Comparison of the effects of poultry manure and its biochar on barley growth in petroleum-contaminated soils

This study was conducted to evaluate and compare the effectiveness of two organic amendments [poultry manure (PM) and poultry manure biochar (PMB)] for the degradation of petroleum hydrocarbons in contaminated soils by barley plant at three levels of total petroleum hydrocarbons (TPHs) during 5 months under greenhouse conditions. TPHs removal efficiency and microbial respiration were shown to be higher at soil-cultivated plant than at uncultivated soil and in lowest level of contamination rather than other levels of contamination and at organic amendment treatment than unamended soil. Soil microbial respiration and TPHs degradation in the rhizosphere of barley increased by 15.64 and 12.74% for PM-amended treatment and 28.07 and 26.83% for PMB-amended treatment, respectively, in the 4% TPHs level compared with unamended treatment. Comparison of two amendments showed that in PMB treatment soil, highest dry weight, microbial respiration, and TPHs degradation potential were observed.     

Enhancing Degradation of Total Petroleum Hydrocarbons and Uptake of Heavy Metals in a Wetland Microcosm Planted with Phragmites Communis by Humic Acids Addition

The effects of humic acid (HA) on heavy-metal uptake by plants and degradation of total petroleum hydrocarbons (TPHs) in a wetland microcosm planted with Phragmites communis were evaluated by comparing waterlogged soils and water-drained upland soils. Experiments were conducted on soils artificially contaminated with heavy metals (Pb, Cu, Cd, Ni) and diesel fuel. HA showed a positive influence on biomass increase for all conditions, but more for belowground than aboveground biomass, and lower in contaminated than uncontaminated soil. The bioavailability and leachability factor (BLF) for all heavy metals except Ni increased with HA addition in both the control and the P. communis planted microcosms, suggesting that more heavy metals could be potentially phytoavailable for plant uptake. Microbial activities were not affected by both heavy metals and TPH contamination, and HA effects on stimulating microbial activities were much greater in the contaminated soil than under uncontaminated conditions. HA addition enhanced the degradation of TPH and n-alkane in waterlogged conditions. The results show that HA can increase the remedial performance in P. communis dominated wetlands simultaneously contaminated with heavy metals and petroleum hydrocarbons and thus prevent contamination of groundwater or other adjacent ecosystems.     

Influence of organic and inorganic soil amendments on plant growth in crude oil-contaminated soil

Phytoremediation can be a viable alternative to traditional, more costly remediation techniques. Three greenhouse studies were conducted to evaluate plant growth with different soil amendments in crude oil-contaminated soil. Growth of alfalfa (Medicago sativa L., cultivar: Riley), bermudagrass (Cynodon dactylon L., cultivar: Common), crabgrass (Digitaria sanguinalis, cultivar: Large), fescue (Lolium arundinaceum Schreb., cultivar: Kentucky 31), and ryegrass (Lolium multiflorum Lam., cultivar: Marshall) was determined in crude oil-contaminated soil amended with either inorganic fertilizer, hardwood sawdust, papermill sludge, broiler litter or unamended (control). In the first study, the addition of broiler litter reduced seed germination for ryegrass, fescue, and alfalfa. In the second study, bermudagrass grown in broiler litter-amended soil produced the most shoot biomass, bermudagrass produced the most root biomass, and crabgrass and bermudagrass produced the most root length. In the third study, soil amended with broiler litter resulted in the greatest reduction in gravimetric total petroleum hydrocarbon (TPH) levels across the six plant treatments following the 14-wk study. Ryegrass produced more root biomass than any other species when grown in inorganic fertilizer- or hardwood sawdust + inorganic fertilizer-amended soil. The studies demonstrated that soil amendments and plant species selection were important considerations for phytoremediation of crude oil-contaminated soil.     

Using Maize (Zea mays L.) and Sewage Sludge to Remediate a Petroleum-Contaminated Calcareous Soil

Phyto-stimulation, the use of plants to stimulate activity of microorganisms in a root zone, has been proposed as an approach to promote the degradation of petroleum hydrocarbons and thus the remediation of petroleum-polluted soils. In this study, we investigated the potential use of sewage sludge to enhance phyto-stimulating effects of maize (Zea mays L.) on the elimination of an aged petroleum contamination in a calcareous soil. In a pot experiment, maize was grown on the experimental soil for two months at three levels of sewage sludge application (0, 20, and 50 g dry matter of sludge per kg soil). The amendments increased root and shoot growth of the experimental plants approximately by a factor of two at the lower sludge treatment level and by a factor of five at the higher sludge treatment level. In a separate incubation experiment, sludge application also led to an immediate stimulation of soil respiration, which then further increased over time. The initial stimulation was three times larger at the higher than at the lower treatment level, but the rate of subsequent increase was similar in both treatments. The two sludge treatments also accelerated TPH elimination in the contaminated soil, and again the effect was approximately three times stronger at the higher than at the lower treatment level. The sludge effect on TPH elimination was much stronger than the effect of the plants. More than half of the initial contamination was reduced in combined treatment with maize and sludge application at the highest rate. The results show that sewage sludge can substantially enhance the remediation of petroleum-contaminated soil, especially when applied in conjunction with a suitable plant such as maize.     

番茄吸收和积累Cd能力的品种间差异

旨在筛选和利用在受污染土壤中可食部位污染物积累水平在食品卫生标准允许范围内的农作物品种(pollution-safecultivar,简称为PSC),以降低水土环境污染物经食物链危害人类健康的风险。研究对象为番茄,探讨了在一定Cd污染土壤中其果实Cd含量低于国际食品法典委员会(CAC)Cd最高限值的品种(Cd-PSC)存在的可能性。通过盆栽试验研究了36个番茄品种(包括普通番茄和樱桃番茄两个变种)不同器官在受不同程度镉污染的土壤中吸收和积累Cd能力的差异。结果表明:(1)在土壤Cd重度胁迫(13.3mg.kg-1)和轻度Cd胁迫(1.1mg.kg-1)下,番茄所有器官Cd含量的变异在统计学上均存在极显著意义(p<0.01),其中果实Cd含量的范围分别为0.08~0.33mg.kg-1和0.00~0.09mg.kg-1,变异系数分别达到28.5%和77.9%;(2)在重度胁迫下,所有供试品种的果实Cd含量均超出CAC标准,而轻度胁迫下的Cd含量超标率达到19.4%,说明番茄是易受Cd污染的农作物种类,其中樱桃番茄的受污染风险特别高,在轻度胁迫下6个供试品种中有5个超标,而30个普通番茄品种仅1个超标。因此,仅在轻度胁迫条件下,存在着番茄的Cd-PSC,其中有5个普通番茄品种的果实中未检出Cd,属于比较安全的PSC,包括品种No.121、6、24、35和No.36,可以推荐在土壤Cd污染程度略超国家三级标准或具有潜在受污染风险的区域应用;(3)尽管大多数情况下各营养器官的Cd含量间均呈显著的正相关,但营养器官与果实Cd含量间均无明显的相关性,推测Cd在番茄营养器官间和营养器官与果实间的转运途径是不一样的。组织中Cd含量的显著相关性说明该特性可以作为农作物品种的种性特征;(4)与轻度胁迫相比,重度胁迫下的所有供试品种平均果实生物量上升了7.5%,一半以上的品种表现出较强的耐Cd能力,这一特性可能导致因不易察觉土壤的Cd污染状况而增加番茄产品受Cd污染风险。     

Organic Bulking Agents for Enhancing Oil Bioremediation in Soil

Soil contaminated with oil is bioremediated by optimizing conditions for microbial activity. Often the question arises about the benefits of bulking with organic materials to improve soil conditions to enhance degradation of the less biodegradable or less bioavailable components. An investigation was undertaken in the laboratory with the objective of measuring the influence of bulking with dried plant material, bermudagrass, and alfalfa on the degradation of oily sludge added to soil. The oily sludge was diluted 50:50 on a weight basis with soil to achieve a final concentration of 100 g oil and grease kg^-1 of final soil mixture. Bulking agents were added 40 d after dilution of the sludge and optimization of environmental conditions to allow time for the readily decomposable fraction to be degraded before amendment with bulking agents. Populations of heterotrophic microorganisms increased approximately ten times by 40 and 80 d after addition of bulking agents, but the numbers of hydrocarbon-degrading microorganisms did not significantly increase above the number in the nonbulked control. Bulking agents increased the quantity of total petroleum hydrocarbons degraded by approximately 20% during the first 40 d after being added. Disappearance of hydrocarbons for bulked treatments was much slower during the next 40 d, such that the total petroleum hydrocarbon content for both bulked and nonbulked treatments generally was not significantly different at the end. It appears that adding bulking agents may enhance the rate of decomposition of total petroleum hydrocarbons by stimulating the general heterotrophic population of microorganisms, but the influence may not be sustained to influence the extent of decomposition.     

The response of maize (Zea mays L.) plant assisted with bacterial consortium and fertilizer under oily sludge

The objective of this study was to evaluate the role of PGPR consortium and fertilizer alone and in combination on the physiology of maize grown under oily sludge stress environment as well on the soil nutrient status. Consortium was prepared from Bacillus cereus (Acc KR232400), Bacillus altitudinis (Acc KF859970), Comamonas (Delftia) belonging to family Comamonadacea (Acc KF859971) and Stenotrophomonasmaltophilia (Acc KF859973). The experiment was conducted in pots with complete randomized design with four replicates and kept in field. Oily sludge was mixed in ml and Ammonium nitrate and Diammonium phosphate (DAP) were added at 70 ug/g and 7ug/g at sowing. The plant was harvested at 21 d for estimation of protein, proline and antioxidant enzymes superoxide dismutase (SOD) and peroxidase (POD). To study the degradation, total petroleum hydrocarbon was extracted by soxhelt extraction and extract was analyzed by GC-FID at different period after incubation. Combined application of consortium and fertilizer enhanced the germination %, protein and, proline content by 90,130 and 99% higher than untreated maize plants. Bioavailability of macro and micro nutrient was also enhanced with consortium and fertilizer in oily sludge. The consortium and fertilizer in combined treatment decreased the superoxide dismutase (SOD), peroxidase dismutase (POD) of the maize leaves grown in oily sludge. Degradation of total petroleum hydrocarbon (TPHs) was 59% higher in combined application of consortium and fertilizer than untreated maize at 3 d. The bacterial consortium can enhanced the maize tolerance to oily sludge and enhanced degradation of total petroleum hydrocarbon (TPHs). The maize can be considered as tolerant plant species to remediate oily sludge contaminated soils.     

Phytoremediation Effect and Growth Responses of Cynodon spp. and Agropyron desertorum in a Petroleum-Contaminated Soil

In order to compare the petroleum tolerance and phytoremediation ability of a native grass, Agropyron desertorum (desert Wheatgrass) with Cynodon spp. (Bermuda grass) in a petroleum hydrocarbon-contaminated soil, a 7-month greenhouse experiment was performed. There were 4 soil treatments with 0% (uncontaminated soil), 2%, 4%, and 12% (w_oil/w_soil) petroleum concentration. Parameters including shoot and root fresh weight and dry weight, root penetration depth and root density depth, soil respiration, and total petroleum hydrocarbons (TPH) degradation were measured during and after experiments. The results showed an increase in shoot fresh weight of A. desertorum in soil polluted with 2% petroleum sludge compared to the uncontaminated soil, whereas the growth of Bermuda grass significantly decreased in corresponding treatment. Root growth of A. desertorum was decreased in 2% and 4% petroleum sludge, whereas it was increased in Bermuda grass species. Overall, root fresh weight of Bermuda grass was higher than that of A. desertorum in all treatments. Significant increase in microorganisms' activity was observed in the presence of petroleum sludge and plants in soil compared with uncontaminated soil without plants, and the highest soil respiration (37.6 mg C-CO₂/kg soil day) has been observed in the rhizosphere of Bermuda grass in treatment with 12% petroleum sludge. Plants had a significant role in the degradation of soil contaminants as TPH degradation in planted soils was significantly higher than that in unplanted soil (TPH degradation (%) was 30.4 and 38.9 in A. desertorum and Bermuda grass, respectively, whereas it was just 13.3 in unplanted soil). The rhizosphere of Bermuda grass had significantly less residual TPHs compared to A. desertorum. The results indicated that both Cynodon spp. and A. desertorum had a peculiar tolerance to petroleum pollution. Therefore, as Bermuda grass has already been suggested to be a typical and efficient species for phytoremediating petroleum-contaminated sites, A. desertorum may also prove to be a suitable native alternative.     

Bioremediation of Oil-contaminated Soil Using Chicken Manure

In less developed countries, the prevalence of soil contaminated with used lubricating oil is high and the situation worsens with the economic advancement. The contamination has been shown to adversely affect the environment and human health. To mitigate, bioremediation could be adopted to tackle the problem of hydrocarbon-contaminated soil. Thus, this experimental research carried out the bioremediation using chicken manure in soils contaminated with 5%, 10% and 20% w/w used lubricating oil for a 42-day composting period. To compare, this research also experimented with the 5%, 10% and 20% oil-contaminated soils untreated with chicken manure. The results showed that the highest total petroleum hydrocarbons (TPHs) reduction efficiency of >60% was achieved in the 5% oil-contaminated compost remediated with chicken manure. The highest biodegradation rate of lubricating oil of 0.023–0.0025 day^?1 as measured by the first-order kinetics could also be achieved under the 5% oil contamination condition with the application of chicken manure. The findings highlight the prospect of chicken manure as a proper nutrient for enhanced remediation of hydrocarbon-contaminated soils, particularly of low contamination concentrations.     

Biotreatment of Total Petroleum Hydrocarbons from an oily Sludge Using Co-Composting Approach

Discharging of non-treated oily sludge from oil refineries has undesirable impacts on the environment. In this research, the biotreatment of total petroleum hydrocarbons (TPHs) from Abadan Petroleum Refinery, Iran was done using co-composting method. A 5 kg mixture of oily sludge and compost (oily sludge: compost ratios 1:0, 1:0.1, 1:0.3, 1:0.5, and 1:0.7 w/w) and a bulking agent of wheat straw were used as treatments. All treatments were placed in a wooden box inside a laboratory and aerated every 2–3 days by mixing during two months of the experiments. The variation of TPHs concentration, bacterial density, C/N ratio, pH, and temperature were assessed during the 63 days- experiments. Results showed that the majority of TPHs of samples was removed at first 30 days. The maximum TPHs removal (65%) was obtained in the sludge: compost = 1:0.5 at the operation time of 63 day. The significant different between removal efficiency of oily sludge: compost ratios and the control sample (p value < 0.05) indicated the appropriate density of TPHs degrader-bacteria in treated samples. The highest variation in C/N ratio was observed 15± 0.58 in the oily sludge: compost = 1:0.1. At the beginning of experiments, the pH values of all treatments was alkaline, but became neutral at the end of the reaction time. The TPHs degradation kinetic for all oily sludge: compost ratios followed pseudo second-order model. In general, co-composting has high efficiency in oily sludge remediation and can be potentially applied as a simple and cost-effective approach for remediation of oily sludge.     

Screening of herbaceous plants for peat-enhanced rehabilitation of contaminated soil with oily sludge

A batch pot experiment using nine herbaceous species were conducted for peat enhanced rehabilitation of contaminated soil with oily sludge in the initial contents of 0%, 1.3%, 7.4%, and 12.2%, respectively. The results showed that petroleum hydrocarbons removal, plant growth indices and enzyme activities varied depending on plant species and oil contents. Cotton, ryegrass and tall fescue were effective in the rehabilitation of oily sludge contaminated soils. The total petroleum hydrocarbon (TPH) removal ranged from 30.0% to 40.0% after 170 days of treatment. Plant biomass was shown to be the preferred indicator for screening phytoremediation plant because it was closely correlated with TPH removal and enzyme activities. Peat-enhanced plant rehabilitation could be a good strategy for the treatment of oily sludge contaminated saline soils.     

石油污染土壤水改旱田后污染物组份及微生物群落结构变化

对沈抚灌区水改旱田不同年限土壤的石油污染物浓度及组分进行了分析,并采用变性梯度凝胶电泳和磷脂脂肪酸分析方法,分析了污染土壤微生物群落结构的变化.结果表明：1)石油污染土壤水改旱田后,年限越长,总多环芳烃在总石油烃中所占的比重越大,高分子量多环芳烃在总多环芳烃中所占比重也越大;2)总磷脂脂肪酸量与总石油烃呈显著正相关,与总多环芳烃相关性不显著;3)两种方法对土壤微生物群落结构的分析得出的结论一致,石油污染土壤微生物群落结构主要与其相对地理位置有关,当污染物的浓度达到一定程度时,土壤微生物群落结构会发生明显的改变.     

Inorganic materials as ameliorants for soil remediation of metal toxicity to wild mustard (Sinapis arvensis L.)

The ameliorating effects of different inorganic materials were investigated on a soil originating from a zinc smelter dumping site contaminated by toxic metals. Wild mustard (Sinapis arvensis L.) was used as a test plant. The soil was amended with different doses of mining sludge, Perferric Red Latosol (LVj), steel shots, cyclonic ash, silifertil, and superphosphate. The most effective amendments improved plant growth with 45% and reduced metal uptake by over 70% in comparison to untreated soil. Reductions in availability as estimated by BaCl2-extractable metals reached up to 90% for Zn and 65% for Cd as compared to unamended soil. These reductions were associated with lower shoot and root metal contents. Shoot Zn content was reduced from 1,369 microg g(-1) in plants grown on untreated soil to 377 microg g(-1) when grown on cyclonic ash amended soil while Cd decreased from 267 to 44 microg g(-1) in steel shots amended soil. Superphosphate addition had no ameliorating effect. On the contrary, it increased BaCl2-extractable amounts of Zn. Considering all parameters we determined, steel shots, cyclonic ash and silifertil are the most promising for remediating metal contaminated soil in the tropics. Further studies evaluating impacts, cost-effectiveness and durability of effects will be conducted.     

Response of two tomato cultivars differing in salt tolerance to inoculation with mycorrhizal fungi under salt stress

Effects of arbuscular mycorrhizal fungi (AMF) and salt stress on nutrient acquisition and growth of two tomato cultivars exhibiting differences in salt tolerance were investigated. Plants were grown in a sterilized, low-P (silty clay) soil-sand mix. Salt was applied at saturation extract (EC_e) values of 1.4 (control), 4.9 (medium) and 7.1 dS m^–1 (high salt stress). Mycorrhizal colonization occurred irrespective of salt stress in both cultivars, but AMF colonization was higher under control than under saline soil conditions. The salt-tolerant cultivar Pello showed higher mycorrhizal colonization than the salt-sensitive cultivar Marriha. Shoot dry matter (DM) yield and leaf area were higher in mycorrhizal than nonmycorrhizal plants of both cultivars. Shoot DM and leaf area but not root DM were higher in Pello than Marriha. The enhancement in shoot DM due to AMF inoculation was 22% and 21% under control, 31% and 58% under medium, and 18% and 59% under high salinity for Pello and Marriha, respectively. For both cultivars, the contents of P, K, Zn, Cu, and Fe were higher in mycorrhizal than nonmycorrhizal plants under control and medium saline soil conditions. The enhancement in P, K, Zn, Cu, and Fe acquisition due to AMF inoculation was more pronounced in Marriha than in the Pello cultivar under saline conditions. The results suggest that Marriha benefited more from AMF colonization than Pello under saline soil conditions, despite the fact that Pello roots were highly infected with the AMF. Thus, it appears that Marriha is more dependent on AMF symbiosis than Pello. Accepted: 22 January 2001     

Leucanthemum Vulgare Lam. Germination,Growth and Mycorrhizal Symbiosis Under Crude Oil Contamination

Oil contamination of soil limits plants’ access to water and nutrients. Leucanthemum vulgare colonized by mycorrhizae could provide an effective tool in remedying oil contamination. Seeds of L. vulgare were planted in pots containing soil mixed with petroleum at 0, 2.5, 5, 7.5, and 10% w/w and propagules of mycorrhizal fungi. Plants were grown under ambient conditions for 16 weeks. Seed germination data were collected weekly for three weeks. Mycorrhizal percentage, spore counts, length and weight of roots and shoots were determined after harvesting. Results showed significant differences in seed germination rates between oil-treated, mycorrhizal and non-mycorrhizal plants. The overall germination rate was greater at 7.5% w/w crude oil contamination (ρ = 0.05) in mycorrhizal and non-mycorrhizal pots with significant differences between their respective Root:Shoot ratios (both length and weight). Results of this research showed L. vulgare could be germinated and grown in crude oil contaminated soils and could be used to augment plant establishment as part of phytoremediation practices.     

The effect of Piriformospora indica on the root development of maize (Zea mays L.) and remediation of petroleum contaminated soil

As the depth of soil petroleum contamination can vary substantially under field conditions, a rhizotron experiment was performed to investigate the influence of endophyte, P. indica, on maize growth and degradation of petroleum components in a shallow and a deep-reaching subsurface layer of a soil. For control, a treatment without soil contamination was also included. The degree in contamination and the depth to which it extended had a strong effect on the growth of the plant roots. Contaminated soil layers severely inhibited root growth thus many roots preferred to bypass the shallow contaminated layer and grow in the uncontaminated soil. While the length and branching pattern of these roots were similar to those of uncontaminated treatment. Inoculation of maize with P. indica could improve root distribution and root and shoot growth in all three contamination treatments. This inoculation also enhanced petroleum degradation in soil, especially in the treatment with deep-reaching contamination, consequently the accumulation of petroleum hydrocarbons (PAHs) in the plant tissues were increased.     

Enhanced bioremediation of n-alkane in petroleum sludge using bacterial consortium amended with rhamnolipid and micronutrients

The purpose of the present study was to investigate possible methods to enhance the rate of biodegradation of oil sludge from crude oil tank bottom, thus reducing the time usually required for bioremediation. Enhancement of biodegradation was achieved through bioaugmentation and biostimulation. About 10% and 20% sludge contaminated sterile and non-sterile soil samples were treated with bacterial consortium (BC), rhamnolipid biosurfactant (RL) and nitrogen, phosphorus and potassium (NPK) solution. Maximum n-alkane degradation occurred in the 10% sludge contaminated soil samples. The effects of treatment carried out with the non-sterile soil samples were more pronounced than in the sterile soils. Maximum degradation was achieved after the 56th day of treatment. n-Alkanes in the range of nC8-nC11 were degraded completely followed by nC12-nC21, nC22-nC31 and nC32-nC40 with percentage degradations of 100%, 83-98%, 80-85% and 57-73% respectively. Statistical analysis using analysis of variance and Duncan's multiple range test revealed that the level of amendments, incubation time and combination of amendments significantly influenced bacterial growth, protein concentration and surface tension at a 1% probability level. All tested additives BC, NPK and RL had significant positive effects on the bioremediation of n-alkane in petroleum sludge.     

Effect of petrochemical sludge concentrations on microbial communities during soil bioremediation

Qualitative and quantitative changes of microbial communities in soil microcosms during bioremediation were determined throughout one year. The soil was contaminated with 0%, 2.5%, 5%, 10% (wt/wt) of petrochemical sludge containing polynuclear aromatic hydrocarbons. We analyzed the hydrocarbon concentration in the microcosms, the number of cultivable bacteria using CFU and most probable number assays, the community structure using denaturing gradient gel electrophoresis, and the metabolic activity of soil using dehydrogenase activity and substrate-induced respiration assays. After one year of treatment, the chemical analysis suggested that the hydrocarbon elimination process was over. The biological analysis, however, showed that the contaminated microcosms suffered under long-term disturbance. The number of heterotrophic bacteria that increased after sludge addition (up to 10(8)-10(9) cells ml(-1)) has not returned to the level of the control soil (2-6 x 10(7) cells ml(-1)). The community structure in the contaminated soils differed considerably from that in the control. The substrate-induced respiration of the contaminated soils was significantly lower (approximately 10-fold) and the dehydrogenase activity was significantly higher (20-40-fold) compared to the control. Changes in the community structure of soils depended on the amount of added sludge. The species, which were predominant in the sludge community, could not be detected in the contaminated soils.     

Somatic embryogenesis and plant regeneration of turf-type bermudagrass: Effect of 6-benzyladenine in callus induction medium

In order to optimize tissue culture conditions for bermudagrass, an important warm-season turfgrass species, tissue culture responses of young inflorescences of a hybrid bermudagrass cultivar `Tifgreen' (Cynodon dactylon×Cynodon transvaalensis) and a common bermudagrass cultivar `Savannah' (Cynodon dactylon) were investigated. When cultured on Murashige and Skoog medium with 4.52 to 13.57 μM (1–3 mg l^-1) 2,4-D, young inflorescence segments yielded non-embryogenic calli which were unorganized and had loosely associated, long tubular cells on the surface. However, inclusion of 6-benzyladenine (BA) in callus induction medium at a level of 0.044 μM (0.01 mg l^-1) induced formation of a compact, nodular embryogenic structure on approximately 20% of the calli. Calli with such a compact embryogenic structure were highly regenerable. When young inflorescences smaller than 0.75 cm were cultured, the embryogenic structure yielded green plantlets with regeneration rates of 79.5% and 83.3%, respectively for the two cultivars. All 96 plants regenerated from calli induced in the BA-containing medium were green and morphologically normal. The embryogenic nature of the compact structure was confirmed by scanning electron microscopy. This revised version was published online in June 2006 with corrections to the Cover Date.     

Differences in responses to iron deficiency among various cultivars of mungbean (Vigna radiata (L.) Wilczek)

Ten mungbean cultivars were evaluated for their resistance to iron deficiency in view of chlorosis symptoms, plant growth and seed yield under field conditions on a calcareous soil in Thailand. The KPS2 cultivar was highly susceptible; the KPS1, PSU1 and Pag-asa 1 cultivars were somewhat susceptible; the VC1163B cultivar was moderately tolerant; the CN36, CN60, UT1 and CNM-I cultivars were tolerant; and the CNM8509B cultivar was very tolerant to iron deficiency. Foliar application of a solution of 5 g L^-1 ferrous sulphate was effective in correcting chlorosis that was induced by iron deficiency, and it enhanced both the growth and the yield of susceptible cultivars. Compared with the susceptible cultivar KPS2, the tolerant cultivar UT1 had a greater ability to lower the pH of the nutrient solution in response to iron deficiency. The root-associated Fe³⁺-reduction activity of UT1 that had been grown in -Fe medium was similar to that of the plants grown in +Fe medium when the acidification of the medium occurred. Acidification of the medium in response to iron deficiency might contribute to the efficient solubilization of iron from calcareous soils, and it related more closely to the resistance to iron deficiency than Fe³⁺ reduction by roots in mungbean cultivars.