Imrul Mosaddek Ahmed

Comparative Proteomic Analysis of 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 rich in genetic diversity and may provide elite genes for crop Al tolerance improvement. The hydroponic-experiments were performed to compare proteomic and transcriptional characteristics of two contrasting Tibetan wild barley genotypes Al- resistant/tolerant XZ16 and Al-sensitive XZ61 as well as Al-resistant cv. Dayton. Results showed that XZ16 had less Al uptake and translocation than XZ61 and Dayton under Al stress. Thirty-five Al-tolerance/resistance-associated proteins were identified and categorized mainly in metabolism, energy, cell growth/division, protein biosynthesis, protein destination/storage, transporter, signal transduction, disease/defense, etc. Among them, 30 were mapped on barley genome, with 16 proteins being exclusively up-regulated by Al stress in XZ16, including 4 proteins (S-adenosylmethionine-synthase 3, ATP synthase beta subunit, triosephosphate isomerase, Bp2A) specifically expressed in XZ16 but not Dayton. The findings highlighted the significance of specific-proteins associated with Al tolerance, and verified Tibetan wild barley as a novel genetic resource for Al tolerance.

Genotypic differences in physiological characteristics in the tolerance to drought and salinity combined stress between Tibetan wild and cultivated barley

Greenhouse pot experiments were conducted to investigate genotypic differences in response to individual and combined stresses of drought and salinity between Tibetan wild barley genotypes (XZ5, drought-tolerant; XZ16, salinity/aluminum tolerant) and cv. CM72 (salinity-tolerant). Either drought (D) or salinity (S) alone and in combination (D + S) stresses significantly decreased plant growth, chlorophyll content, net photosynthetic rate (Pn), maximal photochemical efficiency of PSII (Fv/Fm), water potential and osmotic potential, with the largest suppression under combined stress, and two wild genotypes showing more tolerance than CM72. Water use efficiency (WUE) increased significantly in XZ5 and XZ16 after D + S, but no significant change in CM72. XZ5 and XZ16 showed 30.9% and 12.1% higher K(+) level and 30.5% and 24.1% lower Na(+)/K(+) ratio in plants, compared with CM72, with increased metal nutrients as Ca, Fe and Mn under D + S. The peak accumulation in proline and glycine-beatine was recorded in combined stress with larger accumulation in two wild genotypes. Moreover, larger increases in the level of ASA and GSH, and the activities of Ca(2+)Mg(2+)-ATPase, and superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), guaiacol peroxidase (POD) and glutathione reductase (GR) under D + S vs control were observed in XZ5 and XZ16 than CM72, with less accumulation of H(2)O(2) and malondialdehyde. These results suggest that high tolerance to D + S stress of XZ5 and XZ16 is closely related to lower Na(+)/K(+) ratio and enhanced Ca(2+)Mg(2+)-ATPase, proline, glycine-beatine and WUE, and improved capacity of antioxidative performance to scavenge reactive oxygen species and thus suppressed level of lipid peroxidation.

Difference in Yield and Physiological Features in Response to Drought and Salinity Combined Stress during Anthesis in Tibetan Wild and Cultivated Barleys

Soil salinity and drought are the two most common and frequently co-occurring abiotic stresses constraining crop growth and productivity. Greenhouse pot experiments were conducted to investigate the tolerance potential and mechanisms of Tibetan wild barley genotypes (XZ5, drought-tolerant; XZ16, salinity/aluminum tolerant) during anthesis compared with salinity-tolerant cv CM72 in response to separate and combined stresses (D+S) of drought (4% soil moisture, D) and salinity (S). Under salinity stress alone, plants had higher Na+ concentrations in leaves than in roots and stems. Importantly, XZ5 and XZ16 had substantially increased leaf K+ concentrations; XZ16 was more efficient in restricting Na+ loading in leaf and maintained a lower leaf Na+/K+ ratio. Moreover, a significant decrease in cell membrane stability index (CMSI) and an increase in malondialdehyde (MDA) were accompanied by a dramatic decrease in total biomass under D+S treatment. We demonstrated that glycine-betaine and soluble sugars increased significantly in XZ5 and XZ16 under all stress conditions, along with increases in protease activity and soluble protein contents. Significant increases were seen in reduced ascorbate (ASA) and reduced glutathione (GSH) contents, and in activities of H+K+-, Na+K+-, Ca++Mg++-, total- ATPase, and antioxidant enzymes under D+S treatment in XZ5 and XZ16 compared to CM72. Compared with control, all stress treatments significantly reduced grain yield and 1000-grain weight; however, XZ5 and XZ16 were less affected than CM72. Our results suggest that high tolerance to D+S stress in XZ5 and XZ16 is closely related to the lower Na+/K+ ratio, and enhanced glycine-betaine and soluble protein and sugar contents, improved protease, ATPase activities and antioxidative capacity for scavenging reactive oxygen species during anthesis. These results may provide novel insight into the potential responses associated with increasing D+S stress in wild barley genotypes.

Genotypic and environmental variation in cadmium, chromium, lead and copper in rice and approaches for reducing the accumulation

The field scale trials revealed significant genotypic and environmental differences in grain heavy metal (HM) concentrations of 158 newly developed rice varieties grown in twelve locations of Zhejiang province of China. Grain Pb and Cd contents in 5.3% and 0.4% samples, respectively, were above the maximum permissible concentration (MPC); none of samples had Cr/Cu exceeding MPC. Stepwise multiple linear regression analysis estimated soil HM critical levels for safe rice production. Low grain HM accumulation cultivars such as Xiushui817, Jiayou08-1 and Chunyou689 were recommended as suitable cultivars for planting in slight/medium HM contaminated soils. The alleviating regulator (AR) of (NH₄)₂SO₄ as N fertilizer coupled with foliar spray of a mixture containing glutathione (GSH), Si, Zn and Se significantly decreased grain Cd, Cr, Cu and Pb concentrations grown in HM contaminated fields with no effect on yield, indicating a promising measurement for further reducing grain HM content to guarantee safe food production.

HvEXPB7, a novel β-expansin gene revealed by the root hair transcriptome of Tibetan wild barley, improves root hair growth under drought stress

Highlight A novel root hair development related gene, HvEXPB7, was identified and cloned from the identified drought tolerance-associated genes. BSMV-VIGS of HvEXPB7 confirmed that this gene was involved in root hair growth under drought.

Differential changes in grain ultrastructure, amylase, protein and amino acid profiles between Tibetan wild and cultivated barleys under drought and salinity alone and combined stress

Grain phytochemical profiles were compared in Tibetan wild barley XZ5 (drought-tolerant), XZ16 (salinity/aluminum-tolerant) and cv CM72 (salinity-tolerant) in response to drought and salinity alone and combination (D+S) during anthesis. Total antioxidant capacity assessed by determining ferric-reducing antioxidant potential (FRAP) in grains increased significantly as follows: D+S>drought>salinity, and XZ5>XZ16>CM72. A marked increase in the total phenol (TP) from individual and combined stresses was observed in XZ5, while a decrease occurred in CM72. Moreover, the activity of α-/β-amylase in the grains under combined stress was 81.8%/16.9% in XZ5 and 48.6%/18.7% in XZ16 higher than that of CM72. Increases in amino acids, protein content and protein fractions of albumin, globulin, hordein and glutelin were maximised under D+S, with larger values in the Tibetan wild genotypes. Observation with a scanning electron microscopy showed a distinct genotypic difference under D+S; for example, XZ5 and XZ16 maintained a relatively integral starch granule with a greater protein deposit/matrix, while CM72 degraded by pitting. This research expands our understanding of barley drought and salt-tolerance mechanisms and provides possibility of Tibetan wild barley in developing barley cultivars with both tolerance to drought and salinity.

The changes in physiological and biochemical traits of Tibetan wild and cultivated barley in response to low phosphorus stress

Abstract Low phosphorus (LP) limits crop growth and productivity in the majority of arable lands worldwide. Here, we investigated the changes in physiological and biochemical traits of Tibetan wild barleys (Hordeum vulgare L. ssp. spontaneum) XZ99 (LP tolerant), XZ100 (LP sensitive), and cultivated barley ZD9 (moderately LP tolerant) under two phosphorus (P) levels during vegetative stage. These genotypes showed considerable differences in the change of biomass accumulation, root/shoot dry weight ratio, root morphology, organic acid secretion, carbohydrate metabolism, ATPase (Adenosine triphosphatase) activity, P concentration and accumulation under LP in comparison with CK (control) condition. The higher LP tolerance of XZ99 is associated with more developed roots, enhanced sucrose biosynthesis and hydrolysis of carbohydrate metabolism pathway, higher APase (Acid phosphatase) and ATPase activity, and more secretion of citrate and succinate in roots when plants are exposed to LP stress. The results prove the potential of Tibetan wild barley in developing barley cultivars with high tolerance to LP stress and understanding the mechanisms of LP tolerance in plants.

Differences in grain ultrastructure, phytochemical and proteomic profiles between the two contrasting grain Cd-accumulation barley genotypes.

To reveal grain physio-chemical and proteomic differences between two barley genotypes, Zhenong8 and W6nk2 of high- and low- grain-Cd-accumulation, grain profiles of ultrastructure, amino acid and proteins were compared. Results showed that W6nk2 possesses significantly lower protein content, with hordein depicting the greatest genotypic difference, compared with Zhenong8, and lower amino acid contents with especially lower proportion of Glu, Tyr, Phe and Pro. Both scanning and transmission electron microscopy observation declared that the size of A-type starch molecule in W6nk2 was considerably larger than that of Zhenong8. Grains of Zhenong8 exhibited more protein-rich deposits around starch granules, with some A-type granules having surface pits. Seventeen proteins were identified in grains, using 2-DE coupled with mass spectrometry, with higher expression in Zhenong8 than that in W6nk2; including z-type serpin, serpin-Z7 and alpha-amylase/trypsin inhibitor CM, carbohydrate metabolism, protein synthesis and signal transduction related proteins. Twelve proteins were less expressed in Zhenong8 than that in W6nk2; including barley trypsin inhibitor chloroform/methanol-soluble protein (BTI-CMe2.1, BTI-CMe2.2), trypsin inhibitor, dehydroascorbate reductase (DHAR), pericentrin, dynein heavy chain and some antiviral related proteins. The data extend our understanding of mechanisms underlying Cd accumulation/tolerance and provides possible utilization of elite genetic resources in developing low-grain-Cd barley cultivars.

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⁺-, Ca²⁺Mg²⁺- 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.

Identification of the differentially accumulated proteins associated with low phosphorus tolerance in a Tibetan wild barley accession.

Low phosphorus (LP) in soil is a widely-occurred limiting factor for crop production in the world. In a previous study we identified a highly LP-tolerant Tibetan wild barley accession (XZ99). Here, a comparatively proteomic analysis was conducted using three barley genotypes differing in LP tolerance to reveal the mechanisms underlying the LP tolerance of XZ99. Totally, 31 differentially accumulated proteins were identified in the roots and leaves of the three genotypes using 2-dimensional gel electrophoresis coupled with mass spectrometry. They were involved in the various biological processes, including carbon and energy metabolism, signal transduction, cell growth and division, secondary metabolism, and stress defense. In comparison with XZ100 (LP sensitive) and ZD9 (LP moderately-tolerant), XZ99 had a more developed root system, which is mainly attributed to enhanced carbohydrate metabolizing proteins under LP conditions. The current results showed that Tibetan wild barley XZ99 and cultivated barley cultivar ZD9 differ in the mechanism of LP tolerance. The changes of the proteins associated with carbohydrate metabolism could account for the difference between the LP-tolerant and LP-sensitive genotypes. In addition, the mRNA expression levels of 9 LP responsive proteins were verified by qRT-PCR. The current results may open a new avenue of understanding the LP tolerance in plants on the proteomic basis.