两株镉抗性奇异变形杆菌对龙葵修复镉污染土壤的强化作用

以广东省大宝山尾矿区植物根际土壤为分离样品,人工制备镉污染LB培养基,从中分离筛选出2株具有较强镉耐受能力的细菌,分别为TL3和DBS2,最高镉耐受浓度达300mg／L。结合其形态学特征、生理生化特性及16SrDNA序列分析,初步确定2株菌均为奇异变形杆菌（Proteusmirabilis）。随后以龙葵为实验植株,分别接种TL3、DBS2及TL3与DBS2混菌的培养液,通过盆栽实验检验2株菌对龙葵吸收土壤中镉的强化作用。结果显示,2株菌对龙葵吸收镉具有显著的提升作用,且能够促进植株的生长,3个实验组龙葵根部镉含量分别比对照组（122．7mg／kg）增加了17．2％、85．6％和130．1％。     

Inhibition of Alternaria stem canker on tomato by essential oils from Baccharis species

Abstract

The toxicity of four essential oils extracted from Baccharis articulata, Baccharis ochracea, Baccharis psiadioides and Baccharis trimera was tested against the phytopathogen Alternaria alternata, which causes Alternaria stem canker on tomatoes. Diseases caused by Alternaria fungi are responsible for great economic losses in terms of production and are controlled by synthetic fungicides; however, essential oils offer an alternative, since they have been proven to be effective for controlling against various plant pathogens. In this way, the antifungal activity of Baccharis essential oils was tested using potato dextrose agar medium with concentrations ranging from 0.1 to 20.0?µL mL^?1. Baccharis trimera and Baccharis ochracea essential oils presented 100% mycelial growth inhibition of A. alternata and were also able to control Alternaria stem canker disease under greenhouse conditions. Tomato plants treated with these essential oils exhibited area under the disease progress curve (AUDPC) values of 230.10 and 241.42, differing from the control condition, which showed an AUDPC value of 268.92. The essential oils of B. trimera and B. ochracea can be an alternative for controlling Alternaria stem canker disease of tomatoes and should be formulated as a potential fungicide against the A. alternata pathogen.     

Application of manure and compost to contaminated soils and its effect on zinc accumulation by Solanum nigrum inoculated with arbuscular mycorrhizal fungi

Zn accumulation in Solanum nigrum grown in naturally contaminated soil in the presence of different types of organic amendments was assessed. Under the same conditions, the response of the plant to inoculation with two different isolates of arbuscular mycorrhizal fungi (AMF) (Glomus claroideum and Glomus intraradices) was also evaluated. S. nigrum grown in the non-amended soil always presented higher Zn accumulation in the tissues, with the addition of amendments inducing reductions of up to 80 and 40%, for manure and compost, respectively, and enhancing plant biomass yields. The establishment of S. nigrum in the Zn contaminated soil combined with the application of amendments led to a 70-80% reduction in the amount of Zn leached through the soil. The use of S. nigrum in combination with manure appeared as an effective method for reducing the effects of soil contamination, diminishing Zn transfer to other environmental compartments via percolation.     

外源菌剂联合柠檬酸强化龙葵修复土壤镉污染

设置对照（CK）、摩西球囊霉菌（GM）、摩西球囊霉菌+柠檬酸（GM+CA）、摩西球囊霉菌+巨大芽孢杆菌（GM+BM）和摩西球囊霉菌+巨大芽孢杆菌+柠檬酸（GM+BM+CA）这5个处理,通过测定土壤全Cd、有效Cd和植株Cd吸收量及微生物群落变化,分析了外源菌剂和柠檬酸添加对龙葵修复Cd污染效果的影响.结果表明,相对于CK处理,GM处理龙葵根、茎和叶生物量显著增加35.67%、41.35%和65.38%,GM+BM+CA处理龙葵根和茎生物量显著增加73.38%和75.38%.GM处理提高了龙葵各部位Cd含量但差异不显著,GM+BM+CA处理龙葵叶部的Cd含量显著提高79.34%.GM处理龙葵茎和叶Cd累积量显著提高47.51%和89.58%.GM+BM+CA处理龙葵叶部Cd累积量显著提高226.84%.GM+BM+CA处理显著增加了Cd由龙葵茎向叶的转运系数,增幅为52.47%.GM+BM+CA处理显著增加了叶的富集系数,增幅为120.53%.此外,联合修复对根际微生物群落结构也产生了影响,特别体现在诱导某些关键微生物类群,如变形菌门、放线菌门、球囊菌门和油壶菌门相对丰度增加2.00%~5.77%、0.76%~9.96%、2.11%~3.63%和0.54%~2.98%.RDA分析发现变形菌门和放线菌门与土壤全Cd呈显著负相关,球囊菌门和油壶菌门与土壤全Cd呈负相关.关键微生物的变化增强了龙葵吸收根际养分和抗Cd胁迫的能力,提高龙葵Cd累积能力,有效降低土壤全Cd含量.综上,摩西球囊霉菌、柠檬酸与巨大芽孢杆菌通过共接种的方式活化了土壤中的难溶态Cd,有助于龙葵富集更多的Cd,同时也与摩西球囊霉菌产生联合修复的效果.富集植物-微生物联合修复Cd污染土壤有较好的应用潜力.     

Leaf surface compounds of the potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis>) and their influence on Colorado potato beetle (<Emphasis Type="Italic">Leptinotarsa decemlineata</Emphasis>) feeding

Summary.  The sampling behavior of the Colorado potato beetle Leptinotarsa decemlineata Say (CPB) involves examination of the surface of potato leaves. It has been suggested that leaf surface compounds (volatiles and cuticular waxes) may be involved in host-plant recognition, acceptance or discrimination. Here we report on the effect of leaf surface extracts of six Polish commercial potato varieties on CPB feeding. We tested the influence of potato leaf surface extracts on CPB adult and larval feeding, then separated the extracts with HPLC, and finally tested the effect of the HPLC-separated fractions on CPB feeding. The bioassays were performed using potato leaf discs deprived of their original surface compounds. Applied to test discs at concentrations ten times higher than natural (10 leaf area equivalent), the extracts deterred CPB adults and larvae from feeding. HPLC-separated fractions composed of alkanes, sesquiterpene hydrocarbons, wax esters, benzoic acid esters, fatty acid methyl, ethyl, isopropyl and phenylethyl esters, aldehydes, ketones, methyl ketones, fatty acids, primary alcohols, β-amyrin and sterols did not affect adult CPB feeding. Similarly, alkanes, sesquiterpene hydrocarbons, wax esters, methyl ketones, sesquiterpene alcohols and secondary alcohols had no effect on larval CPB feeding. The sterol fraction (cholesterol, β-sitosterol and stigmasterol) acted as a phagostimulant to CPB larvae. We isolated a fraction demonstrating a phagodeterrent effect on CPB adults and larvae. The qualitative composition of the deterrent fraction was quite similar in all potato extracts, but there were quantitative differences between the varieties. Much further work is needed to identify the compounds that can produce the deterrent effect.     

Analysis and characterization of cultivable heavy metal-resistant bacterial endophytes isolated from Cd-hyperaccumulator Solanum nigrum L. and their potential use for phytoremediation

This study investigates the heavy metal-resistant bacterial endophytes of Cd-hyperaccumulator Solanum nigrum L. grown on a mine tailing by using cultivation-dependent technique. Thirty Cd-tolerant bacterial endophytes were isolated from roots, stems, and leaves of S. nigrum L. and classified by amplified ribosomal DNA-restriction analysis into 18 different types. Phylogenetic analysis based on 16S rDNA sequences showed that these isolates belonged to four groups: Actinobacteria (43%), Proteobacteria (23%), Bacteroidetes (27%) and Firmicutes (7%). All the isolates were then characterized for their plant growth promoting traits as well as their resistances to different heavy metals; and the actual plant growth promotion and colonization ability were also assessed. Four isolates were re-introduced into S. nigrum L. under Cd stress and resulted in Cd phytotoxicity decrease, as dry weights of roots increased from 55% to 143% and dry weights of above-ground from 64% to 100% compared to the uninoculated ones. The total Cd accumulation of inoculated plants increased from 66% to 135% (roots) and from 22% to 64% (above-ground) compared to the uninoculated ones. Our research suggests that bacterial endophytes are a most promising resource and may be the excellent candidates of bio-inoculants for enhancing the phytoremediation efficiency.     

4种茄科植物对矿区污染土壤重金属的吸收和富集

采集了湖南及江苏8个矿区或冶炼厂周边4种茄科(Solanaceae)植物及其根际土壤,分析了植物及土壤样品中Cd、Cu、Pb、Mn和Zn 5种重金属浓度.结果表明:番茄(Lycopersicon esculentum)对5种重金属可能有着较强的耐性,但不具有超积累的潜力;刺天茄(Solanum indicum)对Mn、...     

Induction of host specificity in larvae of Manduca sexta: chemical dependence controlling host recognition and developmental rate

Summary. The tobacco hornworm, Manduca sexta L. (Lepidoptera: Sphingidae) is a specialist on Solanaceae. This host specificity is induced as the larva feeds on solanaceous foliage, so that solanaceous-reared larvae will refuse to feed on other plants. Experiments were designed to determine the role of dietary constituents on the induction of host specificity and the effects of these on development of M. sexta. Choice assays using leaf discs of cowpea, Vigna sinensis (Fabaceae), were used to monitor the isolation of relevant chemical cues from foliage of potato. An aqueous extract of potato foliage strongly stimulated feeding. This extract was partitioned with n-butanol under alkaline conditions to obtain a highly active butanol extract. Reversed phase flash chromatography with a water-methanol gradient gave an active fraction that was used as a supplement for wheat germ-based artificial diet. Larvae reared on this s-diet became dependent on potato allelochemicals for initiation and continuation of feeding activity. These larvae also developed faster than larvae reared on the control p-diet, but no effect on adult mass was detected. Further flash chromatography of the active fraction under alkaline conditions provided a highly active sub-fraction, and semi-preparative HPLC using gradients of water and acetonitrile resulted in the isolation of a single compound. Bioassays indicated that this compound alone can account for host recognition by solanaceous-reared larvae. The results suggest that the mechanism of induced host specificity in M. sexta involves development of dependence on this compound. Received 21 December 1999; accepted 14 March 2000     

Different phagostimulants in potato foliage for Manduca sexta and Leptinotarsa decemlineata

Summary. In the past decades, several studies have focused on the identification of feeding stimulants for specialists that feed on solanaceous plants, especially potato (Solanum tuberosum). In the 1950's, a phagostimulant was isolated from potato and tomato for Manduca sexta and characterised as a glycoside. It was suggested that the phagostimulant for M. sexta and Leptinotarsa decemlineata is identical. We tested whether these insects indeed share the same recognition factor. Previous bioassays for L. decemlineata larvae and adults were found to be unsatisfactory, so a new assay system was developed, using starch and wheat flour-based wafers as a neutral substrate. An aqueous extract of potato foliage that was highly active as a stimulant was fractionated by reverse phase medium pressure liquid chromatography (MPLC). Both insects were stimulated by a MPLC fraction that eluted with 25% methanol in water. Further separation of this and the following fraction (35% methanol, stimulatory for the beetle only) by semi-preparative high performance liquid chromatography (HPLC), using gradients of water and acetonitrile, resulted in the isolation of two distinct fractions that stimulated feeding by L. decemlineata. None of the HPLC fractions was active for M. sexta. However, fractions of the flash chromatography with less polarity (45–75% methanol) had a stimulatory effect only on M. sexta. Thus, the two insects do not use a common feeding stimulant, and for both at least two compounds of different polarity are active. Received 3 July 2000; accepted 24 October 2000     

植被恢复对刺萼龙葵根际土壤细菌群落结构与功能的影响

入侵植物可以改变入侵地土壤微生物群落,从而有助于其入侵,在前期研究中发现,植被恢复措施可有效控制刺萼龙葵（Solanum rostratum）的入侵,但植被恢复前后刺萼龙葵根际土壤细菌群落结构与功能尚未清楚.选取了前期研究中的2个植被组合：沙打旺（Astragalus adsurgens）+披碱草（Elymus dahuricus）+无芒雀麦（Bromus inermis）（T1）;沙打旺+苇状羊茅（Festuca arundinacea）+冰草（Agropyron cristatum）+羊草（Leymus chinensis）（T2）,并选取刺萼龙葵（SR）及本地植被（NR）作为对照,采用16S rDNA MiSeq高通量测序技术研究刺萼龙葵入侵及植被恢复后刺萼龙葵根际细菌群落组成,同时采用PICRUSt功能预测分析其功能.结果表明,刺萼龙葵入侵（SR）后Simpson指数和Chao1指数均高于本地植被（NP）,但未达到显著水平,而植被恢复（T1和T2）后,Shannon指数和Chao1指数显著降低（P<0.05）.刺萼龙葵（SR）显著降低了变形菌门（Proteobacteria）中的微枝形杆菌属（Microvirga）、斯科曼氏菌属（Skermanella）、鞘氨醇单胞菌属（Sphingomonas）及酸杆菌门（Acidobacteria）的Bryobacter属相对丰度（P<0.05）,而植被恢复后,这些菌属丰度也随之上升.RDA分析结果显示,土壤有机质、总氮、总磷、总钾和速效钾是影响细菌群落组成的重要因素.PICRUSt功能预测分析表明,刺萼龙葵入侵显著提高了氨基酸合成（biosynthesis of amino acids）、嘌呤代谢（purine metabolism）、嘧啶代谢（pyrimidine metabolism）、核糖体（ribosome）和氨酰-tRNA合成（aminoacyl-tRNA biosynthesis）等方面的功能,而植被恢复以后其相对丰度显著降低.本文探讨了刺萼龙葵入侵及植被恢复后根际细菌群落和功能,为刺萼龙葵的入侵机制及生态恢复提供理论依据.