Effect of root extracts of resistant pasture plants on the feeding and survival of black beetle larvae,Heteronychus arator (Scarabaeidae)

Abstract

Crude root extracts of the black beetle-resistant legumes red clover, white clover, lupin, Lotus pedunculatus, and lucerne significantly reduced feeding by 3rd-instar black beetle larvae when incorporated in an artificial medium containing a strong feeding stimulant. The same extracts were toxic when administered orally. Lucerne and L. pedunculatus contain particularly active feeding deterrents and toxins. The root of the black beetle-resistant grass Phalaris aquatica (= P. tuberosa), like that of the susceptible perennial ryegrass, had no effect on larval feeding or survival. Lotus pedunculatus was very much more active against black beetle larvae than L. corniculatus or L. corniculatus × pedunculatus.     

Olfactory response of Costelytra zealandica (Coleoptera: Melolonthinae) to the roots of several pasture plants

The influence of natural plant odours on the locomotory behaviour of 3rd‐instar larvae of Costelytra zealandica (White) was studied by observing their movement in glass‐sided test chambers. Through an analysis of the paths followed by individual larvae in single‐option choice‐chamber tests it was possible to evaluate the responses of the larvae to different plant materials. The odour of fresh young perennial ryegrass (Loliumperenne) root was more attractive to the insects than was that of older plants. Larvae were also more strongly attracted to the root of lucerne (Medicago sativa), Lotus pedunculatus, red clover (Trifolium pratense), and white clover (T. repens) than to that of perennial ryegrass. The possible role of volatile chemical factors in plant resistance to grass grub attack is discussed.     

Root turnover in pasture species: chicory,lucerne, perennial ryegrass and white clover

For pastures, root turnover can have an important influence on nutrient and carbon cycling, and plant performance. Turnover was calculated from mini‐rhizotron observations for chicory (Cichorium intybus), lucerne (Medicago sativa), perennial ryegrass (Lolium perenne) and white clover (Trifolium repens) grown in the Manawatu, New Zealand. The species were combined factorially with four earthworm species treatments and a no‐earthworm control. Split plots compared the effects of not cutting and cutting the shoots at intervals. Observations were made c. 18 days apart for 2.5 years. This article concentrates on differences between plant species in root turnover in the whole soil profile to 40 cm depth. At this scale, earthworm effects were generally small and short lived. For ryegrass and white clover, root length and mass were linearly related (R² = 0.82–0.99). For chicory and lucerne, the relationships were poorer (R² = 0.38–0.77), so for those species length turnover may be a poor indicator of mass turnover. Standing root length, total growth and death generally decreased in the sequence ryegrass > lucerne > chicory = white clover. In length terms, scaled turnover (growth divided by average standing root length) generally followed the sequence lucerne > white clover > perennial ryegrass = chicory. Across species the scaled turnover rate averaged 3.4 per year or 0.9% per day. Cutting shoots reduced standing root length, growth and death, but increased scaled turnover. These results indicate fast and prolonged root turnover. For ryegrass and white clover, at least there is need to reappraise how to measure and model shoot : root ratios, dry matter production and carbon cycling.     

高毒力昆虫病原线虫共生细菌菌株的筛选及其杀虫活性的检测

昆虫病原线虫共生细菌是寄生在昆虫病原线虫肠道的一种细菌,二者互惠共生。实验采用6个不同种的菌株为筛选材料。共生细菌菌株的培养液经85%饱和度的(NH₄)_2SO4盐析,浓缩冻干得到杀虫粗提物。以粗提物注射大蜡螟Galleria mellonella、饲喂玉米螟Ostrinia furnacalis和棉铃虫Helicoverpa armigera,发现Xenorhabdus nematophilus D43、X.bovienii A54、Photorhabdus luminescens HZL和CB-8等4个菌株发酵液的粗提物对昆虫有高的血腔毒性,菌株A54对昆虫又有高的胃毒效果。由此确立A54为高毒力的菌株,其杀虫活性表现为：注射大蜡螟48 h的死亡率为80%,96 h为93.3%;粗提物饲喂玉米螟,72 h死亡率为53.3%,120 h死亡率为100%;饲喂棉铃虫,72 h死亡率为80.1%,120 h死亡率为90%。杀虫粗提物经DEAE-52柱层析分离,得到一个穿透峰和三个盐的梯度洗脱峰,其中穿透峰对昆虫有很好的胃毒效果,但没有血腔毒性;三个盐峰均有很高的血腔毒性,但没有胃毒作用。穿透峰样品饲喂2龄、3龄棉铃虫也有很好的杀虫活性,96 h 2龄棉铃虫的死亡率为65%,3龄棉铃虫的死亡率为30%;处理96 h的棉铃虫同处理前相比体重下降,未死棉铃虫体重明显低于对照。     

Nitrate reductase and nitrite reductase activity in free-living cells and bacteroids of Rhizobium loti

Cells of Rhizobium loti strains T1 and U226 cultured in defined medium with glutamate as the only nitrogen source and bacteroids isolated from root nodules of Lotus corniculatus, L. pedunculatus and L. tenuis did not show constitutive (non-nitrate induced) nitrate reductase activity (NRA). In contrast, nitrite reductase activity (NiRA) was present in both free-living cells and bacteroids of either strain T1 or U226. Constitutive NRA and NiRA were detected in the cytosol fraction from nodules of all three symbioses examined. An induced NRA was expressed in bacteroids after a 10 h incubation in the presence of nitrate.     

Effectiveness of Lotus Root Nodules: II. RELATIONSHIP BETWEEN ROOT NODULE EFFECTIVENESS AND 'IN VITRO' SENSITIVITY OF FAST-GROWING LOTUS RHIZOBIA TO FLAVOLANS

An extract from the roots of Lotus pedunculatus plants was foundto contain a compound toxic towards fast-growing Lotus rhizobia.This compound was identified as a flavolan, which has a prodeiphinidin:procyanidin ratio of 75:25. A fast-growing strain of Rhizobium(NZP2213) which forms ineffective root nodules on L. pedunculatuswas four times more sensitive to this flavolan (ED₅₀ = 25 ?gml^–1) than another strain (NZP2037, ED₅₀ = 100 ?g ml^–1)which forms effective root nodules on this species. The rootsof another Lotus species, L. tenuis, on which both strains ofRhizobium form effective root nodules, also contained a flavolan( 95% procyanidin) but both strains were relatively insensitiveto this flavolan (EDED₅₀ = 350 to 500 ?g ml^–1) L. pedunculatusplants bearing ineffective root nodules contained two to threetimes more flavolan in their roots (5–7 mg g^–1 fr.wt.)than uninoculated control plants. Experiments with seven otherLotus species and with hybrid plants developed between L. pedunculatusand L. tenuis showed a relationship between the prodeiphinidin:procyanidin ratio of the flavolan in their roots and the effectivenessof root nodules formed on these plants by NZP2213. Quantitativebinding studies of the flavolan from L. pedunculatus to NZP2037and NZP2213 indicated that, while the affinity constants forbinding were similar for both strains, the surface of strainNZP2037 contained four times more binding sites than NZP2213,possibly correlating with this strain's ability to toleratehigher concentrations of this flavolan. It is suggested thatthe differential sensitivity of these two strains of Rhizobiumto flavolans is related to their ability to form effective rootnodules on Lotus species.     

Larvicidal potentials of leaf and bark extracts of Eucalyptus camaldulensis (Schlect) and Eucalyptus citriodora (Hook) on Culex quinquefasciatus (Say) larvae

The larvicidal activity of leaf and bark extracts of Eucalyptus camaldulensis and Eucalyptus citriodora on juveniles of Culex quinquefasciatus was determined. Larvae were successfully bred in the laboratory to obtain a “pure” colony. One hundred and sixty‐two batches of 25 larvae each were treated with 3.90, 15.63, 62.50, 250 and 1000 mg/L of plant extract in three replicates, for 24 h to record mortality and for up to 96 h to record feeding and fecundity. There was an increase in percentage mortality with increase in concentration of the petroleum ether plant extracts. There was a statistically significant difference (P < 0.05) between extracts used in the bioassay and mortality of larvae, but there was no significant difference (P > 0.05) between the two plant species. Statistical analysis of average mortality figures using Probit analysis for the leaf extracts of E. citriodora and E. camaldulensis, respectively, had LC₅₀ of 245.47 (26.89 ± 10.13 [SE]) and 316.23 (22.89 ± 8.95) for the crude extracts; 97.72 (44.95 ± 12.89) and 223.87 (29.17 ± 9.56) for the petroleum ether extracts; and 162.18 (30.7 ± 11.16) and 257.04 (26.47 ± 9.28) for the methanol extracts. The bark extracts of E. citriodora and E. camaldulensis, respectively, had LC₅₀ of 251.19 (27.78 ± 8.39) and 398.11 (20.7 ± 7.17) for methanol; and 302.0 (21.56 ± 8.11) and 630.96 (13.56 ± 6.4) for hexane. Both plant species showed anti‐feeding properties against larvae of Culex quinquefasciatus.     

唐古特大黄药材提取物对小麦和垂穗披碱草种子萌发和幼苗生长的影响

测定了唐古特大黄（Rheum tanguticum Maxim. ex Balf.）药材水提液、蒽醌粗提液和多糖粗提液对小麦(Triticum aestivum Linn.)和垂穗披碱草（Elymus nutans Griseb.）种子萌发及幼苗生长的影响。结果表明：唐古特大黄药材水提液存在明显的化感作用,水提液在高浓度下对小麦种子萌发和芽长及垂穗披碱草种子萌发、根长和芽长均有明显的抑制作用,而在低浓度下对小麦的根长和芽长具有促进作用。蒽醌粗提液仅对垂穗披碱草幼苗生长有抑制作用,其作用随浓度的提高而增强。多糖处理液抑制了小麦、垂穗披碱草根的生长,同时高浓度的处理液延迟了垂穗披碱草种子发芽。     

Evaluation of quorum sensing modulation by plant extracts originating from Turkey

Quorum sensing (QS) is a widespread mechanism utilized by both Gram (+) and Gram (?) bacteria for communication and regulation of specific virulence traits and phenotypes due to population density. Plants are known to produce a number of compounds that can inhibit this communication and most of them have been discovered through well-known Chromobacterium violaceum biomonitor strains assays. In this study, we have analyzed 36 extracts, from 26 Turkish plant species, for their effects on bacterial growth and inhibition or induction of QS in the said biomonitoring assay. Four of the crude plant extracts from Tanacetum balsamita L. subsp. balsamitoides (Compositaceae), Epilobium angustifolium L. (Onograceae), Quercus frainetto Ten. (Fagaceae) and Quercus robur L. (Fagaceae) showed QS inhibitory activity and significantly reduced violacein production in C. violaceum. Particularly, ethyl acetate soluble compounds extracted from leaves of Quercus spp. were efficient in QS inhibition without any apparent increase in bacterial growth. On the other hand, extracts from Mentha longifolia subsp. longifolia and Hypericum orientale showed enhanced violacein production, thus increasing QS-dependent behaviour. Our results clearly demonstrate the QS-inhibitory effects of Q. frainetto Ten., E. angustifolium and T. balsamita L. extracts, while two other species showed QS-inducing effects, for the first time.     

The Effects of Seaweed Secondary Metabolites on Biofouling

Antifouling activity is one poorly investigated property of seaweed natural products. To determine, in the field, whether seaweeds contain chemicals able to influence the settlement of fouling organisms, crude organic extracts from Stypopodium zonale, Dictyota menstrualis (Phaeophyceae) and Laurencia obtusa (Rhodophyceae) were incorporated at natural volumetric concentrations, into hard stable gels that served as substrata for fouling in the experiments. Fouling organisms settled at a significantly higher rate on plates treated with S. zonale extracts than on control gels, while settlement was strongly inhibited on gels containing L. obtusa extracts. Fouling on gels treated with the D. menstrualis extract was not significantly different from the fouling found on control gels. The findings suggest that the broad antifouling properties of the crude extract of L. obtusa inhibit the settlement of fouling as well as hinder the development of settled fouling species, thereby reducing the richness of species. The results imply that L. obtusa possibly harbours powerful agents that can be explored for the development of antifouling technology.     

Differentiation in DNA fingerprinting among species of the genus Acer L. in Campania (Italy)

Abstract

Natural populations of species in the Acer genus occurring in Campania (southern Italy) were surveyed by screening seven microsatellite loci. Primer pairs for Acer pseudoplatanus L. microsatellite loci were analysed in six different species: Acer lobelii Ten., Acer campestre L., Acer pseudoplatanus L., Acer obtusatum W. et K., Acer neapolitanum Ten. and Acer monspessulanum L. The aim of the present study was to survey the genetic variability and genetic structure of natural populations of the Acer genus in Campania. The high degree of polymorphism observed in six different species of Acer makes these markers useful for investigating genetic variation at various spatial scales, and for the analysis of gene flow and of the mating system.     

小桐子枝叶提取物对植物病原真菌的生物活性

采用菌丝生长抑制法,测定了小桐子枝叶6种不同溶剂提取物对小麦赤霉病菌、稻瘟病菌、烟草疫霉菌和辣椒疫霉菌的抑制作用,从中筛选出抑制作用最强的粗提物进行进一步的活性组分分离和抑菌活性测定。结果表明小桐子枝叶的乙醇提取物对4种植物病原菌抑制活性最高,在浓度为0.8 g·L^-1时,小桐子枝叶的乙醇提取物对小麦赤霉病菌、稻瘟病菌、烟草疫霉菌、辣椒疫霉菌菌丝生长抑制率分别为：87.1%、90.3%、86.4%、77.9%,其抗菌活性与农药世高均无显著差异;在小桐子枝叶乙醇提取物的不同溶剂萃取物对稻瘟病菌和烟草疫霉病菌进行生物活性追踪测试中发现,石油醚和水萃取物都具有较好的活性,当浓度为0.8 g·L^-1时,石油醚和水萃取物对两种病菌抑制率都达50%以上。表明小桐子枝叶含有丰富的抗植物病原真菌活性物质,且主要存在于小桐子枝叶乙醇提取物的石油醚相和水相中。     

Root dynamics in barley,lucerne and meadow fescue investigated with a mini-rhizotron technique

Root development, including depth distribution, was followed in pure barley stands (Hordeum distichum, L.) with or without nitrogen fertilization and in barley undersown with lucerne (Medicago sativa L.) or meadow fescue (Festuca pratensis, Huds.). The number of roots per 5 cm depth level down to 1 m was counted frequently during the growing season using mini-rhizotrons, i.e., transparent tubes inserted into the soil. Root biomass at different depths down to 1 m was estimated from soil cores taken one month before harvest. The results from the two methods were compared and root counts in the different treatments were compared with the above-ground growth and production. Nitrogen-fertilized barley in pure stand had the highest biomass both above and below ground. According to the mini-rhizotron observations this treatment also had a deeper and denser root system, until barley harvest, than the other treatments. After barley harvest, roots from the undersown lucerne continued to increase, whereas the number of roots in the undersown meadow fescue remained the same. The root system in barley/meadow fescue did not penetrate into the subsoil, where more than 60% of the number of roots in barley undersown with lucerne were found. In general, the mini-rhizotron results indicated a higher relative abundance of roots in the deeper layers than the root biomass estimated with the soil coring method.     

Transient expression of human papillomavirus type 16 virus-like particles in tobacco and tomato using a tobacco rattle virus expression vector

The major capsid protein L1 of human papillomavirus type 16 (HPV16) was transiently expressed in tobacco (Nicotiana benthamiana and Nicotiana tabacum) and tomato (Lycopersicon esculentum) leaves using Agrobacterium tumefaciens. The expression vector pTV00 was derived from tobacco rattle virus (TRV). The highest L1 expression 15 μg g⁻¹(f.m.) was achieved when the coding sequence of L1 was optimized for expression in humans that caused an increase of the guanine and cytosine (GC) content from 38.2 % in wild type HPV16 to 64.1 % in optimized sequence. L1 monomers readily self-assembled into capsomeres and further into virus like particles (VLPs). Immunological characterization and electron microscopy showed that 89 % of L1 retained VLP structure also in extracts prepared from freeze-dried leaves. Plant expressed L1 in crude extracts was highly immunogenic without any additional adjuvant as vaccinated mice developed strong humoral and cellular immune response, comparable to that elicited by purified VLPs derived from insect cells. Further, the induced antibodies effectively neutralized infection of 293TT cells with pseudovirions. This finding demonstrates that the TRV expression system is comparable to other plant expression systems and due to the broad host range of TRV is particularly attractive when expression in plants with low content of toxic alkaloids is desired. Moreover, a monoclonal anti-L1 antibody E2 raised in the course of immunization with crude extract from freeze-dried leaves expressing L1 is specific preferentially against HPV VLPs and could be used in direct ELISA for monitoring of VLPs assembly and VLP purification protocols.     

Sources of resistance to stem nematode, Ditylenchus dipsaci (Kühn) Filipjev, in species of Medicago and Trifolium

The putatively resistant lucerne cv. Euver was as susceptible to 11 English and 11 French populations of ‘lucerne race’ stem nematode, Ditylenchus dipsaci, as the susceptible cv. Europe. The ‘resistant’ cv. Vertus showed some resistance to five of the English and four of the French populations but was just as susceptible as Europe to three English and four French populations. Twelve annual species of Medicago were very resistant to a mixture of two English and two French populations of the nematode. None of 13 perennial spp. of Medicago was very resistant to the same mixture of nematodes but resistant plants were identified in M. borealis, M. dzhawakhetica, M. glutinosa and M. romanica. Twenty six spp. of Trifolium were resistant to a mixture of two English and two French populations of ‘red clover race’ stem nematode.     

Salt effects on functional traits in model and in economically important Lotus species

A common stress on plants is NaCl‐derived soil salinity. Genus Lotus comprises model and economically important species, which have been studied regarding physiological responses to salinity. Leaf area ratio (LAR), root length ratio (RLR) and their components, specific leaf area (SLA) and leaf mass fraction (LMF) and specific root length (SRL) and root mass fraction (RMF) might be affected by high soil salinity. We characterised L. tenuis, L. corniculatus, L. filicaulis, L. creticus, L. burtii and L. japonicus grown under different salt concentrations (0, 50, 100 and 150 mm NaCl) on the basis of SLA, LMF, SRL and RMF using PCA. We also assessed effects of different salt concentrations on LAR and RLR in each species, and explored whether changes in these traits provide fitness benefit. Salinity (150 mm NaCl) increased LAR in L. burtii and L. corniculatus, but not in the remaining species. The highest salt concentration caused a decrease of RLR in L. japonicus Gifu, but not in the remaining species. Changes in LAR and RLR would not be adaptive, according to adaptiveness analysis, with the exception of SLA changes in L. corniculatus. PCA revealed that under favourable conditions plants optimise surfaces for light and nutrient acquisition (SLA and SRL), whereas at higher salt concentrations they favour carbon allocation to leaves and roots (LMF and RMF) in detriment to their surfaces. PCA also showed that L. creticus subjected to saline treatment was distinguished from the remaining Lotus species. We suggest that augmented carbon partitioning to leaves and roots could constitute a salt‐alleviating mechanism through toxic ion dilution.     

Evaluation of in vitro antifungal activity of medicinal plants against phytopathogenic fungi

Fourteen medicinal plants belonging to 13 families were collected and extracted with petroleum ether (PE), chloroform, methanol and water to yield 60 crude extracts. Using agar diffusion method, these extracts were evaluated for antifungal activity on the growth of five phytopathogenic fungi. Among all the extracts tested, PE, chloroform and methanol extracts of Piper betle L. and PE and chloroform extracts of Allamanda cathartica exhibited promising antifungal activity. Minimum inhibitory concentration (MIC) values of the above promising extracts were determined using broth dilution technique and observed that chloroform extract of P. betle L. exhibited the least MIC value ranging from 280 to 1130 μg ml^?1. In this study, we report chloroform extract of P. betle L. to be thermally stable even when steam sterilised for the first time and that it could be stored at 4°C with almost no change in its activity for a period of 180 days.     

Genetic and physical localization of the root-knot nematode resistance locus Mi in tomato

As part of a map-based cloning strategy designed to isolate the root-knot nematode resistance gene Mi, tomato F2 populations were analyzed in order to identify recombination points close to this economically important gene. A total of 21 089 F2 progeny plants were screened using morphological markers. An additional 1887 F2 were screened using PCR-based flanking markers. Fine-structure mapping of recombinants with newly developed AFLP markers, and RFLP markers derived from physically mapped cosmid subclones, localized Mi to a genomic region of about 550 kb. The low frequency of recombinants indicated that recombination was generally suppressed in these crosses and that crossovers were restricted to particular regions. To circumvent this problem, a population of Lycopersicon peruvianum, the species from which Mi was originally introgressed, that was segregating for resistance was developed. Screening of this population with PCR, RFLP and AFLP markers identified several plants with crossovers near Mi. Recombination frequency was approximately eight-fold higher in the Mi region of the L. peruvianum cross. However, even within the wild species cross, recombination sites were not uniformly distributed in the region. By combining data from the L. esculentum and L. peruvianum recombinant analyses, it was possible to localize Mi to a region of the genome spanning less than 65 kb. Received: 15 July 1997 / Accepted: 1 October 1997     

Nematicidal activity of <Emphasis Type="Italic">Paecilomyces</Emphasis> spp. and isolation of a novel active compound

Many species of Paecilomyces are entomogenous fungi and several are efficacious toward nematodes. To study the potential of Paecilomyces species in controlling nematodes, fungal extracts of 40 Paecilomyces spp. were evaluated for their nematicidal activity against Bursaphelenchus xylophilus and Panagrellus redivivus. The extracts of six Paecilomyces spp. exhibited the nematicidal activity against P. redivivus, and 11 species exhibited the nematicidal activity against B. xylophilus. The methanol extract of strain 1.01761 incubating on Czapek solid medium killed more than 95% P. redivivus in 24 h at 5 mg/ml, and the filtrate of strain 1.01788 cultured in Sabouraud’s broth medium resulted in 90% mortality of B. xylophilus in 24 h at 5 mg/ml. A novel nematicidal compound 4-(4′-carboxy-2′-ethyl-hydroxypentyl)-5,6-dihydro-6-methylcyclobuta[b]pyridine-3,6-dicarboxylic acid, was isolated from Paecilomyces sp. YMF1.01761. The LD₅₀ value of the compound within 24 h against P. redivivus was 50.86 mg/L, against Meloidogyne incognita was 47.1 mg/L, and against B. xylophilus was 167.7 mg/L.     

高通量测序分析甘肃高寒草原瑞香狼毒根内生菌群落结构特征研究

【背景】瑞香狼毒是我国天然草地退化的标志性植物之一,在入侵的过程中其根内微生物发挥着重要的作用,探究瑞香狼毒根内微生物群落对有益微生物资源的利用及生物防控瑞香狼毒具有重要意义。【目的】探究甘肃省祁连山东麓不同高寒草原毒性杂草瑞香狼毒根部内生真菌和细菌的组成、多样性等群落结构特征。【方法】通过Illumina MiSeq高通量测序技术对甘肃祁连山东麓不同高寒草原的瑞香狼毒根部内生菌组成及多样性进行分析。【结果】不同高寒草原瑞香狼毒根内子囊菌门(Ascomycota)在真菌中占优势地位;蓝细菌(Cyanobacteria)、变形菌门(Proteobacteria)、放线菌门(Actinobacteria)在根内生细菌中属于优势类群。G4样点的真菌和细菌OTU(OperationalTaxonomic Unit)数量和Shannon多样性指数均高于其他4个采样点。主坐标分析(Principal Coordinate Analysis,PCoA)表明,不同高寒草原瑞香狼毒根内生菌群落之间存在一定的差异。One-Way ANOVA分析结合Pearson相关性分析显示,不同样地瑞香狼毒真菌群落结构在属水平上相关性较小,而细菌在属水平存在一定的相关性。【结论】本研究为进一步开发利用瑞香狼毒内生真菌和细菌资源、阐明瑞香狼毒内生菌群的群落结构和生态功能提供理论参考。