However, the dynamics of OP utilization in the barley rhizosphere remain unclear. In this study, phytin was screened out from six OP carriers, which could reflect the difference in OP utilization between a P-inefficient genotype Baudin and a P-efficient genotype CN4027. The phosphorus utilization efficiency (PUE), root morphological traits, and expression of genes associated with P utilization were assessed under P deficiency or phytin treatments. P deficiency resulted in a greater root surface area and thicker roots. In barley fed with phytin as a P carrier, the APase activities of CN4027 were 2-3-fold lower than those of Baudin, while the phytase activities of CN4027 were 2-3-fold higher than those of Baudin. The PUE in CN4027 was mainly enhanced by activating phytase to improve the root absorption and utilization of Pi resulting from OP mineralization, while the PUE in Baudin was mainly enhanced by activating APase to improve the shoot reuse capacity. A phosphate transporter gene HvPHT1;8 regulated P transport from the roots to the shoots, while a purple acid phosphatase (PAP) family gene HvPAPhy_b contributed to the reuse of P in barley.

Genome-Wide Association Study on Total Starch, Amylose and Amylopectin in Barley Grain Reveals Novel Putative Alleles.

The content and composition of starch in cereal grains are closely related to yield. Few studies have been done on the identification of the genes or loci associated with these traits in barley. This study was conducted to identify the genes or loci controlling starch traits in barley grains, including total starch (TS), amylose (AC) and amylopectin (AP) contents. A large genotypic variation was found in all examined starch traits. GWAS analysis detected 13, 2, 10 QTLs for TS, AC and AP, respectively, and 5 of them were commonly shared by AP and TS content. qTS-3.1, qAC-6.2 and qAP-5.1 may explain the largest variation of TS, AC and AP, respectively. Four putative candidate genes, i.e., HORVU6Hr1G087920, HORVU5Hr1G011230, HORVU5Hr1G011270 and HORVU5Hr1G011280, showed the high expression in the developing barley grains when starch accumulates rapidly. The examined 100 barley accessions could be divided into two groups based on the polymorphism of the marker S5H_29297679, with 93 accessions having allele GG and seven accessions having AA. Moreover, significantly positive correlation was found between the number of favorable alleles of the identified QTLs and TS, AC, AP content. In conclusion, the identified loci or genes in this study could be useful for genetic improvement of grains starch in barley.

Pivotal role for root cell wall polysaccharides in cultivar-dependent cadmium accumulation in Brassica chinensis L.

Polysaccharides are the main components of plant cell walls in which they make an important contribution to cadmium (Cd) fixation. However, knowledge regarding the role of root cell wall polysaccharides in Cd accumulation in low-Cd cultivars is limited. Here, we compared the differences in root cell wall polysaccharides between two cultivars of Brassica chinensis L. (pakchoi) with different Cd accumulation abilities. A hydroponic experiment was conducted using low- (Huajun 2) and high-Cd (Hanlv) pakchoi cultivars. We investigated Cd subcellular distribution and Cd accumulation in cell wall polysaccharides and examined polysaccharide modifications in root cell walls by Fourier transform infrared spectroscopy. A Cd adsorption kinetics experiment was conducted to examine the connection between Cd-induced polysaccharide modifications and Cd fixation by cell walls. Amounts of Cd were significantly higher and more Cd was bound to cell walls in the roots of Huajun 2 than in those of Hanlv. These results indicated that the greater Cd retention capacity of the root cell wall in Huajun 2 accounted for the low Cd accumulation in the shoot. Up to 79.4% and 32.1% of cell-wall-bound Cd was found in the pectin and hemicellulose 1, respectively, and higher amounts of Cd were found in these cell wall components of Huajun 2 than in those of Hanlv. Exposure to Cd significantly increased amounts of pectin and hemicellulose 1 in both pakchoi cultivars, but the pectin levels were significantly higher in Huajun 2 than in Hanlv. Huajun 2 had higher pectin methylesterase (PME) activity and a lower degree of pectin methyl-esterification (DM) than Hanlv, although Cd treatments resulted in increased PME activity and decreased DM in both cultivars. The higher Cd treatment (44.5 µM) resulted in enhanced Cd-binding capacity in root cell walls of the two cultivars with higher Cd adsorption levels in the root cell wall of Huajun 2. These results indicate that differences in the amount of cell wall polysaccharide and DM play key roles in establishing the genotypic differences underlying Cd accumulation in pakchoi. These findings conduce to a better understanding of the physiological mechanisms underlying low Cd accumulation in pakchoi and the breeding of new, low-Cd pakchoi cultivars.

Effect of combined application of lead, cadmium, chromium and copper on grain, leaf and stem heavy metal contents at different growth stages in rice.

Most studies on plants' response to heavy metal toxicity have been focusing on single metals. However, soils are always contaminated by several kinds of heavy metals. In this study, pot experiments were carried out to investigate the effects of combined toxicity on two rice genotypes differing in Cd accumulation (Xiushui817, a low-grain-Cd-accumulation and Zheda821, a high-grain-Cd-accumulation genotype). Yield, heavy metal concentrations of grain and leaf/stem at different growth stages were measured under combined application of Cd, Cr, Pb and Cu. Yield was significantly decreased under higher Pb and Cd treatment in both genotypes with Xiushui817 showing greater reduction. Increasing soil Cu level showed no significant effect on grain yield. Zheda821 consistently showed a higher grain Cd content than Xiushui817. The application of Pb, Cd, Cr and Cu significantly affected grain Cd, Cr and Cu accumulations. Similar trends were also observed in leaves and stems at harvest stage. The critical levels of leaf/stem Cd and Cr for safe rice production were also estimated. Alleviation measures should be taken to decrease Cd or Cr accumulations in grain of rice if leaf or stem Cd or Cr concentrations at different growth stages exceed the critical levels.

Comparison of boron uptake and translocation in two vetiver genotypes and evaluation of boron removal efficiency of vetiver floating islands.

The objectives of this study were to identify genotypic differences in the uptake and translocation of boron (B) in vetiver (Chrysopogon zizanioides L.), a promising species in B phytoremediation, and to determine the efficiency of vetiver floating island system in B phytoremediation. Changes in plant biomass and B uptake and translocation were determined in two vetiver genotypes, Sierra and Sunshine, cultured hydroponically in a nutrient solution and exposed to six different B concentrations for 7 d and 14 d. The efficiency of B removal by Sierra (a high B accumulative genotype) grown in floating islands was also determined. Shoot B concentration differed significantly between the two genotypes but differences in root concentration were not significant. Root to shoot B transfer and B uptake ability were higher in Sierra than in Sunshine. The uptake and translocation of B was affected by B concentration and time of exposure. Sierra plants grown in floating islands were more efficient in B removal at lower B concentrations. Most of the B removed accumulated in the middle-upper sections of old leaves. Sierra is more suitable for the removal of B from wastewaters than Sunshine, especially in vetiver floating island system.

Physiological and molecular analysis on root growth associated with the tolerance to aluminum and drought individual and combined in Tibetan wild and cultivated barley.

MAIN CONCLUSION: The drought-stimulated gene expression of NCED, SUS, and KS - DHN and ABA signal cross-talk with other phytohormones maintains barley root growth under drought stress at pH 4.0 plus polyethylene glycol plus aluminum. Aluminum (Al) toxicity and drought are two major factors that limit barley production. In this work, the individual and combined effects of Al/acid and polyethylene glycol (PEG 6000) induced drought stress that suppressed root growth and caused oxidative damage as characterized by increased H2O2 and O2(.-) accumulation. The wild-barley genotypes, XZ5 and XZ29, exhibited a higher tolerance than the two cultivars Dayton (Al tolerant) and Tadmor (drought tolerant) under combined stress (pH 4.0 + PEG + Al). The oxidative damage induced by PEG was more severe at pH 4.0 than at pH 6.0. In XZ29, the highest root secretion of malate and citrate was recorded, and the least Al uptake in the four genotypes. In XZ5, a peak accumulation of ABA and minor synthesis of zeatin riboside and ethylene were found being essential in maintaining primary root elongation and root hair development. PEG-induced drought stress repressed Al uptake in root tips, with a lower increase in callose formation and HvMATE (Hordeum vulgare multidrug and toxic compound exudation) expression compared to Al-induced callose production. Stress by pH 4.0 + PEG + Al up-regulated 9-cis-epoxycarotenoid dioxygenase (NCED) which is involved in ABA biosynthesis. Such treatment stimulated the regulation of ABA-dependent genes sucrose synthase (SUS) and KS-type dehydrin (KS-DHN) in root tips. Our results suggest that the tolerance ranking to pH 4.0 + PEG + Al stress in Tibetan wild barley by gene expression is closely correlated to physiological indices. The results show that acclimatisation to pH 4.0 + PEG + Al stress involves specific responses in XZ5 and XZ29. The present study provides insights into the effects of Al/acid and drought combined stress on the abundance of physiological indices in the roots of barley varieties.

Low-temperature leaf photosynthesis of a Miscanthus germplasm collection correlates positively to shoot growth rate and specific leaf area.

BACKGROUND AND AIMS: The C4 perennial grass miscanthus has been found to be less sensitive to cold than most other C4 species, but still emerges later in spring than C3 species. Genotypic differences in miscanthus were investigated to identify genotypes with a high cold tolerance at low temperatures and quick recovery upon rising temperatures to enable them to exploit the early growing season in maritime cold climates. Suitable methods for field screening of cold tolerance in miscanthus were also identified. METHODS: Fourteen genotypes of M. sacchariflorus, M. sinensis, M. tinctorius and M. × giganteus were selected and grown under warm (24 °C) and cold (14 °C) conditions in a controlled environment. Dark-adapted chlorophyll fluorescence, specific leaf area (SLA) and net photosynthetic rate at a photosynthetically active radiation (PAR) of 1000 µmol m(-2) s(-1) (A1000) were measured. Photosynthetic light and CO2 response curves were obtained from 11 of the genotypes, and shoot growth rate was measured under field conditions. KEY RESULTS: A positive linear relationship was found between SLA and light-saturated photosynthesis (Asat) across genotypes, and also between shoot growth rate under cool field conditions and A1000 at 14 °C in a climate chamber. When lowering the temperature from 24 to 14 °C, one M. sacchariflorus exhibited significantly higher Asat and maximum photosynthetic rate in the CO2 response curve (Vmax) than other genotypes at 14 °C, except M × giganteus 'Hornum'. Several genotypes returned to their pre-chilling A1000 values when the temperature was increased to 24 °C after 24 d growth at 14 °C. CONCLUSIONS: One M. sacchariflorus genotype had similar or higher photosynthetic capacity than M × giganteus, and may be used for cultivation together with M × giganteus or for breeding new interspecies hybrids with improved traits for temperate climates. Two easily measured variables, SLA and shoot growth rate, may be useful for genotype screening of productivity and cold tolerance.

Genome-wide association mapping of cadmium accumulation in different organs of barley.

The threshold value of cadmium (Cd) concentration in grains of barley (Hordeum vulgare) is the lowest among cereal crops; however, it is poorly understood how Cd accumulation in barley grain is genetically controlled. We investigated genotypic variation in Cd accumulation of different organs in 100 accessions from a subset of the barley core collection using both hydroponic and Cd-contaminated soil culture. We also performed a genome-wide association (GWA) mapping for Cd accumulation in different organs. A large genotypic variation in the Cd concentration was found in all organs. There was a good correlation between shoot Cd of solution and soil culture, the shoot Cd and grain Cd, but no correlation between the root Cd and grain Cd. GWA mapping detected 9 quantitative trait loci (QTL) for root Cd, 21 for shoot Cd, 14 for root-to-shoot translocation and 15 for grain Cd. A common QTL for the shoot Cd and root-to-shoot translocation was found at 132.6 cM on chromosome 5H. Two major QTL for grain Cd were identified on chromosome 2H and chromosome 5H. The genetic variation in Cd accumulation and major QTL detected provide useful information helpful for cloning candidate genes for Cd accumulation and breeding low-Cd barley cultivars in future.

Differential expression of Nrat1 is responsible for Al-tolerance QTL on chromosome 2 in rice.

Although rice (Oryza sativa) is the most Al-tolerant species among small-grain cereal crops, there is wide genotypic variation in its tolerance to Al toxicity. A number of quantitative trait loci (QTLs) for Al tolerance have been detected, but the responsible genes have not been identified. By using chromosome segment substitution lines, this work found that Nrat1, a gene encoding an Al transporter, is responsible for a QTL previously detected on chromosome 2. Substitution of the chromosome segment containing Nrat1 from Koshihikari (Al-tolerant variety) by that from Kasalath (Al-sensitive variety) decreased Nrat1 expression and Al uptake and tolerance, but increased binding of Al to the cell wall. Nrat1 in Kasalath showed tissue localization similar to Koshihikari in the roots. Although Koshihikari and Kasalath differed in four amino acids in Nrat1 protein, Nrat1 from Kasalath also showed transport activity for Al. Analysis with site-directed mutagenesis revealed that these differences did not affect the Al-transport activity much. Furthermore, there was no correlation between Al tolerance and the open-reading-frame sequence differences in other rice varieties. On the other hand, there was good correlation between Nrat1 expression and Al tolerance; however, sequence comparison of the promoter region up to 2.1kb did not give a clear difference between the Al-tolerant and -sensitive varieties. Taken together, these results indicate that differential expression of Nrat1 is responsible for the QTL for Al tolerance on chromosome 2, although the mechanism controlling Nrat1 expression remains to be examined.

Morpho-physiological, Genomic, and Transcriptional Diversities in Response to Potassium Deficiency in Rapeseed (Brassica napus L.) Genotypes.

Variations in the resistance to potassium (K) deficiency among rapeseed genotypes emphasize complicated regulatory mechanisms. In this study, a low-K-sensitivity accession (L49) responded to K deficiency with smaller biomasses, severe leaf chlorosis, weaker photosynthesis ability, and deformed stomata morphology compared to a low-K resistant accession (H280). H280 accumulated more K+ than L49 under low K. Whole-genome resequencing (WGS) revealed a total of 5,538,622 single nucleotide polymorphisms (SNPs) and 859,184 insertions/deletions (InDels) between H280 and L49. RNA-seq identified more differentially expressed K+ transporter genes with higher expression in H280 than in L49 under K deficiency. Based on the K+ profiles, differential expression profiling, weighted gene coexpression network analysis, and WGS data between H280 and L49, BnaC4.AKT1 was proposed to be mainly responsible for root K absorption-mediated low K resistance. BnaC4.AKT1 was expressed preferentially in the roots and localized on the plasma membrane. An SNP and an InDel found in the promoter region of BnaC4.AKT1 were proposed to be responsible for its differential expression between rapeseed genotypes. This study identified a gene resource for improving low-K resistance. It also facilitates an integrated knowledge of the differential physiological and transcriptional responses to K deficiency in rapeseed genotypes.

Differential Morpho-Physiological, Ionomic, and Phytohormone Profiles, and Genome-Wide Expression Profiling Involving the Tolerance of Allohexaploid Wheat (Triticum aestivum L.) to Nitrogen Limitation.

Common wheat (Triticum aestivum L.) is a global staple food, while nitrogen (N) limitation severely hinders plant growth, seed yield, and grain quality of wheat. Genetic variations in the responses to low N stresses among allohexaploid wheat (AABBDD, 2n = 6x = 42) genotypes emphasize the complicated regulatory mechanisms underlying low N tolerance and N use efficiency (NUE). In this study, hydroponic culture, inductively coupled plasma mass spectrometry, noninvasive microtest, high-performance liquid chromatography, RNA-seq, and bioinformatics were used to determine the differential growth performance, ionome and phytohormone profiles, and genome-wide expression profiling of wheat plants grown under high N and low N conditions. Transcriptional profiling of NPFs, NRT2s, CLCs, SLACs/SLAHs, AAPs, UPSs, NIAs, and GSs characterized the core members, such as TaNPF6.3-6D, TaNRT2.3-3D, TaNIA1-6B, TaGLN1;2-4B, TaAAP14-5A/5D, and TaUPS2-5A, involved in the efficient transport and assimilation of nitrate and organic N nutrients. The low-N-sensitivity wheat cultivar XM26 showed obvious leaf chlorosis and accumulated higher levels of ABA, JA, and SA than the low-N-tolerant ZM578 under N limitation. The TaMYB59-3D-TaNPF7.3/NRT1.5-6D module-mediated shoot-to-root translocation and leaf remobilization of nitrate was proposed as an important pathway regulating the differential responses between ZM578 and XM26 to low N. This study provides some elite candidate genes for the selection and breeding of wheat germplasms with low N tolerance and high NUE.

Physiological and biochemical characteristics of high and low Cd accumulating Brassica napus genotypes.

Phytoremediation is a widely used and cost-effective technique for in situ remediation of heavy metals. Brassica napus L. genotype with high Cd accumulation and strong Cd tolerance is an ideal candidate for phytoremediation. In this study, a hydroponic experiment was conducted to select a Brassica napus genotype with either high or low Cd accumulation from a panel of 55 genotypes. The physiological mechanisms governing Cd accumulation and Cd tolerance were then explored. BN400 and BN147 were identified as the high and low Cd accumulating genotypes, respectively. Additionally, BN400 exhibited greater tolerance to Cd stress compared to BN147. Root morphology analysis revealed that BN400 exhibited longer root length, smaller root surface area and root volume, and less root tips but bigger root diameter than BN147. Subcellular Cd distribution showed that the Cd concentrations in the cell wall and vacuole in shoot were significantly higher in BN400 than in BN147, whereas the opposite trend was observed in the roots.. Pectate/protein-integrated Cd was found to be the predominant form of Cd in both shoots and roots, with significantly higher levels in BN400 compared to BN147 in the shoot, but the opposite trend was observed in the roots. These results suggest that the long fine roots play a role in Cd accumulation. The high Cd accumulating genotype was able to retain Cd in leaf cell walls and vacuoles, and Cd was mainly present in the form of pectate/protein-integrated Cd, which contributes to its strong Cd tolerance. These findings have important implications for the screening and breeding of Brassica napus genotypes with high Cd accumulation for phytoremediation purposes.

Identification of low grain cadmium accumulation genotypes and its physiological mechanism in maize (Zea mays L.).

Soil cadmium (Cd) contamination poses adverse impacts on crop yield and quality. Maize is a widely cultivated cereal throughout the world. In this study, field and hydroponic experiments were conducted to investigate the genotypic difference in Cd accumulation and tolerance in maize. There were significant genotypic differences in grain Cd concentrations among 95 genotypes. From these 95 genotypes, L42 which showed a higher grain Cd concentration and L63 which showed a lower grain Cd concentration was selected for further study. Under Cd stress, L63 showed much less reduction in plant growth than L42 compared with the control. Seedlings of L63 recorded higher Cd concentration in roots, but lower in shoots L42, indicating that the low grain Cd concentration in L63 is mainly due to the low rate of transportation of Cd from roots to shoots. Most Cd accumulated in epidermis and xylem vessels of L63, while the green fluorescent was found across almost the entire cross-section of root in L42. Obvious ultrastructural damage was observed in L42 under Cd stress, especially in mesophyll cells, while L63 was less affected. These findings could contribute to developing low Cd accumulation and high tolerance maize cultivars.

Expression Level of Transcription Factor ART1 Is Responsible for Differential Aluminum Tolerance in Indica Rice.

Rice is the most aluminum (Al)-tolerant species among the small grain cereals, but there are great variations in the Al tolerance between subspecies, with higher tolerance in japonica subspecies than indica subspecies. Here, we performed a screening of Al tolerance using 65 indica cultivars and found that there was also a large genotypic difference in Al tolerance among indica subspecies. Further characterization of two cultivars contrasting in Al tolerance showed that the expression level of ART1 (ALUMINUM RESISTANCE TRANSCRIPTION FACTOR 1) encoding a C2H2-type Zn-finger transcription factor, was higher in an Al-tolerant indica cultivar, Jinguoyin, than in an Al-sensitive indica cultivar, Kasalath. Furthermore, a dose-response experiment showed that ART1 expression was not induced by Al in both cultivars, but Jinguoyin always showed 5.9 to 11.4-fold higher expression compared with Kasalath, irrespectively of Al concentrations. Among genes regulated by ART1, 19 genes showed higher expression in Jinguoyin than in Kasalath. This is associated with less Al accumulation in the root tip cell wall in Jinguoyin. Sequence comparison of the 2-kb promoter region of ART1 revealed the extensive sequence polymorphism between two cultivars. Whole transcriptome analysis with RNA-seq revealed that more genes were up- and downregulated by Al in Kasalath than in Jinguoyin. Taken together, our results suggest that there is a large genotypic variation in Al tolerance in indica rice and that the different expression level of ART1 is responsible for the genotypic difference in the Al tolerance.

Variations in cadmium and nitrate co-accumulation among water spinach genotypes and implications for screening safe genotypes for human consumption.

Vegetables are important constituents of the human diet. Heavy metals and nitrate are among the major contaminants of vegetables. Consumption of vegetables and fruits with accumulated heavy metals and nitrate has the potential to damage different body organs leading to unwanted effects. Breeding vegetables with low heavy metal and nitrate contaminants is a cost-effective approach. We investigated 38 water spinach genotypes for low Cd and nitrate co-accumulation. Four genotypes, i.e. JXDY, GZQL, XGDB, and B888, were found to have low co-accumulation of Cd (<0.71 mg/kg dry weight) and nitrate (<3100 mg/kg fresh weight) in the edible parts when grown in soils with moderate contamination of both Cd (1.10 mg/kg) and nitrate (235.2 mg/kg). These genotypes should be appropriate with minimized risk to humans who consume them. The Cd levels in the edible parts of water spinach were positively correlated with the concentration of Pb or Zn, but Cd, Pb, or Zn was negatively correlated with P concentration. These results indicate that these three heavy metals may be absorbed into the plant in similar proportions or in combination, minimizing the influx to aerial parts. Increasing P fertilizer application rates appears to prevent heavy metal and nitrate translocation to shoot tissues and the edible parts of water spinach on co-contaminated soils.

Genotypic differences in cadmium transport in developing barley grains.

Genotypic differences in cadmium (Cd) transport in developing grains of barley (Hordeum vulgare L.) were investigated using detached ears cultured in a nutrient solution containing 0.5 and 5 µM Cd. Cd concentration in each part of the ear in W6nk2 (a low-grain-Cd-accumulation genotype) was much less than in Zhenong8 (a high accumulator) with 0.5 µM Cd treatment. However, Cd concentration in W6nk2 grains increased with an increase in external Cd level and was similar to Zhenong8 with 5 µM Cd treatment. Awn removal, a high relative humidity (RH, 90%) and addition of sucrose markedly decreased grain Cd concentration in Zhenong8 but less affected Cd transport to grain in W6nk2. Stem girdling reduced Cd transport to developing grains with 5 µM Cd treatment, especially for W6nk2, whereas no effect was found in either genotype with low Cd treatment. Our results suggested that higher grain Cd in Zhenong8 is closely related to a larger capacity for xylem transport and is connected with Cd translocation in xylem and phloem sap.

Genome-wide identification of chromium stress-responsive micro RNAs and their target genes in tobacco (Nicotiana tabacum) roots.

Tobacco easily accumulates certain heavy metals in leaves and thus poses a potential threat to human health. To systematically dissect Cr-responsive microRNAs (miRNAs) and their targets at the global level, 4 small RNA libraries were constructed from the roots of Cr-treated (Cr) and Cr-free (control) for 2 contrasting tobacco genotypes,Yunyan2 (Cr-sensitive) and Guiyan1 (Cr-tolerant). Using high-throughput-sequencing-technology, the authors identified 53 conserved and 29 novel miRNA families. Comparative genomic analysis of 41 conserved Cr-responsive miRNA families revealed that 11 miRNA families showed up-regulation in Guiyan1 but unaltered in Yunyan2, and 17 miRNA families were up-regulated only in Yunyan2 under Cr stress. Only 1 family, miR6149, was down-regulated in Yunyan2 but remained unchanged in Guiyan1. Of the 29 novel miRNA families, 14 expressed differently in the 2 genotypes under Cr stress. Based on a high-throughput degradome sequencing homology search, potential targets were predicted for the 41 conserved and 14 novel Cr-responsive miRNA families. Clusters of Orthologous Groups functional category analysis revealed that some of these predicted target transcripts of miRNAs are responsive to biotic and abiotic stresses. Furthermore, the expression patterns of many Cr-responsive miRNAs were validated by stem-loop real-time transcription polymerase chain reaction. The results of the present study provide valuable information and a framework for understanding the function of miRNAs in Cr tolerance.

Differences in physiological features associated with aluminum tolerance in Tibetan wild and cultivated barleys.

Aluminum (Al) toxicity is a major limiting factor for plant production in acid soils. Wild barley germplasm is a treasure trove of useful genes and offers rich sources of genetic variation for crop improvement. Al-stress-hydroponic-experiments were performed, and the physiochemical characteristic of two contrasting Tibetan wild barley genotypes (Al-resistant XZ16 and Al-sensitive XZ61) and Al-resistant cv. Dayton were compared. Ultrastructure of chloroplasts and root cells in XZ16 was less injured than that in Dayton and XZ61. Moreover, XZ16 secreted significantly more malate besides citrate and exhibited less Al uptake and distribution than both of XZ61 and Dayton in response to Al stress, simultaneously maintained higher H⁺-, Ca²âºMg²âº- and total-ATPase activities over XZ61. The protein synthesis inhibitor cycloheximide reduced citrate secretion from XZ16, but not from Dayton. In Tibetan wild barley, our findings highlight the significant correlations between Al tolerance, ATPase activity and citrate secretion, providing some insights into the physiological basis for Al-detoxification.