Influence of adenine,isopentyl alcohol and Azotobacter chroococcum on the vegetative growth of Zea mays

Glasshouse experiments were conducted to evaluate the influence of pretested cytokinin precursors, adenine (ADE) and isopentyl alcohol (IA), and a cytokinin producing bacterium, Azotobacter chroococcum added to soil, on the vegetative growth of maize (Zea mays L.). The combination of 2.0 mg ADE kg^-1 soil, 13 mg IA kg^-1 soil, plus an inoculum of A. chroococcum was the most effective in enhancing the vegetative growth of maize compared with the application of ADE plus IA, ADE plus A. chroococcum, or ADE, IA or A. chroococcum alone. The dry weight of root and shoot tissues was increased up to 5.57- and 5.01-fold, respectively, in comparison to the controls; however, the root/shoot ratios were similar. The increases in shoot height, internodal distance, stem and leaf width over the controls under the optimum treatment were: 2.07-, 2.81-, 1.46-, and 2.11-fold, respectively. The improvement in plant yield was primarily attributed to A. chroococcum production of cytokinins in the rhizosphere.     

Influence of the fungicide pentachloronitrobenzene on VA-mycorrhizal and total root length and phosphorus uptake of oats (Avena sativa)

The effect of application of the fungicide pentachloronitrobenzene (PCNB) at levels between 2 and 50 mg kg^–1 soil on root growth, mycorrhizal infection and P uptake was studied in pot culture with oats (Avena sativa cv. Alfred) growing in a rendzina soil low in available P. The soil had been partially sterilized by X-ray, and half of the pots were inoculated with spores of the VAM-fungusGlomus mosseae (indigenous species).Soil irradiation (0.5 Mrad) did not decrease the levels of infection by VAM. Application of PCNB decreased the VAM-infected root length, at 50 mg PCNB kg^–1 soil VAM-infected root length was about 12% of the controls. Total root length, however, increased to about 126% of control values at PCNB rates up to 20 mg kg^–1 soil, but decreased to 89% of the controls at 50 mg kg^–1 soil. Total P-uptake decreased with increasing levels of PCNB and was linearly correlated with infected root length (r=0.92).The stimulation of root growth by PCNB at rates up to 20 mg kg^–1 soil is regarded as an indirect effect, brought about by suboptimal P-supply due to inhibition of VA-mycorrhiza. Conversely, the reduction of total root length at 50 mg PCNB kg^–1 soil is most likely a direct effect. Due to the phytotoxicity of the fungicide, the contribution of the indigenous VA-mycorrhiza to plant P uptake under field conditions cannot be determined by soil application of PCNB at rates sufficient for complete inhibition of VAM.As inhibition or absence of VAM may lead to compensatory root growth, mycorrhizal dependency ought to be calculated from the amounts of P taken up per unit root length in mycorrhizal and nonmycorrhizal plants, respectively.     

Effect of salinity on mycorrhizal onion and tomato in soil with and without additional phosphate

Summary Vesicular-arbuscular mycorrhizal fungi (VAM) are known to increase plant growth in saline soils. Previous studies, however, have not distinguished whether this growth response is due to enhanced P uptake or a direct mechanism of increased plant salt tolerance by VAM. In a glasshouse experiment onions (Allium cepa L.) were grown in sterilized, low-P sandy loam soil amended with 0, 0.8, 1.6 mmol P kg^–1 soil with and without mycorrhizal inoculum. Pots were irrigated with saline waters having conductivities of 1.0, 2.8, 4.3, and 5.9 dS m^–1. Onion colonized withGlomus deserticola (Trappe, Bloss, and Menge) increased growth from 394% to 100% over non-inoculated control plants when soil P was low ( 0.2 mmol kg^–1 NaHCO₃-extractable P) at soil saturation extract salinities from 1.1 dS m^–1 to 8.8 dS m^–1. When 0.8 and 1.6 mM P was added no dry weight differences due to VAM were observed, however, K and P concentrations were higher in VAM plants in saline treatments.Glomus fasciculatum (Gerdeman and Trappe) andGlomus mosseae (Nicol. and Gerd.) isolates increased growth of VAM tomato 44% to 193% in non-sterilized, saline soil (10 dS m^–1 saturation extract) despite having little effect on growth in less saline conditions when soil P was low. Higher tomato water potentials, along with improved K nutrition by VAM in onion, indicate mechanisms other than increased P nutrition may be important for VAM plants growing under saline stress. These effects appear to be secondary to the effects of VAM on P uptake.     

Influence of L-tryptophan and auxins applied to the rhizosphere on the vegetative growth of Zea mays L.

Glasshouse experiments were conducted to evaluate the influence of L-TRP in comparison with indole-3-acetamide (IAM), tryptophol (TOL) and indole-3-acetic acid (IAA) on the growth of Zea mays L. var. Early Sunglow. L-TRP (25 to 2.5×10^–5 mg kg^–1 soil), IAM (22 to 2.2×10^–5 mg kg^–1 soil), TOL (20 to 2.0×10^–5 mg kg^–1 soil), and IAA (22 to 2.2×10^–5 mg kg^–1 soil) were applied as a soil drench to established uniform seedlings. All treatments were applied in a completely randomized design with 10 replicates. IAM had no significant effect on the plant growth parameters. Shoot height, uppermost leaf collar base distance, internodal distance, and shoot dry and fresh weights were significantly improved upon the addition of TOL (2.0×10^–2 mg kg^–1 soil), however, the highest concentration (20 mg kg^–1 soil) caused a 14.6% reduction in leaf width. L-TRP (2.5×10^–3 mg kg^-1 soil) also had a significant influence on shoot height, uppermost leaf collar base distance, internodal distance and fresh weight of shoot compared with the control. The highest concentration of L-TRP (25=mg kg^–1 soil) had a negative effect on leaf width and dry weight of the shoot. The most pronounced response on the corn growth parameters was observed with the application of IAA at lower concentrations (2.2×10^–5 to 2.2×10^–2 mg kg^–1 soil) specifically improving root growth. The highest concentration (22 mg kg^–1 soil) of IAA had a significant negative effect on plant height, leaf width, stem diameter, shoot fresh and dry weight. These findings indicate that L-TRP applied at the appropriate concentrations can have positive effects on corn growth comparable to pure auxins (TOL and IAA).     

Suppression of the root rot–root knot disease complex by Pseudomonas aeruginosa in tomato: The influence of inoculum density,nematode populations,moisture and other plant-associated bacteria

The influence of different application rates of the plant growth-promoting rhizobacterium, Pseudomonas aeruginosa, population densities of the root-knot nematode, Meloidogyne javanica, moisture and other plant-associated bacteria in the suppression of root rot–root knot disease complex of tomato are described. The impact of these factors on bacterial rhizosphere and inner root and shoot establishment are also presented. The highest inoculum level of P. aeruginosa (7.4 × 10⁸ cfu ml^–1) in the presence of the lowest population density of M. javanica (500 J2/plant) caused the greatest reduction in gall formation due to M. javanica. The number of root–knot nematodes recovered from soil and roots treated with P. aeruginosa were also significantly reduced. Root infection caused by the soilborne root-infecting fungi Fusarium oxysporum, F. solani and Rhizoctonia solani was also effectively suppressed following application of P. aeruginosa. A P. aeruginosa-Bacillus subtilis treatment was the most effective in the suppression of root-rot disease complex with enhancement of plant growth. Biocontrol and growth promoting potential of the bacterium was enhanced when soil was kept at 50% or 75% moisture holding capacity, whereas a 25% MHC reduced bacterial efficacy. Rhizosphere population of P. aeruginosa declined drastically in P. aeruginosa-Bradyrhizobium japonicum treatments. Rhizosphere colonisation by P. aeruginosa seems to be governed by two factors: Initial inoculum size of the bacterium and severity of the root-knot disease. Endoroot and endoshoot colonisation of the bacterium was dependent on degree of root-colonisation by Fusarium oxysporum. An inoculum level 2.5 × 10⁸ cfu/ml of P. aeruginosa was optimal for the enhancement of plant growth, whereas inoculum below this level reduced plant growth.     

Effects of soil cadmium on Pinus sylvestris L. seedlings

The effects of Cd on the growth and distribution of Cd and mineral nutrients within plant tissues were investigated for Pinus sylvestris L. seedlings grown in mineral forest soil with increasing levels of Cd addition (0–100 mg kg^–1). Approximately 20% of added Cd was found to be extractable from sandy loam forest soil. Root growth was less affected by Cd than shoot growth, which showed a significant reduction in the 100 mg Cd kg^–1 treatment. Cadmium accumulated in roots up to 325 mg kg^–1. Decreased concentrations of K in needles and Ca in stems with increasing Cd levels suggest a disturbance of mineral nutrition as a result of Cd addition.     

Minimizing the effect of mineral nitrogen on biological nitrogen fixation in common bean by increasing nutrient levels

Although common bean (Phaseolus vulgaris L.) has good potential for N₂ fixation, some additional N provided through fertilizer usually is required for a maximum yield. In this study the suppressive effect of N on nodulation and N₂ fixation was evaluated in an unfertile soil under greenhouse conditions with different levels of soil fertility (low=no P, K and S additions; medium = 50, 63 and 10 mg kg^–1 soil and high = 200, 256 and 40 mg kg^–1 soil, respectively) and combined with 5, 15, 60 and 120 mg N kg^–1 soil of ¹⁵N-labelled urea. The overall average nodule number and weight increased under high fertility levels. At low N applications, nitrogen had a synergistic effect on N₂ fixation, by stimulating nodule formation, nitrogenase activity and plant growth. At high fertility and at the highest N rate (120 mg kg^–1 soil), the stimulatory effect of N fertilizer on N₂ fixation was still observed, increasing the amounts of N₂ fixed from 88 up to 375 mg N plant^–1. These results indicate that a suitable balance of soil nutrients is essential to obtain high N₂ fixation rates and yield in common beans.     

Varying phosphorus supply and development,growth and seed yield in narrow-leafed lupin

Narrow-leafed lupin (Lupinus angustifolius L.) is usually grown in sandy, acidic and phosphorus (P) deficient soil with low yield and variable harvest index. This study aimed to examine the effects of varying P supply on lupin growth, seed yield and harvest index. Non-abscission plants (cv. Danja) were grown in Lancelin sand at seven rates of P supply (5, 10, 15, 20, 25, 30 or 40 mg kg^–1) in a naturally-lit glasshouse. The rate of leaf emergence, flowering time and flower number were decreased or delayed by low P supply (5, 10 or 15 mg kg^–1), with no differences at P rates higher than 20 mg kg^–1. High P supply (25, 30 or 40 mg kg^–1) increased plant seed yield and harvest index largely by increasing the number of pods and consequently yield on the lateral branches, but had less effect on the number of seeds per pod and seed size. Seed yield and seed P concentration continued to increase up to 40 mg P kg^–1but harvest index plateaued at 25 mg P kg^–1, indicating that low P supply decreased reproductive growth more than vegetative growth in narrow-leafed lupin.     

Biotechnological aspects of the production of the anticancer drug podophyllotoxin

The natural lignan podophyllotoxin, a dimerized product of two phenylpropanoid moieties which occurs in a few plant species, is a pharmacologically important compound for its anticancer activities. It is used as a precursor for the chemical synthesis of the anticancer drugs etoposide, teniposide and etopophose. The availability of this lignan is becoming increasingly limited because of the scarce occurrence of its natural sources and also because synthetic approaches for its production are still commercially unacceptable. Biotechnological production using cell culture may be considered as an alternative source. Selection of the best performing cell line, its maintenance and stabilization are necessary prerequisites for its production in bioreactors and subsequent scale-up of the cultivation process to the industrial level. Scale-up of growth and product yield depends on a multitude of factors, such as growth medium, physicochemical conditions, seed inoculum, type of reactor and processing conditions. The composition of the growth medium, elicitors and precursors, etc. can markedly influence the production. Optimum levels of parameters that facilitate high growth and product response in cell suspensions of Podophyllum hexandrum have already been determined by statistical design. P. hexandrum cells have successfully been cultivated in a 3-l stirred-tank bioreactor under low shear conditions in batch and fed-batch modes of operation. The batch kinetic data were used to identify the mathematical model which was then used to develop nutrient-feeding strategies for fed-batch cultivation to prolong the productive log phase of cultivation. An improvement in the production of podophyllotoxin to 48.8 mg l^–1 in a cell culture of P. hexandrum was achieved, with a corresponding volumetric productivity of 0.80 mg l^–1 day^–1, when the reactor was operated in continuous cell-retention mode. Efforts are being made to further enhance its production levels by the development of hairy root culture or by varying the channeling of precursors towards the desired biosynthetic pathway by molecular approaches.     

Removal characteristics of high load ammonia gas by a biofilter seeded with a marine bacterium, Vibrio alginolyticus

When the cells of the newly isolated marine bacterium, Vibrio alginolyticus, were inoculated on to an inorganic packing material in biofilter, and a load of ammonia of 2.4–22.5 g-N kg^–1dry packing material was introduced continuously under non-sterile conditions, the average amount of NH₃removed exceeded 85% over 61-d operation. The maximum removal capacity and the complete removal capacity were 22.8 g-N kg^–1dry packing material dand 18.6 g-N kg^–1dry packing material d, respectively, which were about four times larger than those obtained in autotrophic nitrifying sludge inoculated on the same packing material.     

Dynamics of phosphorus fractions during fallow with natural vegetation and planted Pueraria phaseoloides in south-western Nigeria

A study was conducted in 1998 and 1999 on a long-term fallow management trial, established in 1989 at the International Institute of Tropical Agriculture (IITA), Ibadan, in the derived savanna of southwestern Nigeria, to quantify P fractions under natural fallow (NF) and a Pueraria cover crop fallow. Plots with previous 1:1, 1:2, and 1:3 crop/fallow ratios before reverting to fallow in either 1998 or 1999 were considered. Biomass accumulation under the two fallow types was comparable and increased linearly with fallow age, reaching slightly above 7 t dry matter ha^–1 after 8 months of fallow. Phosphorus accumulation in the fallow vegetation ranged from 2.1 – 9.1 kg ha^–1 for natural fallow (NF) and from 1.5 – 6.6 kg ha^–1 for Pueraria. Magnesium was also higher under NF (9.1 – 21 kg ha^–1) than under Pueraria (4.4 – 13 kg ha^–1), whereas N, Ca, and K contents were higher in Pueraria biomass than under NF at 1 year after fallow. Pueraria fallow tended to lower soil pH compared with NF. However, plots with less frequent cropping (1:3 crop/fallow ratio) did not have significantly different pH irrespective of the fallow vegetation type. Olsen extractable soil P increased as fallow length increased irrespective of the fallow system and previous crop/fallow ratio. For example, under NF (0–5 cm depth, 1:1 crop/fallow ratio in1998) Olsen P increased from 12 mg kg^–1 to 17 mg kg^–1 after 1 year of fallow and under Pueraria, it increased from 8 mg kg^–1 to 15 mg kg^–1. Fallow type and previous crop/fallow ratio had no significant and consistent effects on soil P fractions. However, NaOH- and concentrated HCl- extractable organic P fractions increased with fallow length. In 1998, under NF, NaOH- extractable organic P increased from 12 to 21 mg kg^–1 (1:1 crop/fallow ratio) and from 10 to 19 mg kg^–1 for both 1:2 and 1:3 crop/fallow ratio. HCl- extractable organic P increased from 11 to 30 mg kg^–1 (1:1 crop/fallow ratio), from 13 to 27 mg kg^–1 (1:2 crop/fallow ratio) and from 18 to 35 mg kg^–1 (1:3 crop/fallow ratio). Similar trend was observed under Pueraria fallow. These results suggest that P was reallocated to non-readily available organic P fractions irrespective of fallow type and previous land use. These organic P fractions, which are usually more stable, reflect the overall change in soil organic P levels when the soil was stressed by cultivation and then reverted to fallow. These pools may thus represent an active reservoir (source and sink) of P in shifting cultivation under tropical conditions without inorganic fertilizer application.     

Uptake of cadmium from an experimentally contaminated calcareous soil by arbuscular mycorrhizal maize (<Emphasis Type="Italic"> Zea mays </Emphasis>L.)

We investigated uptake of Cd by arbuscular mycorrhizal (AM) maize inoculated with Glomus mosseae from a low-P sandy calcareous soil in two glasshouse experiments. Plants grew in pots containing two compartments, one for root and hyphal growth and one for hyphal development only. Three levels of Cd (0, 25 and 100 mg kg^–1) and two of P (20 and 60 mg kg^–1) were applied separately to the two compartments to assess hyphal uptake of Cd. Neither Cd nor P addition inhibited root colonization by the AM fungus, but Cd depressed plant biomass. Mycorrhizal colonization, P addition and increasing added Cd level led to lower Cd partitioning to the shoots. Plant P uptake was enhanced by mycorrhizal colonization at all Cd levels studied. When Cd was added to the plant compartment and P to the hyphal compartment, plant biomass increased with AM colonization and the mycorrhizal effect was more pronounced with increasing Cd addition. When P was added to the plant compartment and Cd to the hyphal compartment, plant biomass was little affected by AM colonization, but shoot Cd uptake was increased by colonization at the low Cd addition rate (25 mg kg^–1) and lowered at the higher Cd rate (100 mg kg^–1) but with no difference in root Cd uptake. These effects may have been due to immobilization of Cd by the fungal mycelium or effects of the AM fungus on rhizosphere physicochemical conditions and are discussed in relation to possible phytostabilization of contaminated sites by AM plants.     

Nematicidal and allelopathic potential of Argemone mexicana,a tropical weed

Argemone mexicana L. (Papaveraceae), a tropical annual weed, is phytotoxic to many crop species. This study was designed to examine the allelochemical and nematicidal potential of A. mexicana and to better understand the role of this weed in the ecosystem. A methanol-soluble extract of the leaf material caused greater juvenile mortality of Meloidogyne javanica than did ethyl acetate or hexane extracts indicating the polar nature of the toxins. Decomposing tissues of A. mexicana in soil at 50 g kg^–1 were highly deleterious causing 80% mortality of tomato plants. At 10 g kg^–1 plant growth was enhanced, while at 30 g kg^–1 plant growth was substantially retarded. M. javanica population densities in the rhizosphere and in roots, and gall formation were significantly suppressed when 10, 30 or 50 g kg^–1 A. mexicana was allowed to decompose in the soil. To establish whether decomposition was necessary to produce phytotoxic symptoms, or whether the shoot extract alone could interfere with plant growth, an aqueous shoot extract was applied to soil. Whereas a 50% extract promoted plant growth, a 100% (100 g/500 mL distilled water) concentration significantly reduced plant height, and fresh weights of shoot and root. In general, decomposing plant material caused greater phytotoxicity compared to the aqueous extract. Addition of N as NH₄NO₃ partially alleviated the phytotoxic action of A. mexicana,and also reduced severity of root-knot disease. Adding Pseudomonas aeruginosa to soil amended with A. mexicana resulted in decreased density of M. javanicain the rhizosphere and in tomato roots, suppressed galling rates and enhanced plant growth.     

Achillea millefolium (yarrow) cell suspension cultures: Establishment and growth conditions

Summary Friable calli were obtained fromAchillea millefolium L. hypocotyls, in Gamborg B5 medium, supplemented with 1.5mg.1^–1 2,4-D / 0.1mg.1^–1 Kin, and used for the production of cell suspension cultures in the same liquid medium. The growth pattern of the cultures was determined in permanent light or dark conditions and with different inoculum densities, basal media, growth regulators and sucrose concentrations. Different sources and nitrogen amounts were assayed to study the effect on yarrow cell growth. The conditions found to be optimal for growth of yarrow cell suspension cultures were: 70g (f.w.).1^–1 of initial inoculum in Gamborg B5 medium, supplemented with 1.5mg. 1^–1 2,4-D / 0.1mg.1^–1 Kin, NO₃ ^–/NH₄ ⁺ (30/lmM), and 2% sucrose, in darkness. In these culture conditions the cell suspensions showed a doubling time of 35–40h.Abbreviations 2,4-D dichlorophenoxyacetic acid - NAA naphtalenacetic acid - BA benzyladenine - Kin Kinetin     

Experimental and mathematical simulation of plant growth promoting rhizobacteria and plant interaction under cadmium stress

Bacterial inoculants of the commercially available plant growth promoting rhizobacteria (PGPR) Arthrobacter mysorens 7, Flavobacterium sp. L30, and Klebsiella mobilis CIAM 880 were selected to obtain ecologically safe barley crop production on cadmium (Cd) polluted soils. All the PGPR immobilized 24–68% soluble cadmium from soil suspension. A. mysorens 7 and K. mobilis CIAM 880 were highly resistant to Cd and grew in up to 1 and 3 mmol CdCl₂ on DAS medium respectively. All PGPR were able to fix nitrogen (276–1014 nmol mg^–1 bacterial DW) and to produce indole acetic acid (IAA) (126–330 nmol mg^–1 bacterial DW) or ethylene (4.6–13.5 nmol bacterial DW). All the PGPR actively colonized barley root system and rhizosphere and significantly stimulated root elongation of barley seedlings (up to 25%), growing on soil containing 5 or 15 mg Cd kg^–1 of soil. Created in the simulation mathematical model confirms our hypothesis that PGPR beneficial effect on barley growing under Cd-stress is a complex process. One of mechanisms underlying this effect might be increase of bacterial migration from rhizoplane to rhizosphere, where PGPR bind soluble free Cd ions in biologically unavailable complex forms. Among the studied PGPR K. mobilis CIAM 880 was the most effective inoculant. Inoculation with K. mobilis CIAM 880 of barley plants growing on Cd contaminated soil (5 mg Cd kg^–1 of soil) under field conditions increased by 120% grain yield and 2-fold decreased Cd content in barley grain. The results suggest that the using K. mobilis CIAM 880 is an effective way to increase the plant yield on poor and polluted areas.     

Heavy metal and PCB contamination of Bryozoan colonies in the River Meuse (Belgium)

In the Meuse River (Liège area, Belgium), large amounts of three species of Bryozoans, Fredericella sultana (BLUM.), Plumatella emarginata ALL. and Plumatella fungosa (PALL.) occur. They cover 3 to 40% of the bank walls and their biomass ranges from 12 to 293 g m^–2 (dry weight). In the heated waters of a nuclear power plant (Tihange) and of industries lined up along the river, colony development occurs 3 to 4 weeks earlier than at an upstream station. The heavy metal content of living colonies ranges from 4 to 21 mg kg^–1 Cd, 45 to 182 mg kg^–1 Cu, 803 to 2232 mg kg^–1 Zn, 150 to 483 mg kg^–1 Pb and 21 to 138 mg kg^–1 Cr (DW). The mean concentration of PCBs was 925 µg kg^–1 (DW). Heavy metal and PCB concentrations in the sediments were close to those of colonies, suggesting that most of the pollutants found in the Bryozoans is in fact in the sediment trapped by the colonies.     

The influence of gibberellic acid and sulfur fertilization rate on growth and S-use efficiency of mustard (<Emphasis Type="Italic">Brassica juncea</Emphasis>)

Earlier research has shown that exogenous gibberellic acid (GA₃) application increases shoot growth, photosynthesis and soil nitrogen (N) utilisation in mustard (Brassica juncea L. Czern & Coss.). Mustard has a high sulfur (S) requirement. Its assimilatory pathway is well coordinated with N and dependent on photosynthesis. Thus, the higher photosynthate production and an efficient use of N with the use of GA₃ could result in an increase in S-use efficiency of the crop. The research was, therefore, carried out to study the effects of 10~M GA₃ spray on specific leaf area, plant dry mass, leaf carbon dioxide exchange rate (CER), plant growth rate (PGR), relative growth rate (RGR), net assimilation rate (NAR) and S-use efficiency (SUE) of mustard treated with 0, 100 or 200 mg S kg^–1 soil levels. Plants treated with 100~mg S kg^–1 soil and receiving GA₃ treatment showed increased specific leaf area and dry mass accumulation compared to the control. At 0~mg S kg^–1 soil, N and S concentrations were reduced. They increased with increasing S supply. GA₃ application significantly increased N and S concentrations further. A two-fold increase in SUE in GA₃-treated plants at 100~mg S kg^–1 soil was noted in comparison to the control. SUE was not increased under excess S conditions beyond 100~mg S kg^–1 soil. The increase in SUE was through increase in the growth, CER and use efficiency of N by the crop due to GA₃ application.     

Cell suspension cultures ofOnonis natrix L. subspeciesramosissima: Establishment and culture conditions

Summary Explants from petioles, folioles or hypocotyls ofOnonis natrix have been used for calli initiation. Hypocotyls inoculated on MS medium supplemented with 2% sucrose and 0.5 mg.1^–1 2,4-D / 1 mg.1^–1 Kin showed to be the best primary explant. Cell suspension cultures were established in MS basal medium supplemented with 2% sucrose, 0.5 mg.1^–1 NAA or 2,4-D and 1 mg.1^–1 Kin. Different subculturing periods, inoculum density, hormonal supplementation and sucrose concentration were assayed in order to obtain the best culture growth conditions. The optimal conditions were achieved with cultures initiated with 40 g.1^–1 of initial inoculum, growing in MS basal medium supplemented with 4% sucrose, 0.5 mg.1^–1 NAA and 1 mg.1^–1 Kin subcultured every twelve days. Under these experimental conditions, the cultures showed a doubling time of 36.3 hours.     

Comparison of soil nitrogen mineralization and nitrification in a mixed grassland and forested ecosystem in central Taiwan

We used the buried bag incubation method to study temporal patterns of net N mineralization and net nitrification in soils at Ta-Ta-Chia forest in central Taiwan. The site included a grassland zone, (dominant vegetation consists of Yushania niitakayamensis and Miscanthus transmorrisonensis Hayata) and a forest zone (Tsuga chinensis var. formosana and Yushania niitakamensis). In the grassland, soil concentration NH₄ ⁺ in the organic horizon (0.1–0.2 m) ranged from 1.0 to 12.4 mg N kg^–1 soil and that of NO₃ ^– varied from 0.2 to 2.1 mg N kg^–1 soil. In the forest zone, NH₄ ⁺ concentration was between 2.8 and 25.0 mg N kg^–1 soil and NO₃ ^–varied from 0.2 to 1.3 mg N kg^–1 soil. There were lower soil NH₄ ⁺ concentrations during the summer than other seasons. Net N mineralization was higher during the summer while net nitrification rates did not show a distinct seasonal pattern. In the grassland, net N mineralization and net nitrification rates were between –0.1 and 0.24 and from –0.04 to 0.04 mg N kg^–1 soil day^–1, respectively. In the forest zone, net N mineralization rates were between –0.03 and 0.45 mg N kg^–1 soil day^–1 and net nitrification rates were between –0.01 and 0.03 mg N kg^–1 soil day^–1. These differences likely result from differing vegetation communities (C₃ versus C₄ plant type) and soil characteristics.     

Growth promotion of Prunus rootstocks by root treatment with specific bacterial strains

A collection of bacterial strains obtained from a wide-range origin was screened for ability to promote growth in two types of Prunus rootstocks in a commercial nursery. Only few strains promoted growth significantly and consistently, and a strong specificity for the rootstock cultivar was observed. Irrigation of plants with Pseudomonas fluorescens EPS282 and Pantoea agglomerans EPS427 significantly increased plant height and root weight of the plum Marianna 2624 and the peach–almond hybrid GF-677, respectively. Plant height showed a higher rate of growth in early stages of development (2.6–3.5 times the non-treated controls), but the effect decreased with plant age. However, in aged plants growth promotion was more significant on root weight (1.9 times the non-treated controls) than on plant height. The efficacy of growth promotion and the persistence of strains in the root environment were dependent on the bacterial inoculum concentration applied. Increases in root development were maximum at inoculum concentrations of up to 8 log₁₀ CFU ml^–1 (ca 10 log₁₀ CFU L^–1 of potting mix). Population levels at the optimum inoculum concentration were around 7 log₁₀ CFU g f.w.^–1 root material at early stages of development and decreased to 4 log₁₀ CFU g f.w.^–1 after several months of development. The best plant growth-promoting strains were very diverse in secondary metabolite production and antagonistic ability against several plant pathogens.