Weed–barley interactions affect plant acceptance by aphids in laboratory and field experiments

Increased botanical diversity can lead to suppression of insect pests. One route by which botanical diversity is increased in crops is through the occurrence of weeds, which increasingly interact with crop plants as organic production expands. However, the mechanisms by which this might affect insect herbivores are poorly understood. This study examined whether volatile chemical interactions between weeds and barley, Hordeum vulgare L. (Poaceae), can affect plant acceptance by the bird cherry oat aphid, Rhopalosiphum padi L. (Hemiptera: Aphididae). In laboratory experiments, exposure of barley to volatiles from Chenopodium album L. (Amaranthaceae) and Solanum nigrum L. (Solanaceae) resulted in significantly reduced aphid acceptance compared with unexposed plants. In a series of field experiments in which the occurrence of weeds was manipulated in plots of barley, significantly lower aphid acceptance was recorded on barley plants grown in plots with C. album compared with barley plants in weedless plots. The results indicate that interaction between weeds and barley can affect aphid–plant interactions in the field as well as in the laboratory and provide further evidence that the effects of chemical interactions between visibly undamaged plants can extend to higher trophic levels.     

Barley exposed to aerial allelopathy from thistles (Cirsium spp.) becomes less acceptable to aphids

Abstract.  1. Recent studies have shown that plant–plant interaction via chemicals (allelopathy) can affect insects. Here the effects on aphid acceptance of barley after exposure to volatiles and root exudates from two common weeds, the thistles Cirsium arvense and Cirsium vulgare , were investigated.
2. Settling by bird cherry-oat aphid, Rhopalosiphum padi , was significantly reduced on barley plants that had been exposed to volatiles from Cirsium species for 5 days. Settling by Sitobion avenae was also reduced on Cirsium -exposed plants, whereas settling by Metopolophium dirhodum was not.
3. In olfactometer tests, Cirsium -exposed barley was significantly less attractive to R. padi than was unexposed barley, indicating that exposure causes a change in the volatile profile of barley.
4. Exposure of barley to root exudates from Cirsium species had no effect on R. padi settling.
5. The results lend weight to the theory that the effects of plant–plant allelopathy can extend to higher trophic levels.     

Effect of Herbivore Density, Timing of Attack and Plant Community on Performance of Creeping Thistle Cirsium arvense (L.) Scop. (Asteraceae)

Creeping thistle, Cirsium arvense (L.) Scop. (Asteraceae), is one of the most serious weeds in ecological compensation areas (within-field or border refugia) in Europe. Since conventional weed control measures are restricted in compensation areas, augmenting indigenous agents for biological control of the weed may be a feasible alternative. In this paper, we studied the effect of density of the shield beetle, Cassida rubiginosa Muller (Coleoptera, Chrysomelidae), and two vegetation types typical for ecological compensation areas, on the performance of creeping thistle plants in an open field experiment. Early-season larval feeding had no measurable effect on creeping thistle growth, while late-season feeding significantly reduced shoot growth. These findings were attributed to higher feeding rates of the herbivores at higher ambient temperatures late in the season. Defoliation had a strong effect on the above-ground performance of C. arvense plants, but not on the below-ground performance. In contrast, the plant community affected all below-ground performance parameters measured, but only some of the above-ground performance parameters of creeping thistle. A combination of high levels of plant competition and herbivory by C. rubiginosa larvae led to 50% mortality in C. arvense plants during the growing season. We conclude that augmentation of indigenous herbivores of C. arvense in combination with breaking up the root system by tillage and the establishment of a highly competitive plant community of beneficial herbs may be a feasible way to control this problematic weed in ecological compensation areas.     

Dropping behaviour of larvae of aphidophagous ladybirds and its effects on incidence of intraguild predation: interactions between the intraguild prey, Adalia bipunctata (L.) and Coccinella septempunctata (L.), and the intraguild predator, Harmonia axyridis Pallas

Abstract.  1. Two experiments were performed in the laboratory to assess the behaviour of dropping from a host plant as a defence against intraguild predation in aphidophagous ladybird larvae.
2. In the first experiment, encounters were observed on bean plants between fourth instars of the intraguild predator species, Harmonia axyridis , and first instars of two other ladybird species, Adalia bipunctata (L.) and Coccinella septempunctata (L.). The percentages of first instars of the latter two species that dropped from the plant in response to attack differed dramatically, with 47.5% of C. septempunctata first instars dropping vs. 0% of A. bipunctata .
3. In the second experiment, first instars of A. bipunctata or C. septempunctata and a fourth instar of H. axyridis were allowed to forage together on bean plants for 3 h. During this time, 44.3% of C. septempunctata larvae dropped from the plant, but less than 2% of A. bipunctata larvae did so. In contrast, 95.0% of A. bipunctata larvae fell victim to intraguild predation by H. axyridis vs. only 54.5% of C. septempunctata larvae.
4. The significance of dropping behaviour of ladybird larvae as a defence against intraguild predation, and the relationship of dropping behaviour to species-specific habitat affinity of ladybirds, is discussed.     

Above‐ and Belowground Trophic Interactions on Creeping Thistle (Cirsium arvense) in High‐ and Low‐Diversity Plant Communities: Potential for Biotic Resistance?

The capacity of local communities to control introduced plants is called biotic resistance. Biotic resistance has been almost exclusively tested for plant competition and above ground herbivores and pathogens, while neglecting root herbivores and soil pathogens. Here, we present biotic resistance by above- and below ground herbivores in concert, and relate the abundance of the plant enemies to the species diversity of the local plant communities. The study was carried out in a 7-year-old biodiversity field experiment. We used creeping thistle (Cirsium arvense) as a model, and quantified sap-sucking herbivores: above ground aphids, their antagonists, and root-feeding nematodes. As plant diversity treatments, we used field plots sown with high (15 plant species, HSD) or low (4 plant species, LSD) diverse seed mixtures in 1996 and that were not weeded. Creeping thistle became established spontaneously at the start of the experiment. In 2002, in HSD plots, 90 % of the plant community was made up by 11 species, compared to seven species in LSD plots. No differences were found for C. arvense abundance or biomass. Above ground, three aphid species were found on C. arvense-Uroleucon cirsii, Aphis fabae, and Macrosiphum euphorbiae, but the latter was found only in low densities. Significantly more aphid species were found on individual plants in HSD plots. Moreover, in HSD plots, on average 10 % of aphids were parasitized, while no parasitism was observed in LSD plots. In the root zone of C. arvense, significantly more nematodes were found in HSD than in LSD plots, and a significantly higher proportion of those nematodes were plant parasites. The dominant plant parasitic nematode in both treatments was Paratylenchus. We conclude that biotic resistance by natural enemies may be enhanced by plant species diversity, but that above- and below ground sap-sucking herbivores do not necessarily have to respond similarly to the diversity of the surrounding plant community.     

Approaches to reconstruction of early Celtic land-use in the cent ral Neckar region in southwestern Germany

In four early Celtic settlements in the central Neckar region of Baden-Württemberg (Germany), 250 archaeobotanical samples were collected and studied for their plant macrofossils.Hordeum vulgare L. (hulled barley) andTrilicum spella L. (spelt wheat) could be recognized as the main crops, as well asPanicum miliaceum L. (millet) in some places. Legumes andLinum usitatissimum L. (flax) were important crops. Typical field weeds dominated the wild plant remains, but some present-day grassland taxa were abundantly and frequently found. The analysis of the data set of finds of the cereal remains, field weeds and grassland plants was done by Correspondence Analysis, but no significant differences between the various groups were found. Samples dominated by remains of grassland taxa, which could indicate that there were hay meadows, were absent. Consideration of plant macrofossil results as well as archaeological, osteological and pollen analytical data shows that there is likely to have been a land management system of arable fields and grassland. The densely settled fertile central Neckar region could therefore have been intensively utilised, using all available resources. Permanent grasslands in the form of pasture on river floodplains and on dry soils were certainly present. Hay meadows, on the other hand, were not identified. Cultivated plants such asFicus carica L. (fig) andVicia ervilia (L.) Willd. (bitter vetch) as well as some field weeds suggest connections with the Mediterranean lands.     

Studies in the diagnosis of mineral deficiency; the composition of weed leaves in relation to potassium deficiency in barley

Leaves of five weed species (Brassica sinapis, Cirsium arvense, Polygonum aviculare, P. convolvulus and Potentilla reptans) from fertilizer trials on barley at four sites in Hampshire were analysed with a view to using their composition in forecasting the response of barley to fertilizers. The samples were gathered from as many of the 108 plots as possible, and were analysed spectrographically for calcium, iron, magnesium, manganese, potassium and sodium.
Marked differences in composition between the five species were recorded, the most noteworthy being the high sodium content of Brassica sinapis and Cirsium arvense and the high manganese content of Potentilla reptans. There were also marked differences between the four sites, but these were not uniform as between the different species, and often failed to agree with those observed for barley.
Superphosphate applications decreased the manganese content of the weeds in many cases, and increased their calcium content. Muriate of potash increased their potassium content, but tended to decrease that of magnesium and sodium. The only general effect of sulphate of ammonia on the composition of the weeds was a decrease in iron content.
Except in Cirsium arvense , the potassium content of weed leaves was correlated with that of barley on the same plot if differences within a site only were considered. Differences between sites were not correlated in this way. The correlation between potassium content of weed leaves and the response of barley to muriate of potash application was worthy of note only in Polygonum convolvulus , and even in this case the correlation of site differences did not reach significance. It is tentatively suggested that increases in the grain yield of barley as a result of muriate of potash application are likely to occur only where the leaves of P. convolvulus contain less than 1.83% potassium.     

Interactions between introduced and native predatory ladybirds (Coleoptera, Coccinellidae): factors influencing the success of species introductions

Abstract.  1. Why some exotic species invade successfully while others do not is poorly understood. This study focuses on the inability of the exotic predatory ladybird Adalia bipunctata (L.) (Coleoptera, Coccinellidae) to establish and spread in Japan. In particular, this study considers the interactions between A. bipunctata and three native Japanese ladybird species, Harmonia axyridis Pallas, Coccinella septempunctata Linn., and Propylea japonica Mulsant.
2. The number of aphids occurring on the host plants declined dramatically over the course of larval development in the presence of H. axyridis or C. septempunctata . In contrast, many aphids remained at the end of the experiment when only A. bipunctata or P. japonica was present.
3. The survivorship of A. bipunctata from second instar to adult was significantly reduced in the presence of H. axyridis , but not in the presence of C. septempunctata or P. japonica . Most deaths were the result of intra-guild predation when H. axyridis or C. septempunctata larvae were present. Larvae of H. axyridis and C. septempunctata suffered very high mortality in this experiment, but survived better when co-occurring with A. bipunctata than when occurring only with conspecifics. In contrast, P. japonica survived better when occurring with conspecifics than with A. bipunctata .
4. The finding that larvae of the exotic species A. bipunctata died at high rates from intra-guild predation by the native Japanese ladybirds H. axyridis and C. septempunctata may bear on the question of why A. bipunctata has not established more successfully in Japan whereas H. axyridis and C. septempunctata have been very successful in establishing themselves in North America.     

Crop presence,but not genetic diversity,impacts on the rare arable plant Valerianella rimosa

Background: Intensive farming affects farmland biodiversity, and some arable plants in particular. Increasing crop genetic diversity can increase crop productivity or resilience and could also benefit rare arable plants.

Aims: We examined whether barley presence, sowing density and genetic diversity impacted the rare plant Valerianella rimosa and explored possible underlying mechanisms.

Methods: In a field study near Dundee, Scotland, we sowed plots of five single barley genotypes, and all five genotypes combined, at three densities; we also had barley-free plots. Valerianella seeds were sown into half of all plots. Measured responses included early-season cover and harvest biomass of barley and common weeds, abiotic parameters (soil moisture, light) and establishment, biomass and seed production by V. rimosa.

Results: Barley presence promoted V. rimosa establishment early in the growing season, but without barley density or genetic diversity effects. By harvest, the impact of barley presence on V. rimosa abundance was lost; there were no effects on Valerianella seed production. Barley negatively impacted common weeds, but V. rimosa did not benefit from any indirect facilitation by barley, being bigger without barley.

Conclusions: Early beneficial effects of barley on V. rimosa abundance appear offset by late-season competition. However, limited impacts of barley on V. rimosa reproductive success, and negative impacts on common weeds, indicate crops might play a role in conservation management of rare arable plants by creating space in the weed community.     

Herbicidal potential of catechol as an allelochemical

Catechol is an allelochemical which belongs to phenolic compounds synthesized in plants. Its herbicidal effects on weed species; field poppy (Papaver rhoeas), creeping thistle (Cirsium arvense), henbit (Lamium amplexicaule) and wild mustard (Sinapis arvensis) were investigated using wheat (Triticum vulgare) and barley (Hordeum vulgare) species as control plants. In comparison to 2,4-D (a common synthetic herbicide), 13.64 mM of catechol have been found to have a strong herbicidal effect, as effective as 2,4-D on field poppy weed by killing it, and a suppressive herbicidal effect on the other weeds by inhibiting their growth significantly. Concerning all the weeds, in general, elongation of the shoot was affected more negatively than that of the root. Fresh weights of the weeds were decreased by catechol significantly only in field poppy but not in other weeds. The study reveals that catechol is a potent inhibitor of growth of the weeds and therefore it can be evaluated as a herbicide for future weed management strategies.     

Host Specificity of Altica carduorum Guer. Chrysomelidae: Coleoptera , a Defoliator of Cirsium arvense L. Scop. Asteraceae from North-western China

Traditionally, insects are not approved for release in North America as agents for the biological control of weeds if the larvae complete development on desirable plants in no-choice tests. This criterion excludes the use of the leaf-feeding beetle Altica carduroum Guer. since it can complete development on all Cirsium spp., including rare North American species, although in nature it is only known from the target weed Cirsium arvense. Host suitability is the result of a sequence of selection factors, and this study shows that most discrimination between thistle species is the result of host finding and egg production, and not larval developmental ability. In no-choice tests, the larvae completed development on 18 Cirsium species in five sections of the genus that were tested and on Silybum marianum, but not on other genera in the tribe Cardueae or more distantly related Asteraceae. The adult food consumption on C. arvense was two to eight times greater than on other Cirsium species, many of which, such as C. pitcheri and C. drummondii, received no feeding by over 50 of the beetles in the first 24 h. The best North American thistle for egg production was C. scariosum, but this was only 28 of that on C. arvense. The preference for C. arvense in multiple-choice tests increased with the size 2 of the arena. In the largest, an area of 6 m , 80 of the beetles and eggs were found on C. arvense and none were found on plants outside the subtribe Carduineae. Thus, as theory predicts, host discrimination between thistle species takes place at earlier stages than suitability for larval development, and this should be considered when determining the safety of candidate agents.     

Nutritional resources used by the invasive maize pest Diabrotica virgifera virgifera in its new South-east-European distribution range

Food utilization by adults of the invasive maize (Zea mays L.) (Poaceae) pest western corn rootworm (WCR; Diabrotica virgifera virgifera LeConte) (Coleoptera: Chrysomelidae) was investigated in the south‐eastern part of its new European distribution range. At weekly intervals over a 10‐week period, 10 beetles per field were collected from six fields that had a high abundance of flowering weeds and six fields with a low abundance of flowering weeds, with the aim of understanding adult feeding behaviour in Europe and comparing this behaviour with North American WCR. Gut content analysis was performed to determine the use of maize tissue and weed pollen with regard to maize phenology. Furthermore, all pollen found within the gut was quantified and identified to plant species level. The use of maize tissue by adult WCR changed over time according to maize phenology. Pollen originating from plants other than maize was used more frequently as the maize matured. Adults fed on pollen from 19 of 25 different plant species found in maize fields and showed a preference for the pollen of specific weeds. Pollen from weed species was found more often in beetles from fields with a high abundance of weeds compared to beetles from fields with a low abundance of weeds. Females consumed greater amounts of pollen than males, the latter feeding on a wider diversity of host plants. The pollen resources used by adult WCR in Hungary were more diverse compared to WCR in the USA, which may contribute to the invasion success of WCR in Europe.     

Determination of fungal pathogens of common weed species in the vicinity of Tokat, Turkey

This study was carried out to determine the fungal pathogens on Chenopodium album L., Cirsium arvense (L.) Scop., Convolvulus arvensis L., Cynodon dactylon (L.) Pers., Delphinium consolida L., Portulaca oleracea L., Rumex crispus L., Solanum nigrum L., Sorghum halepense (L.) Pers. and Xanthium strumarium L. which were common weed species of agricultural areas. Surveys were conducted in May-June and August-September in 2004-2005 growing seasons. During the surveys density and frequency of the above mentioned weed species were also determined and number of infected plants was counted in each sampling area. Infected weed samples were collected from each sampling point and brought to the laboratory in polyethylene bags and the pathogens were identified at genus or species level. As a result of two year surveys, ten fungal pathogens were determined on eight weed species. The most important fungal pathogens determined on common weed species were as follow; Peronospora farinosa (Fr.) Fr. on C. album, and Septoria convolvuli DC., Erysiphe convolvuli DC., and Puccinia punctiformis (Strauss) Roehrl. on C. arvensis. These fungal diseases were observed mainly on the weeds located at the borders of fields. Infection rates of these pathogens reached up to 21.2% in some of the survey areas. Further studies should be conducted to evaluate the efficacy of these pathogen under in vitro and in vivo conditions.     

中国香港新记录的十个分类单元

中国香港位于热带北缘,其植被在历史上曾遭到大规模破坏,近几十年来逐渐恢复。香港植物标本采集有很长的历史,迄今为止已记录的维管植物超过2 170种,然而新种和新分布仍在不断增加。在最近的野外工作中,作者发现10种在香港新记录到的维管植物,分别是唇边书带蕨[Haplopteris elongata (Swartz) E. H.Crane]、雷公连[Amydrium sinense (Engl.) H. Li]、百足藤[Pothos repens (Lour.) Druce]、四川轮环藤(Cycleasutchuenensis Gagnep.)、山黑豆(Dumasia truncata SieboldZucc.)、倒心叶珊瑚[Aucuba obcordata (Rehder) Fu ex W. K. Hu et Soong]、竹叶榕(Ficus stenophylla Hemsl.)、长叶冠毛榕[F. gasparriniana var. esquirolii (H. Lév.Vaniot) Corner]、硬叶冬青(Ilex ficifolia C. J. Tseng ex S. K. Chen et Y. X. Feng)、假福王草[Paraprenanthes sororia(Miq.) C. Shih.]。雷公连属(Amydrium Schott)、山黑豆属(Dumasia DC.)和假福王草属(Paraprenanthes C. C.Chang ex C. Shih)为香港的新记录属。以上的发现不仅丰富了香港的植物区系,而且表明保护残存植被非常重要,且极小种群的保护也值得特别关注。     

Effects of an exotic plant and habitat disturbance on pollinator visitation and reproduction in a boreal forest herb

The invasion of exotic species into natural habitats is considered to be a major threat to biodiversity, and many studies have examined how exotic plants directly affect native plant species through competitive interactions for abiotic resources. However, although exotics can have potentially great ecological and evolutionary consequences, very few researchers have studied the effect of exotics on the interactions between plants and their mutualistic partners, such as pollinators, and none have reported on such impacts in logged and undisturbed boreal forest ecosystems. Here we show how experimental introductions of an exotic plant species (Phacelia tanacetifolia Bentham) affect pollinator visitation and female reproductive success of a native plant (Melampyrum pratense L.) in recently disturbed (i.e., logged) and in undisturbed boreal forest habitats. The presence of Phacelia significantly increased the number of bumble bees entering plots in both habitat types. However, the exotic species had a strong negative impact on the visitation rate to the native species in both habitat types. Despite this negative impact on pollinator visitation, the exotic had no effect on female reproductive success of the native species in any habitat. Our results show that seed production may be more robust than pollinator visitation to exotic invasion, irrespective of habitat disturbance history.     

Attractiveness of Michigan native plants to arthropod natural enemies and herbivores

The use of plants to provide nectar and pollen resources to natural enemies through habitat management is a growing focus of conservation biological control. Current guidelines frequently recommend use of annual plants exotic to the management area, but native perennial plants are likely to provide similar resources and may have several advantages over exotics. We compared a set of 43 native Michigan perennial plants and 5 frequently recommended exotic annual plants for their attractiveness to natural enemies and herbivores for 2 yr. Plant species differed significantly in their attractiveness to natural enemies. In year 1, the exotic annual plants outperformed many of the newly established native perennial plants. In year 2, however, many native perennial plants attracted higher numbers of natural enemies than exotic plants. In year 2, we compared each flowering plant against the background vegetation (grass) for their attractiveness to natural enemies and herbivores. Screening individual plant species allowed rapid assessment of attractiveness to natural enemies. We identified 24 native perennial plants that attracted high numbers of natural enemies with promise for habitat management. Among the most attractive are Eupatorium perfoliatum L., Monarda punctata L., Silphium perfoliatum L., Potentilla fruticosa auct. non L., Coreopsis lanceolata L., Spiraea alba Duroi, Agastache nepetoides (L.) Kuntze, Anemone canadensis L., and Angelica atropurpurea L. Subsets of these plants can now be tested to develop a community of native plant species that attracts diverse natural enemy taxa and provides nectar and pollen throughout the growing season.     

Morphological variation in different populations of <Emphasis Type="Italic">Aceria anthocoptes</Emphasis> (Acari: Eriophyoidea) associated with the Canada thistle, <Emphasis Type="Italic">Cirsium arvense</Emphasis>, in Serbia

The russet mite, Aceria anthocoptes (Nal.), is the only eriophyid that has been recorded on Canada thistle, Cirsium arvense (L.) Scop. It has been noted in several European countries and recently in the USA. With its apparent host specificity and because of the damage it causes to its host plant, A. anthocoptes is being studied as a potential candidate for classical biological control. The aim of the present study was to examine quantitative morphological traits in four populations of A. anthocoptes living on two infraspecific host plant taxa (C. arvense var. arvense and C. arvense var. vestitum) in two geographically separate areas of Serbia in order to test the hypothesis of absence of the possible host plant impact on mite morphology. MANOVA analysis revealed significant differences between populations from different localities in Serbia. Populations of A. anthocoptes inhabiting two thistle varieties in the vicinity of Belgrade differed significantly from mites inhabiting the same two host varieties in the vicinity of the town of Ivanjica. Canonical discriminant analysis showed that the trait which best discriminates the populations of A. anthocoptes is the number of dorsal annuli. It was not possible to ascribe morphological differences to the impact of the host plant.     

Parasitic hymenoptera fauna on agromyzidae (diptera) colonizing weeds in ecological compensation areas in northern Italian agroecosystems

Parasitoids (Hymenoptera) associated with agromyzid leafminers (Diptera: Agromyzidae) were studied in three rural farms located in northern Italy. The parasitoids were reared from mined foliage of weeds growing in field margins. We reared 998 Hymenoptera specimens, representing five families, 23 genera, and 53 species, from leafminers infesting weeds. Eulophidae was the most abundant family (67.64%), followed by Braconidae (28.86%), Eucoilinae (1.40%), Tetracampidae (1.40%), and Pteromalidae (0.7%). Braconids was the most species rich family, accounting for 28 species; eulophids were represented by 19 species, pteromalids by four species, and eucoilins and tetracampids by one species each. The dominant parasitoid was the eulophid Pediobius metallicus (Nees), representing 18.17% of the total, followed by Diglyphus isaea (Walker) (12.73%), and Neochrysocharis formosa (Westwood) (10.82%). The most abundant braconid parasitoid was Dacnusa maculipes Thomson (9.62%). More than 80% of parasitoids were recovered from 10 plant species: Cirsium arvense (L.) Scopoli, Plantago lanceolata L., Sonchus asper (L.) Hill, Papaver rhoeas L., Picris echioides L., Lactuca serriola L., Myagrum perfoliatum L., Ranunculus velutinus Tenore, Arctium lappa L., and Medicago sativa L. The retention and the management of wild plants within field margins can be crucial tools to enhance the populations of biological control agents of agromyzids and to conserve rare parasitic wasp species.     

Effects of Lavandula officinalis and Equisetum arvense dry extracts and isoquercitrin on the fermentation of diets varying in forage contents by rumen microorganisms in batch culture

The short-term actions of Lavandula officinalis and Equisetum arvense dry extracts, and of isoquercitrin, flavonoid present in Equisetum arvense, on in vitro fermentation by rumen microbes were studied in batch culture. The orchard grass hay:barley ratios in the three experimental diets were 100:0, 75:25, 50:50 on a DM basis. The production rates of all volatile fatty acids except isobutyrate were strongly influenced by the composition of the diet and to a lesser extent, by plant extracts, with significant interactions between both factors. When hay was the only substrate, the addition of L. officinalis and E. arvense enhanced the fermentation rate by 50%, through an increased release of acetate and propionate. On the contrary, with the two other diets, the fermentation rate was strongly lowered by isoquercitrin. Gas outputs were not significantly influenced by plant extracts.