Ethylene insensitivity impairs resistance to soilborne pathogens in tobacco and Arabidopsis thaliana

Transgenic ethylene-insensitive tobacco (Tetr) plants spontaneously develop symptoms of wilting and stem necrosis when grown in nonautoclaved soil. Fusarium oxysporum, F. solani, Thielaviopsis basicola, Rhizopus stolonifer, and two Pythium spp. were isolated from these diseased Tetr plants and demonstrated to be causal agents of the disease symptoms. Pathogenicity of the two Pythium isolates and four additional Pythium spp. was tested on ethylene-insensitive tobacco and Arabidopsis seedlings. In both plant species, ethylene insensitivity enhanced susceptibility to the Pythium spp., as evidenced by both a higher disease index and a higher percentage of diseased plants. Based on the use of a DNA probe specific for Pythium spp., Tetr plants exhibited more pathogen growth in stem and leaf tissue than similarly diseased control plants. These results demonstrate that ethylene signaling is required for resistance to different root pathogens and contributes to limiting growth and systemic spread of the pathogen.     

Ethylene sensitivity affects changes in growth patterns, but not stem properties, in response to mechanical stress in tobacco

Plant responses to mechanical stress (e.g. wind or touch) involve a suite of physiologic and developmental changes, collectively known as thigmomorphogenesis, including reductions in height increment, Young's modulus of stems, shoot growth, and seed production, and increased stem girth and root growth. A role of the phytohormone ethylene in thigmomorphogenesis has been proposed but the extent of this involvement is not entirely clear. To address this issue, wild-type (WT) and ethylene-insensitive transgenic (Tetr) tobacco ( Nicotianum tabacum ) plants were subjected to three levels of mechanical stress: 0, 25 and 75 daily flexures. Flexed plants produced shorter, thicker stems with a lower Young's modulus than non-flexed ones, and these responses occurred independently of genotype. This suggests that ethylene does not play a role in thigmomorphogenesis-related changes in stem characteristics in tobacco. The effect of mechanical stress on dry mass increment (growth), on the other hand, differed between the genotypes: in the WT plants, shoot growth but not root growth was reduced under mechanical stress, resulting in reduced total growth and increased root mass fractions. In the Tetr plants, neither shoot nor root growth were affected. This suggests that ethylene is involved in the inhibition of tobacco shoot growth under mechanical stress.     

Interactions between ethylene and gibberellins in phytochrome-mediated shade avoidance responses in tobacco

Plants respond to proximate neighbors with a suite of responses that comprise the shade avoidance syndrome. These phytochrome-mediated responses include hyponasty (i.e. a more vertical orientation of leaves) and enhanced stem and petiole elongation. We showed recently that ethylene-insensitive tobacco (Nicotiana tabacum) plants (Tetr) have reduced responses to neighbors, showing an important role for this gaseous plant hormone in shade avoidance. Here, we investigate interactions between phytochrome signaling and ethylene action in shade avoidance responses. Furthermore, we investigate if ethylene acts in these responses through an interaction with the GA class of hormones. Low red to far-red light ratios (R:FR) enhanced ethylene production in wild-type tobacco, resulting in shade avoidance responses, whereas ethylene-insensitive plants showed reduced shade avoidance responses. Plants with inhibited GA production showed hardly any shade avoidance responses at all to either a low R:FR or increased ethylene concentrations. Furthermore, low R:FR enhanced the responsiveness of hyponasty and stem elongation in both wild-type and Tetr plants to applied GA(3), with the stem elongation process being more responsive to GA(3) in the wild type than in Tetr. We conclude that phytochrome-mediated shade avoidance responses involve ethylene action, at least partly by modulating GA action.     

Nitrate and phosphate uptake by Agropyron desertorum and Artemisia tridentata from soil patches with balanced and unbalanced nitrate and phosphate supply

To test the ability of plants to integrate small-scale imbalances in soil nitrate and phosphate patches, plant growth and acquisition of nitrate and phosphate were measured for the perennial grass Agropyron desertorum (Fisch. ex Link) Schult. and the shrub Artemisia tridentata Nutt. ssp. vaseyana (Rydb.) Beetle in soil where the principal supply of nitrate and phosphate came from two enriched patches. The soil was calcareous loamy-skeletal Typic Haploxerolls. These patches were applied in two treatments: either nitrate and phosphate were applied in both patches (balanced treatment) or one patch contained only nitrate and the other only phosphate (unbalanced treatment). The same total quantity of nutrients was applied in both treatments and these included ¹⁵N and ³²P tracers. The plants were in large pots in open field conditions. There were no significant differences in total biomass production and nitrogen concentration between the two treatments, indicating that both species had the physiological ability to integrate soil nutrient resources. Artemisia was able to acquire more phosphate in the unbalanced treatment, probably due to the high local solution phosphate concentration. Generally Artemisia acquired more N and P than did Agropyron .     

Nutrient foraging traits in 10 co-occurring plant species of contrasting life forms

1 Responses to spatial heterogeneity of soil nutrients were tested in 10 plant species that differ in life form and successional status, but which co-occur in the South Carolina coastal plain. The morphological responses of the root system were tested by assessing scale (represented by root mass and root length densities), precision (preferential proliferation of roots in nutrient-rich patches compared with less fertile patches) and discrimination (ability to detect and proliferate within the richest patches when patches vary in nutrient concentration). We also investigated sensitivity (growth benefits gained as spatial heterogeneity of nutrients increases, measured as total biomass).
2 Ten individuals of each species were grown in pots under four treatments that had differing nutrient distribution but the same overall nutrient addition. Plants were harvested when roots reached pot edge.
3 We observed high variation between species in scale, precision and sensitivity. No significant discrimination responses were observed, although greatest root mass density occurred at intermediate fertility levels for all species.
4 We rejected the hypothesis that scale and precision are negatively correlated. Indeed, in herbaceous species alone, scale and precision were positively correlated.
5 Sensitivity was not closely related to precision, indicating that proliferation of roots in fertile patches does not always yield growth benefits in heterogeneous soils. Further, some sensitive species had very low precision, suggesting that a positive growth response in heterogeneous environments may be related to plasticity in physiology or root life span, rather than morphology.
6 Plant life form was not correlated with precision or sensitivity. However, scale of response was greater in herbs than in woody plants, possibly because the two life forms develop root systems at different rates.     

Canopy studies on ethylene-insensitive tobacco identify ethylene as a novel element in blue light and plant-plant signalling

Plants growing at high densities express shade avoidance traits as a response to the presence of neighbours. Enhanced shoot elongation is one of the best researched shade avoidance components and increases light capture in dense stands. We show here that also leaf movements, leading to a more vertical leaf orientation (hyponasty), may be crucial in the early phase of competition. The initiation of shade avoidance responses is classically attributed to the action of phytochrome photoreceptors that sense red:far-red (R:FR) ratios in light reflected by neighbours, but also other signals may be involved. It was recently shown that ethylene-insensitive, transgenic (Tetr) tobacco plants, which are insensitive to the gaseous plant hormone ethylene, have reduced shade avoidance responses to neighbours. Here, we report that this is not related to a reduced response to low R:FR ratio, but that Tetr tobacco plants are unresponsive to a reduced photon fluence rate of blue light, which normally suppresses growth inhibition in wild-type (WT) plants. In addition to these light signals, ethylene levels in the canopy atmosphere increased to concentrations that could induce shade avoidance responses in WT plants. Together, these data show that neighbour detection signals other than the R:FR ratio are more important than previously anticipated and argue for a particularly important role for ethylene in determining plant responses to neighbours.     

Root Formation in Ethylene-Insensitive Plants

Experiments with ethylene-insensitive tomato (Lycopersicon esculentum) and petunia (Petunia x hybrida) plants were conducted to determine if normal or adventitious root formation is affected by ethylene insensitivity. Ethylene-insensitive Never ripe (NR) tomato plants produced more below-ground root mass but fewer above-ground adventitious roots than wild-type Pearson plants. Applied auxin (indole-3-butyric acid) increased adventitious root formation on vegetative stem cuttings of wild-type plants but had little or no effect on rooting of NR plants. Reduced adventitious root formation was also observed in ethylene-insensitive transgenic petunia plants. Applied 1-aminocyclopropane-1-carboxylic acid increased adventitious root formation on vegetative stem cuttings from NR and wild-type plants, but NR cuttings produced fewer adventitious roots than wild-type cuttings. These data suggest that the promotive effect of auxin on adventitious rooting is influenced by ethylene responsiveness. Seedling root growth of tomato in response to mechanical impedance was also influenced by ethylene sensitivity. Ninety-six percent of wild-type seedlings germinated and grown on sand for 7 d grew normal roots into the medium, whereas 47% of NR seedlings displayed elongated tap-roots, shortened hypocotyls, and did not penetrate the medium. These data indicate that ethylene has a critical role in various responses of roots to environmental stimuli.     

Nitrate and phosphate availability and distribution have different effects on root system architecture of Arabidopsis

Plant root systems can respond to nutrient availability and distribution by changing the three-dimensional deployment of their roots: their root system architecture (RSA). We have compared RSA in homogeneous and heterogeneous nitrate and phosphate supply in Arabidopsis. Changes in nitrate and phosphate availability were found to have contrasting effects on primary root length and lateral root density, but similar effects on lateral root length. Relative to shoot dry weight (DW), primary root length decreased with increasing nitrate availability, while it increased with increasing phosphate supply. Lateral root density remained constant across a range of nitrate supplies, but decreased with increasing phosphate supply. In contrast, lateral root elongation was suppressed both by high nitrate and high phosphate supplies. Local supplies of high nitrate or phosphate in a patch also had different effects. Primary root growth was not affected by a high nitrate patch, but growth through a high phosphate patch reduced primary root growth after the root left the patch. A high nitrate patch induced an increase in lateral root density in the patch, whereas lateral root density was unaffected by a high phosphate patch. However, both phosphate- and nitrate-rich patches induced lateral root elongation in the patch and suppressed it outside the patch. This co-ordinated response of lateral roots also occurs in soil-grown plants exposed to a nutrient-rich patch. The auxin-resistant mutants axrl, axr4 and aux1 all showed the wild-type lateral root elongation responses to a nitrate-rich patch, suggesting that auxin is not required for this response.     

Root proliferation and seed yield in response to spatial heterogeneity of below-ground competition

Here, we tested the predictions of a 'tragedy of the commons' model of below-ground plant competition in annual plants that experience spatial heterogeneity in their competitive environment. Under interplant competition, the model predicts that a plant should over-proliferate roots relative to what would maximize the collective yield of the plants. We predict that a plant will tailor its root proliferation to local patch conditions, restraining root production when alone and over-proliferating in the presence of other plants. A series of experiments were conducted using pairs of pea (Pisum sativum) plants occupying two or three pots in which the presence or absence of interplant root competition was varied while nutrient availability per plant was held constant. In two-pot experiments, competing plants produced more root mass and less pod mass per individual than plants grown in isolation. In three-pot experiments, peas modulated this response to conditions at the scale of individual pots. Root proliferation in the shared pot was higher compared with the exclusively occupied pot. Plants appear to display sophisticated nutrient foraging with outcomes that permit insights into interplant competition.     

Competitive interactions between Nardus stricta L. and Calluna vulgaris (L.) Hull: the effect of fertilizer and defoliation on above- and below-ground performance

1 The role of nutrient supply and defoliation on the competitive interactions between pot-grown Calluna vulgaris and Nardus stricta plants was investigated.WP leading adjustment
2 Young plants were grown alone and together in pots under a combination of fertilizer and defoliation treatments. After 18 months, parameters reflecting both above- and below-ground performance were measured, namely: total above-ground biomass, shoot nitrogen and phosphorus content, root length and the extent of mycorrhizal infection of the roots.
3 In the pots that received fertilizer, the shoot nutrient content and above-ground biomass of Nardus plants increased to a greater extent than those of Calluna plants; this effect was more marked for Nardus plants growing with Calluna plants than for those growing with other Nardus plants. In contrast , Calluna plants growing in competition with Nardus failed to respond to the addition of nutrients. However, in unfertilized pots, Calluna gained more above-ground biomass during the experimental period than Nardus.
4 Calluna had greater root length than Nardus , but Nardus had a higher proportion of its root length infected by mycorrhizal fungi. In both plants, the addition of fertilizer reduced the mycorrhizal infection and increased the root length. Nardus root length was decreased when grown in competition with Calluna only in pots where no nutrients were added. Defoliation decreased the extent of mycorrhizal infection in Calluna roots but not in those of Nardus; defoliation decreased the shoot nutrient content in Calluna plants, but not in Nardus plants.
5 These results suggest that the competitive balance between Nardus and Calluna may be altered by the addition of nutrients, and by defoliation, which may have serious implications for the future dominance of Calluna in heathland ecosystems, particularly those where nutrient inputs are increasing significantly or where grazing pressures are high.     

Signaling of systemic acquired resistance in tobacco depends on ethylene perception

The hypersensitive interaction between Tobacco mosaic virus (TMV) and tobacco results in accumulation of salicylic acid (SA), defense gene expression, and development of systemic acquired resistance (SAR) in uninfected leaves. The plant hormones SA and ethylene have been implicated in SAR. From a study with ethylene-insensitive (Tetr) tobacco, we concluded that ethylene perception is required to generate the systemic signal molecules in TMV-infected leaves that trigger SA accumulation, defense gene expression, and SAR development in uninfected leaves. Ethylene perception was not required for the responses of the plant to the systemic signal that leads to SAR development.     

Ethylene insensitivity results in down-regulation of rubisco expression and photosynthetic capacity in tobacco

Little is known about the effect of hormones on the photosynthetic process. Therefore, we studied Rubisco content and expression along with gas exchange parameters in transgenic tobacco (Nicotiana tabacum) plants that are not able to sense ethylene. We also tested for a possible interaction between ethylene insensitivity, abscisic acid (ABA), and sugar feedback on photosynthesis. We measured Rubisco content in seedlings grown in agar with or without added sugar and fluridone, and Rubisco expression in hydroponically grown vegetative plants grown at low and high CO(2). Furthermore, we analyzed gas exchange and the photosynthetic machinery of transformants and wild-type plants grown under standard conditions. In the presence of exogenous glucose (Glc), agar-grown seedlings of the ethylene-insensitive genotype had lower amounts of Rubisco per unit leaf area than the wild type. No differences in Rubisco content were found between ethylene-insensitive and wild-type seedlings treated with fluridone, suggesting that inhibition of ABA production nullified the effect of Glc application. When larger, vegetative plants were grown at different atmospheric CO(2) concentrations, a negative correlation was found between Glc concentration in the leaves and Rubisco gene expression, with stronger repression by high Glc concentrations in ethylene-insensitive plants. Ethylene insensitivity resulted in plants with comparable fractions of nitrogen invested in light harvesting, but lower amounts in electron transport and Rubisco. Consequently, photosynthetic capacity of the insensitive genotype was clearly lower compared with the wild type. We conclude that the inability to perceive ethylene results in increased sensitivity to Glc, which may be mediated by a higher ABA concentration. This increased sensitivity to endogenous Glc has negative consequences for Rubisco content and photosynthetic capacity of these plants.     

Effect of mutual shading on the emergence of nodal roots and the root/shoot ratio of maize

The effect of mutual shading on the root/shoot ratio and on the number of nodal roots of maize was studied. Plants of two varieties (Dea and LG2281) were grown in individual pots of 9 L, at three plant densities: 7.5, 11 and 15 plants m^–2. A control experiment was carried out in order to study if root growth was affected by the small size of the pots. Maize plants (cv Dea) were grown at a low plant density (7.5 plants m^–2) in pots of two different volumes (9 and 25 L respectively). In both experiments plants were watered every two hours with a nutrient solution. Some plants were sampled at five dates in the main experiment and the following data were recorded: foliar stage; root, stem and leaf dry weight; number of root primordia and number of emerged roots per phytomer. The final sampling date occurred at silking.Results of the control experiment showed that the root biomass was lower in small pots but the number of nodal roots per phytomer was not affected.Results of the main experiment showed that the total plant biomass and the root/shoot ratio were lower at high plant density. The number of emerged roots was strongly reduced on the upper phytomer (P₈). This reduction was mainly due to a lower percentage of root primordia which elongated. A proposed interpretation is that the number of roots which emerge on upper phytomers is controlled by carbohydrate availability.     

Growth Consequences of Soil Nutrient Heterogeneity for two Old-field Herbs,Ambrosia artemisiifoliaandPhytolacca americana, Grown Individually and in Combination

Plant species can respond to small scale soil nutrient heterogeneityby proliferating roots or increasing nutrient uptake kineticsin nutrient-rich patches. Because root response to heterogeneitydiffers among species, it has been suggested that the distributionof soil resources could influence the outcome of interspecificcompetition. However, studies testing how plants respond toheterogeneity in the presence of neighbours are lacking. Inthis study, individuals of two species,Phytolacca americanaL.andAmbrosia artemisiifoliaL. were grown individually and incombination in soils with either a homogeneous or heterogeneousnutrient distribution. Above-ground biomass of individuallygrown plants of both species was greater when fertilizer waslocated in a single patch than when the same amount of fertilizerwas distributed evenly throughout the soil. Additionally, bothspecies proliferated roots in high-nutrient patches.A. artemisiifoliaexhibitedlarger root:shoot ratios, increased nitrogen depletion fromnutrient patches, and a higher growth rate thanP. americana,suggestingA. artemisiifoliais better suited to find and rapidlyexploit nutrient patches. In contrast to individually grownplants, soil nutrient distribution had no effect on final above-groundplant biomass for either species when grown with neighbours,even though roots were still concentrated in high nutrient patches.This study demonstrates that increased growth of isolated plantsas a consequence of localized soil nutrients is not necessarilyan indication that heterogeneity will affect interspecific encounters.In fact, despite a significant below-ground response, soil nutrientheterogeneity was inconsequential to above-ground performancewhen plants were grown with neighbours.Copyright 1999 Annalsof Botany Company Phytolacca americana, pokeweed,Ambrosia artemisiifolia, ragweed, nutrient heterogeneity, root proliferation, plasticity, foraging, nutrient patches.     

Selectivity and Kinetics of Ion Uptake by Barley Plants Following Nutrient Deficiency

Barley plants grown without an external supply of phosphorus,sulphur, chlorine or nitrogen subsequently absorbed these nutrients,as phosphate, sulphate, chloride and nitrate, more rapidly thandid nutrient, sufficient control plants under similar conditions.With phosphorus, sulphur and chlorine, increased absorptionwas restricted to the nutrient which had been deficient, orto close chemical analogues of it, the uptake of other anionsbeing unaffected or decreased. The selectivity of enhanced nitrateuptake by nitrogen-deficient plants was not examined. The differencesin the rates of phosphate, sulphate and chloride absorptionby plants of differing nutrient status were due principallyto changes in the maximum transport capacity for these anionsper unit weight of root, although in plants grown without externalchloride there was some evidence that the roots also developedan increased affinity for that ion. Hordeum vulgare, barley, mineral nutrient deficiency, ion absorption, kinetics of ion uptake, phosphate, sulphate, chloride, arsenate, bromide, selenate     

土壤养分空间异质性与植物根系的觅食反应

植物在长期进化过程中,为了最大限度地获取土壤资源,对养分的空间异质性产生各种可塑性反应．包括形态可塑性、生理可塑性、菌根可塑性等．许多植物种的根系在养分丰富的斑块中大量增生,增生程度种间差异较大,并受斑块属性(斑块大小、养分浓度)、营养元素种类和养分总体供应状况的影响．植物还通过调整富养斑块中细根的直径、分枝角、节问距以及空间构型来实现斑块养分的高效利用．根系的生理可塑性及菌根可塑性可能在一定程度上影响其形态可塑性．生理可塑性表现为处于不同养分斑块上的根系迅速调整其养分吸收速率,从而增加单位根系的养分吸收,对在时间上和空间上变化频繁的空间异质性土壤养分的利用具有重要意义,可在一定程度上弥补根系增生反应的不足．菌根可塑性目前研究较少,一些植物种的菌根代替细根实现在富养斑块中的增生．菌根增生的碳投入养分吸收效率较高、根系增生对增加养分吸收的作用较复杂,取决于养分离子在土壤中的移动性能以及是否存在竞争植物;对植物生长(竞争能力)的作用因种而异,一些敏感种由此获得生长效益,而其它一些植物种受影响较小．植物个体对土壤养分空间异质性反应能力和生长差异,影响其在群落中的地位和命运,最终影响群落组成及其结构．     

Decreased Ethylene Biosynthesis, and Induction of Aerenchyma, by Nitrogen- or Phosphate-Starvation in Adventitious Roots of Zea mays L

Plants of Zea mays L. cv TX5855 were grown in a complete, well oxygenated nutrient solution then subjected to nutrient starvation by omitting either nitrate and ammonium or phosphate from the solution. These treatments induced the formation of aerenchyma close to the apex of the adventitious roots that subsequently emerged from the base of the shoot, a response similar to that shown earlier to be induced by hypoxia. Compared with control plants supplied with all nutrients throughout, N- or P-starvation consistently depressed the rates of ethylene release by excised, 25 mm apical segments of adventitious roots. Some enzymes and substrates of the ethylene biosynthetic pathway were examined. The content of 1-amino cyclopropane-1-carboxylic acid (ACC) paralleled the differences in ethylene production rates, being depressed by N or P deficiency, while malonyl-ACC showed a similar trend. Activity of ACC synthase and of ethylene forming enzyme (g⁻¹ fresh weight) was also greater in control roots than in nutrient starved ones. These results indicate that much of the ethylene biosynthetic pathway is slowed under conditions of N- or P-starvation. Thus, by contrast to the effects of hypoxia, the induction of aerenchyma in roots of Zea mays by nutrient starvation is not related to an enhanced biosynthesis and/or accumulation of ethylene in the root tips.