Toxicity of 2,3,5,6-tetrachlorophenol to willow trees (Salix viminalis)

Chlorinated phenols have been intensively investigated from an eco-toxicological point of view, however almost nothing is known about toxicity of tetrachlorophenol (TeCP) to higher terrestrial plants. This article applied the willow tree acute toxicity test to study the toxicity of 2,3,5,6-TeCP to willows Salix viminalis (S. viminalis) at neutral and acidic conditions (roughly pH 7 and 4) with inhibition of transpiration as toxic endpoint. At neutral pH the EC₅₀ was >10 mg L^?1 while the EC₅₀ at acidic conditions was 0.32 ± 0.17 mg L^?1, clearly indicating that toxicity is exerted by the non-ionic chemical fraction. Standard tests running at neutral pH are therefore not capturing the full toxicity of weak acids and bases.     

Use of wastewater as a nutrient solution in a closed gravel hydroponic culture of giant reed (Arundo donax)

The objective of this experimental work was to examine the efficiency of giant reed (Arundo donax L.), as a source of biomass production and as a biofiltering device for sewage effluents. Two giant reed populations were cultivated in a closed gravel hydroponic system, where pig's waste was used as a nutrient solution. The results showed that stem biomass production varied from 12 to 23 kg DM m(-2) yr(-1), more than the ordinary production in the soil. According to stem analysis, for the first two years, there was an average infiltration rate of 31 g m(-2) yr(-1) total N, 7.5 g m(-2) yr(-1) total P, 18.8 g m(-2) yr(-1) K, 2.1 g m(-2) yr(-1) Ca, 2.1 g m(-2) yr(-1) Mg, 0.27 g m(-2) yr(-1) Fe, 0.02 g m(-2) yr(-1) Mn, 0.14 g m(-2) yr(-1) Zn and 0.08 g m(-2) yr(-1) Cu. During the third year, when a nutrient solution with added P was used, the average infiltration rate for most elements increased by 46% and for P by 169%.     

The development of two flanking SCAR markers linked to a sex determination locus in Salix viminalis L

Most studies of sex determination systems in plants involve dioecious annuals that have known sex chromosomes. Despite the absence of such structures in the majority of dioecious plants, gender seems to be under relatively strict genetic control in some species. Genetic markers linked to a female sex-determination locus in Salix viminalis L. have been discovered through bulked segregant analysis of three full-sib families using approximately 1,000 arbitrary primers. Two RAPD markers that were present in the common female parent as well as in predominantly female progeny of these families were subsequently sequenced and converted to sequence characterized amplified region (SCAR) markers. The two SCAR markers are correlated with gender in the three full-sib families and are present in 96.4% of the female progeny and 2.2% of the males, providing evidence of linkage to a putative female-specific locus associated with gender determination in S. viminalis. Estimates of recombination suggest that the two markers are flanking a putative sex determination locus, SDL-II, in certain families of S. viminalis.     

Development of a hydroponic screening technique to assess heavy metal resistance in willow (Salix)

A nutrient thin film hydroponic system has been developed which allows rapid screening of willow (Salix) clones for their resistance to heavy metals, and hence their use in phytoremediation. Two clones known to be different in their resistance to heavy metals (Salix burjatica (Germany) and S. triandra x viminalis (Q83)), could be distinguished on the basis of leaf biomass, root biomass and stem height after 6 weeks. There were also differences in the uptake of heavy metals between the two clones.     

A genetic linkage map of willow ( Salix viminalis) based on AFLP and microsatellite markers

The genus Salix (willow) contains a number of species of great value as biomass crops. Efforts to breed varieties with improved biomass yields and resistances to pests and diseases are limited by the lack of knowledge on the genetic basis of the traits. We have used AFLP and microsatellite markers to construct a genetic map of willow from a full-sib cross of the diploid species Salix viminalis (2n = 38). In accordance with a double pseudo-testcross approach, separate parental maps were constructed and merged to produce a consensus map comprising 291 AFLP and 39 willow microsatellite markers. Nineteen poplar microsatellites were also tested in willow. Five of these amplified loci, of which two were mapped. Linkage groups of the consensus map that could be identified in the parental maps are presented here and spanned 1,256.5 cM with an average interval between markers of 4.4 cM.     

Periodicity of response to abscisic acid in lateral buds of willow (Salix viminalis L.)

Aseptically cultured lateral buds of Salix viminalis L. collected from field-grown trees exhibited a clear periodicity in their ability to respond to exogenous abscisic acid (ABA). Buds were kept unopened by ABA only when the plants were dormant or entering dormancy. Short days alone did not induce bud dormancy in potted plants but ABA treatment following exposure to an 8-h photoperiod prevented bud opening although ABA treatment of buds from long-day plants did not. Naturally dormant buds taken from shoots of field-grown trees and cultured in the presence of ABA opened following a chilling treatment. In no cases were the induction and breaking of dormancy and response to ABA correlated with endogenous ABA levels in the buds.Abbreviations ABA abscisic acid - GA₃ gibberellic acid - HPLC high-performance liquid chromatography - LD long day - MeABA methyl ABA - PAR photosynthetically active radiation - SD short day     

Response of Organ Structure and Physiology to Autotetraploidization in Early Development of Energy Willow Salix viminalis

The biomass productivity of the energy willow Salix viminalis as a short-rotation woody crop depends on organ structure and functions that are under the control of genome size. Colchicine treatment of axillary buds resulted in a set of autotetraploid S. viminalis var. Energo genotypes (polyploid Energo [PP-E]; 2n = 4x = 76) with variation in the green pixel-based shoot surface area. In cases where increased shoot biomass was observed, it was primarily derived from larger leaf size and wider stem diameter. Autotetraploidy slowed primary growth and increased shoot diameter (a parameter of secondary growth). The duplicated genome size enlarged bark and wood layers in twigs sampled in the field. The PP-E plants developed wider leaves with thicker midrib and enlarged palisade parenchyma cells. Autotetraploid leaves contained significantly increased amounts of active gibberellins, cytokinins, salicylic acid, and jasmonate compared with diploid individuals. Greater net photosynthetic CO₂ uptake was detected in leaves of PP-E plants with increased chlorophyll and carotenoid contents. Improved photosynthetic functions in tetraploids were also shown by more efficient electron transport rates of photosystems I and II. Autotetraploidization increased the biomass of the root system of PP-E plants relative to diploids. Sections of tetraploid roots showed thickening with enlarged cortex cells. Elevated amounts of indole acetic acid, active cytokinins, active gibberellin, and salicylic acid were detected in the root tips of these plants. The presented variation in traits of tetraploid willow genotypes provides a basis to use autopolyploidization as a chromosome engineering technique to alter the organ development of energy plants in order to improve biomass productivity.Energy security and climate change as global problems urge increased efforts to use plants as renewable energy sources both for power generation and transportation fuel production. Selected wood species, such as willows (Salix spp.), can be cultivated as short-rotation coppice for the rapid accumulation of biomass and reduction of CO₂ emission. Coppicing reinvigorates shoot growth, resulting in a special woody plant life cycle that differs from natural tree development, which takes decades. In this cultivation system, small stem cuttings are planted at high densities (15,000–25,000 ha⁻¹). In the soil, these dormant wood cuttings first produce roots and shoots that emerge from reactivated buds. During the first year, the growing shoots mature to woody stems. In the winter, these stems are cut back, and in the following spring, the cut stumps develop multiple shoots. The short-rotation coppice plantations are characterized by a very short, 2- to 3-year rotation, and the most productive varieties can produce up to 15 tons of oven-dried wood per hectare per year (Cunniff and Cerasuolo, 2011). The high-density willow plantations can also be efficiently used for heavy metal or organic phytoremediation, as reviewed by Marmiroli et al. (2011).The biomass productivity of shrub willows is largely dependent on coppicing capability, early vigorous growth, shoot growth rate and final stem height, root system size, photosynthetic efficiency, formation and composition of woody stems, water and nutrient use, as well as abiotic and biotic stress tolerance. Genetic improvement of all these traits can be based on broad natural genetic resources represented by more than 400 species in the genus Salix. More than 200 species have hybrid origins, and ploidy levels vary from diploid up to dodecaploid (Suda and Argus, 1968; Newsholme, 1992). In addition to molecular marker-assisted clone selection, intraspecific and interspecific crosses have been shown to further extend genetic variability in breeding programs for biomass yield (Karp et al., 2011).During natural diversification and artificial crossings of Salix spp., the willow genomes frequently undergo polyploidization, resulting in triploid or tetraploid allopolyploids. In triploid hybrids, both heterosis and ploidy can contribute to the improved biomass yield (Serapiglia et al., 2014). While the alloploid triploids have attracted considerable attention in willow improvement, the potentials of autotetraploid willow genotypes have not been exploited so far. As shown for other short-rotation wood species (poplar [Populus spp.], black locust [Robinia pseudoacacia], Paulownia spp., and birch [Betula spp.]), doubling the chromosome set by colchicine treatment can cause significant changes in organ morphology or growth parameters (Tang et al., 2010; Cai and Kang, 2011; Harbard et al., 2012; Mu et al., 2012; Wang et al., 2013a, 2013b). In several polyploidization protocols, the in vitro cultured tissues are exposed to different doses of colchicine or other inhibitors of mitotic microtubule function, and plantlets are differentiated from polyploid somatic cells (Tang et al., 2010; Cai and Kang, 2011). Alternatively, seeds or apical meristems of germinating seedlings can be treated with a colchicine solution (Harbard et al., 2012). Allotetraploids of poplar were produced by zygotic chromosome doubling that was induced by colchicine and high-temperature treatment (Wang et al., 2013a).Since tetraploid willow plants with 2n = 4x = 76 chromosomes are expected to represent novel genetic variability, especially for organ development and physiological parameters, a polyploidization project was initiated that was based on a highly productive diploid energy willow (S. viminalis var. Energo). Colchicine treatment of reactivated axillary buds of the in vitro-grown energy willow plantlets resulted in autotetraploid shoots and, subsequently, plants. For comparison of diploid and tetraploid variants of willow plants, digital imaging of green organs and roots was used for phenotyping. Among the tetraploid lines, genotypes were identified with improved biomass production, better photosynthetic parameters, and altered organ structure and hormone composition. The new tetraploid willow variants produced can serve as a unique experimental material to uncover key factors in biomass production in this short-rotation energy plant. In the future, these plants can also serve as crossing partners of diploid lines for the production of novel triploid energy willow genotypes.     

Expression of a cytokinin synthesis gene from Agrobacterium tumefaciens T-DNA in basket willow (Salix viminalis)

Willow cells transformed with an ipt gene from Agrobacterium tumefaciens grow in tissue culture as undifferentiated callus without shoot induction. We show that the transformed calluses contained high levels of the cytokinins 9-β- d -ribofuranosyl zeatin and its monophosphate, demonstrating the presence of a functional isopentenyl transferase enzyme. The ipt gene was transcribed at different levels in different transformed callus lines. The absence of shoot differntiation is apparently not due to a lack of zeatin-type cytokinins in the transformed callus.     

The root growth of Salix viminalis and Eucalyptus nitens in response to dairy farm pond effluent irrigation.

The spatial root distribution after two years of three energy crops was investigated, with the influence of two rates of dairy pond effluent application, applied every fortnight. The roots of Salix viminalis PN 386 and NZ 1295 and Eucalyptus nitens were studied using soil cores. It was found that spatial distribution was influenced by effluent rate, with greater quantities of both fine and coarse roots in the top soil horizons with the higher effluent rate of 300 m3 ha(-1) compared to 150 m3 ha(-1). The study has implications on the harvesting of such crops using heavy machinery which could cause root damage and lessen vigour in the second rotation.     

Transport of 14C-labelled Assimilates and 32P-labelled Phosphate in Salix viminalis in Relation to Phyllotaxis and Leaf Age

Autoradiographic techniques have been used to investigate thetransport of ¹⁴C-assimilates and ³²P-phosphate in relation tophyllotaxis and age of leaves in Salix viminalis. The leavesborne on the young shoots of S. viminalis are arranged in fiveorthostichies. Three of these orthostichies share one main transportchannel, a second channel being shared by the leaves on theremaining two orthostichies. Where two orthostichies are separatedby an angular distance of 72°, transport may occur betweenleaves on these orthostichies. In addition to the phyllotaxis of the leafy shoot, it has beendemonstrated that the age of the leaf is a factor which determinesthe distribution of materials between leaves.     

Purification of nitrate-rich agricultural runoff by a hydroponic system

The purification of nitrate-rich agricultural runoff by a floating-raft (FR) hydroponic system was investigated at 3-, 2- and 1-d hydraulic retention times (HRTs) with particular emphasis on nitrogen conversion and removal through the system. The FR system has a dissolved oxygen (DO) environment similar to the horizontal subsurface flow system, generally 0.00mgL(-1), that facilitates denitrification. An efficient nitrate-nitrite-nitrogen (NO(x)-N) removal, 91%, 97% and 71% on average at 3-, 2- and 1-d HRT, respectively, was frequently achieved. The mean retentions were 17-47% for chemical oxygen demand, 31-64% for total nitrogen, and 8-15% for total phosphorus for the FR system. Mass balance analysis implied that the detectable DO concentration in the reactor, as low as 0.7mgL(-1), played a very important role in the conversion and removal of NH(3)-N and NO(x)-N, which finally affected the NO(x)-N removal at 3-d HRT.     

Roots of willow (Salix viminalis L.) show marked tolerance to oxygen shortage in flooded soils and in solution culture

Responses to soil flooding and oxygen shortage were studied in field, glasshouse and controlled environment conditions. Established stools ofSalix viminalis L., were compared at five field sites in close proximity but with contrasting water table levels and flooding intensities during the preceding winter. There was no marked effect of site on shoot extension rate, time to half maximum length or final length attained. When rooted cuttings were waterlogged for 4 weeks in a glasshouse, soil redox potentials quickly decreased to below zero. Shoot extension was slowed after a delay of 20 d, while, in the upper 100 mm of soil, formation and outgrowth of unbranched adventitious roots with enhanced aerenchyma development was promoted after 7 d. At depths of 100–200 mm and 200–300 mm, extension by existing root axes was halted by soil flooding, while adventitious roots from above failed to penetrate these deeper zones. After 4 weeks waterlogging, all arrested root tips recommenced elongation when the soil was drained; their extension rates exceeding those of roots that were well-drained throughout. Growth in fresh mass was also stimulated. The additional aerenchyma found in adventitious roots in the upper 100 mm of soil may have been ethylene regulated since gas space development was inhibited by silver nitrate, an ethylene action inhibitor. The effectiveness of aerenchyma was tested by blocking the entry of atmospheric oxygen into plants with lanolin applied to lenticels of woody shoots of plants grown in solution culture. Root extension was halved, while shoot growth remained unaffected. H Lambers Section editor     

Screening for fetal alcohol syndrome in primary schools: a feasibility study

BACKGROUND: This project was undertaken as a feasibility study to determine the possibility of screening for fetal alcohol syndrome (FAS) in early school-age children for epidemiological and interventional purposes. METHODS: All elementary schools in two counties in Washington State were asked to screen first graders for possible FAS. A child was screen positive if found to be growth deficient, to have certain specific facial abnormalities, or have a known history of substantial alcohol exposure in gestation. All screen-positive children were invited to "special diagnostic clinics" for final diagnosis and treatment planning. RESULTS: In County A, virtually all students were screened. In County B only about 25% of children were screened. This difference was related to the number of schools that agreed to participate in the project and the methods employed by each county to obtain parental permission. In each county, only about one-half of the screen-positive children were seen in the special clinics for diagnostic considerations. Only one of the seven children found to have FAS had been diagnosed previously. The minimal prevalence of FAS in County A was 3.1 in 1,000 students. The minimal prevalence of FAS in County B could not be calculated. The most efficient component in the screening process leading to a diagnosis of FAS was finding the specific facial features of the disorder. The diagnosis of FAS was generally helpful in improving educational planning. CONCLUSIONS: This study demonstrated that population-based FAS screening within a school system may be possible, but participation is dependent on local trust and understanding of the project before its inception within the schools and the community at large.     

植物物种丰富度对水培微宇宙中硝态氮去除的影响

在沙基质的人工湿地中植物多样性能够提高污水去除效果,但无基质水生系统中植物多样性对除氮效应的影响还未知.本研究在45个水培微宇宙(53 cm×37.5 cm×18.5cm)中配置了4个物种丰富度梯度(1、2、3和4),并定期供给硝氮为唯一氮形态的模拟污水,氮载荷率为548.5 gN· m-2 ·a-1.结果表明:物种丰富度对出水中氮去除有显著效应,4个种系统出水总无机氮浓度(54.3 mg·L-1)明显低于单种系统(129.0 mg·L-1);物种丰富度显著提高群落生物量,4个种微宇宙系统群落总生物量为1621.6 g·m-2,高于单种群落的1032.7 g·m-2.水培微宇宙的氮平均移除速率为466.8 g N·m-2·a-1,不低于已有报道的全尺度人工湿地的去除能力,同时,4个种的系统比单种系统约高13％,因而可以通过物种多样性配置提高人工湿地效能.     

Seasonal variation in the ultrastructure of the cambium in young stems of willow (Salix viminalis) in relation to phenology

The growth period of Salix viminalis L. (clone 683) plants near Stockholm, Sweden, (59.5°N, 18.3°E) started in April with flowering and ended in October with abscission of the shoot tips. Cell divisions in the vascular cambium started almost two months before sprouting and ceased at about the same time as the elongation growth of the shoots. Phloem cells were apparently produced before flowering, while new xylem production started at the time of flushing. Cytodifferentiation in immature xylem continued until November. Thick-walled cells with protoplasm were observed adjacent to xylem mother cells in the cambium during the winter. The number of radially arranged cells in the cambial zone increased from 3–4 during dormancy to about 18 during the mitotic maximum in July. Seasonal variation was apparent in vacuolization, wall thickness and presence of storage material in the cells. Lipid bodies and protein bodies occurred in both fusiform and ray initials, while starch was observed in ray initials, ray cells and in the phloem. In September the ultrastructure of the cambium showed anatomical features characteristic for both active and dormant cells. Dictyosomes with vesicles and rough ER were present in thick-walled cells that contained lipid bodies and starch granules. Nuclear divisions in the cambium ended in October.     

The concurrent growth of plants and chemical purification of wastewater used as a hydroponic unit

In this study the seedling of a variety of plants were successfully grown hydroponically on raw wastewater obtained from one of the main sewer outfalls in Beirut. In the first phase, a series of experiments was run on a batch system in glass or plastic containers provided with aeration facilities. A continuous-flow system with recirculation was adopted in the second phase. Iron supplementation was applied in all cases to compensate for its deficiency in the raw wastewater used. The immediate and ultimate objectives of the project were threefold: (a) to demonstrate the feasibility of utilizing as a hydroponic medium untreated municipal wastewater having relatively high mean values for BOD and mineral content; (b) to achieve the growth of useful plants on such readily available hydroponic media, thereby saving on fertilizers and scarce water resources; and (c) reclamation of the wastewater through biological purification leading to the gradual depletion of the nutritive constituents. Experimental conditions are described, and the data presented leads to the conclusion that the system is practicable on a laboratory scale. It has great potential for trial on a pilot scale prior to field applications in developing countries suffering from water shortage and hard currency expended on imported fertilizers and wastewater purification facilities.     

A feasibility study of a filtration type autotransfusion device

This paper describes a feasibility study of a disposable autotransfusion device for blood salvage during surgery. The goal was to concentrate hemolyzed blood at 20% hematocrit to 50% while reducing the plasma free hemoglobin concentration from 10 to 1.5 g/l. The device should have a total membrane area of less than 0.6 m² and should be able to process ten 500 ml blood bags. The processing time for each blood bag should not exceed 5 min. The basic idea was to use several polypropylene hollow fibre plasma filters of 0.1 m² in series with saline addition between them. Since the mean pore size is 0.5μ m, anticoagulant and plasma hemoglobin can pass freely across the membrane and their concentration is reduced by dilution. The process was first modelled using mass balance equations for red blood cells and plasma hemoglobin in order to find the best device configuration (number of filters and dilutions). It was found that a three filter system could theoretically meet the requirements, if the last filter had a larger surface area (0.3 m²). Some experiments permitted us to prove the validity of this model and to define fully the third filtration stage. Finally, it was shown that the treatment of a 500 ml blood bag required three filtration stages (whose surface areas were respectively 0.1, 0.1 and 0.3 m²) and the use of 750 ml of saline solution added between the filters. This configuration also offers the possibility of using a vacuum driving force instead of pumps, so that the device becomes completely disposable.     

Removal of nitrate and phosphorus from hydroponic wastewater using a hybrid denitrification filter (HDF)

A laboratory-scale hybrid-denitrification filter (HDF) was designed by combining a plant material digester and a denitrification filter into a single unit for the removal of nitrate and phosphorus from glasshouse hydroponic wastewater. The carbon to nitrate (C:N) ratio for efficient operation of the HDF was calculated to be 1.93:1 and the COD/BOD₅ ratio was 1.2:1. When the HDF was continuously operated with the plant material replaced every 2 days and 100% internal recirculation of the effluent, a high level of nitrate removal (320–5 mg N/L, >95% removal) combined with a low effluent sBOD₅ concentration (<5 mg/L) was consistently achieved. Moreover, phosphate concentrations in the effluent were maintained below 7.5 mg P/L (>81% reduction). This study demonstrates the potential to combine a digester and a denitrification filter in a single unit to efficiently remove nitrate and phosphate from hydroponic wastewater in a single unit.