Aluminium (Al) toxicity is one of the major limiting factors for barley production on acid soils It inhibits root cell division and elongation, thus reducing water and nutrient uptake, consequently resulting in poor plant growth and yield Plants tolerate Al either through external resistance mechanisms, by which Al is excluded from plant tissues or internal tolerance mechanisms, conferring the ability of plants to tolerate Al ion in the plant symplasm where Al that has permeated the plasmalemma is sequestered or converted into an innocuous form Barley is considered to be most sensitive to Al toxicity among cereal species Al tolerance in barley has been assessed by several methods, such as nutrient solution culture, soil bioassay and field screening Genetic and molecular mapping research has shown that Al tolerance in barley is controlled by a single locus which is located on chromosome 4H Molecular markers linked with Al tolerance loci have been identified and validated in a range of diverse populations This paper reviews the (1) screening methods for evaluating Al tolerance, (2) genetics and (3) mechanisms underlying Al tolerance in barley

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Aluminium tolerance in barley ( Hordeum vulgare L.): physiological mechanisms, genetics and screening methods

To better understand the mechanisms involved in aluminum toxicity and tolerance in plants, microarray technology was used to evaluate changes in gene expression in Arabidopsis thaliana under Al stress. With the use of Affymetrix Arabidopsis ATH1 Genechip, a comparison of RNA expression profiles was made between control and Al-treated Arabidopsis seedlings. A total of 256 genes were identified as Al-responsive. Ninety-four genes were shown to be up-regulated and 162 were down-regulated; comprising 1.1 % of the 24 000 Arabidopsis genes. Real-time RT-PCR was used to confirm the microarray data. The analysis showed that a large number of transcription factors and several putative signaling components were up-regulated by aluminum. Chloroplast structural and photosynthetic genes were, in general, down-regulated. A number of previously identified Al-responsive genes, e.g. GST, Auxin-regulated, Peroxidase, and Chitinase, were up-regulated by Al-stress, whereas Wali 3 and Wali 4 were down-regulated. We also identified several up-regulated genes involved in vacuolar signaling, sorting and docking. Three genes were also up-regulated by Al-stress, Ras GTP-binding protein, ABC-cassette binding, and the AtELP1 receptor genes, have previously been documented as responsive to drought and/or oxidative stress and may play important roles the detoxification of Al ions by transportation and storage into root vacuoles. Ultrastructural changes in the roots tips cells of Arabidopsis were evaluated using transmission electron microscopy and energy-dispersive X-ray analysis with scanning electron microscopy and results showed Al accumulation in the root tips of Arabidopsis.

Microarray analysis of Arabidopsis genome response to aluminum stress

Barley is one of the cereals that are most sensitive to aluminum (Al). Al in acid soils limits barley growth and development and, as a result, its productivity. The inhibition of root growth is a widely accepted indicator of Al stress. Al toxicity is affected by many factors including the culture medium, pH, Al concentration and the duration of the treatment. However, Al can act differently in different species and still Al toxicity in barley deserves study. Since the mechanism of Al toxicity is discussed we cytogenetically describe the effects of different doses of bioavailable Al on the barley nuclear genome—mitotic activity, cell cycle profile and DNA integrity. At the same time, we tested an established deep-water culture (DWC) hydroponics system and analyzed the effects of Al on the root system parameters using WinRHIZO software. We demonstrated the cytotoxic and genotoxic effect of Al in barley root cells. We showed that Al treatment significantly reduced the mitotic activity of the root tip cells and it also induced micronuclei and damaged nuclei. The DNA-damaging effect of Al was observed using the TUNEL test. We define the inhibitory influence of Al on DNA replication in barley. Analysis with the labelling and detection of 5-ethynyl-2‘-deoxyuridin (EdU) showed that the treatment with Al significantly decreased the frequency of S phase cells. We also demonstrated that Al exposure led to changes in the cell cycle profile of barley root tips. The delay of cell divisions observed as increased frequency of cells in G2/M phase after Al treatment was reported using flow cytometry.

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Analysis of aluminum toxicity in Hordeum vulgare roots with an emphasis on DNA integrity and cell cycle

Wild barley (Hordeum sp.) germplasm is rich in genetic diversity and provides a treasure trove of useful genes for crop improvement. We carried out a comprehensive program combining short-term hydroponic screening via hematoxylin-staining of root-regrowth procedure and filter paper–based evaluation of diverse germplasm in response to Al/acid stress using 105 annual Tibetan wild barley and 45 cultivated barley genotypes. Root elongation among the 105 Tibetan wild barley genotypes varied significantly after Al exposure, ranging from 62.9% to 80.0% in variation coefficients and 4.35 to 4.45 in diversity index. These genotypic differences in Al resistance were fairly consistent in both the hydroponic and filter paper–based evaluations: XZ16, XZ166, and XZ113 were selected as Al-resistant genotypes, and XZ61, XZ45, and XZ98 as Al-sensitive wild genotypes. Furthermore, significantly lower Al concentrations in roots and shoots were detected in the three selected Al-resistant genotypes than in the three sensitive genotypes in the filter paper–based experiment. Meanwhile, XZ16 was the least affected by Al toxicity in regard to reduced SPAD value (chlorophyll meter readings), plant height, root length, dry biomass, tillers per plant, and chlorophyll fluorescence (Fv/Fm) in the long-term hydroponic experiment compared with the Al-resistant cultivated barley cv. Dayton, while XZ61 had the severest stress symptoms.

Difference in response to aluminum stress among Tibetan wild barley genotypes

This study tested the hypothesis that lucerne (Medicago sativa L.) populations selected in solution culture high in aluminium (Al) would increase seedling root growth when grown in an acid soil high in exchangeable Al. Root growth of six elite populations (Aurora C2, UQL-1 C2, T02-011 C1, T02-011 C2, A513 C3 and Sardi 7 C2) selected in high-Al solution culture (SHASC) was compared with that of corresponding parent as well as the Georgia acid soil-tolerant populations in an acid soil in pots grown for 8 days under controlled environmental conditions. Lime was added to the soil to provide contrasts in the severity of stress imposed by low pH and high Al. Averaged across six SHASC populations, total root length increased 19% at pH 4.34 in CaCl2 (35% exchangeable Al) and 26% at pH 5.26 (<1% exchangeable Al) compared with the control populations. At all pH levels SHASC populations showed increased tap root length, total root length (includes lateral roots), root weight and root surface area, but decreased average root diameter compared with the six control populations. A large amount of variability was observed both between and within lucerne populations with three SHASC populations (Aurora C2, UQL-1 C2 and Sardi 7 C2) exhibiting increased root growth at lower pH levels, but little increase in root length at higher pH, consistent with increased tolerance to Al toxicity. This was in contrast to three other SHASC populations (T02-011 C1, T02-011 C2 and A513 C3), which exhibited increased root length at all pH levels, consistent with increased seedling vigour. The Sardi 7 C2 population exhibited the greatest increase in tap root growth with tap root length increasing by 40 and 30% at pH 4.34 and 4.48, respectively, compared with its parent population Sardi 7. This study provides evidence that seedlings of lucerne populations selected in high-Al solution culture can confer significantly improved root and shoot growth in acid soil. It is recommended that such screening be incorporated into lucerne breeding programs to reduce costs in space and time.

Seedling validation of acid soil tolerance of lucerne populations selected in solution culture high in aluminium

Aluminium (Al) toxicity in the soil is an important factor that limits the production of barley in areas with acid soil. Selection and breeding of barley cultivars tolerant to Al toxicity is one of the most useful approaches to increase productivity. A reliable screening system is very important for selecting Al-tolerant plants in a breeding program. Using a hydroponic culture technique in which all the treatments were isolated in order to minimise complex interaction between genotypes, experiments were conducted to distinguish between susceptible and tolerant cultivars. Three different methods were investigated. Two previously reported methods could not provide consistent results or detect the difference between tolerant and susceptible cultivars. A new method was developed as follows: pre-germinated seedlings (2 days at 22°C) were cultured for 3 days in nutrient solution (Al free) followed by 24 h growing in a solution with 50 or 100 µm Al, and then 48 h regrowth in Al free nutrient solution. Following this method, seminal root regrowth length (SRRL) and relative seminal root regrowth length (RSRRL) showed significant differences between tolerant and sensitive cultivars. The SRRL of the most tolerant cultivar, Dayton, was 4–8 times greater than of the sensitive cultivars and about twice as long as of the other tolerant cultivars, FM404 and Brindabella. All the sensitive cultivars showed significantly shorter SRRL or RSRRL. Both SRRL and RSRRL were found to be closely correlated with plant height, plant dry weight, and grain weight in a soil-based experiment. This method was also used to evaluate F2 populations from crosses between tolerant and susceptible cultivars. Both SRRL and RSRRL gave results consistent with the hypothesis that the tolerance was controlled by a single dominant gene.

Mamun Hossain

Papers

A reliable screening system for aluminium tolerance in barley cultivars