Aluminum-mycorrhizal interactions in the physiology of pitch pine seedlings

Aluminum (Al) in the rhizosphere adversely affects plant nutrition and growth. Although many conifer species, and pitch pine (Pinus rigida) in particular, have evolved on acidic soils where soluble Al is often high, controlled environment studies often indicate that Al interferes with seedling growth and nutrient relations. Under normal field conditions, conifer roots grow in a symbiotic relationship with ectomycorrhizal fungi, and this association may modulate the effects of Al on root physiology. To investigate the influence of mycorrhizal infection on Al toxicity, pitch pine seedlings were grown with or without the ectomycorrhizal symbiont Pisolithus tinctorius and were exposed to low levels of Al in sand culture. Aluminum at 50 μM reduced nonmycorrhizal seedling growth and increased foliar Al concentrations, but did not alter photosynthetic gas exchange or other aspects of seedling nutrition. Nonmycorrhizal seedlings exposed to 200 μM Al exhibited decreased growth, increased transpiration rates, decreased water use efficiency, increased foliar Al and Na levels, and reduced foliar P concentrations. Seedlings inoculated with P. tinctorius exhibited unaltered growth, physiological function, and ionic relations when exposed to Al. The fungal symbiont evidently modulated ionic relations in the rhizosphere, reducing Al-P precipitation reactions, Al uptake, and subsequent root and shoot tissue Al exposure.     

Effects of aluminium and mineral nutrition on growth and chemical composition of hydroponically grown seedlings of five different forest tree species

Forest die-back and impaired tree vitality have frequently been ascribed to Al-toxicity and Al-induced nutritional disorders due to increased acidification of forest soils. Therefore, in this experiment effects of Al were studied on growth and nutrient uptake with seedlings of five different forest tree species. During growth in culture solutions with and without Al all five species proved to be very Al-tolerant, despite high accumulation of Al in roots. In the coniferous evergreens Douglas-fir and Scots pine shoot as well as root Al concentrations were significantly higher than in the deciduous broad-leaved species oak and birch. Larch showed intermediate Al levels. In none of the five species did Al reduce nutrient concentrations or the Ca/Al ratio to values below the critical level. Besides differences in Al accumulation, coniferous and broad-leaved species also differed with respect to uptake and assimilation of nitrogen. Due to extra NH ₄ ⁺ uptake, oak and birch showed a much higher N uptake and higher NH ₄ ⁺ preference than the coniferous species. Especially with oak this high NH ₄ ⁺ preference in combination with a low specific root surface area resulted in a high root proton efflux density. In comparison to both broad-leaved trees and Scots pine the NO ₃ ⁻ reduction capacity of larch and Douglas-fir was extremely low. This may have important consequences for both species if grown in NO ₃ ⁻ -rich soils.     

Phytotoxic, clastogenic and bioaccumulation effects of the environmental endocrine disruptor bisphenol A in various crops grown hydroponically

The effects of the endocrine disruptor bisphenol A (BPA) at concentrations of 10 and 50 mg l⁻¹ were evaluated on the germination and morphology, micronuclei (MN) content in root tip cells and BPA bioaccumulation of hydroponic seedlings of broad bean (Vicia faba L.), tomato (Lycopersicon esculentum Mill.), durum wheat (Triticum durum Desf.) and lettuce (Lactuca sativa L.) after 6 and 21 days of growth. In general, BPA at any dose used did not inhibit germination and early growth (6 days) of seedlings of the species examined, with the exception of primary root length of tomato which decreased at the higher BPA dose. In contrast, an evident phytotoxicity was induced by BPA in all species after 21 days of growth with evident morphological anomalies and significant reductions of the lengths and fresh and dry weights of shoots and roots of seedlings. With respect to the nutrient medium without seedlings, BPA concentration decreased markedly during the growth period in the presence of broad bean and tomato seedlings, and limitedly in the presence of durum wheat and, especially, lettuce. Further, the presence of BPA measured in roots and shoots of broad bean and tomato after 21-day growth indicated that bioaccumulation of BPA had occurred. The number of MN in broad bean and durum wheat root tip cells increased markedly by treatment with BPA at both concentrations, thus suggesting a potential clastogenic activity of BPA in these species.     

Effects of elevated soil solution Al concentrations on fine roots in a middle-aged Norway spruce (Picea abies (L.) Karst.) stand

Aluminium (Al), mobilized by acidic deposition, has been claimed to be a major threat to forest vitality. Fine root mortality, decreased root growth and reduced nutrient uptake have been observed in controlled laboratory experiments where roots of tree seedlings were exposed to elevated concentrations of Al. Yet, evidence for Al-induced root damage from forest stands is scarcely reported. Nevertheless, Al dissolved in soil water has received a key role in the critical load concept for forests. Here, we present effects of artificially elevated concentrations of Al in the soil solution on fine roots in a middle-aged stand of Norway spruce (Picea abies (L.) Karst.). Although the inorganic Al concentrations about 200 µM and Ca:Al ratio about 0.7 that were established in the soil solution within this experiment have been associated with reduction of root growth and root mortality for spruce seedlings in hydroponic studies, no acute damage on fine roots was observed. Three years of treatment did not cause visual root damage, nor were effects on fine root necromass observed. Fine root necromass made up about 10% of fine root biomass for all treatments. However, significantly lower molar Ca:Al and Mg:Al ratios in living and dead fine roots were found in the plots where Al concentrations were highest and ratios of Ca to Al in the soil solution were lowest. The lack of response on fine root biomass suggests that forest stands tolerate higher Al levels than results from laboratory experiments indicate. We conclude that effect studies in the laboratory have limited value for field conditions. The key role of Al toxicity, expressed as the Ca/Al ratio, in critical load calculations for forests may have to be reconsidered.     

The effect of aluminium on the distribution of calcium,magnesium and phosphorus in mycorrhizal and non-mycorrhizal seedlings of Eucalyptus rudis: a cryo-microanalytical study

The distribution of Al, Ca, Mg and P in the lateral roots and leaves of mycorrhizal and non-mycorrhizal seedlings of Eucalyptus rudis grown with and without Al was analysed using energy-dispersive X-ray microanalysis on a cryo-scanning electron microscope. Al accumulated in all tissues of nonmycorrhizal plants: the endodermis was not a barrier to the translocation of Al. In mycorrhizal roots, Al was concentrated within the sheath. The presence of Al reduced the levels of Ca and Mg in both mycorrhizal and non-mycorrhizal roots and shoots in comparison with control plants. The presence of mycorrhizas increased the levels of Ca and Mg in plants grown with Al in comparison with non-inoculated plants, although there was no evidence that mycorrhizas increased the levels of P in plants grown in Al-amended soils. P levels were higher in the mycorrhizal sheath of plants grown with Al than the controls.     

Aluminium causes nutrient imbalance and structural changes in the needles of Scots pine without inducing clear root injuries

The effects of aluminium chloride (AICI3) treatments (50 and 150 mg/l) on 3-year-old Scots pine (Pinus sylvestris L.) seedlings were studied in a sand culture during 2 growing periods in an open field experiment. Even by the end of the first growing period, a decline was observed in the concentrations of Ca, Mg and P within the needles, and of Ca and Mg in the roots. After the second growing period, increased N and K concentrations were observed in the needles of Al-treated seedlings. Both the needles and roots of Al-treated seedlings showed, after the second growing period, a decline in growth and increased concentrations of AI as the amount of AICI3 in the nutrient solution increased. Al-induced changes in needle structure were found to be symptomatic of a nutrient imbalance, particularly of Mg and P. Al-stress did not result in any observable changes in root anatomy or in the number of mycorrhizas. Scots pine proved to be rather resistant to Al-stress, indicating that direct Al-injuries are not likely in the field, though Al-stress may be a contributing factor in the formation of nutrient imbalances.     

Seed source variation in 32P uptake in inus taeda L. seedlings: A possible regulation by efflux

Short-term ³²P uptake experiments were conducted with intact seedlings of loblolly pine (Pinus taeda L.) to examine possible seed source variation in net accumulation of ³²P in roots and shoots, and in rates of unidirectional influx. Seed source had a highly significant effect on biomass and P concentrations of shoots and roots. Seedlings from two seed sources representing fast-growing populations (a broadly-adapted and wet-site seed source) accumulated over 60% more total seedling P than smaller seedlings from a drought-hardy seed source, reflecting higher biomass and root P concentrations. Rates of unidirectional ³²P influx in seedlings from the drought-hardy seed source were more than twice the rates of the seedlings from the broadly-adapted seed source. However, after 24 h in labeled uptake solution, net accumulation of ³²P was similar, suggesting that rates of unidirectional efflux from roots of the drought-hardy seed source were also high. Although there were no significant differences in biomass and tissue P concentrations between the two fast-growing seed sources, rates of unidirectional influx in seedlings from the broadly-adapted seed source were 42% lower than rates in seedlings from the wet-site source. Yet, after 24 h in labeled uptake solution, net accumulation of ³²P in seedlings from the broadly-adapted seed source was 50% higher. Unidirectional efflux out of the root may regulate net uptake of P as much, if not more, than influx in loblolly pine seedlings-at least under high-P growth conditions. The results in this study do not support previous studies with herbaceous plants suggesting that fast-growing species typically exhibit higher rates of nutrient uptake than slow-growing species.     

Effect of zinc supply on growth of three species of Eucalyptus seedlings and wheat

Summary Effects of zinc supply on shoot and root dry weight, root length, zinc concentrations and carbonic anhydrase activity were measured in 52 day old seedlings ofEucalyptus maculata, E. marginata, E. patens and wheat grown in a zinc deficient soil in the glasshouse.Symptoms of zinc deficiency in the eucalyptus and wheat appeared within 20 to 35 days. Eucalypt seedlings had short internodes and small necrotic leaves, reduced dry weight of shoots and roots, root length and zinc concentrations in young leaves; the measurable level of leaf carbonic anhydrase activity decreased to zero. Similar responses also occurred in wheat.The level of zinc fertilizer required for normal growth of Eucalyptus seedlings is therefore likely to be similar to that used for wheat and other agricultural crops.     

Influence of aluminum and phosphorus on growth and xylem sap composition in Melastoma malabathricum L.

Melastoma (Melastoma malabathricum L.) is an aluminum-accumulating woody plant that accumulates more than 10 000 mg kg^–1 of aluminum (Al) in mature leaves. The influence of Al and phosphorus (P) applications on plant growth and xylem sap was examined in the present study in order to elucidate the interaction between Al-induced growth enhancement and P nutrition, and to determine the form of Al for translocation from roots to shoots. Although the Al application significantly increased the growth of Melastomaseedlings with the high P pre-treatment, and P concentrations in the leaves and Pi concentrations in the xylem sap regardless of the P pre-treatment, we could not come to the conclusion that a primary cause of the Al-induced growth enhancement in Melastoma is the stimulation of P uptake. The degree of Al-induced growth enhancement corresponded not with the P concentrations but with the Al concentrations in the plant tissue, suggesting that the Al-induced growth enhancement in Melastoma is primarily caused by Al itself in the plant tissue rather than by the stimulation of P uptake. Through the analysis of organic acids and Al in the xylem sap and plant tissue, the form of Al for translocation from roots to shoots was shown to be an Al-citrate complex that was transformed into Al-oxalate complex for Al storage in the leaves. In addition, the xylem sap of Melastoma seedlings grown in the absence of Al contained higher concentrations of malate. In the presence of Al, however, higher concentrations of citrate were found, indicating that Melastoma changes its organic acid metabolism in the presence or absence of Al; more specifically, it increases the synthesis of citrate.     

Effect of soil temperature on nutrient allocation and mycorrhizas in Scots pine seedlings

We studied the effect of soil temperature on nutrient allocation and mycorrhizal development in seedlings of Scots pine (Pinus sylvestris L.) during the first 9 weeks of the growing season. One-year-old seedlings were grown in Carex-peat from a drained and forested peatland at soil temperatures of 5, 9, 13 and 17 °C under controlled environmental conditions. Fourteen seedlings from each temperature treatment were harvested at intervals of three weeks and the current and previous year's parts of the roots, stems and needles were separated. Mineral nutrient and Al contents in all plant parts were determined and the tips and mycorrhizas of the new roots were counted. Microbial biomass C and N in the growth medium were determined at the end of the experiment. None of the elements studied, except Fe, were taken up from the soil by the seedlings during the first three weeks. Thereafter, the contents of all the elements increased at all soil temperatures except 5 °C. Element concentrations in needles, stems and roots increased with soil temperature. Higher soil temperature greatly increased the number of root tips and mycorrhizas, and the numbers of mycorrhizas increased more than did the length of new roots. Cenococcum geophilum was relatively more abundant at lower soil temperatures (5 and 9 °C) than at higher ones (13 and 17 °C). A trend was observed for decreased microbial biomass C and N in the peat soil at higher soil temperatures at the end of the experiment.     

干旱条件下接种AM真菌对小马鞍羊蹄甲幼苗根系的影响

为了探讨岷江干旱河谷丛枝菌根真菌(AMF)对寄主植物幼苗根系的影响,通过接种购买的AMF摩西球囊霉菌(Funneliformis mosseae)到优势乡土灌木小马鞍羊蹄甲(Bauhinia faberi var.microphylla)幼苗,在重度、中度和轻度干旱条件下培养3个月,研究不同干旱条件下AMF对幼苗根系形态特征、结构特征、功能性状的影响。方差分析结果表明:(1)3种干旱胁迫条件下,接菌均显著增加了幼苗的根总长、根表面积、根分枝数、根尖数(P0.001),在中度胁迫和轻度胁迫下,接菌显著促进根鲜重、根体积的增加(P0.001),轻度胁迫条件下接菌幼苗的根鲜重、根总长、根表面积、根体积、根尖数最高并显著高于其它处理,但接菌与未接菌的根平均直径之间没有显著差异;(2)接菌幼苗根系趋向于叉状分支结构,在重度胁迫时,叉状分支趋势更显著(P0.001);(3)接菌幼苗的根比例都显著小于未接菌的,但幼苗比根长不存在显著差异。相关分析结果表明:菌根侵染率与根鲜重、根总长、根表面积、根体积、根分枝数、根尖数呈极显著正相关(P0.001),与拓扑指数、根比例呈极显著负相关(P0.001)。研究表明,在干旱条件下,AMF虽然没有提高生长初期的根系的吸收效率,但接种AMF显著影响幼苗根系形态特征和结构特征,更利于植物适应干旱环境,并且AMF对幼苗根系的促生作用随着干旱胁迫程度减轻而提高。     

Root regeneration as an indicator of aluminium toxicity in loblolly pine

Loblolly pine (Pinus taeda L.) seedlings were grown in sand culture and exposed to solution Al concentrations of 0, 5, 10, 20, 40, and 80 mg.l⁻¹ in two separate studies. Root regeneration potential (RRP) of loblolly pine was found to be very sensitive to Al. Both the total number and total length of new white roots produced during a 24-day period declined with as little as 5 mg Al.l⁻¹ in solution.     

Lignosulfonate promotes the interaction between Scots pine and an ectomycorrhizal fungus Pisolithus tinctorius in vitro

Lignosulfonate (LS) is a lignin-based polymer obtained as a by-product from paper industry, which may have potential as an amendment with macronutrients. We studied effects of LS on the interaction between Scots pine (Pinus sylvestris L.) seedlings and hypocotyl cuttings and the ectomycorrhizal (ECM) fungusPisolithus tinctorius (Pers.) Coker and Couch. The experiments were performed in vitroon the MMN agar medium containing Fe–LS chelate at the concentrations of 0, 5, 10 and 25 mg/L. Inoculation with P. tinctoriusincreased root growth of the seedlings. Fe–LS enhanced P. tinctorius induced formation of lateral roots and had a dose-dependent positive effect on the establishment of mycorrhizas on the seedlings. The growth of the fungal mycelium was improved by Fe–LS, which might cause faster and more intensive contact with the roots and, thus, better root growth and mycorrhiza formation. P.tinctorius enhanced also adventitious root formation and subsequent root growth of the hypocotyl cuttings but without any synergistic effect with Fe–LS. Our study with P. tinctorius and Scots pine in vitro indicates that a low-cost by-product Fe–LS, obtained from paper industry, may be a potential tool to improve the efficiency of fungal inoculations, thus, facilitating the early interaction between an ECM fungus and host seedling.     

Relationship of aluminium and calcium to net CO2 exchange among diverse Scots pine provenances under pollution stress in Poland

Light-saturated net photosynthesis (A_sat), dark respiration (RD), and foliar nutrient content of eight European Scots pine (Pinus sylvestris L.) provenances were measured at experimental sites in western Poland. Two-year-old seedlings were planted in 1984 at two sites with similar soils in areas of contrasting air pollution. One site was near a point source of SO₂ and other pollutants, and another 12 km to the southeast in an area free of acute air pollution was treated as a control. The eight provenances were from a large north-tosouth latitudinal range (60 to 43° N). At the heavily polluted site Scots pine trees exhibited lower growth rates and crown dieback and deformation. Soil pH, Ca and Mg were at least 10 times lower, and Al 10 times higher at the polluted than the control site. In 1991, concentrations of Al, P, Ca, S, Mn, Fe, and Zn in oneyear old Scots pine foliage were higher and Mg lower at the polluted than control site. At both sites foliar Mg levels were within the range considered deficient (0.6 mg g^-1), and at the polluted site, Al concentrations were very high (670 to 880 g g^-1). In all provenances, RD of one-year-old needles was higher (by 22% on average) and A_sat was lower (by 37% on average) at the polluted than the control site. The ratio of A_sat: RD was half as great in all provenances at the polluted (4 to 6) than control site (8 to 11). Provenances of southern origin had greater increases in RD and water-use efficiency at the polluted site than other provenances. Within the polluted site alone, or across both sites, A_sat in Scots pine was negatively correlated to the Al: Ca ratio (p<0.001, r=–0.93). Across sites RD increased with needle N and Al (multiple regression, p<0.001). The data suggest that at the polluted site there is excessive soil Al and deficient Mg availability, low needle Mg and high Al concentrations and high Al: Ca ratios, and that these have resulted in reduced photosynthetic capacity and increased respiration.     

Interactive effects of potassium and sodium on root growth and expression of K/Na transporter genes in rice

Sufficient supply of potassium (K) can alleviate the adverse effects of excess sodium (Na) on plant growth. However, it remains unclear if such a beneficial function is related to regulation of root growth and/or expression of K/Na transporters. Herein we report the responses of a rice cultivar, which was pretreated with normal nutrient solution for 1 month, to three levels of Na (0, 25, and 100 mM) without or with supply of K for 9 days. High Na (100 mM) significantly decreased plant growth, root activity, and total K uptake, and increased biomass ratio of roots to shoots. Short-term removal of K supply (9 days) did not affect root morphology and biomass ratio of roots to shoots, but decreased root activity of seedlings grown in high Na solution. K deficiency increased uptake of Na and transport of K from roots to shoots. Moreover, expression of OsHAK1, a putative K transporter gene, was upregulated by low Na (25 mM) and downregulated by high Na (100 mM) in roots. In leaves, its expression was suppressed by the Na treatments when K supply was maintained. Expression of OsHKT2;1, which encodes a protein that acts mainly as a Na transporter, was downregulated by high Na, but was enhanced by K deficiency both in roots and leaves. Expression of five other putative K/Na transporter or Na⁺/H⁺ genes, OsHKT1;1, OsHKT1;2, OsHKT2;3, OsNHX1, and OsSOS1, was not affected by the treatments. The results suggest that OsHAK1 and OsHKT2;1 were involved in the interactive effects of K and Na on their uptake and distribution in rice. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Response of understorey vegetation and Scots pine root systems to fertilization at multiple deficiency stress

The stump and root systems of Scots pine (Pinus sylvestris L.) and field-layer vegetation were sampled before and three growing seasons after drainage and fertilization of a low-shrub pine bog in SE Norway. Although the understorey vegetation roots responded significantly to nutrient application with higher concentrations of Ca and P, root biomass weights did not change. The fine and small pine roots responded with higher N, Ca, P and S concentrations, while those of Mn and Zn were significantly reduced. The NPK-application resulted in significantly higher pine root biomass. Relative to the total stores in the root zone the amounts of most elements in roots shifted to higher ratios at NPK-application. High figures for K, B and Mn indicate tight biochemical cycles of these elements. Compared to totals in above and below ground biomass, major parts of Fe and Pb were held by the roots. In contrast the field layer roots kept a very small per cent of total K, while the pine roots were low in Mn. The understorey vegetation was primarily restricted by P-deficiency, while the pine trees were also restricted by low supply of N. The field and the tree layer species seem to differ with respect to required nutrient concentrations in the root zone. These characteristics are important for direction and extent of successional changes after fertilization in low-shrub pine bog ecosystems.     

<Emphasis Type="Italic">In vitro</Emphasis> organogenesis and antioxidant enzymes activity in <Emphasis Type="Italic">Acanthophyllum sordidum</Emphasis>

The effect of various hormonal combinations on callus formation and regeneration of shoot and root from leaf derived callus of Acanthophyllum sordidum Bunge ex Boiss. has been studied. Proteins and activity of antioxidant enzymes were also evaluated during shoot and root organogenesis from callus. Calli were induced from leaf explants excised from 30-d-old seedlings grown on Murashige and Skoog medium containing 4.52 μM 2,4-dichlorophenoxyacetic acid + 4.65 μM kinetin. Maximum growth of calli and the most efficient regeneration of shoots and roots occurred with 2.69 μM 1-naphthalene acetic acid (NAA), 2.69 μM NAA + 4.54 μM thidiazuron and 2.46 μM indole-3-butyric acid. Protein content decreased in calli and increased significantly during regeneration of shoots from callus. Superoxide dismutase activity decreased in calli comparing to that of seedlings, then increased in regenerated shoots and roots. High catalase activity was detected in seedlings and regenerated shoots, whereas high peroxidase activity was observed in calli and regenerated roots.     

Role of manganese in low-pH-induced root hair formation in<Emphasis Type="Italic"> Lactuca sativa</Emphasis> cv. Grand Rapids seedlings

Root hair formation is induced by low pH in lettuce (Lactuca sativa L. cv. Grand Rapids) seedlings cultured in mineral medium. The role of mineral concentrations in this phenomenon was investigated, especially for manganese. When lettuce seedlings were cultured in media that were deficient in calcium (Ca), manganese (Mn), boron (B) or molybdenum (Mo), morphological changes were induced in roots. Deficiency of other nutrients had little effect on root hair formation. Ca or B deficiency inhibited the growth of the main root and the formation of root hairs, regardless of pH. Mn or Mo deficiency increased root hair formation at pH 6 and suppressed main root growth slightly. In contrast, increasing the Mn concentration suppressed low-pH-induced root hair formation. The Mn content of roots grown at pH 4 was only about 15% of that at pH 6. In contrast, the Mo content of roots grown at low pH was about six times that of roots grown at neutral pH. These results suggest that root hair formation induced by low pH is at least partly mediated by decreased Mn uptake in root cells.     

Short-term effects of manipulated increase in acid deposition on soil, soil solution chemistry and fine roots in Scots pine (Pinus sylvestris) stand on a podzol

A manipulated increase in acid deposition (15 kg S ha⁻¹), carried out for three months in a mature Scots pine (Pinus sylvestris) stand on a podzol, acidified the soil and raised dissolved Al at concentrations above the critical level of 5 mg l⁻¹ previously determined in a controlled experiment with Scots pine seedlings. The induced soil acidification reduced tree fine root density and biomass significantly in the top 15 cm of soil in the field. The results suggested that the reduction in fine root growth was a response not simply to high Al in solution but to the depletion of exchangeable Ca and Mg in the organic layer, K deficiency, the increase in NH₄:NO₃ ratio in solution and the high proton input to the soil by the acid manipulation. The results from this study could not justify the hypothesis of Al-induced root damage under field conditions, at least not in the short term. However, the study suggests that a short exposure to soil acidity may affect the fine root growth of mature Scots pine.     

Calcium and calmodulin inhibitor effects on the growth of carrot (Daucus carota L.) and radish (Raphanus sativus L.) in nutrient culture

Growth of carrot and radish seedlings in nutrient culture was inhibited by pretreatment with three calmodulin inhibitors. There was little selective effect on specific organs, shoots, tap root and fibrous roots over a range of concentrations. Although pretreatment with CaCl₂ (0.5 mM) did not affect growth of untreated seedlings, it partially reduced the inhibitory effects of trifluoperazine over the concentration range 0.01–0.05 mM. Trifluoperazine reduced the growth of GA₃-treated seedlings but did not overcome the modifying effect of GA₃ in favouring shoot/root ratio; ABA exacerbated its inhibitory effect on overall seedling growth and particularly on tap root development.Abbreviations GA₃ gibberellic acid - ABA abscisic acid - CaCl₂ calcium chloride - GAs gibberellins - Tfp trifluoperazine