Factors influencing uptake of contaminated particulate matter in leafy vegetables

The contents of cadmium, iron, lead and zinc in the biomass of two species of leafy vegetables after urban particulate matter (PM) application was investigated in lettuce (Lactuca sativa var. capitata) and chard (Beta vulgaris var. cicla). The experimental design consisted of four variables: i) two different soil types, ii) two vegetables, iii) two size fractions of contaminated particulate matter (PM) (0.063–0.119 mm, and <0.063 mm), and iv) foliar and soil application of the PM. The aliquots of the PM samples were applied to the soil before the experiment and as a foliar suspension during plant growth. The element uptake by plant biomass was significantly higher via foliar application, simulating the atmospheric deposition, than via the roots from the soil application treatment. The content in plants increased rapidly compared to the control treatment for the elements iron, lead and zinc. Gently washing the leaves only slightly reduced the amounts of Fe and Zn. However, the majority of Pb was removed by washing with the concentration dropping from 3000% to 500%. The effect of PM application on Cd contents in plant leaves was negligible, in most cases. Lettuce exhibited higher element uptake compared to chard. No adverse effects of PM application on growth parameters of the vegetables were observed. No significant differences were reported for particle size fractions of PM. Fluvisol soils had a higher element uptake via roots than Chernozem soils. Moreover, the addition of PM into the Fluvisol altered the sorption properties of the soil resulting in a lower Cd uptake by plants growing in PM amended Fluvisols.     

Transgenic tobacco plants expressing a rice cysteine synthase gene are tolerant to toxic levels of cadmium

Plants tolerate heavy metals through sequestration with cysteine-rich peptides, phytochelatins. In this reaction, the rate limiting step is considered to be the supply of cysteine, which is synthesized by cysteine synthase (CS, EC 4.2.99.8) from hydrogen sulfide andO-acetylserine. In this study, we transformed tobacco (Nicotiana tabacum) plants withRCS1, a cytosolic cysteine synthase gene of rice (Oryza sativa), and examined their sensitivity to cadmium. The transgenic plants had up to 3-fold higher activity of cysteine synthase than wild-type plants. Upon exposure to cadmium, they exhibited obvious tolerance with much greater growth than wild-type plants. The level of phytochelatins in shoots was higher in transgenic than in wild-type plants after cadmium treatment, suggesting that cadmium was actively trapped by phytochelatins. However, the cadmium concentration per g fresh weight of whole transgenic plants was 20 percnt; lower than that of wild-type plants, suggesting cadmium to be either actively excreted or diluted by fast growth. Genetic analysis of progenies clearly showed segregation of cadmium tolerance, indicating that the trait resulted from the introduced gene. These results suggest that introduction of a cysteine synthase gene into tobacco plants resulted not only in high level production of sulfur-containing compounds that detoxify cadmium, but also in active elimination of cadmium toxicity from plant bodies.     

Dual inoculation with VA mycorrhiza and Rhizobium is beneficial to Leucaena

Summary Response ofLeucanea leucocephala to inoculation withGlomus fasciculatum and/or Rhizobium was studied in a phosphorus deficient unsterile soil.G. fasciculatum only inoculation improved nodulation by native rhizobia and Rhizobium only treatment improved colonization of roots by native mycorrhizal fungi. Dual inoculation with both the organisms improved nodulation, mycorrhizal colonization, dry weight, nitrogen and phosphorus content of the plants compared to single inoculation with either organism. Contribution of U.A.S. Research Project DR/AMB-1.     

Effects of inoculation with plant growth promoting rhizobacteria (PGPRs) andSinorhizobium fredii on biological nitrogen fixation, nodulation and growth ofGlycine max cv. Osumi

We investigated the effects of three plant growth promoting rhizobacteria (PGPR), on Biological Nitrogen Fixation (BNF), nodulation and growth promotion by soybean (Glycine max) var. Osumi plants. The strains, Aur 6, Aur 9 and Cell 4, belong toPsedomonas fluorescens, Chryseobacterium balustinum andSerratia fonticola, respectively. Inoculation modes for the PGPRs andSinorhizobium fredii (carried out through irrigation), were examined. In the first mode, PGPRs andS. fredii were co-inoculated. In the second mode, we first inoculatedS. fredii and after the PGPRs, which were added 5 or 10 days later (each inoculation being an independent treatment). In the third mode, the PGPRs were inoculated first, and theS. fredii was inoculated 5 days later. We also included treatments inoculated with only the PGPRs (one PGPR per treatment) and only withS. fredii. Plants were maintained in a greenhouse under controlled environmental conditions, and were sampled 3 months after sowing. The results obtained showed the effects of the inoculation sequence. The most significant effects on growth parameters (stem plus leaf weight and fresh root weight) were found when inoculations with PGPR andS. fredii were at different times or when we inoculated only with PGPR and the plants were watered with nitrogen. Co-inoculation had no positive effects on any parameter, probably due to competition between the PGPR andS. fredii. Our results indicate that the inoculation modes with PGPR and rhizobia play a very important role in the effects produced. Thus, although plant growth promoting rhizobacteria may interact synergistically with root-nodulating rhizobia, plant growth promoting rhizobacteria selected for one crop should be assessed for potentially hazardous effects on other crops before being used as inoculants.     

甜菜属厚皮菜SWTY-1细胞质VNTRs多态性分析

利用数目可变串联重复序列(Variable Number of Tandem Repeats,VNTRs)微卫星标记方法,对重庆厚皮菜甜菜材料SWTY-1群体中100个单株的细胞质线粒体DNA片段中TR2位点VNTRs片段多态性进行分析。结果显示97个单株线粒体TR2位点微卫星串联重复序列均为3拷贝,与普通糖甜菜一致;3个单株线粒体TR2位点微卫星串联重复序列为6拷贝,发现甜菜属厚皮菜细胞质TR2位点VNTRs存在多态性,在该群体中发现了不同于甜菜栽培种新的细胞质单株。对该群体材料100个单株的抽薹及结籽进行观测,结果显示微卫星串联重复序列为6拷贝的变异植株中2个单株花期未抽苔开花,1株抽苔晚未形成正常种子;细胞质TR2位点VNTRs片段拷贝数为3的植株中2个单株未能正常抽薹,其他植株均正常抽薹结籽。     

Cadmium biosorption by <Emphasis Type="Italic">Bacillus</Emphasis><Emphasis Type="Italic"> circulans</Emphasis> strain EB1

Summary A heavy metal resistant bacterium, Bacillus circulans strain EB1 showed a high cadmium biosorption capacity coupled with a high tolerance to this metal when grown in its presence. Bacillus circulans EB1 cells grown in the presence of 28.1 mg cadmium/l were capable of removing cadmium with a specific biosorption capacity of 5.8 mg Cd/g dry wt biomass in the first 8 h. When the cells were pre-conditioned with low concentrations of cadmium in pre-grown medium, the uptake was increased to 6.7 mg Cd/g dry wt biomass. The maximum uptake of␣cadmium was during mid-logarithmic phase of growth. The resting cells (both wet and dry) of EB1 were also able to biosorb cadmium. Specific biosorption capacities of wet and dry biomass were 9.8 and 26.5 mg Cd/g dry wt biomass, respectively. Maximum cadmium removals by both wet and dry cells were at pH 7.0. The results showed that the cadmium removal capacity of resting cells was markedly higher than that of growing cells. Since both growing and resting cells had a high biosorption capacity for cadmium, EB1 cells could serve as an excellent biosorbent for removal of cadmium from natural environments.     

Growth and Cadmium Phytoextraction by Swiss Chard,Maize, Rice,Noccaea caerulescens,and Alyssum murale in pH Adjusted Biosolids Amended Soils

Past applications of biosolids to soils at some locations added higher Cd levels than presently permitted. Cadmium phytoextraction would alleviate current land use constraints. Unamended farm soil, and biosolids amended farm and mine soils were obtained from a Fulton Co., IL biosolids management facility. Soils contained 0.16, 22.8, 45.3 mg Cd kg^–1 and 43.1, 482, 812 mg Zn kg^–1 respectively with initial pH 6.0, 6.1, 6.4. In greenhouse studies, Swiss chard (Beta vulgaris var. cicla), a Cd-accumulator maize (inbred B37 Zea mays) and a southern France Cd-hyperaccumulator genotype of Noccaea caerulescens were tested for Cd accumulation and phytoextraction. Soil pH was adjusted from ～5.5–7.0. Additionally 100 rice (Oryza sativa) genotypes and the Ni-hyperaccumulator Alyssum murale were screened for potential phytoextraction use.

Chard suffered phytotoxicity at low pH and accumulated up to 90 mg Cd kg^–1 on the biosolids amended mine soil. The maize inbred accumulated up to 45 mg Cd kg^–1 with only mild phytotoxicity symptoms during early growth at pH > 6.0. N. caerulescens did not exhibit phytotoxicity symptoms at any pH, and accumulated up to 235 mg Cd kg^–1 in 3 months. Reharvested N. caerulescens accumulated up to 900 mg Cd kg^–1 after 10 months. Neither Alyssum nor 90% of rice genotypes survived acceptably.

Both N. caerulescens and B37 maize show promise for Cd phytoextraction in IL and require field evaluation; both plants could be utilized for nearly continuous Cd removal. Other maize inbreds may offer higher Cd phytoextraction at lower pH, and mono-cross hybrids higher shoot biomass yields. Further, maize grown only for biomass Cd maximum removal could be double-cropped.     

Soybean biofertilization in sandy soils of Egypt using15N tracer technique

The effect of inoculating soybean plants withBradyrhizobium japonicum andAzospirillum brasilense either solely or in mixture, and of using different N fertilizer levels was studied in pot experiments. The nodulation of soybean grown in a sandy soil was enhanced by the inoculation while the highest nodule numbers and fresh mass, recorded at a N-fertilizer dose of 20 kg N/hm², decreased when the fertilizer quantity increased to 40 kg N/hm². By contrast, the dry mass of above-ground parts and the N uptake was increased with increasing N fertilizer level. A similar effect was observed for inoculation as compared with the uninoculated variant. Although the nodulating and non-nodulating soybeans has nearly the same dry mass, the nodulating isoline accumulated more N than the non-nodulating. The percentage of nitrogen derived from air (%, Ndfa) and estimated by isotope dilution (ID) or N difference method (DM) dropped with increasing N level from 10 to 40 kg N/hm². Dual inoculation resulted in a high per cent of fixed N₂ (42.5%) at 10 kg N/hm². Correlation between the ID and DM methods was found to be dependent on inoculation treatments. The amount of nitrogen utilized by nodulating soybean (FUE %) was enhanced as a function of inoculation withB. japonicum.     

秋华柳和枫杨幼苗对镉的积累和耐受性

以秋华柳和枫杨当年实生幼苗为研究对象,采用向土壤添加外源镉(CdCl₂ · 2.5H₂O)的形式设置了0(对照组)、10 、20 、50、100 mg/kg 5个处理,研究了镉胁迫下秋华柳和枫杨幼苗的生长、生物量变化和根茎叶镉含量,并评价了两树种的耐性指数(Ti)、转移系数(Tf)和生物富集系数(BCF)。结果表明:(1)在镉含量为10 mg/kg时,秋华柳和枫杨幼苗基于生长和生物量参数的耐性指数(Ti)分别为91.72和91.62,与对照组相比无显著变化,其余各组(20、50、100 mg/kg)则显著低于对照植株(P<0.05);(2) 土壤镉浓度小于20mg/kg时,秋华柳植株茎、叶镉积累量分别高达61.73 mg/kg、163.04 mg/kg,根镉积累量为91.05 mg/kg;枫杨植株茎、叶镉积累量最高分别为7.9 mg/kg、5.25 mg/kg,仅为秋华柳茎、叶的12.8%和3.2%,根镉积累量高达190.68 mg/kg;(3) 除对照外,秋华柳幼苗各部分镉含量为叶＞根＞茎,转移系数(Tf)介于0.789-1.513之间,枫杨幼苗各部分镉含量为根＞茎＞叶,转移系数(Tf)介于0.037-0.044之间,远远小于秋华柳Tf;(4)秋华柳和枫杨幼苗在土壤镉浓度为10 mg/kg时具有很高的生长适应性和耐性,秋华柳根吸收的镉向地上部分转移能力、地上部分积累镉的能力都远远大于枫杨,生物富集系数(BCF)进一步证实了这一特性。研究证明,秋华柳植株具有很高的镉耐性、镉转移能力及地上部分积累镉的能力,适合于镉污染严重区域的植物修复。     

Biological Control of Crown Gall: Seed and Root Inoculation

S ummary . Experiments on the control of crown gall by inoculating susceptible plants with a non-pathogenic strain of Agrobacterium radiobacter have continued. In all experiments, highly significant disease control was achieved. In one experiment, 42% of untreated plants growing in soil heavily infested with A. radiobacter var. tumefaciens died; inoculation of seed with the non-pathogenic strain reduced this to nil. Combined seed and root inoculation was more efficient than seed inoculation alone. In naturally infested soil, combined seed and root inoculation at transplanting gave 99% control of gall formation (as dry weight). A significant increase in plant growth resulted from combined seed and root inoculation. At transplanting, roots should probably be inoculated within 2 h of lifting. This method of biological control is now widely practised by commercial growers in South Australia.     

Influence of soil pH on the soybean-endomycorrhiza symbiosis

Summary Soybean (Glycine max {L.} Merr.) cultivars were inoculated withGigaspora gigantea andGlomus mosseae to determine mycorrhizal: cultivar relationships as affected by soil pH. The specific cultivarfungal response was dependent on soil pH. Overall cultivar responses in unlimed soil (pH 5.1) were greater forG. gigantea thanG. mosseae. The Bossier —G. gigantea combination was particularly responsive in unlimed soil and showed an increase of 10% in shoot length, 35% in shoot dry weight. 75% in root dry weight, and 397% in nodule dry weight over uninoculated controls. Little cultivar response was observed withG. mosseae inoculation in unlimed soil. In limed soil (pH 6.2), the larger responses were obtained withG. mosseae inoculated plants, although inoculation with eitherG. mosseae orG. gigantea appeared effective. In general, nodulation was greater on mycorrhizal roots than on control roots.     

Interaction of yeasts and some nitrogen fixing bacteria on nodulation of legumes

Summary Combined inoculation ofRhizobium trifolii withSaccharomyces cerevisiae and other yeasts generally enhanced the number of nodules, length of plants and dry weight of Egyptian clover (Trifolium alexandrinum) seedlings grown on agar slopes. Similar effects were observed when seedlings were inoculated withR. trifolii in the presence of dialyzed culture filtrate ofS. cerevisiae.     

Cd-Resistant Strains of B. cereus S5 with Endurance Capacity and Their Capacities for Cadmium Removal from Cadmium-Polluted Water

The goal of this study was to identify Cd-resistant bacterial strains with endurance capacity and to evaluate their ability to remove cadmium ions from cadmium-polluted water. The Bacillus cereusS5 strain identified in this study had the closest genetic relationship with B. cereus sp. Cp1 and performed well in the removal of Cd²⁺ions from solution. The results showed that both the live and dead biomasses of the Cd²⁺-tolerant B. cereus S5 strain could absorb Cd²⁺ ions in solution but that the live biomass of the B. cereus S5 strain outperformed the dead biomass at lower Cd²⁺concentrations. An analysis of the cadmium tolerance genes of B. cereus S5 identified ATPase genes that were associated with cadmium tolerance and involved in the ATP pumping mechanism. The FTIR spectra revealed the presence of amino, carboxyl and hydroxyl groups on the pristine biomass and indicated that the cadmium ion removal ability was related to the structure of the strain. The maximum absorption capacity of the B. cereus S5 strain in viable spore biomass was 70.16 mg/g (dry weight) based on a pseudo-second-order kinetic model fit to the experimental data. The Langmuir and Langmuir-Freundlich isotherm adsorption models fit the cadmium ion adsorption data well, and the kinetic curves indicated that the adsorption rate was second-order. For Cd²⁺ concentrations (mg/L) of 1–109 mg/L, good removal efficiency (>80%) was achieved using approximately 3.48–10.3 g/L of active spore biomass of the B. cereus S5 strain. A cadmium-tolerant bacteria-activated carbon-immobilized column could be used for a longer duration and exhibited greater treatment efficacy than the control column in the treatment of cadmium-polluted water. In addition, a toxicity assessment using mice demonstrated that the biomass of the B. cereus S5 strain and its fermentation products were non-toxic. Thus, the isolated B. cereus S5 strain can be considered an alternative biological adsorbent for use in emergency responses to severe cadmium pollution and in the routine treatment of trace cadmium pollution.     

Pink root rot,a revised name of brown root rot of gentian,and the causal fungi,Pyrenochaeta gentianicola sp. nov. andP. terrestris in Japan

Cultivated gentian (Gentiana scabra var.buergeri) has been seriously damaged by pink root rot since the 1970s. Diseased plants finally collapse after wilting and stunting. More than 40 fungal genera were found to be associated with roots of mature and immature diseased plants or seeds. Among these fungi,Pyrenochaeta gentianicola sp. nov. andP. terrestris were almost always associated with the diseased plants. Their morphologies and temperature responses were compared, and their pathogenicity was also demonstrated by artificial inoculation tests.Part of this work was conducted at the National Institute of Agricultural Sciences (presently National Institute of Agro-Environmental Sciences), Tsukuba S.C., Ibaraki 305, Japan by T. Watanabe.     

Genetically engineered resistance to potato virus X in four commercial potato cultivars

The genes for the capsid protein (CP) and the 8K movement protein of PVX were introduced into potato (Solanum tuberosum L.) and expressed under the control of CaMV 35S promoter using a binary vector andAgrobacterium tumefaciens. Four commercial potato cultivars (Russet Burbank, Shepody, Desirée and Bintje) have been efficiently transformed. Eleven independent transgenic clones, with CP expression levels higher than 0.05% of the soluble leaf proteins, were analyzed for resistance to inoculation with PVX (5 and 50µg/ml). The resistance of the transgenic plants to PVX was observed with the lower titer of virus inoculation (5 µg/ml) but not with higher titer (50 µg/ml). A significant reduction in the accumulation of virus in the inoculated transgenic potato plants has been observed under greenhouse and field conditions. Furthermore, the CP gene is very stable and is transferred to new plants originated from stem cuttings or from tubers. The transgenic plants appeared to be phenotypically identical to the nontransformed controls.Abbreviations BAP benzyl-aminopurine - BCIP 5-bromo-4-chloro-3-indolylphosphate p-Toluidine salt - CaMV cauliflower mosaic virus - CP capsid protein - GA₃ gibberellic acid - Kbp kilobase pair - NAA naphthalene acetic acid - NBT nitroblue tetrazolium chloride - NOS nopaline synthase - NPT II neomycin phosphotransferase II - PMSF phenyl methyl sulfonyl fluoride - PVX potato virus X - PVY potato virus Y     

Cadmium adsorption by the non-living biomass of microalgae grown in axenic mass culture

The freeze-dried (extracted and non-extracted) biomass of 15 microalgal species grown in axenic mass culture and belonging to the Cyanobacteria, Chloro-, Eustigmato-, Phaeo-, Rhodo- and Tribophyceae were investigated for their ability to adsorb cadmium (Cd) ions from aqueous solutions. For comparison, other standard adsorbing materials (activated carbon, silica gel, siliceous earth) were included in the studies. The biomass of 11 microalgae exhibited a higher Cd adsorption than the standard materials. Extraction of the algal biomass increased the Cd adsorption capability of some, but not all microalgae. High Cd adsorption was found inAnabaena lutea, Nodularia harveyana, andNostoc commune (Cyanobacteria),Chlamydomonas sp. (Chlorophyceae),Bumilleriopsis filiformis (Tribophyceae), and inEctocarpus siliculosus, Halopteris scoparia andSpermatochnus paradoxus (Phaeophyceae). The specific surface (m² cm^–3) of the various microalgae was determined by means of laser diffractometry.Anabaena inaequalis andA. lutea (Cyanobacteria) and the Phaeophyceae had especially high Cd adsorption per surface unit. Most of the Cd adsorbed to these various materials could be desorbed subsequently with diluted mineral acid (pH 2).     

Morphological variation among members of theStellaria longipes complex:S. longipes,S. longifolia,andS. porsildii (Caryophyllaceae)

Morphological relationships were investigated among diploidStellaria porsildii, polyploidS. longipes, and diploidS. longifolia. Canonical discriminant analysis, based on a priori assumptions to maximize differences among groups, showed thatS. longipes clusters equally distant between the two diploid species along an axis connecting the diploids' centroids, but it differs along an axis perpendicular to this axis. The intermediacy along the former axis is evidence thatS. longipes is an amphiploid derived fromS. longifolia andS. porsildii. The divergence along the latter axis may be attributable to adaptively valuable heterotic traits which were retained following amphidiploidization. The only morphological discontinuity occurred between the two diploids, whereas the morphological range ofS. longipes overlapped the range of both diploids forming a continuum. The lack of discrete clusters is likely due to hybridization and introgression withS. longifolia on one hand, and convergence of traits betweenS. longipes var.monantha andS. porsildii on the other. High a posteriori assignments in classificatory discriminant analysis supports the separation ofS. longipes var.monantha from otherS. longipes specimens. AlthoughS. longipes var.monantha grouped close toS. porsildii, the two groups separate based on leaf shape traits. Overall results support, firstly, the hypothesis thatS. porsildii is a diploid parent species which by hybridizing withS. longifolia gave rise to polyploidS. longipes. Secondly, results suggest thatS. longipes var.monantha converged morphologically towardsS. porsildii relatively recently due to ecological specialization, and merits distinction at least as a variety ofS. longipes.     

Evaluation of the symbiotic nitrogen-fixing potential of soils by direct microbiological means

A method for estimating the nitrogen-fixing capacity of a population of rhizobia resident in soil is presented. legume test plants, growing under microbiologically-controlled conditions in test tubes packed with a vermiculite substrate moistened with a nitrogen-free plant nutrient solution, are inoculated directly with a suspension of the soil under examination. Rhizobia in the soil nodulate the test plants, and the amount of foliage dry matter produced in the 28 days after inoculation is regarded as an index of their effectiveness. An inoculum of at least 30, and preferably 100, rhizobia is needed to ensure that nitrogen fixation is not masked by delayed nodulation. The new method is tentatively described as the ‘whole-soil inoculation’ technique. Appraisals were made withTrifolium subterraneum L. andRhizobium trifolii and withMedicago sativa L. andR. meliloti. Soil-borne pathogens did not interfere with plant growth. The whole-soil inoculation technique was less tedious and time-consuming than an alternative method which involved extracting representative isolates from the soil and testing their effectiveness individually, and appeared to give more realistic values for the nitrogen-fixing capacity of the soil as a whole. Used in association with a field experiment, the whole-soil inoculation technique confirmed microbiologically that there had been an agronomic response to surface application of inoculant to poorly-nodulatedT. subterraneum pasture. It is submitted that this technique for determining the effectiveness of rhizobia in soil, combined with a plant-infection method for counting rhizobia, can be a reliable guide to the need for inoculation in the field.     

Field inoculation of sorghum and rice withAzospirillum spp. andHerbaspirillum seropedicae

Two field experiments were carried out at the UAPNPBS experimental station, Seropédica, with two sorghum and one rice cultivars. The establishment, and inoculation effects, ofAzospirillum spp. andHerbaspirillum strains marked with antibiotic resistance were investigated. One grain sorghum (BR 300) and one sugar sorghum (Br 505) cultivar were used.Azospirillum lipoferum strain S82 (isolated from surface sterilized roots of sorghum) established in both cultivars and comprised 40 to 80% of theAzospirillum spp. population in roots and stems 60 days after plant emergence (DAE).Azospirillum amazonense strain AmS91 (isolated from surface-sterilized roots of sorghum) reached only 50%. At 90 DAE, S82 almost disappeared (less than 30% of establishment) while the establishment of AmS91 remained constant in roots and stems. No establishment ofH. seropedicae strain H25 (isolated from surface-sterilized roots of sorghum) orA. lipoferum strain S65 (isolated from the root surface of sorghum) could be observed on inoculated roots. Inoculation with S82, AmS91 or S65 but not withH. seropedicae H25, increased plant dry weight of both cultivars and total N in grain of the grain sorghum. In rice,A. lipoferum Al 121 andA. brasilense Sp 245 (isolated from surface sterilized rice and wheat roots respectively) established in the roots but there was no increase inAzospirillum spp. numbers due to inoculation. None of the strains affected plant growth or rice grain yield.Azospirillum amazonense, A82 andH. seropedicae Z95, which did not establish in roots, significantly enhanced seed germination.