Effect of Leafy Spurge (Euphorbia esula) Genotype on Feeding Damage and Reproduction of Aphthona spp.: Implications for Biological Weed Control

North American leafy spurge (Euphorbia esula L.) is genetically diverse and composed of multiple genotypes introduced from several areas of Europe and Asia. Five species of leafy spurge flea beetle (Aphthona spp.) have been introduced as biological control agents for leafy spurge, but were collected in a relatively small region of Europe. Greenhouse and field experiments were conducted to determine if observed variation in feeding preference and reproduction of Aphthona spp. on North American leafy spurge may be due in part to leafy spurge genotype. Leafy spurge genotypes were collected from Austria; Manitoba in Canada; and the states of Montana, Nebraska, North Dakota, South Dakota, and Wyoming in the United States. Leafy spurge genotype affected feeding but not egg laying by Aphthona spp. adults. Aphthona czwalinae/lacertosa fed slightly less in a free-choice test on a genotype from Manitoba (7%) compared to genotypes from Nebraska and North Dakota (14%). Aphthona flava tended to feed less on the Nebraska genotype than any other genotype evaluated. Reproduction of Aphthona spp. was greatly affected by leafy spurge genotype. For instance, A. czwalinae/lacertosa produced 72 adults per plant from a Nebraska genotype compared to 11 to 32 adults per plant from all other genotypes evaluated. Thus, some observed variation in establishment and reproduction of Aphthona spp. can be attributed to leafy spurge genotype.     

Post hoc assessment of an operational biocontrol program: efficacy of the flea beetle Aphthona lacertosa Rosenhauer (Chrysomelidae: Coleoptera), an introduced biocontrol agent for leafy spurge

Mixed populations of Aphthona lacertosa and Aphthona czwalinae were released at more than 50 locations in Alberta in 1997. Two and 3 years post-release, beetle populations were primarily A. lacertosa, with A. czwalinae forming less than 0.5% of the sampled populations. Beetle densities were moderate (10–70 beetles per m²) or high (>70 beetles per m²) at 14% and more than 60% of the sampled sites in 1999 and 2000, respectively. Larger beetles had greater instantaneous egg loads (r²=0.424,P=0.003). In 2000, the largest beetles were found at moderate density sites and there was a significant negative relationship between beetle size and the time taken to accumulate a degree day threshold of 1230 (for females: r²=0.678,P=0.001). Sites with the most rapid accumulation of degree days have the greatest potential for beetle population growth based on potential fecundity. Changes in leafy spurge percent cover, stem density, and canopy height from 1997 to 2000 were assessed across sites with low (<10 beetles per m²), moderate, and high beetle densities in 2000. Sites with high beetle densities had significantly greater reductions of leafy spurge within 5 m of the release point than sites with low beetle densities (P<0.017). Damage caused by the beetles at high-density sites was often visible as a halo-shaped patch of dead leafy spurge stems. The significant overall reduction of leafy spurge within release patches makes A. lacertosa a promising biocontrol agent for leafy spurge in Alberta.     

Phenology of first and peak emergence of Aphthona lacertosa and A. nigriscutis: Two flea beetles introduced for biological control of leafy spurge, Euphorbia esula L.

Nonlinear models were used to estimate first emergence and peak abundance dates for Aphthona lacertosa Rosenhauer and A. nigriscutis Foudras, two flea beetles introduced to control leafy spurge, Euphorbia esula L., in North America. For model development, 26 field sites were sampled for flea beetle abundance at weekly intervals for eight weeks in three western Minnesota counties in 2000, 2001, and 2002. A three-parameter Weibull function, fit to observed cumulative probability distributions, were used to predict accumulated degree-days (ADD) to first emergence. Bias testing indicated the Weibull function provided a useful estimate of first emergence for A. lacertosa (304 ADD, lower developmental threshold 7.5 °C), but failed to produce a useful estimate for A. nigriscutis. A third-order polynomial was used to approximate seasonal abundance and predict peak abundance for each species. Estimated ADD to peak abundance of A. lacertosa was 594 ± 24 (DD > 7.5 °C) and 670 ± 15 (DD > 9.3 °C) for A. nigriscutis. Models were validated with additional data sets from Minnesota, Montana, and North Dakota. Estimated date of peak emergence provided useful predictions of peak emergence for Minnesota and North Dakota, but failed to predict peak emergence in Montana. We speculate that variation in climate and environmental conditions between Midwestern states and Montana were responsible for differing emergence patterns. We conclude that phenology models should be developed regionally to provide useful predictions of peak emergence for land managers. Maps were developed for Minnesota to spatially display predicted dates of peak abundance for A. lacertosa and A. nigriscutis.     

Attack behavior and host utilization ofAphthona chinchihi (Col.: Chrysomelidae), a potential biological control agent ofEuphorbia esula (Euphorbiaceae, leafy spurge) in North America

Aphthona chinchihi Chen was collected in China feeding on leafy spurge (Euphorbia esula L.). Studies were conducted on its host specificity in the laboratory, using field collected adults and their progeny.Aphthona chinchihi can effectively complement the impact of the other natural enemies of leafy spurge established from Europe in the U.S.A. and Canada. The adults feed on leaves and shoots and the larvae, which cause the main damage to the plant, feed on the hypogeous portion of the plant, seriously stressing the plant and preventing its vegetative spread. The host range ofA. chinchihi was studied with tests on adult feeding and oviposition, larval survival and host suitability, using 40 plant species or varieties distributed in 12 families. The experiments demonstrated that it has a high level of specificity. This flea beetle completed its life cycle only on leafy spurge. Also, because of its ecological valence,A. chinchihi has a very good potential as a biocontrol agent in North America.     

The influence of matrix habitat on Aphthona flea beetle immigration to leafy spurge patches

Variation in movement ability by insects among different non-habitat (matrix) types may have important implications for both metapopulation dynamics and weed biocontrol practices. We used a mark-recapture experiment to explore the effects of two different matrix habitats (grass vs shrub) on the ability of two species of Aphthona (Chrysomelidae: Coleoptera) flea beetle to immigrate to patches of the invasive weed, leafy spurge. Using generalized linear models, we compared effects of the matrix habitat types, species and sex on observed immigration probabilities. Our analyses demonstrated that one species (A. nigriscutis) had a much higher immigration probability when moving through a grass-dominated matrix than a shrub-dominated matrix whereas immigration probabilities for the second species (A. lacertosa) were similar in both matrix habitats but significantly lower overall than for A. nigriscutis. Furthermore, A. nigriscutis females were more likely to immigrate to spurge patches embedded in a grass matrix than in shrub, whereas the opposite occurred for males. Our results suggest that metapopulation dynamics may be strongly affected by the type(s) of matrix habitat present on a landscape. These effects also suggest that release strategies for weed biocontrol should be tailored according to the structure of the landscape into which releases are planned. In addition, even closely related species can have significantly different movement abilities which will also affect release strategies.     

Is two company or a crowd: How does conspecific density affect the small-scale dispersal of a weed biocontrol agent?

To predict the growth and spread of an insect population introduced for the biological control of weeds, one must first understand the factors affecting the movement of individuals in the population. The purpose of this study was to determine how the dispersal rate of Aphthona lacertosa (Rosenhauer) (Chrysomelidae) was affected by conspecific density and by the characteristics of leafy spurge (Euphorbia esula L.: Euphorbiaceae) in patches where these beetles feed. In 2002 in Manitoba and in 2003 in Alberta, Canada, between 200 and 2500 insects were released in small patches (<10 m²) of spurge. The number and location of beetles within patches was monitored over subsequent days. In 1 m² plots within patches, spurge ramet density, the proportions of vegetative and reproductive ramets, and ramet height were measured. In both years, beetle movement within patches and emigration from patches, was not affected by conspecific density. In Manitoba in 2002, beetles aggregated non-randomly on either vegetative or reproductive ramets within plots, but plot characteristics were not related to the formation of aggregations. In Alberta in 2003, plots in which beetles aggregated had significantly higher spurge density but did not differ in either the proportion of vegetative ramets or in the amount of non-spurge vegetation. These results suggest that density-dependent dispersal does not limit the population's ability to reach densities up to 2500 beetles/m².     

Evidence for the influence of conspecific chemical cues on <Emphasis Type="Italic">Aphthona nigriscutis</Emphasis> (Coleoptera: Chrysomelidae) behaviour and distribution

Although the distribution of biological control agents may have a significant effect upon their impacts, the mechanisms regulating these distributions are often unknown. Such is the case with Aphthona nigriscutis, a classical biological control agent of leafy spurge in North America. These beetles assume aggregated distributions at some sites but disperse rapidly at others. The potential influence of plant and insect-factors upon aggregation and dispersal was investigated to try to explain these observations. Male beetles produce a putative aggregation pheromone. Responses of conspecifics to male-associated cues are greater when beetles are feeding on host plants. Densities of beetle groups greatly impact their attractiveness. Males are more sensitive to dispersal cues and females are more sensitive to congregation cues.     

Biology and host specificity ofChamaesphecia hungarica andCh. Astatiformis (Lep.: Sesiidae) two candidates for the biological control of leafy spurge,Euphorbia esula (Euphorbiaceae) in North America

Leafy spurge (Euphorbia esula (s.1.)) is an herbaceous perennial and serious weed of Eurasian origin that has been accidentally introduced into North America. The two European root-boring mothsChamaesphecia hungarica andCh. astatiformis are univoltine and overwinter as mature larvae. Both species have a lower survival rate on leafy spurge than on their field hosts, and thus are not optimal candidates for the biological control of leafy spurge. However, the rate of larval development and larval growth on the target weed and on the two field hosts is nearly the same. The experimental host range of both species is restricted to a few species in the subgenusEsula within the genusEuphorbia. The two species occupy different habitats in the steppe biome and are targeted for similar leafy spurge habitats in North America.     

Energy potential of leafy spurge (Euphorbia esula)

Leafy spurge (Euphorbia esula) is a noxious, perennial weed that infests pastures, rangeland and waste areas in the northern Great Plains. The objective of this study was to determine the productive potential of this species when grown under optimum agronomic conditions. Plants were fertilized and irrigated. Oil, hydrocarbon, total protein, and dry-weight production were measured on 3 harvest dates. Calorimetric analyses were performed to determine the potential of leafy spurge as a fuel crop. The hydrocarbon content of 12 strains of leafy spurge was determined to measure genetic variability for this trait. The addition of fertilizer doubled dryweight production but did not affect percent oil or hydrocarbon content. Oil and hydrocarbon production averaged 6.8 and 0.6% on a plant dry-weight basis. Maximum production of plant biomass, protein, and hydrocarbon was obtained from a mid-July harvest. Oil content increased later in the growing season. The total protein content of leafy spurge averaged 12%. Whole-plant biomass had a caloric value of 4,407 cal/g while the oils contained 10,019 cal/g. Leafy spurge hay can produce 4 times more energy per year than wheat straw; therefore, the immediate potential of leafy spurge whole-plant biomass as a locally grown fuel crop for home-heating purposes is suggested.     

Phytotoxic Allelochemicals From Roots and Root Exudates of Leafy Spurge (Euphorbia esula L.)

Invasive plants are a widespread problem but the mechanisms used by these plants to become invasive are often unknown. The production of phytotoxic natural products by invasive weeds is one mechanism by which these species may become successful competitors. Here we conducted a bioactivity-driven fractionation of root extracts and exudates from the invasive plant leafy spurge (Euphorbia esula L.), and structurally characterized jatrophane diterpenes and ellagic acid derivatives. Ellagic acid derivatives and one of the jatrophane diterpenes, esulone A, have been previously reported from leafy spurge, but another of the jatrophane diterpenes, kasuinine B, has not. We show that these compounds are phytotoxic but affect plants in different ways, either inducing overall plant necrosis or reducing root branching and elongation.Key Words: phytotoxicity, allelochemicals, roots, root exudates, jatrophane diterpenes, kansuinine B, ellagic acid derivatives, leafy spurge, Euphorbia esula, Arabidopsis thaliana     

Nature,nurture, and vegetation management: Studies with sheep and goats

Diet selection and preference by grazing animals are determined by genetic and environmental factors (i.e., nature and nurture) that interact and affect their efficacy for managing vegetation as targeted grazers. The effect of rearing environment on the consumption of leafy spurge by sheep and goats was investigated. We hypothesized that although rearing environment will affect the preference for chemically defended plants ultimately, the inherent ability to detoxify or eliminate phytotoxins will limit an animal’s preference for them. The objective of this study was to determine if sheep would consume more of the invasive weed leafy spurge (Euphorbia esula) if they were raised by goat compared to sheep raised by sheep and goat raised by goat. Sheep were raised on leafy spurge-infested pastures by either their ewe (S) or a goat doe (FS) on which they were fostered within 24 hours of birth and parturition of lamb and doe, respectively. Does that fostered lambs also raised their own goat offspring (G) such that the same doe raised the FS and G animals. The rearing environment’s effect on leafy spurge consumption was tested the following growing season by simultaneously grazing all animals on the same leafy spurge-infested rangeland and estimating percentage leafy spurge in their diet with either fecal near-infrared spectroscopy (f.NIR) or bite count. Goats consumed more leafy spurge as determined by either f.NIR (62.8%, P < 0.06) or bite count (71.9%, P < 0.01) than FS (35.2 % f.NIR, 39.3% bite count) or S (10.1 % f.NIR, 18.2% bite count). The FS consumed over twice as much leafy spurge as S and were numerically intermediate to G and S for leafy spurge consumption but not significantly different from the S sheep, most likely because one FS sheep did not eat leafy spurge during the evaluation period. Because leafy spurge is aversive to sheep but not goats, higher leafy spurge consumption by FS sheep is hypothesized to result from inoculation of their rumen microbes with microbes from the does capable of denaturing aversive phytotoxins in leafy spurge. The higher consumption of leafy spurge by G compared to FS shows that genetically determined physiological differences influence an animal’s ability to ameliorate phytotoxins and determine the upper limit of an animal’s preference for a chemically defended plant. It also indicated that in addition to the animal’s genome, the genome of an animal’s microbiome, which the mother may influence, can play an important role in diet selection.     

Hierarchical Bayesian methods estimate invasive weed impacts at pertinent spatial scales

Invasive weed impact estimates are needed to determine whether or not weeds warrant costly control measures. Typically, land managers seek local weed impact estimates (e.g. ranches, parks) and policy-makers want to know how weeds are impacting entire regions. Our goal was to provide local and regional impact estimates for a ubiquitous invasive weed: leafy spurge (Euphorbia esula L.). The specific impacts we looked at related to desired species biomass production, livestock carrying capacities, and grazing land values. Our basic approach was to use an empirical model that characterizes weed biomass across the landscape in combination with another empirical model that predicts weed impact from weed biomass. Our investigation revealed that, without on-site plant biomass data, site-specific leafy spurge impacts are highly uncertain. Supplementing our general predictive model with small quantities of on-site data increased precision considerably. For the 17-state region we considered, 95% Bayesian credibility intervals indicated leafy spurge reduces cattle carrying capacities by 50–217 thousand animals a year and reduces grazing land values by 8–34 million dollars a year. Additional plant biomass data from randomly selected, leafy spurge-infested sites would shrink these fairly wide intervals.     

Effects of ecotypes and morphotypes in feedstock composition of switchgrass (Panicum virgatum L.)

Switchgrass (Panicum virgatum L.) is a C4 grass with high biomass yield potential and is now a model species for the Bioenergy Feedstock Development Program. Two distinct ecotypes (e.g., upland and lowland) and a range of plant morphotypes (e.g., leafy and stemmy) have been observed in switchgrass. The objective of this study was to determine the influence of ecotype and morphotype on biomass feedstock quality. Leaf and stem tissues of leafy and stemmy morphotypes from both lowland and upland ecotypes were analyzed for key feedstock traits. The leaf : stem ratio of leafy morphotype was more than 40% higher than the stemmy morphotype in both upland and lowland ecotypes. Therefore, the stemmy morphotype has significant advantages over leafy morphotype during harvesting, storage, transportation and finally the feedstock quality. Remarkable differences in feedstock quality and mineral composition were observed in switchgrass genotypes with distinct ecotypic origins and variable plant morphotypes. Lignin, hemicelluloses and cellulose concentrations were higher in stems than in the leaves, while ash content was notably high in leaves. A higher concentration of potassium was found in the stems compared to the leaves. In contrast, calcium was higher and magnesium was generally higher in the leaves compared to stems. The upland genotypes demonstrated considerably higher lignin (14.4%) compared with lowland genotypes (12.4%), while hemicellulose was higher in lowland compared with upland. The stemmy type demonstrated slightly higher lignin compared with leafy types (P < 0.1). Differences between the ecotypes and morphotypes for key quality traits demonstrated the potential for improving feedstock composition of switchgrass through selection in breeding programs.     

Field and laboratory biology of the stem-feeding beetle Thamnurgus euphorbiae (Küster) (Col., Scolytidae) in Italy, a potential biological control candidate of leafy spurge in the USA and Canada

Thamnurgus euphorbiae overwinters as adults in dry stems of Euphorbia characias until the end of March before exiting through circular holes cut with their mandibles. Females and males appeared in the field at the same time, and the first pairs were observed on 28 March on new stems of E. characias having the open‐flower at the beginning of its development. Throughout the rest of April it was possible to find and collect the fairly abundant adults. Mating occurs on new stems of the host plant and females walk up towards the top of the stem and start to mine the centre of it in order to penetrate. Oviposition occurs in new stems and eggs are laid singly along the interior of the stem; females laid 35–85 eggs. Upon hatching, larvae started feeding on vascular bundles and the inner cortex. There are three larval instars, and pupation occurred in the stem of the host plant; this insect is univoltine. Thamnurgus euphorbiae has been accepted by the Technical Advisory Group for Biological Control Agents of Weeds for release as a biological control agent of leafy spurge in the US. Prior to this study the biology of T. euphorbiae was unknown.     

Changes in miombo woodland structure under different land tenure and use systems in central Zambia

Aim Population pressure and communal land ownership are often perceived as serious threats to forest conservation in savanna woodlands of central and southern Africa. I aimed at testing the hypothesis that the rate of miombo woodland recovery after clearing and re‐growth structure are determined by land tenure and use. Location Miombo woodland under customary, leasehold, forest reserve and national park on ten permanent and temporary sites was studied in central Zambia. Two sites were in mature woodland and eight sites were in re‐growth miombo ranging in age from 1 to 30 years. Methods I enumerated and measured girth at breast height (1.3 m above ground) of trees/stems in sixty‐four 20 × 10 m plots in 1982, 1986 and 2000 at six sites and annually from 1990 to 2001 at four sites to determine stem density and status (live, dead or cut) and wood biomass. A total of 239 trees were cut, wood biomass measured and the data used to develop equations for estimating wood biomass on study plots. Distance between each study site and the nearest human settlement was estimated during each sampling period using aerial photographs, topographical maps and the global positioning system. Results Land tenure was responsible for significant differences in stem density, wood biomass and rate of biomass accumulation in re‐growth following clearing of mature miombo woodland. Although stem density was highest on customary land, wood biomass and accumulation rate were lowest. The highest biomass was on plots in forest reserves, with intermediate values for leasehold and national park. Fire was responsible for tree mortality at all the study sites and its impact was highest at a site in a national park. Sites close to human settlements had the highest density of cut stems but this activity did not significantly reduce wood biomass. Rate of woodland recovery was higher on sites cleared in the 1970s than on sites cleared in the 1990s, irrespective of age of re‐growth. The development of the first, second and third re‐growths following successive woodland clearing in 1972, 1981 and 1990, respectively, was not significantly different, except for stem density which was highest in the second re‐growth. Analysis of interactions between five land tenure and use factors (independent variables) and re‐growth structure revealed that 52% (P=0.0000) of the variation in stem density was because of re‐growth age and decade in which the woodland was cleared while distance to human settlements and age of re‐growth explained 42% (P=0.0000) of the variation in wood biomass. Individually, distance to human settlements explained 25% (P=0.0000) of the variation in wood biomass accumulation rate. Conclusion The results supported the hypothesis that rate of miombo woodland recovery and structure were influenced by land tenure and use. However, analysis of interactions between factors revealed that use related factors (i.e. decade in which woodland was cleared and distance to human settlements) and re‐growth related factors (age and type of re‐growth) were more important than land tenure per se in explaining variation in miombo recovery. The conclusion from these results is that regulation of land use is more important than change in land tenure to the proper management of miombo woodland.     

Characterization, expression and function of DORMANCY ASSOCIATED MADS-BOX genes from leafy spurge

DORMANCY ASSOCIATED MADS-BOX (DAM) genes are related to AGAMOUS-LIKE 24 and SHORT VEGETATIVE PHASE genes of arabidopsis and are differentially regulated coordinately with endodormancy induction and release in buds of several perennial plant species. DAM genes were first shown to directly impact endodormancy in peach where a deletion of a series of DAM resulted in loss of endodormancy induction. We have cloned and characterized several MADS box genes from the model perennial weed leafy spurge. Leafy spurge DAM genes are preferentially expressed in shoot tips and buds in response to cold temperatures and day length in a manner that is relative to the level of endodormancy induced by various environmental conditions. Over-expression of one DAM gene in arabidopsis delays flowering. Additionally, we show that at least one DAM gene is differentially regulated by chromatin remodeling. Comparisons of the DAM gene promoters between poplar and leafy spurge have identified several conserved sequences that may be important for their expression patterns in response to dormancy-inducing stimuli.     

An efficient method for <Emphasis Type="Italic">in vitro</Emphasis> regeneration of leafy spurge (<Emphasis Type="Italic">Euphorbia esula</Emphasis> L.)

Leafy spurge (Euphorbia esula L.) is a perennial, invasive weed used as a model to study invasive plant behavior, because molecular tools (such as a deep expressed sequence tag database and deoxyribonucleic acid microarrays) have been developed for this species. However, the lack of effective in vitro regeneration and genetic transformation systems has hampered molecular approaches to study leafy spurge. In this study, we describe an efficient in vitro regeneration system. Three highly regenerative lines were selected by screening the in vitro regeneration capabilities of stem explants of 162 seedlings. The effects of various culture conditions on in vitro regeneration were then evaluated based on explant competence to form calluses and shoots. High rates of shoot regeneration can be obtained using a growth medium containing 1× woody plant basal medium and 1× Murashige and Skoog (MS) basal salts, 1× MS vitamins, 1.11 μM 6-benzylaminopurine, 1.97 μM indole-3-butyric acid, and 3% sucrose, pH 5.6–5.8. After 30 d culture, multiple shoots formed either directly from the stem or indirectly from the callus. This method is a requisite for the development of genetic transformation systems for leafy spurge and may be used to develop in vitro regeneration techniques for other species in the Euphorbiaceae.