In vitro physical mutagenesis of giant reed (Arundo donax L.)

Giant reed (Arundo donax L.) is a C₃ perennial, warm‐season, rhizomatous grass of emerging interest for bioenergy and biomass derivatives production, and for phytoremediation. It only propagates vegetatively and very little genetic variation is found among ecotypes, basically precluding breeding efforts. With the objective to increase the genetic variation in this species, we developed and applied a mutagenesis protocol based on γ‐irradiation of in vitro cell cultures from which regenerants were obtained. Based on a radiosensitivity test, the irradiation dose reducing to 50% the number of regenerants per callus (RD₅₀) was estimated at 35 Gy. A large mutagenic experiment was carried out by irradiating a total of 3120 calli with approx. 1×, 1.5× and 2× RD₅₀. A total of 1004 regenerants from irradiated calli were hardened in pots and transplanted to the field. Initial phenotypic characterization of the collection showed correlated responses of biomass‐related quantitative traits to irradiation doses. Approx. 10% of field‐grown clones showed remarkable morphological aberrations including dwarfism, altered tillering, abnormal inflorescence, leaf variegation and others, which were tested for stability over generations. Clone lethality reached 0.4%. Our results show for the first time that physical mutagenesis can efficiently induce new genetic and phenotypic variation of agronomic and prospective industrial value in giant reed. The methodology and the plant materials described here may contribute to the domestication and the genetic improvement of this important biomass species.     

Growth and physiological response of Arundo donax L. to controlled drought stress and recovery

Soil water deficit constrains crop growth more than any other abiotic stress, both per se and in combination with other factors, making drought resistance a key element for the successful exploitation of marginal areas. Arundo donax (Poaceae), is a mesophyte that can adapt to a wide variety of ecological conditions, although its growth can be retarded by a shortage of water. The objective of the present research is to draw a comprehensive picture of the integrated response mechanisms of A. donax to controlled drought conditions, and characterize its ability to recover upon rewatering in terms of photochemical efficiency. Plants were subjected to a gradually applied drought stress for a period of three weeks, after which they were returned to fully hydrated soil conditions for one week. Overall, plant dry weight and key growth parameters were not significantly affected. However, detrimental effects were visible in the form of impaired leaf gas exchange, which influences the performance of photosynthesis, and pre-dawn leaf water potential (pdΨ_W), ?92.1% of net CO₂ assimilation rate and ?0.36 MPa, respectively at the cessation of stress. Nonetheless, a rapid restoration of A. donax physiological functions was observed upon rewatering, testifying to the environmental plasticity of this species.     

Screening of giant reed (Arundo donax L.) ecotypes for biomass production under salt stress

Given its high biomass and plasticity, Arundo donax L. is a promising ligno-cellulosic crop for cultivation in marginal lands in temperate climates. In order to test for adaptation to salinity, growth parameters of several A. donax clones were evaluated under two salt regimes in hydroponics. Mild NaCl stress (50 mM NaCl, 5.6 mS cm^?1 EC, for 10 days) failed to discriminate between ecotypes, while a more severe NaCl treatment (150 mM, 18.8 mS cm^?1 EC, for 21 days) enabled the identification of ecotypes maintaining plant growth under high salinity. Among several biometric parameters, 4th leaf width, and shoot and root DW consistently highlighted differences between ecotypes. Gas-exchange parameters also responded to severe NaCl treatment, while the photosystem efficiency was good, regardless of treatment. The results confirm that A. donax can be considered moderately tolerant to NaCl stress, with variation between ecotypes. Our screening protocol identified ecotypes with higher biomass production under severe NaCl treatment and can be useful for preliminary evaluation of NaCl tolerant clones for increasing productivity under salinity. The detected inter-ecotype variability could also be investigated to identify suitable clones for different environments.     

Origin,diffusion and reproduction of the giant reed (Arundo donax L.): a promising weedy energy crop

Giant reed (Arundo donax) is a promising energy crop of the Mediterranean areas. It has long been associated with humans and has been cultivated in Asia, southern Europe, North Africa and the Middle East for thousands of years. It is a perennial herbaceous plant (Poaceae) found in grasslands and wetlands throughout a wide range of climatic zones. Amplified fragment length polymorphism (AFLP) analysis was used to assess genetic inter and intrarelationships between A. donax and other Arundo species. Furthermore, the development of the sexual apparatus was analysed to understand the basis of sterility in the accession examined. The dendrograms obtained by phenetic and cladistic analysis support the monophyletic origin of giant reed and suggest that it originated in Asia and began to spread into the Mediterranean without traces of hybridisation with the other Arundo species. In particular, samples from Mediterranean areas are characterisd by a lower gene diversity and incidence of rare AFLP fragments indicating that these populations are recent in origin. Moreover, results indicate the occurrence of post-meiotic alterations in the ovule and pollen developmental pathway. Thus, the success of giant reed can be attributed mainly to its rapid clonal spread by rhizome extension, flood dispersal of rhizome and culm fragments.     

Xylose production from giant reed (Arundo donax L.): Modeling and optimization of dilute acid hydrolysis

Statistical modeling and optimization of dilute sulfuric acid hydrolysis of potential energy crop giant reed (Arundo donax L.) has been performed using response surface methodology. Central composite rotatable design was applied to assess the effect of acid concentration, reaction time and temperature on efficiency and selectivity of xylan polysaccharide conversion to xylose. Second-order polynomial model was fitted to experimental data to find the optimum reaction conditions by multiple regression analysis. The monomeric xylose recovery ca. 94% (vs. 93% predicted) was achieved under optimized hydrolysis conditions (1.27% acid concentration, 141.6 °C and 36.4 min), confirming the high validity of the developed model. The low content of glucose (2.7%) and monosaccharide degradation products (0.9% furfural and 0.7% 5-hydroxymethylfurfural) provided a high quality xylose-rich subtract, ready for subsequent biochemical conversion to value-added products. The solid xylan-free residue was easily converted to fermentable sugars resulting in cellulose digestibility of 70% vs. 9% for untreated biomass.     

Evaluation of the phytoremediation potential of Arundo donax L. for nickel-contaminated soil

This study investigates the accumulation and distribution of nickel in Arundo donax L. parts to assess the potential use of this plant in phytoremediation of Ni-contaminated soils. The effect of ethylene diamine tetra-acetic acid (EDTA) and nutrient solution containing NPK on the plant was proped. A 35-day pot experiment was performed in the laboratory and the pots were irrigated with Ni-contaminated solution combined or not with EDTA and NPK. The growth of plants was evaluated at the end of the experiment. The accumulation of Ni was analyzed by atomic absorption spectroscopy (AAS). The obtained results indicate that the plant was able to survive with high Ni content. The growth and the concentrations of Ni in the plant tissues were less affected. In the absence of the amendments, Ni was accumulated in the stems and leaves. However, the addition of NPK significantly reduced Ni concentration in the stems and leaves. The application of EDTA enhanced Ni uptake in roots. The translocation factor (TF) was greater than 1, which categorizes A. donax L. as a great candidate for Ni phytoextraction. A. donax L. is suitable for phytoremediation of Ni. This investigation contributes to the studies on the potential of phytoremediation technologies in Algeria.     

Effect of water deficit on generation time and reproduction of the gall wasp Tetramesa romana,a biological control agent of giant reed (Arundo donax)

Water deficit stress can reduce the reproductive performance of galling insects, but has not previously been studied in the context of mass-rearing of a galling agent on a perennial grass. The effects of water deficit were examined for the wasp Tetramesa romana Walker (Hymenoptera: Eurytomidae), released in the Lower Rio Grande Basin of Texas and Mexico in 2009 and in northern California in 2010 for control of giant reed (Arundo donax (L)) (Poaceae). In one study, water deficit was imposed for 7–10 weeks during gall maturation, ending when adult progeny began to emerge. Aboveground water content was reduced by 1.7% and culm height by 41% in pots receiving one-fourth of normal watering (soil saturation), indicating that water deficit stress occurred. Water deficit did not affect proportion of culms successfully galled or number of progeny produced. However, time to first exit hole appearance was 2–4 days longer and adult wasp generation time 5–7 days longer on galls on water deficit–stressed compared to control culms, thereby reducing the wasp's intrinsic rate of increase. Water deficit imposed only during parent wasp oviposition had no effect on galling success or reproduction. Water deficit thus has a negative effect on rearing by delaying adult emergence and reducing the rate of population increase, even without affecting adult fertility. Mass-rearing should involve well-watered plants, and variable drought conditions in the field should be considered when evaluating T. romana establishment and impact.     

Reduction of riparian arthropod abundance and diversity as a consequence of giant reed (Arundo donax) invasion

The non-indigenous perennial grass, Arundo donax, is an aggressive invader of riparian areas throughout California and many sub-tropical regions of the world, and is hypothesized to provide poorer quality habitat for native wildlife in riparian systems. We sampled aerial and ground-dwelling insects and other terrestrial arthropods associated with Arundo, native willow vegetation (Salix spp.), and mixtures of the two vegetation types during two seasons to determine how Arundo influences invertebrate composition in a low gradient stream in central California. The total number of organisms, total biomass and taxonomic richness of aerial invertebrates associated with native vegetation was approximately twice that associated with Arundo vegetation, while mixed vegetation supported intermediate arthropod levels. Shannon-Weaver (Weiner) diversity associated with native vegetation stands was also higher than that of Arundo vegetation. Ground-dwelling assemblages did not show differences as great as aerial assemblages which are more critical to foraging avian species. These results indicate that vegetation type is a significant factor reducing the abundance and diversity of invertebrates in this, and presumably in many other riparian ecosystems where this invasive species has become a dominant component. Arundo invasion changes the vegetation structure of riparian zones and in turn, may increasingly jeopardize its habitat value for birds and other wildlife whose diets are largely composed of insects found in native riparian vegetation.     

不同生境两种生态型芦苇的抗氧化系统

以分布于甘肃临泽平川乡的两种芦苇生态型——水生芦苇(水芦)和重度盐化草甸芦苇(盐芦)叶片为材料,研究了其抗氧化系统的特征。结果表明,与水芦相比,盐芦中未出现活性氧和MDA(丙二醛)的积累,抗氧化酶SOD(超氧化物歧化酶)、CAT(过氧化氢酶)、POD(过氧化物酶)和APX(抗坏血酸过氧化物酶)的活性显著升高。总抗坏血酸和类胡萝卜素含量在两种生态型芦苇中没有差异,但还原型抗坏血酸和总谷胱甘肽含量在盐芦中显著升高。而且,盐芦的LOX(脂氧合酶)活性比水芦低。这些结果表明,盐芦中有效的抗氧化防御系统对抵抗盐渍胁迫起着重要的作用。此外,盐芦中高活性的Ca^2 -和Mg^2 -ATPase对细胞中过多离子的转运以及避免离子毒害起着重要的作用。     

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%.     

Allocation pattern,ion partitioning,and chlorophyll a fluorescence in Arundo donax L. in responses to salinity stress

Biometric and physiological analyses of salt stress responses were performed in two time-course experiments on giant reed (Arundo donax L). Experiment I evaluated biomass production in plants exposed to 128, 256, 512 mM NaCl for 84 days. For Experiment II, plants grown under 256 mM NaCl were further assessed for chlorophyll a fluorescence, ionic partitioning, and proline content at 14 and 49 days after treatment (DAT). Biomass allocation was affected with all the concentrations of NaCl used from 28 DAT onward. Proline biosynthesis in leaves was more stimulated than that in roots after salt stress. Photosynthetic efficiency of photosystem II (PSII) was not affected by salt stress up to 42 DAT, while 49 DAT plants exhibited a significant reduction of both potential (Φ_PSII) and maximal (F_v/F_m) PSII quantum yield. A. donax resulted a moderately sensitive species in response to 256 and 512 mM NaCl, concentrations that are however higher than that commonly found in most marginal lands (such as 128 mM or lower), where the biomass yield is appreciable, especially in short-term cultivation (56 DAT here). Altogether, this study indicates that A. donax can be considered as a promising and valuable energy crop for exploiting the Mediterranean marginal land.     

Sorption of phosphate onto giant reed based adsorbent: FTIR, Raman spectrum analysis and dynamic sorption/desorption properties in filter bed

A sorption process for the removal of phosphate was evaluated under various conditions using a filter bed packed with giant reed (GR) based adsorbent. FTIR spectrum measurement validated the existence of grafted amine groups in the adsorbent and Raman spectrum displayed the characteristic peaks of different forms of phosphate. The column sorption capacity of the adsorbent for phosphate was 54.67 mg g⁻¹ in comparison with the raw GR of 0.863 mg g⁻¹. Influent pH demonstrated an essential effect on the performance of the filter bed as compared to other influent conditions (flow rates and influent concentrations) and the optimal pH was selected at 5.0-10.0. Eluents of HCl, NaOH and NaCl solutions with concentrations of 0.01-0.1 mol l⁻¹ showed the excellent capacities for desorption of phosphate from the adsorbent, and their elution processes could be finished in 90 min.     

Evaluation of Spanish Arundo scale Rhizaspidiotus donacis (Hemiptera; Diaspididae) survival and fecundity on three new world genotypes of Arundo donax (Poaceae; Arundinoideae)

A pre-release evaluation of survival and fecundity of the arundo scale, Rhizaspidiotus donacis, was conducted on three invasive genotypes of the riparian weed, Arundo donax. The three A. donax genotypes were collected from Laredo, Austin and Balmorhea, TX, which represented the majority of the genotypic diversity found in Texas watersheds. Although R. donacis developed on all three genotypes of the plant, the Austin A. donax genotype, followed by the Laredo genotype, were the most suitable in terms of the size of first-generation immature and adult scale populations that developed after crawler release. Both the Laredo and Austin genotypes of A. donax are likely to be of Spanish origin and are close genetic matches with scale's original host plant genotype in Alicante, Spain. In comparison, survival was lowest on the phylogenetically distant genotype of A. donax from Balmorhea, TX. Although the population size of settled, immature second-generation scales varied in a manner similar to that of the first generation, the fecundity of isolated first-generation females was not significantly different across the three plant genotypes, suggesting that R. donacis is not a genotype specialist in terms of nutrient assimilation for reproduction. Rather, differences in genotype suitability affect rates of success of crawler settling. These results indicate that selection of scale genotype from the native range may have a moderate influence on the success of R. donacis and ultimately the biological control programme.     

Host range of the European leaf sheath mining midge,Lasioptera donacis Coutin (Diptera: Cecidomyiidae), a biological control of giant reed,Arundo donax L.

The fundamental host range of the arundo leafminer, Lasioptera donacis a candidate agent for the invasive weed, Arundo donax was evaluated. L. donacis collects and inserts spores of a saprophytic fungus, Arthrinium arundinis, during oviposition. Larvae feed and develop in the decomposing leaf sheath channel tissue. Thirty-six closely related and economic grass species along with several key habitat associates were evaluated in no-choice tests. L. donacis and its associated saprophyte completed development only on A. donax, in concurrence with published reports from its native range in Mediterranean Europe. The arundo leafminer feeding leads to premature defoliation, constituting a different mode of attack on the host plant as compared to two previously released insects, the arundo wasp and arundo scale, which feed on shoot tips and rhizomes, respectively. Defoliation of A. donax is expected to increase light penetration into stands of A. donax which increases visibility for law enforcement, reduces the survival of cattle fever ticks, and enhance recovery of the native riparian vegetation along the Rio Grande and other habitats where this weed is invasive.     

Impact of the Arundo scale Rhizaspidiotus donacis (Hemiptera: Diaspididae) on the weight of Arundo donax (Poaceae: Arundinoideae) rhizomes in Languedoc southern France and Mediterranean Spain

Arundo donax L. (Poaceae) is native to Mediterranean Europe and invasive in the Rio Grande Basin of North America. Rhizomes from nine sites in France and Spain infested with a candidate control agent, the armoured scale Rhizaspidiotus donacis (Hemiptera: Diaspididae) weighed 50% less than those from nine sites without scale.     

不同Hg浓度下水稻中Hg的分布累积特征

利用土壤盆栽实验方法,研究了土壤中Hg的形态分布,以及不同Hg浓度下,水稻不同生长时期各组织中Hg的分布规律和累积特征.土壤Hg存在的形态为:有机结合态＞残渣态＞＞氧化态＞＞溶解与可交换态≈特殊吸附态,在水稻的生长过程中,土壤中的Hg呈现从残渣态向有机结合态转化的趋势,有机结合态Hg平均占比为61.7％,是土壤Hg最重要的存在形态.Hg在水稻不同部位的浓度分布呈现W根＞W叶＞W茎＞W穗＞W籽粒,分析表明,水稻根、茎和籽粒中的Hg与土壤各形态Hg浓度呈显著或极显著正相关,但在水稻生长后期叶片中Hg与土壤Hg浓度的相关性不显著,叶片Hg与大气Hg进行交换起主要作用,改变了累积状况.随着土壤Hg浓度的增加和生长期的延长,根对Hg的束缚能力逐渐增加,根部Hg累积量增大,所占比例上升,而茎和叶的累积量相对稳定.     

Origin of the invasive Arundo donax (Poaceae): a trans-Asian expedition in herbaria

Background and Aims

The hypothesis of an ancient introduction, i.e. archaeophyte origin, is one of the most challenging questions in phylogeography. Arundo donax (Poaceae) is currently considered to be one of the worst invasive species globally, but it has also been widely utilzed by man across Eurasia for millennia. Despite a lack of phylogenetic data, recent literature has often speculated on its introduction to the Mediterranean region.

Methods

This study tests the hypothesis of its ancient introduction from Asia to the Mediterranean by using plastid DNA sequencing and morphometric analysis on 127 herbarium specimens collected across sub-tropical Eurasia. In addition, a bioclimatic species distribution model calibrated on 1221 Mediterranean localities was used to identify similar ecological niches in Asia.

Key Results

Despite analysis of several plastid DNA hypervariable sites and the identification of 13 haplotypes, A. donax was represented by a single haplotype from the Mediterranean to the Middle East. This haplotype is shared with invasive samples worldwide, and its nearest phylogenetic relatives are located in the Middle East. Morphometric data characterized this invasive clone by a robust morphotype distinguishable from all other Asian samples. The ecological niche modelling designated the southern Caspian Sea, southern Iran and the Indus Valley as the most suitable regions of origin in Asia for the invasive clone of A. donax.

Conclusions

Using an integrative approach, an ancient dispersion of this robust, polyploid and non-fruiting clone is hypothesized from the Middle East to the west, leading to its invasion throughout the Mediterranean Basin.     

Mobilization of minerals and moisture loss during senescence of the energy crops Miscanthus×giganteus,Arundo donax and Phalaris arundinacea in Wales,UK

Miscanthus×giganteus, Arundo donax and Phalaris arundinacea were planted at three sites in Wales, UK, to study the timing of mineral and moisture decline during autumn and winter. The mineral and moisture content at several harvest dates throughout the period were compared with power station threshold levels. All crops showed a highly significant decrease in moisture content in all years with the exception of Arundo which did not lose moisture in the final year. The Miscanthus and Arundo crops remained above the moisture content threshold in all years and would require further drying before combustion. The Phalaris fell below the moisture threshold in January. The Miscanthus leaves fell below the nitrogen threshold before leaf abscission at one site but not at another, although the cane remained mostly within the guideline range. Both its leaves and cane remained above the potassium threshold level. Its leaf sulphur fell within acceptable levels during November and its cane remained below the lowest threshold from November onwards. The Arundo leaves remained above the nitrogen, potassium and sulphur thresholds throughout the period and it did not lose its leaves. The cane nitrogen increased above the threshold during autumn whereas its potassium remained higher than its threshold, with the exception of one sampling date at one site. Its sulphur remained within the guidelines. The Phalaris crop fell to within nitrogen guidelines at one site but increased at the other. Its potassium decreased to below the threshold at both sites in January, and its sulphur content was consistently within the guidelines.     

Movement and aggregation in the fluted giant clam (Tridacna squamosa L.)

Aquaculture has been the traditional focus of tridacnid giant clam research whereas their ecology and behaviour have received much less attention. This study was based on the observation that juvenile fluted giant clams (Tridacna squamosa), when evenly distributed in a tank, will move and aggregate over time. We observed movement in clams ranging from 10 to 313 mm in shell length and ‘climbing’ up the sides of tanks was noted for clams with lengths between 10 and 22 mm. Locomotion also occurred after byssal attachment to the substrate; there was a highly significant association between type of movement (i.e. translation, rotation and no movement) and presence of attachment. Tests for phototaxis were negative. Aggregation was examined by placing clams in regular patterns on grids. After three days in the aquarium and 24 h in the field, their positions were analysed to obtain a statistical parameter for ‘clumpiness’. This was found to be greater in the live clam runs than both random walk and random distribution simulations, suggesting that clams were attracted to conspecifics. The latter was tested by recording clam movement with respect to five types of fixed ‘targets’ (i.e. live clam, fouled clam shell, foul-free clam shell, random inanimate object and none). The test clams moved, non-significantly, towards live clam targets and displayed higher mobility compared to tests with other target objects; a negative correlation between mobility and clam length was also observed. A choice experiment using bidirectional water inflow with clam effluent as one source resulted in clams moving toward the effluent, offering the first direct support for positive chemotaxis among conspecifics in Bivalvia. Together, our results indicate the presence of chemical signalling among clams, leading to movement toward one another and clumping. Aggregation could serve several ecological functions, such as defence against predation, physical stabilisation and facilitation of reproduction. With worldwide decline in natural giant clam densities, the opportunity for conspecific clumping is reduced, and local stocks could be facing increased vulnerability to Allee affects.