Lateral root frequency decreases when nitrate accumulates in tobacco transformants with low nitrate reductase activity: consequences for the regulation of biomass partitioning between shoots and root1

Accumulation of nitrate in the shoot of low-nitrate reductase tobacco transformants leads to an increase of the shoot:root ratio to higher values than in nitrogen-sufficient wild-type plants, even though the transformants are severely deficient in organic nitrogen. In the present paper, wild-type plants and low- nitrate reductase transformants were grown on vertical agar plates to investigate whether this inhibition of root growth by internal nitrate (i) can be reversed by adding sugars to the roots and (ii) is due to slower growth of the main roots or to a decreased number of lateral roots. When grown with a low nitrate supply, the transformants resembled wild-type plants with respect to amino acid and protein levels, shoot-root allocation, lateral root frequency, and rates of growth. When the transformants were grown with a high nitrate supply in the absence of sucrose they grew more slowly and had lower levels of amino acids and protein than wild-type plants, but accumulated more nitrate and developed a high shoot:root ratio. Root length was not affected, but the number of lateral roots per plant decreased. The slower root growth was accompanied by an increase of the concentration of sugars in the roots. Addition of 2% sucrose to the medium partially reversed the high shoot:root ratio in the transformants, but did not increase the frequency of lateral roots. It is concluded that nitrate accumulation in the plant leads to decreased root growth via (i) changes in carbon allocation leading to decreased allocation of sugars to root growth, and (ii) a decrease in the number of lateral roots and a shift in the sensitivity with which root growth responds to the sugar supply. This revised version was published online in June 2006 with corrections to the Cover Date.     

Accumulation of nitrate in the shoot acts as a signal to regulate shoot-root allocation in tobacco†

Mutants and transformants of tobacco (Nicotiania tabacum L. cv Gatersleben 1) with decreased expression of nitrate reductase have been used to investigate whether nitrate accumulation in the shoot acts as a signal to alter allocation between shoot and root growth. (a) Transformants with very low (1–3% of wild-type levels) nitrate reductase activity had growth rates, and protein, amino acid and glutamine levels similar to or slightly lower than a nitrate-limited wild-type, but accumulated large amounts of nitrate. These plants should resemble a nitrate-limited wild-type, except in responses where nitrate acts as a signal. (b) Whereas the shoot:root ratio decreases from about 3.5 in a well-fertilized wild-type to about 2 in a nitrate-limited wild-type, the transformants had a very high shoot:root ratio (8–10) when they were grown on high nitrate. When they were grown on lower nitrate concentrations their shoot:root ratio declined progressively to a value similar to that in nitrate-limited wild-types. Mutants with a moderate (30–50%) decrease of nitrate reductase also had a small but highly significant increase of their shoot:root ratio, compared to the wild-type. The increased shoot:root ratio in the mutants and transformants was due to a stimulation of shoot growth and an inhibition of root growth. (c) There was a highly significant correlation between leaf nitrate content and the shoot:root ratio for eight genotypes growing at a wide range of nitrate supply. (d) A similar increase of the shoot:root ratio in nitrate reductase-deficient plants, and correlation between leaf nitrate content and the shoot:root ratio, was found in plants growing on ammonium nitrate. (f) Split-root experiments, in which the transformants were grown with part of their root system in high nitrate and the other part in low nitrate, showed that root growth is inhibited by the accumulation of nitrate in the shoot. High concentrations of nitrate in the rooting medium actually stimulate local root growth. (g) The inhibition of root growth in the transformants was relieved when the transformants were grown on limiting phosphate, even though the nitrate content of the root remained high. This shows that the nitrate-dependent changes in allocation can be overridden by other signals that increase allocation to root growth. (h) The reasons for the changed allocation were investigated in transformants growing normally, and in split-root culture. Accumulation of nitrate in the shoot did not lead to decreased levels of amino acids or protein in the roots. However, it did lead to a strong inhibition of starch synthesis and turnover in the leaves, and to decreased levels of sugars in the root. The rate of root growth was correlated with the root sugar content. It is concluded that these changes of carbon allocation could contribute to the changes in shoot and root growth.     

天机营养对春小麦抗旱适应性的影响

本文研究了无机营养对春小麦一些抗旱适应性的影响,主要包括：渗透调节的大小和变化过程、可溶性糖的积累、脯氨酸的积累、叶片导度的变化、离体叶片的失水速率、叶面积和耗水量的变化、根系生长和根／植冠值,并且分析了各个处理植株的水分利用效率(WUE)和产量的变异。认为,在干旱条件下,无机营养对于春小麦不同器官、不同生理功能,并不都具有一致的作用。有的利于提高植株的抗旱性,有的可以改变一些适应性产生的时间和发展过程,有的则不利于植株的抗旱性。通过综合分析,提出旱地施肥使作物增产的主要原因是,营养元素满足了作物生长所需,促进了根系发育,利于吸收水分、维持水分平衡和正常生理功能,但对作物自身的耐旱性并没有产生显著影响。     

Shoot growth, root growth and resource capture under limiting water and N supply for two cultivars of lettuce (Lactuca sativa L.)

Background and aims

To improve vegetable crops adapted to low input and variable resource availability, better understanding is needed of root system functioning, including nitrogen and water capture.

Methods

This study quantified shoot and root development and patterns of water and nitrate capture of two lettuce cultivars subjected to temporary drought at two development stages (Trial 1) or to continuous, localized drought and/or nitrate shortage (Trial 2).

Results

In Trial 1, early drought slowed down shoot and root growth, whereas late drought enhanced root proliferation in the top 0.1 m. Nitrate capture during drought was sustained by increased nitrate inflow from deeper layers. Plants did not recover fully from drought after re-watering. In Trial 2, root proliferation was stimulated in the drier soil compartment partially compensating reduced water availability and nitrate mobility. Under nitrate shortage, root proliferation was enhanced in the compartment where nitrate was more abundant, irrespective of water availability.

Conclusions

Changes observed in the root system are ‘feed-forward’ mechanisms to sustain resource capture in a limiting growing environment. The type of stress (drought or nitrate shortage) affects coping strategies; nitrate concentration in the soil solution, combined with the nutritional status of the plant will determine the stress response.     

Translocation of nitrogen in osmotically stressed wheat seedlings

Wheat (Triticum aestivum L., cv. Drabant) seedlings were grown in a ‘split root’ system where either the whole root system or one root half was subjected to osmotic stress for 24 h, using 200 g polyethylene glycol (PEG, molecular weight 4000) dm^?3 nutrient solution. ¹⁵N-Labelled nitrate was fed to one of the root compartments and total N and ¹⁵N-labelling were measured in plant material and xylem sap. Untreated plants translocated 87% of the N taken up to the shoot, and 10% of this was then retranslocated back to the root. Recalculated on a root nitrogen basis, 36% of the label recovered in the root after 24 h had passed through the shoot. Significant labelling of xylem sap collected from non-labelled roots indicated cycling of organic N through the roots. PEG-treatment of the whole root system caused significant water loss in both roots and shoots. Uptake of nitrate and retranslocation of N to roots were inhibited, whereas cycling of organic nitrogen through the root was still measurable. Treatment of half the root system with PEG had minor effects on shoot water content, but reduced the water content of the treated root part. The total uptake of nitrate by the root system was unaffected, and the effect on the treated root half was comparatively small. Nitrate reductase activity (NRA) declined in PEG-treated roots even if high nitrate uptake rates were maintained. Shoot NRA was unaffected by osmotic stress. The data indicate that the reduction in water content of the root per se has only small effects on nitrate uptake. Major inhibition of nitrate uptake was observed only after treatment of a sufficiently large portion of the root system to given an effect on shoot water content.     

Effect of sucrose application,minerals, and irradiance on the <Emphasis Type="Italic">in vitro</Emphasis> growth of <Emphasis Type="Italic">Cistus incanus</Emphasis> seedlings and plantlets

To study the effect of sucrose on the sink-source relationship in in vitro-grown plants, Cistus incanus seedlings and plantlets were grown horizontally in a two-compartment Petri dish (split dish), with the root system in one compartment and the shoot in the other. Shoots and roots were exposed to different sucrose concentrations (0–30 g dm⁻³), two irradiance levels (25 and 160 μmol m⁻²s⁻¹) and the presence or absence of a minimum medium containing minerals and vitamins (M medium). Root and shoot biomass of the seedlings was enhanced by an increase in irradiance when the growth medium was not supplemented with sucrose indicating the role of photosynthesis in biomass production. When sucrose was added to either organ growth was enhanced as well. In the presence of sucrose in the root compartment, sucrose applied to the shoot compartment enhanced growth of both organs under low irradiance, while under high irradiance, sucrose had no further additive effect. In the absence of sucrose in the root compartment, the enhancement of root biomass by sucrose added to the shoot compartment was lower under high irradiance than under low irradiance. The response of Cistus plantlets to sucrose and irradiance differed from that of seedlings, probably reflecting a greater susceptibility of the plantlets to sucrose feedback inhibition on photosynthesis and biomass accumulation. The decrease in root and shoot growth when M medium was added to the shoot compartment and the relatively better growth of these organs when the roots were supplied with minerals and the shoot with sucrose, indicate that growth of the two organs in our experimental set-up was regulated by opposing fluxes of C and nutrients.     

The role of nutrient availability in regulating root architecture

The ability of plants to respond appropriately to nutrient availability is of fundamental importance for their adaptation to the environment. Nutrients such as nitrate, phosphate, sulfate and iron act as signals that can be perceived. These signals trigger molecular mechanisms that modify cell division and cell differentiation processes within the root and have a profound impact on root system architecture. Important developmental processes, such as root-hair formation, primary root growth and lateral root formation, are particularly sensitive to changes in the internal and external concentration of nutrients. The responses of root architecture to nutrients can be modified by plant growth regulators, such as auxins, cytokinins and ethylene, suggesting that the nutritional control of root development may be mediated by changes in hormone synthesis, transport or sensitivity. Recent information points to the existence of nutrient-specific signal transduction pathways that interpret the external and internal concentrations of nutrients to modify root development. Progress in this field has led to the cloning of regulatory genes that play pivotal roles in nutrient-induced changes to root development.     

Adaptation of roots to low water potentials by changes in cell wall extensibility and cell wall proteins

It is common for the root/shoot ratio of plants to increase when water availability is limiting. This ratio increases because roots are less sensitive than shoots to growth inhibition by low water potentials. The physiological and molecular mechanisms that assist root growth under drought conditions are reviewed, with a focus on changes in cell walls. Maize seedlings adapt to low water potential by making the walls in the apical part of the root more extensible. In part, this is accomplished by increases in expansin activity and in part by other, more complex changes in the wall. The role of xyloglucan endotransglycosylase, peroxidase and other wall enzymes in root adaptation to low water potential is evaluated and some of the complications in the field of study are listed.     

Production of Biomass and Ginsenosides from Adventitious Roots of Panax ginseng in Bioreactor Cultures

Organic nutrients play a central role during Panax ginseng adventitious root culture in bioreactor systems. To understand how the nutrient elements were uptaken during the adventitious root growth as well as the production of biomass and natural ginsenosides, a biotechnological approach to identifying the nutritional physiology of ginseng in a commercial‐scale bioreactor was necessary. Normal MS medium nutrient in the bioreactor culture of adventitious roots resulted in slow growth, low biomass, and Rg and Rb ginsenoside contents. When the ginsenoside production increased to higher levels, a group of regulatory nutritional elements that have the potential to interact with biomass was identified. The effects of the salt strength of the medium, of macroelements, metal elements, the ammonia/nitrate ratio, sucrose concentration, and osmotic agents on the growth, the formation of biomass and the production of ginsenosides from adventitious roots were investigated. Appropriate conditions allowed for a maximum ginsenoide production of up to 12.42 [mg/g DW] to be obtained after 5 weeks of culture. The results demonstrated that the key organic nutrients can be regulated to improve the biomass and growth, and increase the ginsenoside yield in bioreactor cultures of P. ginseng adventitious roots.     

Root growth and functioning under atmospheric CO2 enrichment

This paper examines the extent to which atmospheric CO₂ enrichment may influence growth of plant roots and function in terms of uptake of water and nutrients, and carbon allocation towards symbionts. It is concluded that changes in dry matter allocation greatly depend on the experimental conditions during the experiment, the growth phase of the plant, and its morphological characteristics. Under non-limiting conditions of water and nutrients for growth, dry matter partitioning to the root is not changed by CO₂ enrichment. The increase in root/shoot ratio, frequently observed under limiting conditions of water and/or nutrients, enables the plant to explore a greater soil volume, and hence acquire more water and nutrients. However, more data on changes in dry matter allocation within the root due to atmospheric CO₂ are needed. It is concluded that nitrogen fixation is favored by CO₂ enrichment since nodule mass is increased, concomitant with an increase in root length. The papers available so far on the influence of CO₂ enrichment on mycorrhizal functioning suggest that carbon allocation to the roots might be increased, but also here more experiments are needed.Abbreviations LAR leaf area ratio - LWR leaf weight ratio - SWR stem weight ratio - RGR relative growth rate - R/S root/shoot - RWR root weight ratio     

Response of Split-Root Sour Orange Seedlings to NaCl and Polyethylene Glycol Stresses

Soil water cotent and salinity levels are seldom uniform inthe field, particularly with the use of micro-irrigation systemsthat may water only a portion of the root zone. For studyingnon-uniform salinity, a split-root experiment was designed toevaluate growth and water relations when half of the root systemof sour orange (Citrus aurantium) seedlings was stressed withsodium chloride (NaCl) or polyethylene glycol (PEG). This studyalso determined if non-stressed portions of the root systemcompensated for the decrease in water uptake by the stressedportions. One or both halves of the root system were treated for fourmonths with nutrient solution adjusted with NaCl or PEG to osmoticpotentials of –0.10, –0.20, or –0.35 MPa.Shoot dry weight was reduced by only 9% when half of the rootsystem was irrigated with saline solution at –0.10 MPa,but with both halves of the root system at –0.10 MPa,shoot and root dry weights were reduced as much as 45%. Similarly,leaf water and osmotic potentials were also more disturbed underuniform salinity than under non-uniform salinity conditions. Plant growth, leaf water potential, osmotic potential, stomatalconductance, and evapotranspiration decreased with increasingNaCl and PEG concentrations in the nutrient solution. Turgorpotential and leaf thickness increased in response to NaCl treatments.Microscopic examination showed that the increase in leaf thicknesswas due to the development of larger cells in the spongy mesophyll. Shoot growth did not correlate with the average osmotic potentialof the two root halves. Seedlings with one stressed half-rootsystem had shoot dry weight and leaf water potential valuescloser to those of the non-stressed control than to those withthe completely stressed root system. Key words: Non-uniform salinity, water relations, citrus     

Influence of soil water deficits on root growth of cotton seedlings

Summary Cotton (Gossypium hirsutum L. cv. H14) seedlings were raised in soil of differing soil water content in specially designed pots in which the roots had access to freely available water and nutrients located 2.5 cm below the base of the soil core. The time for root emergence from the soil core and the rate of root growth were measured daily from sowing to harvest. The root and shoot dry weight and leaf water potential were measured at the final harvest 16 days after sowing. As soil water content decreased, the root emerged from the soil earlier and the initial rate of root elongation was faster. In spite of the availability of freely available water, the plants in the soil at low water contents had significantly lower leaf water potentials than those in soil at high water contents. The root: shoot ratio increased as the soil water content decreased. This arose from an absolute increase in root weight, with shoot weight not being significantly affected.     

Partitioning of Sugar between Growth and Nitrate Reduction in Cotton Roots

The level of endogenous sugars was inversely related to nitrate availability in young cotton (Gossypium hirsutum L.) plants, with high nitrate causing a greater decline in sugar content of roots than of shoots. High nitrate (low sugar) plants also displayed relatively more shoot growth and less root growth than low nitrate (high sugar) plants. These data are consistent with the theory that roots are poor competitors for sugar, and that sugar supply is a major factor limiting root growth in vivo.

The effects of endogenous sugar level on root growth and on nitrate reductase activity in the root were different. When root sugar level was experimentally controlled by varying nitrate concentration in the nutrient solution, root growth was less sensitive than nitrate reductase activity to sugar deficiency. Also, in sterile root tips cultured on media containing a wide range of sucrose concentrations, growth rate was considerably less sensitive to endogenous sugar deficiency than was nitrate assimilation rate. Similarly, in plants which were detopped or girdled, nitrate reductase activity in the roots declined more rapidly than did root sugars, especially glucose and fructose. These results suggest that when sugar is deficient, cotton roots preferentially use it for growth at the expense of nitrate reduction.

     

Environmental effects on dry matter partitioning between shoot and root of crop plants: relations with growth and shoot protein concentration

The literature on environmental effects on dry matter partitioning in higher plants, in particular crop plants, is reviewed focussing on changes in shoot to root dry weight ratio (S:R). Of particular consistency is the finding that S:R increases with increased nitrogen (N) supply. Relations between nitrogen (N) supply, growth, S:R and tissue N and protein concentration are examined. In some cases, the increase in S:R with increased N supply is likely to have been at least in part an effect on growth and development, but there is unequivocal evidence that N affects S:R independently of growth and development. A positive correlation between S:R and leaf protein concentration is highlighted. It is argued that the N effect on S:R outside the effect on growth and development is related to increased shoot protein concentration. Specifically, shoot and root growth are colimited by local carbon (C) and N (primarily protein) substrate concentrations and shoot growth will increase relative to root growth with increased N substrate availability due to the proximity of the shoot to the C source. It is further argued that results in the literature are consistent with the proposal that macronutrient, water, irradiance, CO₂ and temperature effects on S:R are often primarily mediated through their effects on growth and development, and shoot protein concentration and hence shoot growth.     

Compensatory Changes in the Partitioning of Dry Matter in Relation to Nitrogen Uptake and Optimal Variations in Growth

Equations are derived relating relative growth rate (RGR) toroot:shoot ratio, root length, nitrogen inflow rate, leaf area,photosynthesis and carbon and nitrogen concentrations in theplant. The extents to which changes in specific root lengthand root: shoot ratio can compensate for the effects of lowN availability upon RGR are examined. Such responses could haveseveral compensatory functions: maximizing RGR; maintaininggrowth in which the activities of root and shoot limit RGR equally;and maximizing the efficiency of increase in RGR. Growth, nitrogen, carbon, dry matter, partitioning, root:shoot ratio, relative growth rate     

Review: role of carbon sources for in vitro plant growth and development

In vitro plant cells, tissues and organ cultures are not fully autotrophic establishing a need for carbohydrates in culture media to maintain the osmotic potential, as well as to serve as energy and carbon sources for developmental processes including shoot proliferation, root induction as well as emission, embryogenesis and organogenesis, which are highly energy demanding developmental processes in plant biology. A variety of carbon sources (both reducing and non-reducing) are used in culture media depending upon genotypes and specific stages of growth. However, sucrose is most widely used as a major transport-sugar in the phloem sap of many plants. In micropropagation systems, morphogenetic potential of plant tissues can greatly be manipulated by varying type and concentration of carbon sources. The present article reviews the past and current findings on carbon sources and their sustainable utilization for in vitro plant tissue culture to achieve better growth rate and development.     

Rhizosphere humic acid interacts with root cell walls to reduce hydraulic conductivity and plant development

Humic acids are ubiquitous, organic-end-products of the chemical and microbial degradation of dead biota in soils throughout the world. Humic acids can be transported in soil water as heterogeneous, supra-molecular, colloidal-agglomerates. Humic acid accumulation in the rhizosphere of transpiring plants may chemically stimulate development by increasing root availability of mineral nutrients and/or growth regulatory biomolecules. This report introduces novel, physical mechanisms by which humic acid can also reduce plant development. Effects of humic acid addition to the root media of intact maize plants (Zea mays L.) on their growth, transpiration and resistance to water deficits were assayed, as were the effects of external humic acid on the hydraulic conductivity of excised primary-roots. Humic acid reduced shoot growth, transpiration and resistance to water stress but not root growth. Root hydraulic conductivity was reduced by up to 44% via a time-, concentration- and size-dependent fouling mechanism resulting from humic acid accumulation at root cell-walls. Thus, humic acid is shown, apparently for the first time, to be able to exert novel physical effects in addition to its known chemical effects on plant development.     

A role for shoot protein in shoot-root dry matter allocation in higher plants

BACKGROUND AND AIMS: It is stated in many recent publications that nitrate (NO3-) acts as a signal to regulate dry matter partitioning between the shoot and root of higher plants. Here we challenge this hypothesis and present evidence for the viewpoint that NO3- and other environmental effects on the shoot:root dry weight ratio (S:R) of higher plants are often related mechanistically to changes in shoot protein concentration. METHODS: The literature on environmental effects on S:R is reviewed, focusing on relationships between S:R, growth and leaf NO3- and protein concentrations. A series of experiments carried out to test the proposal that S:R is dependent on shoot protein concentration is highlighted and new data are presented for tobacco (Nicotiana tabacum). KEY RESULTS/EVIDENCE: Results from the literature and new data for tobacco show that S:R and leaf NO3- concentration are not significantly correlated over a range of environmental conditions. A mechanism involving the relative availability of C and N substrates for growth in shoots can explain how shoot protein concentration can influence shoot growth and hence root growth and S:R. Generally, results in the literature are compatible with the hypothesis that macronutrients, water, irradiance and CO2 affect S:R through changes in shoot protein concentration. In detailed studies on several species, including tobacco, a linear regression model incorporating leaf soluble protein concentration and plant dry weight could explain the greater proportion of the variation in S:R within and between treatments over a wide range of conditions. CONCLUSIONS: It is concluded that if NO3- can influence the S:R of higher plants, it does so only over a narrow range of conditions. Evidence is strong that environmental effects on S:R are often related mechanistically to their effects on shoot protein concentration.     

The development of waterlogging damage in wheat seedlings (Triticum aestivum L.)

Summary The effects of waterlogging on concentrations of gases and various solutes dissolved in the soil water were investigated in the laboratory, to determine whether the early disruption to the growth of wheat was most closely associated with depletion of dissolved oxygen, accumulation of toxins, or changes in concentrations of nutrient ions in the soil water. Waterlogging slowed shoot fresh weight accumulation, leaf extension and nodal root growth; it also caused death of the seminal root system and early senescence of the lower leaves. However, the shoot dry weight initially increased above that of the non-waterlogged controls, and thus was not a reliable indicator of the early restriction to plant growth and development. The symptoms of damage to shoots and roots were attributed to the fall in soil oxygen concentrations, rather than to any decrease in concentration of inorganic nutrients in the soil water, or to the accumulation of any other measured solutes to toxic concentrations.