Chlorate as a Transport Analog for Nitrate Absorption by Roots of Tomato

Several studies have indicated that chlorate (ClO3-) and nitrate (NO3-) may share a common transport system in higher plants. Here, we compared the interactions between ClO3- and NO3-uptake by roots of intact tomato (Lycopersicon esculentum cv T5) plants. Exposure to ClO3- for more than 2 h inhibited both net ClO3- and K+ uptake, presumably because of ClO3- toxicity; consequently, subsequent measurements were conducted after short exposures to ClO3-. The apparent affinity and apparent maximum rate of absorption for net ClO3- and NO3- uptake were very similar. Interactions between ClO3- and NO3- transport were complex; 50 [mu]M NO3- acted as a mixed inhibitor of net ClO3- uptake, but 50 [mu]M ClO3- had no significant effect on net NO3- uptake, and 500 [mu]M ClO3- had no significant effect on 15NO3- influx. If the two ions share a single common high-affinity transport system, it is much more selective for NO3- than would be suggested by the similarity of net NO3- and ClO3- uptake kinetics. Our results indicate that, although NO3- may interfere with root ClO3- uptake, ClO3- is not a useful analog for the root high-affinity NO3- transport system.     

Methylammonium as a Transport Analog for Ammonium in Tomato (Lycopersicon esculentum L.)

Methylammonium (CH3NH3+) has been widely used as an analog of ammonium (NH4+) for examining transport in bacteria and fungi. We compared the kinetics of root CH3NH3+ and NH4+ uptake from solution culture in intact tomato (Lycopersicon esculentum cv T5) plants. Efflux of NH4+ and CH3NH3+ was negligible. The apparent maximum rate of absorption (apparent Vmax) was similar for NH4+ and CH3NH3+, but the apparent affinity (apparent Km) was about 10-fold greater for NH4+ than for CH3NH3+. In characterizing the interaction between NH4+ and CH3NH3+ transport, we used [15N]NH4+ and [14C]CH3NH3+ as well as improved methods for analysis of nonisotopic CH3NH3+ and NH4+. CH3NH3+ acted as an inhibitor of NH4+ influx. Relatively low concentrations of NH4+ strongly inhibited CH3NH3+ influx. Treatments with 1 mM methionine sulfoximine that blocked NH4+ assimilation had little influence on NH4+ inhibition of CH3NH3+ influx. These results suggest that the two ions share a common transport system in tomato, but because this transport system has a much greater affinity for NH4+, CH3NH3+ may be used as a transport analog only when ambient concentrations of NH4+ are very low.     

Inoculation of cranberry (Vaccinium macrocarpon) with the ericoid mycorrhizal fungus Rhizoscyphus ericae increases nitrate influx

Despite the ubiquitous presence of ericoid mycorrhizal (ERM) fungi in cranberry (Vaccinium macrocarpon), no prior studies have examined the effect of ERM colonization on NO(3)(-) influx kinetics. Here, (15)NO(3)(-) influx was measured in nonmycorrhizal and mycorrhizal cranberry in hydroponics. Mycorrhizal cranberry were inoculated with the ERM fungus Rhizoscyphus (syn. Hymenoscyphus) ericae. (15)NO(3)(-) influx by R. ericae in solution culture was also measured. Rhizoscyphus ericae NO(3)(-) influx kinetics were linear when mycelium was exposed for 24 h to 3.8 mm NH(4)(+), and saturable when pretreated with 3.8 mm NO(3)(-), 50 microm NO(3)(-), or 50 microm NH(4)(+). Both low-N pretreatments induced greater NO(3)(-) influx than either of the high-N pretreatments. Nonmycorrhizal cranberry exhibited linear NO(3)(-) influx kinetics. By contrast, mycorrhizal cranberry had saturable NO(3)(-) influx kinetics, with c. eightfold greater NO(3)(-) influx than nonmycorrhizal cranberry at NO(3)(-) concentrations from 20 microm to 2 mm. There was no influence of pretreatments on cranberry NO(3)(-) influx kinetics, regardless of mycorrhizal status. Inoculation with R. ericae increased the capacity of cranberry to utilize NO(3)(-)-N. This finding is significant both for understanding the potential nutrient niche breadth of cranberry and for management of cultivated cranberry when irrigation water sources contain nitrate.     

Green Revolution Trees: Semidwarfism Transgenes Modify Gibberellins, Promote Root Growth, Enhance Morphological Diversity, and Reduce Competitiveness in Hybrid Poplar

Semidwarfism has been used extensively in row crops and horticulture to promote yield, reduce lodging, and improve harvest index, and it might have similar benefits for trees for short-rotation forestry or energy plantations, reclamation, phytoremediation, or other applications. We studied the effects of the dominant semidwarfism transgenes GA Insensitive (GAI) and Repressor of GAI-Like, which affect gibberellin (GA) action, and the GA catabolic gene, GA 2-oxidase, in nursery beds and in 2-year-old high-density stands of hybrid poplar (Populus tremula × Populus alba). Twenty-nine traits were analyzed, including measures of growth, morphology, and physiology. Endogenous GA levels were modified in most transgenic events; GA₂₀ and GA₈, in particular, had strong inverse associations with tree height. Nearly all measured traits varied significantly among genotypes, and several traits interacted with planting density, including aboveground biomass, root-shoot ratio, root fraction, branch angle, and crown depth. Semidwarfism promoted biomass allocation to roots over shoots and substantially increased rooting efficiency with most genes tested. The increased root proportion and increased leaf chlorophyll levels were associated with changes in leaf carbon isotope discrimination, indicating altered water use efficiency. Semidwarf trees had dramatically reduced growth when in direct competition with wild-type trees, supporting the hypothesis that semidwarfism genes could be effective tools to mitigate the spread of exotic, hybrid, and transgenic plants in wild and feral populations.Semidwarfism is a valuable trait in many crop species and agricultural environments. In cereal crops, it can result in decreased lodging, increased yield, and improved harvest index (Dalrymple, 1985; Hedden, 2003). Therefore, it was a critical foundation of the “Green Revolution” that resulted in large improvements of yield in wheat (Triticum aestivum) and rice (Oryza sativa; Hargrove and Cabanilla, 1979; Perovic et al., 2008). Semidwarfism has had substantial benefits for fruit tree production, where it enables earlier fruit bearing, higher yields, and easier harvests in orchards (Battisini and Battisini, 2005). Semidwarf woody species are also extensively used in ornamental horticulture, where they allow more compact forms to be fit into small areas around homes and on streets and reduce the need for pruning to avoid interference with structures and transmission lines (Busov et al., 2003).Although against the current orthodoxy of forest tree breeding, where height growth is emphasized, semidwarfism might also have benefits for wood and biomass production (Bradshaw and Strauss, 2001). Such trees could be useful if they were less prone to wind throw due to their shorter, stockier forms and expected greater allocation to roots. Reduced stature could also result in less bending and slanting of trunks in the face of wind and gravity on hillslopes and thus reduce the extent of reaction wood formation, which degrades the performance and value of solid wood and pulp products. Reduced height and increased allocation of growth to roots might enhance stress tolerance, soil nutrient uptake, bioremediation, and carbon sequestration.Semidwarfism can be achieved by the modification of several types of genes and physiological mechanisms, but the most prevalent and advanced forms in agriculture affect GAs or their signaling (for review, see Busov et al., 2008). GAs are endogenous plant hormones that influence several aspects of plant growth and development, including seed germination, leaf expansion, shoot growth, cell division, flower induction, and fruit development (Sun and Gubler, 2004; Fleet and Sun, 2005; Swain and Singh, 2005). With respect to shoot growth, the most obvious effect of GA is its promotion of stem elongation by stimulating both cell elongation and division (Marth et al., 1956). GA modification also has significant effects on plant biochemistry, changing the amounts and distribution of a wide variety of metabolites in shoots and roots (Rossetto et al., 2003; Chen et al., 2004; Busov et al., 2006).Little is known about how semidwarfism affects belowground growth. GA has been shown to play a controlling role in lateral root development (Gou et al., 2010), and GA and ethylene synergistically promote both the initiation and growth of adventitious roots (Osmont et al., 2007). In tomato (Solanum lycopersicum), isogenic GA-deficient mutants (gib) allocate more biomass to roots compared with shoots (Nagel et al., 2001). In poplar (Populus spp.), semidwarf transgenic plants grown in vitro had a lower shoot-to-root ratio, which was at least partly due to proliferation of lateral roots (Busov et al., 2006; Gou et al., 2010).As a domestication trait, semidwarfism has been proposed as a means for mitigating the spread of transgenic plants within and outside of crop environments (Al-Ahmad et al., 2005). The genetic dominance of most semidwarfism transgenes would cause reduced height growth in transgene-containing progeny, reducing their ability to compete for light. Moreover, because of the close linkage of the semidwarfism genes to other genes that were cointroduced on the same plasmid, they would also powerfully retard their spread or introgression, even in cases where the linked transgene would, on their own, impart a selective advantage. However, there have been very few plant species where this concept has been explicitly tested (Al-Ahmad and Gressel, 2006; Gressel and Valverde, 2009), and we know of no examples in woody or perennial plants.To study the effects of semidwarfism genes in a woody plant grown under field conditions, we inserted a number of dominant GA-modifying transgenes into hybrid poplar (Populus tremula × Populus alba), the widely recognized model woody plant for genomics and biotechnology (Herschbach and Kopriva, 2002; Brunner et al., 2004a; Tuskan et al., 2004). Most of the genes studied were overexpressed forms of GA 2-oxidase, GA-Insensitive (GAI), or Repressor of GAI-Like (RGL), all known to cause semidwarfism in other plant species. GA 2-oxidase is a major GA catabolic enzyme in plants, and GAI and RGL are negative regulators of the GA signal transduction pathway (Appleford et al., 2007; Busov et al., 2008). The transgenic trees were first analyzed in the greenhouse (Busov et al., 2006) and then assayed for their effect on height growth in a 2-year field trial (Zawaski et al., 2011), from which we selected 10 transgenic events that grew at approximately three-quarters the rate of wild-type trees. The goal was to select semidwarf trees whose phenotype was not so severe as to be irrelevant to possible crop uses but strong enough to give a clear phenotype in a field study. In this study, we analyzed changes in a number of morphological, physiological, and growth traits and investigated the prospect for semidwarfism to be used as a mitigation tool to reduce the frequency of spread of transgenic and exotic species.     

Nitrogen cycling in poplar stands defoliated by insects

Large-scale outbreaks of defoliating insects are common in temperate forests. These outbreaks are thought to be responsible for substantial cycling of nitrogen (N), and its loss from the system. Gypsy moth (Lymantria dispar) populations within poplar plots were manipulated over 2 years so that the ecosystem-wide consequences of catastrophic defoliation on N cycling could be examined. The quantities of N in leaf litter-fall, ammonia volatilization and soil N pools were estimated across the two seasons. Defoliated leaf biomass was estimated from experimentally derived approximate digestibility factors and added to the mass of senesced leaf to determine total annual leaf production. Throughout the growing season the defoliation treatment peaked at about 40% in year 1 and 100% in year 2. Rapid regrowth after defoliation meant that only 45% of the annual leaf biomass was consumed in the defoliation treatment in year 2, while control plots suffered about 20% consumption each year. In each year, defoliated plots produced 20% more leaf biomass and N than the controls, a phenomenon attributed to compensatory photosynthesis. No substantial losses of N via ammonia volatilization, nitrous oxide emission or nitrate leaching were observed. Neither was there any sustained or substantial gain in the soils microbial biomass or inorganic N pools. These observations suggest that the defoliated poplars were able to compete with soil microbes and N loss mechanisms for soil N as it became available, thereby ameliorating the effects of defoliation on soil nitrogen cycling. We conclude from this study that the N mineralized from defoliation residues was conserved in this plantation ecosystem.     

Repeated insect defoliation effects on growth, nitrogen acquisition, carbohydrates, and root demography of poplars

Large-scale outbreaks of defoliating insects are common in temperate forests. The effects of defoliation on tree physiology are expected to cascade through the entire forest ecosystem, altering carbon, nitrogen, and water fluxes, and subsequently affecting nitrogen cycling and plant-herbivore interactions. If these post-defoliation changes are largely driven by N deficiency, tree root system responses to defoliation should be central to regulating the long-term effects of defoliation; N fertilization should reverse the effects. We examined these phenomena in a 3-year large-scale replicated manipulative field experiment in a hybrid poplar plantation, where we regulated defoliation by gypsy moths as well as nitrogen availability. To our knowledge, this is the first manipulative field experiment at this scale to examine the effects of severe insect defoliation on whole-tree physiology. Defoliation decreased tree growth and increased the rate of top dieback in the stand. Defoliation led to transient declines in carbon allocation to starch in fine roots, trunk, and twigs in the year of heaviest defoliation. Root production and root mortality were unaffected by the heaviest defoliation, but nitrate and ammonium uptake were strongly depressed. N fertilization increased tree growth, but did not alter defoliation effects on starch accumulation or top dieback. Defoliation and fertilization treatments did not interact. In this system, defoliation effects on tree recovery of leaf nitrogen lost to herbivory were primarily driven by effects on nitrogen uptake, rather than effects on root production or mortality.     

Resource competition and suppression of plants colonizing early successional old fields

Early colonizing annual plants are rapidly suppressed in secondary succession on fertile midwestern old fields, while later colonizing perennials persist. Differences in competitive ability for above- and belowground resources may be partly responsible for differences in species persistence during succession, as both light and nutrient availability may change rapidly. We found that, although both above- and belowground competition suppress growth of colonizing plants, belowground competition was the dominant factor in the suppression of the annual Ambrosia artemisiifolia in 2nd-year-old fields near the W.K. Kellogg Biological Station in southwestern Michigan. Despite an ability to persist in later successional fields, seedling transplants of the perennial Achillea millefolium were also suppressed by above- and belowground competition, with belowground competition having the strongest effect. As in many old fields, nitrogen availability is the primary factor limiting plant productivity. There was no clear difference between the species in ability to compete for ¹⁵N from an enriched patch, although there was an indication of greater precision of foraging by Achillea. Life history differences between these species and consequent differences in the phenology of root growth relative to other old-field plants are likely to play a large role in the persistence of Achillea in successional fields where Ambrosia is suppressed. Received: 8 January 1998 / Accepted: 16 September 1998     

The fate of surface roots of citrus seedlings in dry soil

The top portions of the root system of deeply rooted plantsare frequently in dry soil while deeper roots still have accessto water. We expected that many surface roots would be shedwhen subject to localized soil drying. We further hypothesizedthat the cost of fine root construction per unit root lengthwould be negatively correlated with the rate at which root lengthis shed. Seedlings of four citrus root- stocks that varied widelyin specific root length (cm g^–1 root) were used to testthese hypotheses. Plants were grown for 4 months in a split-potsystem divided into a top and bottom pot. After roots were wellestablished in the bottom pot, water was withheld from the toppots of half of the plants; plants were harvested every 2 weeksthereafter. Sufficient water was supplied to the bottom potto prevent shoots of droughted seedlings from experiencing significantwater stress. All plants were labelled with ¹⁴CO₂ 48 h beforeharvesting, and autoradiographs made of the fine roots harvestedfrom the droughted compartment. Comparisons of the autoradiographswith digitized images of the root system allowed us to assessroot mortality and root sink activity. As expected, the proportionof ¹⁴C-labelled photosyrithate allocated to fine roots in thetop pot declined with soil drying in all four genotypes; however,there was no genotypic effect on this decline. Contrary to ourexpectations, extensive root mortality was not apparent forany genotype, even after 60 d of localized soil drying. Apparently,selection for rapid shedding of roots in response to soil dryinghas not occurred in these Citrus species. Key words: Carbon allocation, drought, root death, split root, root autoradiography     

Cost-effectiveness of managing Natura 2000 sites: an exploratory study for Finland,Germany, the Netherlands and Poland

Natura 2000 sites are expected to assure the long-term survival of Europe’s most valuable and threatened species and habitats. It follows that successful management of the sites is of great importance. Next to goal attainment, cost-effectiveness is increasingly recognised as a key requirement for gaining social and political acceptance for costly conservation measures. We identify and qualitatively examine issues of cost-effectiveness related to the design and implementation of management measures in Natura 2000 sites in Finland, Germany, the Netherlands and Poland. Given the wide variety of management design and implementation options within the four countries, our study is purely of an exploratory nature. We derive recommendations for improving the cost-effectiveness of management in Natura 2000 sites and for future research. Examples of policy recommendations include guaranteeing the availability of funds for longer periods, and ensuring the appropriate allocation of funds between the different tasks of designing and implementing management plans. Further research should examine the cost-effectiveness of controversial suggestions such as, for example, more tailored payment schemes for conservation measures that result in higher ecological outputs but are costly to administer. Moreover, more research is needed to better understand how rules for administrations, as well as rules and governance structures for tasks within administrations, should be designed.