Responses of Al-tolerant Dayton and Al-sensitive Kearney barley cultivars to calcium and magnesium during al stress

Summary Two barley cultivars differing in Al tolerance, Kearney (Al-sensitive) and Dayton (Al-tolerant) were exposed to Al stress with varied Ca and Mg concentrations in the nutrient solution. Increase in calcium and magnesium supply protected root meristems and root growth from Al toxicity more effectively in the Al-tolerant cultivar than in the Al-sensitive one. Lateral roots were much more sensitive to Al than adventitious roots. Exposure to 0.33 mM Al with low concentrations of Ca (1.3 mM) and Mg (0.3 mM) caused damage to root tips in both cultivars. Increasing the Ca concentration to 4.3 and 6.3 mM prevented root tip damage in Dayton but not in Kearney. In the Al-tolerant cultivar Dayton, however, the root tips regenerated even at the low Ca concentration of 1.3 mM, whereas 6.3 mM Ca was necessary for this to occur in Kearney. This difference was due to the fact that Dayton's root meristem cells were more resistant to damage. Magnesium responses also varied between the two cultivars. At the lowest Ca concentration an increase in Mg to 6.3 mM permitted regeneration of damaged Kearney root tips and completely prevented any damage in Dayton. It is to be assumed that the different responses of the two cultivars are due to differences in plasma membrane properties.     

Influence of aluminum in nutrient solutions on chemical composition in two rice cultivars at different growth stages

Summary A solution culture experiment was conducted using 2 rice cultivars (EEA 304, and CICA 4) to determine the effects of Al concentrations on chemical composition. The treatments consisted of five Al concentrations: 0, 10, 30, 40 and 60 ppm.Aluminum content in plant tissues way increased with increasing levels of Al in two cultivars. Increased Al concentrations in the nutrient solution exerted an inhibiting effect on the uptake of N, P, K, Ca, Mg, S, Fe, B, Cu, Zn, and Mn. Rice cultivars responded differently to Al treatments with respect to nutrients uptake. Tolerant cultivar, EEA 304, absorbed more phosphorus compared to susceptible cultivar CICA 4. Macro and micronutrients inhibiting effect was much lower in this Al tolerant cultivar. These results suggested that one of the Al tolerance mechanism in rice cultivars is associated with more efficient nutrients uptake.     

Interaction between iron plaque and root border cells ameliorates aluminum toxicity of Oryza sativa differing in aluminum tolerance

Root border cells (RBCs), which are generated during plant growth and surround the root cap, and iron plaque (IP), ubiquitously formed on the root surfaces of rice, are known to alleviate aluminum (Al) toxicity. To verify the interactive effects of IP and RBCs on ameliorating Al toxicity, two rice cultivars differing in Al resistance were used to compare Al tolerance between cultivars. Additionally, root elongation, Al uptake and RBCs viability were measured as indicators of the effects of Al. The amounts of DCB-extractable Fe and Al on the root surfaces were much higher in the presence of IP than the absence. IP presence significantly decreased Al-induced inhibition of root elongation and Al contents in roots and root tips. The removal of RBCs from the root tips caused a more severe inhibition of root elongation and a higher Al accumulation in rice roots and root tips. Furthermore, root growth inhibition and Al contents in roots and root tips were significantly lower in roots with a combination of IP and RBCs than in roots with IP or RBCs only. The formation of IP on the root surface maintained higher RBCs viability and depressed mucilage exudation in an Al-tolerant rice cultivar. The results suggest that both IP and RBCs ameliorate Al toxicity, and IP has a greater capacity for Al resistance. The combination of IP and RBCs exhibited a synergistic effect associated with Al resistance.     

Response and tolerance of root border cells to aluminum toxicity in soybean seedlings

Root border cells (RBCs) and their secreted mucilage are suggested to participate in the resistance against toxic metal cations, including aluminum (Al), in the rhizosphere. However, the mechanisms by which the individual cell populations respond to Al and their role in Al resistance still remain unclear. In this research, the response and tolerance of RBCs to Al toxicity were investigated in the root tips of two soybean cultivars [Zhechun No. 2 (Al-tolerant cultivar) and Huachun No. 18 (Al-sensitive cultivar)]. Al inhibited root elongation and increased pectin methylesterase (PME) activity in the root tip. Removal of RBCs from the root tips resulted in a more severe inhibition of root elongation, especially in Huachun No. 18. Increasing Al levels and treatment time decreased the relative percent viability of RBCs in situ and in vitro in both soybean cultivars. Al application significantly increased mucilage layer thickness around the detached RBCs of both cultivars. Additionally, a significantly higher relative percent cell viability of attached and detached RBCs and thicker mucilage layers were observed in Zhechun No. 2. The higher viability of attached and detached RBCs, as well as the thickening of the mucilage layer in separated RBCs, suggest that RBCs play an important role in protecting root apices from Al toxicity.     

Aluminium toxicity in rye (Secale cereale): root growth and dynamics of cytoplasmic Ca2+ in intact root tips

Aluminium (Al) toxicity in rye (Secale cereale L.), an Al-resistant crop, was examined by measuring root elongation and cytoplasmic free activity of calcium ([Ca2+]cyt) in intact root apical cells. Measurement of [Ca2+]cyt, was achieved by loading a Ca2+-sensitive fluorescent probe. Fluo-3/AM ester, into root apical cells followed by detection of intracellular fluorescence using a confocal laser scanning microscope. After 20 min of exposure to 50 microM Al (pH 4-2) a slight increase in [Ca2+]cyt of root apical cells was observed, while the response of [Ca2+]cyt to 100 microM Al (pH 4.2) was faster and larger ([Ca2+]cyt increased by 46% in 10 min). Increases in [Ca2+]cyt were correlated with inhibition of root growth, generally measurable after 2 h. Addition of 400 microM malic acid (pH 4.2) largely ameliorated the effect of 100 microM Al on [Ca2+]cyt in root apical cells and protected root growth from Al toxicity. These results suggest that an increase in [Ca2+]cyt in root apical cells in rye is an early effect of Al toxicity and is followed by the secondary effect on root elongation.     

Aluminium effects on growth, nutrient net uptake and transport in 3 rice (Oryza sativa) cultivars with different sensitivity to aluminium

Varietal differences in net nutrient uptake rate and transport efficiency in the presence of aluminium have seldom been investigated in rice. Therefore, effects of Al on growth, uptake and transport of macronutrients (K, P, Ca, and Mg) and micronutrients (Fe, Zn, Cu, and Mn) were evaluated in 3 rice cultivars (BG35, DA14 and IR45) with different Al sensitivity. The plants were grown in nutrient solution at pH 4.1. An initial growth was completed in the time interval 1 to 5 days immediately before the addition of Al. The final growth period with Al (0, 140, 280 or 560 μ M ) was completed on day 26. With Al, a comparatively high P accumulation occurred in shoots and roots of the Al tolerant cultivar BG35. In contrast, the Al sensitive cultivar IR45 maintained a relatively high Ca accumulation during the Al treatment. A reduced total net uptake rate of P and Ca by IR45 in the time period 5 to 26 days was due to both a reduced root fresh weight and a reduced net uptake rate per g fresh weight of root. Moreover, net Ca transport to the shoots higher than net uptake rate in DA14 and IR45 at > 140 μ M Al during the test period suggests restricted Ca uptake by the roots in combination with a continuous net loss of Ca from the roots to the shoots as time proceeds. In the case of Mg and Mn, there was a general reduction of net uptake rates, irrespective of Al sensitivity of cultivars. With Al treatment, comparatively high accumulation of Fe, Zn and Cu occurred in the roots of IR45, concomitant with a high net Zn and Cu uptake rate. It is concluded that differences in Al sensitivity among rice cultivars BG35, DA14 and IR45 are not primarily linked to the depressed internal Mg or Mn status of the plants but rather to changes in the uptake and distribution of Ca and P.     

水稻依赖抗坏血酸H2O2清除系统在抗铁毒中的作用

根据营养液培养试验从水稻Ａｚｕｃｅｎａ×ＩＲ６４ 发展的一双单倍体（ＤＨ） 群体中筛选出抗铁毒与敏感品系。在铁毒害处理后,各品系的抗坏血酸过氧化物酶（ＡＰ） 、脱氢抗坏血酸还原酶（ＤＲ） 、谷胱甘肽还原酶（ＧＲ） 的活性均有明显提高;水稻受铁毒后的生物量递减量与ＡＰ、ＤＲ、ＧＲ 活性呈负相关。说明抗坏血酸过氧化物酶Ｈ２Ｏ２ 清除系统在水稻抗铁毒中起着十分重要的作用。     

茶陵野生稻苗期耐冷性研究

以耐冷性强弱不同的栽培稻为参比,通过自然冷胁迫与(或)人工冷处理,比较了茶陵野生稻与不同类型栽培稻经冷胁迫后的秧苗成活率、净光合速率和光系统Ⅱ光化学量子效率的变化,对茶陵野生稻苗期耐冷性作出评估.结果表明:经冷胁迫后,茶陵野生稻上述指标值的变化小于典型籼稻和爪哇稻,大于典型粳稻.说明茶陵野生稻苗期耐冷性强于籼稻和爪哇稻,但弱于粳稻.     

边缘细胞对荞麦根尖铝毒的防护效应和对细胞壁多糖的影响

以2个荞麦(Fygopyrum esculentum Moench)基因型‘江西荞麦’(耐性)和‘内蒙荞麦’(敏感)为材料,采用悬空培养(保持边缘细胞附着于根尖和去除根尖边缘细胞),研究边缘细胞对根尖铝毒的防护效应以及对细胞壁多糖组分的影响。结果表明,铝毒抑制荞麦根系伸长,导致根尖Al积累。去除边缘细胞的根伸长抑制率和根尖Al含量高于保留边缘细胞的根。去除边缘细胞使江西荞麦和内蒙荞麦根尖的酸性磷酸酶(APA)活性显著升高,前者在铝毒下增幅更大。同时,铝毒胁迫下去除边缘细胞的根尖果胶甲酯酶(PME)活性和细胞壁果胶、半纤维素1、半纤维素2含量显著高于保留边缘细胞的酶活性和细胞壁多糖含量。表明边缘细胞对荞麦根尖的防护效应,与其阻止Al的吸收,降低根尖细胞壁多糖含量及提高酸性磷酸酶活性有关,以此缓解Al对根伸长的抑制。     

Direct Evaluation of the Ca2+-Displacement Hypothesis for Al Toxicity

One explanation for Al toxicity in plants suggests that Al displaces Ca2+ from critical sites in the apoplasm. We evaluated the Ca2+-displacement hypothesis directly using near-isogenic lines of wheat (Triticum aestivum L.) that differ in Al tolerance at a single locus. We measured both the growth and total accumulation (apoplasmic plus symplasmic) of 45Ca and Al into roots that had been exposed to Al alone or to Al with other cations. Root growth in the Al-sensitive line was found to be severely inhibited by low activities of Al, even though Ca2+ accumulation was relatively unaffected. In solutions containing the same activity of the Al3+ and Ca2+ ions as above, but also including either 3.0 mM Mg2+, 3.0 mM Sr2+, or 30 mM Na+, growth improved, whereas 45Ca2+ accumulation was significantly decreased. Since most of the 45Ca2+ accumulated by roots during short-term treatments will reside in the apoplasm, these results indicate that displacement of Ca2+ from the apoplasm by Al cannot account for the Al-induced inhibition of root growth and, therefore, do not support the Ca2+-displacement hypothesis for Al toxicity. We also show that total accumulation of Al by root apices is greater in the Al-sensitive genotype than the Al-tolerant genotype and suggest that cation amelioration of Al toxicity is caused by the reduction in Al accumulation.     

Prospect of the QTL-qSB-9^Tq utilized in molecular breeding program of japonica rice against sheath blight

The major QTL-qSB-9Tq conferring partial resistance to rice (Oryza sativa L.) sheath blight (Rhizoctonia solani Kühn) has been verified on chromosome 9 of the indica rice cultivar, Teqing. In this study, the prospect of this QTL utilized in molecular breeding program of japonica rice for sheath blight resistance was investigated. Most of the japonica rice cultivars showed lower level of sheath blight resistance than the indica rice cultivars. At the corresponding site of qSB-9Tq, nine typical japonica rice culfivars from different ecological regions or countries proved to possess the susceptible allele(s). Introgression of qSB-9Tq into these cultivars enhanced their resistance level by decreasing sheath blight score of 1.0 (0.5-1.3), which indicated that qSB-9Tq had a large potential in strengthening the resistance of japonica rice to sheath blight. The use of the three molecular markers, which were polymorphic between Teqing and many japonica rice cultivars, promotes the application of qSB-9Tq in a concrete molecular breeding program.     

Effects of a pregrowth period in Al-free nutrient solution on macronutrient composition of two upland rice cultivars with various Al sensitivity

Effects of Al on growth and macronutrient distribution (K. P. Ca and Mg) were examined in two rice ( Oryza sativa L.) cultivars (BG35 and IR45) pregrown in a nutrient solution without Al for 21 days followed by 14 days at 0.70. 140 or 280 μ M Al (pH 4.2 ± 0.1). Relative growth rate (RGR) of pregrown plants was significantly less hampered by Al compared to their controls, than RGR of plants that were not pregrown. However, a similar ranking of the cultivars for Al sensitivity was obtained irrespective of pregrowth period. The more Al tolerant BG35 consistently outyielded the Al sensitive IR45 under all Al treatments. A comparatively high Al-retention in roots and low Al transport rate to shoots of pregrown IR45 was the opposite of conditions in non-pregrown IR45. In BG35, uptake and transport of nutrients to the shoots were efficient at all Al levels. In IR45, particularly the uptake and distribution of P and Mg were disturbed by Al. It is suggested that a pregrowth or nursery period without Al reduces Al toxicity in both BG35 and IR45 at least up to 280 μ M Al.     

Aluminium effects on growth and Al,Ca, Mg,K and P levels in triticale,wheat and rye

Summary The effects of A1 on the growth and mineral composition of different cultivars of triticale (X Triticosecale, Wittmack), wheat (Triticum aestivum L.) and rye (Secale cereale L.) growing in 1/5 strength Steinberg solutions containing 0 or 6 ppm A1 were evaluated after 32 days. Aluminum increased the concentrations of P and K in the roots and K in the tops of most of the cultivars tested. A1 tolerant triticale retained a lower concentration of Mg in the roots and tops than the A1 sensitive triticale, when subjected to A1 stress. In addition, A1 treatments resulted in smaller increases in root P for the A1 tolerant triticale than for the A1 sensitive cultivars.The concentration of root Ca and P of the A1 tolerant wheat cultivars were significantly below that of the more sensitive plants. Aluminum tolerance in rye appeared to be associated with lower Ca and higher Mg concentrations in the tops. The accumulation of P and A1 in the roots was characteristic of sensitivity in triticale, wheat and rye.     

Boron toxicity in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.). I. Quantitative trait locus (QTL) analysis of tolerance to boron toxicity

Boron toxicity tolerance of rice plants was studied. Modern japonica subspecies such as Koshihikari, Nipponbare, and Sasanishiki were tolerant, whereas indica subspecies such as Kasalath and IR36 were intolerant to excessive application of boron (B), even though their shoot B contents under B toxicity were not significantly different. Recombinant inbred lines (RILs) of japonica Nekken-1 and indica IR36 were used for quantitative trait locus (QTL) analysis to identify the gene responsible for B toxicity tolerance. A major QTL that could explain 45% of the phenotypic variation was detected in chromosome 4. The QTL was confirmed using a population derived from a recombinant inbred line which is heterogenic at the QTL region. The QTL was also confirmed in other chromosome segment substitution lines (CSSLs).     

Mechanical impedance increases aluminium tolerance of soybean (Glycine max) roots

The effect of aluminium (Al) on root elongation was studied in solution culture and sand culture. Compared to solution culture, in sand culture a ten times higher Al supply was necessary to inhibit root elongation to a comparable degree. This was due to a much lower Al uptake into the 5 mm root tips in sand culture. Fe concentrations in root tips were also lower in sand culture. Ca concentrations were higher and less depressed by Al, whereas Mg and K concentrations were not affected by the culture substrate. Regressions of Al concentrations in root tips versus inhibition of root elongation by Al revealed root damage at lower Al concentrations in sand culture. The effect of culture substrate on Al tolerance was independent of N source and could also be shown in flowing solution culture with and without sand. The results indicate that mechanical impedance in sand culture decreased Al uptake. This may be due to enhanced exudation of organic complexors thus reducing activites of monomeric Al species.     

Comparison of techniques for determining the effect of aluminium on the growth of,and the inheritance of aluminium tolerance in wheat

The effect of Al on the growth of plants derived from the F3 generation of a cross between Al tolerant (Waalt) and Al sensitive (Warigal) wheat cultivars, grown in low ionic strength nutrient solutions, were assessed by a number of methods viz; root length and haematoxylin stain after 3 days exposure to Al and plant top and root yields, and root length and visual assessment for Al damage after 4 weeks growth.Of these methods haematoxylin stain (3 days) and visual assessment at 4 weeks identified the same plants as being sensitive or tolerant to Al and clearly segregated the 2 populations. Consequently these 2 methods were used as standard techniques to determine the ability of the other methods to distinguish between tolerant and sensitive plants.The ratio of plant top: root yields clearly segregated the 2 populations. The 2 populations could not be clearly distinguished based on plant top or root yields, or on root length either after 3 days or 4 weeks exposure to Al.Within the population of tolerant plants, root length was significantly correlated with root weight (r²=0.86) and top weight (r²=0.71). None of these relationships were significant for the population of sensitive plants.These techniques were applied in a number of separate experiments on the F2 and F3 populations from a Waalt × Warigal cross. The results indicate that Al tolerance in wheat is inherited by a single gene and that this gene has incomplete dominance.     

Magnesium ameliorates aluminum rhizotoxicity in soybean by increasing citric acid production and exudation by roots

Superior effectiveness of Mg over Ca in alleviating Al rhizotoxicity cannot be accounted for by predicted changes in plasma membrane Al3+ activity. The influence of Ca and Mg on the production and secretion of citrate and malate, and on Al accumulation by roots was investigated with soybean genotypes Young and PI 416937 which differ in Al tolerance. In the presence of a solution Al3+ activity of 4.6 microM, citrate and malate concentrations of tap root tips of both genotypes increased with additions of either Ca up to 3 mM or Mg up to 50 microM. Citrate efflux rate from roots exposed to Al was only enhanced with Mg additions and exceeded malate efflux rates by as much as 50-fold. Maximum citrate release occurred within 12 h after adding Mg to solution treatments. Adding 50 microM Mg to 0.8 mM CaSO4 solutions containing Al3+ activities up to 4.6 microM increased citrate concentration of tap root tips by 3- to 5-fold and root exudation of citrate by 6- to 9-fold. Plants treated with either 50 microM Mg or 3 mM Ca had similar reductions in Al accumulation at tap root tips, which coincided with the respective ability of these ions to relieve Al rhizotoxicity. Amelioration of Al inhibition of soybean root elongation by low concentrations of Mg in solution involved Mg-stimulated production and efflux of citrate by roots.     

Magnesium enhances aluminum-induced citrate secretion in rice bean roots (Vigna umbellata) by restoring plasma membrane H+-ATPase activity

We demonstrated that magnesium (Mg) can alleviate aluminum (Al) toxicity in rice bean [Vigna umbellata (Thunb.) Ohwi & Ohashi] more effectively than is expected from a non-specific cation response. Micromolar concentrations of Mg alleviated the inhibition of root growth by Al but not by lanthanum, and neither strontium nor barium at the micromolar level alleviates Al toxicity. Aluminum also induced citrate efflux from rice bean roots, and this response was stimulated by inclusion of 10 microM Mg in the treatment solution. The increase in the Al-induced citrate efflux by Mg paralleled the improvement in root growth, suggesting that the ameliorative effect of Mg might be related to greater citrate efflux. Vanadate (an effective H+-ATPase inhibitor) decreased the Al-induced citrate efflux, while addition of Mg partly restored the efflux. Mg addition also increased the activity of Al-reduced plasma membrane H+-ATPase, as well as helping to maintain the Mg and calcium contents in root apices. We propose that the addition of Mg to the toxic Al treatment helps maintain the tissue Mg content and the activity of the plasma membrane H+-ATPase. These changes enhanced the Al-dependent efflux of citrate which provided extra protection from Al stress.     

Genetic architecture of aluminum tolerance in rice (Oryza sativa) determined through genome-wide association analysis and QTL mapping

Aluminum (Al) toxicity is a primary limitation to crop productivity on acid soils, and rice has been demonstrated to be significantly more Al tolerant than other cereal crops. However, the mechanisms of rice Al tolerance are largely unknown, and no genes underlying natural variation have been reported. We screened 383 diverse rice accessions, conducted a genome-wide association (GWA) study, and conducted QTL mapping in two bi-parental populations using three estimates of Al tolerance based on root growth. Subpopulation structure explained 57% of the phenotypic variation, and the mean Al tolerance in Japonica was twice that of Indica. Forty-eight regions associated with Al tolerance were identified by GWA analysis, most of which were subpopulation-specific. Four of these regions co-localized with a priori candidate genes, and two highly significant regions co-localized with previously identified QTLs. Three regions corresponding to induced Al-sensitive rice mutants (ART1, STAR2, Nrat1) were identified through bi-parental QTL mapping or GWA to be involved in natural variation for Al tolerance. Haplotype analysis around the Nrat1 gene identified susceptible and tolerant haplotypes explaining 40% of the Al tolerance variation within the aus subpopulation, and sequence analysis of Nrat1 identified a trio of non-synonymous mutations predictive of Al sensitivity in our diversity panel. GWA analysis discovered more phenotype-genotype associations and provided higher resolution, but QTL mapping identified critical rare and/or subpopulation-specific alleles not detected by GWA analysis. Mapping using Indica/Japonica populations identified QTLs associated with transgressive variation where alleles from a susceptible aus or indica parent enhanced Al tolerance in a tolerant Japonica background. This work supports the hypothesis that selectively introgressing alleles across subpopulations is an efficient approach for trait enhancement in plant breeding programs and demonstrates the fundamental importance of subpopulation in interpreting and manipulating the genetics of complex traits in rice.     

Improvement of rice callus regeneration in the presence of NaCl

The effects of abscisic acid (37.8 μM), polyethylene glycol (5%), proline (10 mM), tryptophan (490 μM) and indoleacetic acid (5.7 μM) on rice callus regeneration were studied at various doses of NaCl (0, 50 and 100 mM) on three month-old mature embryo-derived callus of two japonica (I Kong Pao and Aiwu) and two indica (IR 2153 and Nona Bokra) rice cultivars differing in salinity tolerance. NaCl strongly decreased the regeneration frequency of all cultivars but slightly increased the survival of regenerated plantlets. Tryptophan stimulated regeneration and increased subsequent survival rates of regenerated plantlets in all cultivars at all NaCl doses. Abscisic acid and polyethylene glycol, though not affecting the final regeneration percentages, delayed regeneration and reduced the mean number of plantlets produced per regenerating callus in all cultivars, as well as rooting ability and survival of regenerated plantlets in indica genotypes. Proline had no marked effect on regeneration, whatever the NaCl dose or cultivar, while indoleacetic acid reduced shoot regeneration and increased root regeneration. This revised version was published online in June 2006 with corrections to the Cover Date.