The relative performance of white clover genotypes with rhizobial and mineral nitrogen in agar culture and in soil

Vegetative propagules of adult plants of white clover grown in agar culture show a better agreement with the performance of adult plants grown in soil than that previously reported of nodulated seedlings. This suggests that, whilst results obtained from the inoculation of seedlings can only be extrapolated to other situations with great caution, laboratory techniques which allow nitrogen fixation assessments to be made on adult plants may be useful in predicting performance under more natural conditions. The growth of vegetative propagules of mature plants dependent on rhizo-bium fixed nitrogen was also compared with propagules receiving abundant inorganic nitrogen. In agar culture there were no differences in yield between nitrogen sources, indicating that the symbiotic associations could fix sufficient nitrogen for growth under these relatively restrictive conditions. In soil, however, only the slower genotypes came near to providing sufficient nitrogen from symbiosis to sustain their potential yield as measured from their growth with inorganic nitrogen. The more vigorous the genotype the greater was the short-fall in nitrogen supplied by the rhizobium association. Joint regression analysis showed that any apparent interactions between plant genotypes and the source of nitrogen was a function of the genotypes' potential growth rate. The importance of quantifying nitrogen fixation in relation to the growth potential of the host genotype is emphasized, and the need, in legume breeding programmes, for simultaneous selection of host and rhizobium is discussed.     

根瘤菌菌剂的研究与开发现状

根瘤菌与豆科植物共生成为豆科植物固氮的重要方式,它可以为豆科植物提供所需氮量的1/2～1/3。因此,土壤中有效根瘤菌的数量是决定豆科植物产量的重要因素,而根瘤菌菌剂的使用可以有效地提高土壤中根瘤菌数量。本文从根瘤菌菌剂制备中高效菌种的选育及匹配、高密度菌剂的制备、菌剂保存方法等方面进行综述。比较了自然选育、杂交选育和诱变选育等各类选育方法及琼脂试管配对法和水培配对法的优缺点;总结了菌剂制备的一般过程和方法;论述了菌剂保藏过程中冷冻干燥法和各种保护剂的使用对菌剂保藏效果的影响。本文阐述了根瘤菌菌剂的制备工艺和发展方向,为根瘤菌剂的研制提供重要参考。     

Response of cowpea (Vigna unguiculata) to inoculation with VA-mycorrhizal fungi and to rock phosphate fertilization in some unsterilized Nigerian soils

Summary The effects on cowpea of inoculation with vesicular-arbuscular (VA) mycorrhizal fungi and rock phosphate (RP) fertilization were studied in pots using Alagba and Araromi series soils and in the field on Alagba, Apomu and Egbeda series soils. Inoculation of the plants with VA-mycorrhizal fungi caused very rapid infection of the roots. A higher per cent mycorrhizal infection was maintained during subsequent plant growth in the field. RP application reduced the degree of infection without affecting plant growth in the field and in pot experiments. Nodulation, nitrogen fixation and utilization of RP were increased by inoculation with mycorrhizal fungi in the pot experiments but not in the field experiments. In the pot experiments, inoculated plants supplied with RP flowered earlier, and took up more phosphorus than either inoculated plants without RP or uninoculated plants. The largest response to inoculation in terms of shoot dry matter, nodule yield and nitrogen content of shoots was obtained in Alagba soil under both pot and field conditions.IITA Journal Series Paper No. 136.     

The influence of living plants on mineralization and immobilization of nitrogen

Summary Changes in the pattern of distribution of the nitrogen of the soil and seedling grass plants have been investigated when the grass plants were grown in pots of sandy soil, from a pasture, at pH 5.7. Net mineralization of soil nitrogen was not observed during an experimental period of one month in the absence of added nitrogenous fertilizer (Table 2). Addition of labeled nitrogen (as ammonium sulphate) to the soil at the beginning of the experimental period resulted in a negative net mineralization during this period (Table 4b). When none of the fertilizer nitrogen remained in its original form in the soil it was found that approximately 12 per cent of the labeled nitrogen had been immobilized in soil organic compounds. Clipping of the grass at this date was followed by a decrease in the amount of labeled soil organic nitrogen, indicating that mineralization was not depressed by living plants. The application of unlabeled ammonium sulphate subsequent to the utilization of the labeled nitrogen did not decrease the amount of immobilized labeled nitrogen in the soil organic matter, as would be expected if the organic nitrogen compounds of the soil had been decomposed to ammonia. This was thought to be due to the fact that decomposition of organic nitrogen compounds in permanent grassland results in the production of peptides, amino acids etc. which are utilized by microorganisms without deamination taking place. In pots with ageing grass plants, labeled organic nitrogen compounds were found to be translocated from the grass shoots to the soil (Table 7). Net mineralization of soil organic nitrogen was positive in the contents of pots containing killed root systems (Table 3b). About 8 per cent of the labeled nitrogen added to the contents of such pots, in the form of ammonium sulphate, was found to be present in soil organic nitrogen compounds approximately 4 weeks after application, while a total of about twice this amount of soil organic nitrogen was mineralized during that period. From the results obtained in this investigation, it is concluded that the constant presence of living plants is responsible for the accumulation of nitrogen in organic compounds in permanent grassland. No evidence was obtained that the decomposition of such compounds in the soil is inhibited by living plants.     

Selection for Nitrogen Responses in Lolium

Lolium populations from a diallel cross were tested for heritabilityof response to nitrogen in sand culture, in pots of soil andfinally in the field. Heritability of response was found insand culture for several growth parameters including leaf areaand shoot dry weight. Response in pots of soil varied significantlybetween populations, but in a small experiment heritabilitywas not shown to be significant. Selection of individual genotypes within the diallel populationswas then carried out simultaneously for nitrate uptake fromsolutions and for growth in pots. The selected plants were finallytested for nitrogen response in simulated swards in the field.L. perenne plants did not differ after selection for high orlow uptake, but L. multiflorum and interspecific hybrids producedlines which showed significant differences in nitrogen response. A second cycle of selection confirmed the lack of variationin the L. perenne lines chosen, and revealed a complex situationin L. multiflorum whereby the effects of selection were apparentlyreversed. In the L. perenne x L. multiflorum hybrid lines, however,the large differences between high and low response selectionswere maintained. High response plants had broad pale leaveswhile low response plants had rather narrow dark leaves.     

Some effects of combined nitrogen on the nodule symbioses of Casuarina and Ceanothus

Summary 1. The effect of ammonium-nitrogen on the further growth and activity in fixation of nodules already present at the commencement has been studied inCasuarina cunninghamiana growing in water culture and inCeanothus velutinus var.laevigatus in Peralite culture. 2. In Casuarina, at a low level of ammonium-nitrogen nodule growth remained similar to that in plants in nitrogen-free solution, but was stimulated in Ceanothus. In both genera nodule growth was strongly retarded at higher levels. 3. Fixation of nitrogen fell continuously in Casuarina as the level of ammonium-nitrogen was increased, while that in Ceanothus was unaffected at a low level but markedly decreased at a higher one. These effects were compounded from the changes noted in 2 and a tendency for the efficiency of nodule tissues in fixation to fall in the presence of ammonium-nitrogen, though this was not always shown. 4. In both genera but especially in Casuarina the growth of plants entirely dependent on nodule nitrogen was inferior to that of plants additionally supplied with ammonium-nitrogen. The reasons are discussed. 5. The results indicate the effects likely to be produced in the field as the soil nitrogen level rises through the action of the nodules of these species.     

Genotypic variability among peanut (Arachis hypogea L.) in sensitivity of nitrogen fixation to soil drying

Peanuts (Arachis hypogea L.) are often grown on sandy soils and drought stress can be a major limitation on yield. In particular, loss in nitrogen fixation activity associated with soil drying might be limiting due to the need for high nitrogen amounts in both vegetative tissues and seeds of peanut. This study examined the response of nitrogen fixation of intact plants of seventeen peanut genotypes when subjected to soil drying in pots over approximately a 2-wk period. A large range in the sensitivity of nitrogen fixation to soil drying was observed among the seventeen genotypes. Genotype ICGV86015, in particular, was found to have nitrogen fixation that was especially tolerant of soil drying. Significant positive (P?<?0.0001) correlation was found between the soil water content at which nitrogen fixation began decreasing and the amino acid concentration in the leaves of severely stressed plants.     

Prospects for fungicidal control of take-all of wheat

A range of fungicides and herbicides was tested against Gaeumannomyces graminis (causal agent of take-all) on agar plates, and on wheat seedlings in pots and in liquid culture. Benomyl, the standard in all tests, was consistently most effective: like iprodione, nuarimol and KWG 0599 , it diminished infection from inoculum placed just below the seeds more effectively when applied as a drench than as a soil-mix. Benomyl as a soil-mix was most effective in soils with least organic matter. Some compounds toxic to the pathogen on agar plates and in plants grown in liquid culture were ineffective as soil treatments. The practical limitations of soil treatment with conventional fungicides and application methods are discussed.     

Comparison of common bean (Phaseolus vulgaris L.) genotypes for nitrogen fixation tolerance to soil drying

Aims

Common bean is a major source of protein for many people worldwide. However, the crop is often subjected to drought conditions and its advantage in undertaking symbiotic nitrogen fixation can be severely decreased. The primary objective of this study was to compare the resistance of nitrogen fixation of 12 selected genotypes to soil drying.

Methods

Twelve common bean genotypes of diverse genetic background were compared. Plants were grown in pots and subjected to soil drying over about 2 weeks. Nitrogen fixation was measured daily using a flow-through acetylene reduction technique. The plants were exposed to acetylene for only a short time period allowing repeated measures. The acetylene reduction rate of plants on drying soil was normalized against the rates measured for well-watered plants.

Results

Substantial variability was identified among genotypes in the threshold soil water content at which nitrogen fixation was observed to decrease. Genotypes SER 16, SXB 412, NCB 226, and Calima were found to have the greatest delay in their decrease in nitrogen fixation rates based on soil water content. These four genotypes expressed substantial tolerance of nitrogen fixation to soil drying. These experiments also resulted in data on the threshold soil water contents at which transpiration rates decreased. A decrease in transpiration rates at high soil water contents is potentially advantageous since it allows soil water conservation for use as the severity of the drought increases. There was a general trend of those genotypes with sustained nitrogen fixation rates to low soil water contents also expressing decreased transpiration rates at high soil water contents.

Conclusions

This study identified genetic variation among common bean genotypes in their response of nitrogen fixation and transpiration to soil drying. Five genotypes (SER 16, SXB 412, NCB 226, Calima, and SEA 5) expressed the desired traits for water-limited conditions, which might be exploited in breeding efforts.     

Soil moisture and potassium affect the performance of symbiotic nitrogen fixation in faba bean and common bean

Potassium (K) is reported to improve plant's resistance against environmental stress. A frequently experienced stress for plants in the tropics is water shortage. It is not known if sufficient K supply would help plants to partially overcome the effects of water stress, especially that of symbiotic nitrogen fixation which is often rather low in the tropics when compared to that of temperate regions. Thus, the impact of three levels of fertilizer potassium (0.1, 0.8 and 3.0 mM K) on symbiotic nitrogen fixation was evaluated with two legumes under high (field capacity to 25% depletion) and low (less than 50% of field capacity) water regimes. Plants were grown in single pots in silica sand under controlled conditions with 1.5 mM N (¹⁵N enriched NH₄NO₃). The species were faba bean (Vicia faba L.), a temperate, amide producing legume and common bean (Phaseolus vulgaris L.), a tropical, ureide producing species. In both species, 0.1 mM K was insufficient for nodulation at both moisture regimes, although plant growth was observed. The supply of 0.8 or 3.0 mM K allowed nodulation and subsequent nitrogen fixation which appeared to be adequate for respective plant growth. High potassium supply had a positive effect on nitrogen fixation, on shoot and root growth and on water potential in both water regimes. Where nodulation occurred, variations caused by either K or water supply had no consequences on the percentage of nitrogen derived from the symbiosis. The present data indicate that K can apparently alleviate water shortage to a certain extent. Moreover it is shown that the symbiotic system in both faba bean and common bean is less tolerant to limiting K supply than plants themselves. However, as long as nodulation occurs, N assimilation from the symbiotic source is not selectively affected by K as opposed to N assimilation from fertilizer.     

Nodulation and growth response ofAllocasuarina andCasuarina species to phosphorus fertilization

To examine how soil phosphorus status affects nitrogen fixation by the Casuarinaceae —Frankia symbiosis,Casuarina equisetifolia and two species ofAllocasuarina (A. torulosa andA. littoralis) inoculated or fertilized with KNO₃ were grown in pots in an acid soil at 4 soil phosphate levels. InoculatedC. equisetifolia nodulated well by 12 weeks after planting and the numbers and weight of nodules increased markedly with phosphorus addition. Growth ofC. equisetifolia dependent on symbiotically fixed nitrogen was more sensitive to low levels of phosphorus (30 mg kg^–1 soil) than was growth of seedings supplied with combined nitrogen; at higher levels of phosphorus, the growth response curves were similar for both nitrogen fertilized and inoculated plants. The interaction between phosphorus and nitrogen treatments (inoculated and nitrogen fertilized) demonstrated that there was a greater requirement of phosphorus for symbiotic nitrogen fixation than for plant growth when soil phosphorus was low.WithAllocasuarina species, large plant to plant variation in nodulation occurred both within pots and between replicates. This result suggests genetic variation in nodulation withinAllocasuarina species. Nodulation ofAllocasuarina species did not start until 16 weeks after planting and no growth response due toFrankia inoculation was obtained at the time of harvest. Addition of nitrogen starter is suggested to boost plant growth before the establishment of the symbiosis. Growth ofAllocasuarina species fertilized with nitrogen responded to increasing levels of phosphorus up to 90 mg P/kg soil after which it declined by 69% forA. littoralis. The decrease in shoot weight ofA. littoralis, A. torulosa, C. equisetifolia andC. cunninghamiana at high phosphorus was confirmed in a sand culture experiment, and may be atributable to phosphorus toxicity.     

Ara-Rhizotron: An Effective Culture System to Study Simultaneously Root and Shoot Development of Arabidopsis

Studying Arabidopsis thaliana (L.) Heynh. root development in situ at the whole plant level without affecting shoot development has always been a challenge. Such studies are usually carried out on individual plants, neglecting competition of a plant population, using hydroponic systems or Agar-filled Petri dishes. Those both systems, however, present some limitations, such as difficulty to study precisely root morphogenesis or time-limited culture period, respectively. In this paper, we present a method of Arabidopsis thaliana (L.) Heynh. cultivation in soil medium, named “Ara-rhizotron”. It allows the non-destructive study of shoot and root development simultaneously during the entire period of vegetative growth. In this system, roots are grown in 2D conditions, comparable to other soil cultures. Moreover, grouping several Ara-rhizotrons in a box enables the establishment of 3D shoot competition as for plants grown in a population. In comparison to a control culture grown in pots in the same environmental conditions, the Ara-rhizotron resulted in comparable shoot development in terms of dry mass, leaf area, number of leaves and nitrogen content. We used this new culture system to study the effect of irrigation modalities on plant development. We found that irrigation frequency only affected root partitioning in the soil and shoot nitrogen content, but not shoot or root growth. These effects appeared at the end of the vegetative growth period. This experiment highlights the opportunity offered by the Ara-rhizotron to point out tardy effects, affecting simultaneously shoot development and root architecture of plants grown in a population. We discuss its advantages in relation to root development and physiology, as well as its possible applications.     

Use of plant growth-promoting rhizobacteria to control stress responses of plant roots

Ethylene is a key gaseous hormone that controls various physiological processes in plants including growth, senescence, fruit ripening, and responses to abiotic and biotic stresses. In spite of some of these positive effects, the gas usually inhibits plant growth. While chemical fertilizers help plants grow better by providing soil-limited nutrients such as nitrogen and phosphate, over-usage often results in growth inhibition by soil contamination and subsequent stress responses in plants. Therefore, controlling ethylene production in plants becomes one of the attractive challenges to increase crop yields. Some soil bacteria among plant growth-promoting rhizobacteria (PGPRs) can stimulate plant growth even under stressful conditions by reducing ethylene levels in plants, hence the term “stress controllers” for these bacteria. Thus, manipulation of relevant genes or gene products might not only help clear polluted soil of contaminants but contribute to elevating the crop productivity. In this article, the beneficial soil bacteria and the mechanisms of reduced ethylene production in plants by stress controllers are discussed.     

Effects of arbuscular mycorrhizal infection on the growth and reproduction of the annual legumeKummerowia striata growing in a nutrient-poor alluvial soil

A culture experiment was conducted to examine the effects of arbuscular mycorrhizal (AM) fungi on the growth and reproduction ofKummerowia striata, a common annual legume of river floodplains of Japan. The plants were grown from seeds in pots with nutrient-poor sandy soil collected from a fluvial bar. Arbuscular mycorrhizal infection increased the aboveground biomass, nodule weight, leaf nitrogen concentration and seed production. However, flowering occurred earlier in plants without AM fungi. These effects of AM fungi were insignificant in plants supplied with phosphate. These results suggest that AM fungi may influence the establishment ofK. striata in nutrient-poor, disturbed habitats.     

Use of three screening techniques for the evaluation of fungicides to control rhizoctonia root rot of wheat

Twenty fungicides were screened for their abilities to reduce the saprophytic growth of Rhizoctonia solani on agar and in soil, and to control root rot of wheat (Triticum aestivum) in pots of soil. Thirteen showed some activity against the in vitro growth of R. solani, the most promising being Sandoz 619F and flutolanil, although none of these reduced saprophytic growth of the fungus in soil. Only benomyl and thiabendazole, which were ineffective on agar and against disease in pots, reduced saprophytic growth in soil. None of the fungicides reduced the number of plants infected in the pot experiments and only six (flutriafol, diniconazole, Schering 539865, propiconazole, Bayer HWG1608 and flutolanil) of the thirteen active on agar (+ triadimefon which was not) reduced root rot severity. The results of this study indicate the differing reactions towards R. solani of chemicals in the three screening tests and confirm the potential for chemical control of rhizoctonia root rot of wheat.     

Influence of nitrogen on the production of hypericins by St. John’s wort

The effect of nitrogen supply on the production of ‘hypericins’ (hypericin and pseudohypericin) in leaves of St. John’s wort (Hypericum perforatum L.) was examined with plants grown in sand culture and soil. In sand culture, 56-d growth of St. John’s wort plants with decreased nitrogen levels resulted in increased production of hypericins in leaves. A short-term low nitrogen stress in sand culture also resulted in increased production of leaf hypericins. While growth in a low nitrogen-containing soil resulted in elevated levels of hypericins, their production was decreased by supplementation of the soil with additional nitrogen. Increased production of hypericins in St. John’s wort leaves did not require the nitrogen supply to be decreased to levels that resulted in nitrogen deficiency symptoms. Moreover, alteration in the production of leaf hypericins occurring with changes in nitrogen supply did not alter the concentration ratio of pseudohypericin and hypericin. Increased production of leaf hypericins was not associated with any significant changes in the number of dark glands on the leaves and only a weak correlation was observed between leaf dark gland number and levels of leaf hypericins. These results are discussed in terms of the biochemistry of naphthodianthrone production by St. John’s wort plants and implications for growth environment effects during cultivated growth of this medicinal plant.     

Some effects of the flowerpot on plant growth

Summary This investigation has thus shown that while pots from various sources differ but little in appearance, there are considerable differences in the permeability of the walls, due fundamentally to variations in the pore size. The characteristic features of porous pots in that there is a continuous current of soil solution outwards from the soil through the pot wall, the rate of this current increasing with permeability. Air to replace this water enters the soil at its upper surface, thus, the extent to which the soil is aerated is indirectly determined by the permeability of the pot. This increased aeration has been shown to lead to an increased rate of nitrification in the soil, together with an increase in the available potash, resulting in greater plant yields. Good aeration of the soil also had a beneficial effect on root development.Thus, in general, high permeability is advantageous in pots, unless it is so great as to lead to such rapid drying of the soil that the plant suffers drought effects. Such detrimental effects are particularly likely to occur under extreme conditions of high temperature and low humidity.Another important point, emphasized in this study is the large demand of the pot plant for nitrogen in comparison with the small amount of soil explorable by its roots. Addition of nitrogen is therefore desirable in any case, while for impermeable pots in particular, it is advisable that such nitrogen should be directly assimilable rather than in an organic form.     

Nitrogen fixation throughout growth,and varietal differences in nitrogen fixation by the rhizosphere of rice planted in pots

Summary The nitrogen-fixing activity in the rhizospheres of various rices was measured by the acetylene-reduction method throughout plant growth in green-house pots. The activity began to increase 4 weeks after transplanting, increased until heading stage, then decreased. The concentrations of exchangeable ammonium and sugars in the soil were not related to the variation of nitrogen-fixing activity during rice growth.The nitrogen-fixing activities in the rhizospheres of 41 rice varieties in pots were measured to discover varietal differences. Levels of nitrogen fixation were highly correlated with the rices' dry root weight at heading stage.     

Effects of some systemic imidazole and triazole fungicides on white clover and symbiotic nitrogen fixation by Rhizobium trifolii

Effects of prochloraz, imazalil, diclobutrazol, triadimefon and propiconazole were studied. Compared with most systemic fungicides previously tested they were all relatively toxic to R. trifolii on agar. Effects of soil residues of the compounds on the growth and nitrogen fixing capacity of white clover were measured. The lowest concentration of the fungicides used was that which would be present if all the active ingredient from a single application remained unchanged and evenly distributed in the top 5 cm of soil. This concentration of diclobutrazol decreased the size and weight of 11-wk-old clover plants. The rate of nitrogen fixation was reduced by twice this concentration and root nodulation by four times this amount. None of the other fungicides had any significant effect. The possibility of field application of diclobutrazol to cereals causing damage to clover crops is discussed. It is questionable whether harmful soil residues will ever accumulate from normal usage of the fungicide but exceeding the recommended rate of application, or application to cereals undersown with clover, might cause undesirable effects on the clover.     

Growth and uptake of nitrogen by wheat and ryegrass in fumigated and irradiated soil

Summary Wheat and ryegrass were grown in pots containing soil that had either been irradiated, fumigated with methyl bromide, fumigated with formaldehyde, or left untreated. All pots received a basal dressing of potassium, phosphorus and magnesium; response to nitrogen was tested by applying either 0, 0.177 or 0.354 g nitrogen per pot. Irradiation increased the growth of wheat and ryegrass; uptake of nitrogen was also increased in both crops. The amount of fertilizer nitrogen equivalent to the nitrogen supplied by seeds and soil (the “N value”) can be calculated from the efficiency of uptake of fertilizer nitrogen and used to allow for the effect on crop growth of the nitrogen released by irradiated soil. With wheat the increase in growth can be attributed solely to the extra mineral nitrogen released by irradiated soil. However, ryegrass grew a little better than would have been expected if the only effect of irradiation was to increase the release of soil nitrogen. Fumigation with methyl bromide or formaldehyde increased the growth of wheat and ryegrass not given fertilizer nitrogen. However, fumigation with methyl bromide left ionic bromide in the soil, and this depressed the growth of wheat receiving fertilizer nitrogen. Formaldehyde also left residues; these influenced soil metabolism and sometimes depressed the growth of plants given fertilizer nitrogen.