棉花黄萎病生防菌脂肽类抑菌物质分离和鉴定

棉花黄萎病(Cotton Verticillium Wilt)是一种由大丽轮枝菌(Verticillium dahliae Kleb)引起的严重危害棉花生产的土传维管束病害[1]。因大丽轮枝菌菌丝体产生的大量微菌核在土壤中存活时间长以及病原菌变异快等特点,造成培育抗病品种、使用化学药剂和利用新的栽培技术等方法防病效果均不理想。近年来,以生物防治为主的综合防治措施日益受到人们重视,并取得了一定效果[2-4];该措施     

棉花品种黄萎病抗性与免疫反应的关系

棉花黄萎病是由大丽轮枝菌引起的土传真菌病害,不同棉花品种对黄萎病的抗性不同。为了比较不同棉花品种对黄萎病菌的免疫反应程度与抗黄萎病水平间的关系,利用4×106CFU·m L~(-1)的大丽轮枝菌孢子悬浮液处理6个不同棉花品种的根系,进行抗病性评价。结果表明,接种大丽轮枝菌后,棉花植株的根、下胚轴、叶等器官中大丽轮枝菌的生物量随品种抗病性的增强而减少,根部木质素、叶片胼胝质含量随抗病性的增强而增多,L-苯丙氨酸解氨酶(PAL)、多酚氧化酶(PPO)、过氧化物酶(POD)和过氧化氢酶(CAT)活性的升高幅度随抗病性的增强而增大。由此可见,大丽轮枝菌侵染棉花后,植株的免疫反应程度越强其抗病性越强。本研究结果为棉花与黄萎病菌互作机制的进一步研究提供了理论依据。     

陆地棉酵母双杂交cDNA文库及VdSCP7诱饵载体的构建

由大丽轮枝菌(Verticillium dahliae)引起的棉花黄萎病是制约我国棉花生产的首要病害,严重影响棉花的产量与品质。为研究棉花与黄萎病菌互作的分子机制,本研究以国审棉百棉1号为材料,提取被大丽轮枝菌侵染的棉花根部总RNA,检测质量并用DNase Ⅰ处理后,用RNA 5'末端的模板转换方法,即SMART技术合成双链cDNA,再经酶切、过柱纯化去除短片段后,连接至pGADT7载体上构建棉花cDNA文库。VdSCP7是定位于寄主细胞核的大丽轮枝菌效应因子,能激发棉花的免疫反应,增强棉花抗病性。以大丽轮枝菌cDNA为模板扩增VdSCP7,产物纯化后重组到载体pGBKT7上构建诱饵载体pGBKT7-SCP7。诱饵载体经酶切及测序鉴定后,转化到酵母AH109中,鉴定诱饵蛋白毒性并检测诱饵载体是否存在自激活活性。结果表明,获得了库容量为2×10⁶ CFU的棉花cDNA文库,文库片段多样性良好,文库重组率约94%,质粒文库滴度约2.0×10⁹ CFU·mL^-1,满足酵母双杂交cDNA文库构建要求。酶切及测序结果表明,诱饵载体pGBKT7-SCP7构建成功且经过鉴定后无毒性、无自激活活性。本研究结果为棉花抗黄萎病基因的筛选及VdSCP7和棉花互作机制的解析奠定了基础。     

大丽轮枝菌侵染拟南芥的过程及诱导的防御反应研究

为进一步了解大丽轮枝菌的致病机制,本研究以绿色荧光蛋白标记的大丽轮枝菌菌系为材料,研究其对拟南芥的侵染过程和诱导拟南芥产生的防御反应。结果表明,大丽轮枝菌能侵染拟南芥,接菌6h后,分生孢子吸附在根表面,2~3 d到达皮层,9 d到达维管束,随后迅速扩展,接菌后11 d完成系统侵染,产生黄萎病症状。大丽轮枝菌在侵染过程中,能诱导拟南芥胼胝质、活性氧的产生和防御酶活性升高,植物产生的防御反应特征明显。本研究结果为大丽轮枝菌致病机理研究提供了理论依据。     

基于Web of Science对土传病害研究现状和趋势的计量分析

土传病害是限制我国作物产量和品质的重要因素,同时土传病害的田间蔓延也使得作物产量对农药依赖程度越来越大,造成环境污染问题。为了了解土传病害国内外的研究状况,本文利用CiteSpace知识图谱分析工具、VOSviewer可视化分析软件与HistCite引文图谱分析工具,对Web of Science核心合集数据库中土传病害研究领域的发文数量与学科分布、发文的主要国家(地区)与机构、发文的主要学术期刊、主要研究学者、主要研究热点及其变化趋势等进行计量分析。结果表明:(1)土传病害相关研究受到的重视程度越来越高,进一步支持土壤科学工作者对土传病害的研究将有重要意义。(2)美国、中国、澳大利亚、法国等国家在该领域发文较多,且国家之间合作密切。(3)土传病害研究领域的本地引用次数列前二的期刊为Phytopathology和Plant Disease,其中5年影响因子较高的主要发文期刊有:Soil BiologyBiochemistry、Plant Disease、Plant and Soil以及Applied Soil Ecology等。(4)土传病害领域高产学者有Shen Qirong(沈其荣)、Gilligan Christopher A、Mazzola Mark、Weller David M、Cao Aocheng(曹坳程)和Cai Zucong(蔡祖聪)等。(5)生物防控是土传病害的一个研究热点,典型的病原微生物有立枯丝核菌、大丽轮枝菌、尖孢镰刀菌、青枯雷尔氏菌等。(6)土传病害未来研究趋势主要集中于土壤微生物多样性与土传病害间的关系、寄主植物与病原微生物间的关系及土传病害防控机制与技术等方面。     

棉花黄萎病拮抗细菌HMB-1005鉴定及定殖

以大丽轮枝菌(Verticillium dahliae)为指示菌,对从棉田土壤中已经分离筛选到的细菌HMB-1005进行形态、19项生理生化特征及16S rDNA试验,结果表明,HMB-1005为解淀粉芽孢杆菌(Bacillus amyloliquefaciens);通过灭菌土和非灭菌土进行盆栽试验,拮抗菌HMB-1005芽孢液浸种、土壤全部混菌和部分混菌3种处理,经过5个月的盆栽试验表明,拮抗细菌HMB-1005能够在灭菌土和非灭菌土中定殖,且定殖数量达106cfu/g土左右,并仍能保持较高的抑菌活性。通过对棉苗根内细菌的分离,证实拮抗细菌HMB-1005能在棉花根内定殖,数量达到103cfu/g。实验结果显示,灭菌土的拮抗菌数高于非灭菌土,并且3种处理方式差异不明显,在实际生产应用中,建议使用浸种方法。     

棉花内生解淀粉芽孢杆菌489-2-2对棉花黄萎病的防效研究

为更好地防治棉花黄萎病,在冀棉11根系中分离得到一株对大丽轮枝菌抗性明显的内生细菌,经分子鉴定为解淀粉芽孢杆菌489-2-2。本试验以489-2-2为材料,研究该菌株对棉花黄萎病的防效和机理。结果表明,解淀粉芽孢杆菌489-2-2能够抑制黄萎病菌Vd080的生长,可导致Vd080菌丝形态异常。用该菌株发酵液浸泡棉花种子,对棉花黄萎病的防效为54.99%,灌根法的防效为60.31%。解淀粉芽孢杆菌489-2-2能使植株产生防御酶,诱导的免疫反应较强,且该菌株能定殖到棉花幼苗根系内部。本研究结果为解淀粉芽孢杆菌489-2-2的利用提供了理论依据。     

拮抗菌和生物有机肥防治棉花黄萎病及其对土壤酶活性的影响

棉花黄萎病是世界上毁灭性病害之一,它由土传真菌病原菌大丽轮枝菌引起并可以通过特制的生物有机肥进行防治。将生物有机肥、有机肥和三种拮抗细菌S37、S44、S228分别施入土壤,研究其对棉花黄萎病的防治效果及对几种主要土壤酶活性的影响。试验结果显示:防病效果达到5.9%~38.3%,由高到低依次为S37>生物有机肥>S44>S228>灭菌的生物有机肥。两种肥料处理显著提高了土壤蔗糖酶、多酚氧化酶、碱性磷酸酶、蛋白酶和脲酶的活性。3种拮抗菌处理提高了多酚氧化酶、碱性磷酸酶和蛋白酶的活性。除过氧化氢酶之外,其他酶的活性两两之间均呈现极显著正相关,但是病情指数与各种酶活性相关性均不显著。     

大丽轮枝菌RAPD反应组分的优化与应用

根据正交实验理论，设计一种优化RAPD各个反应组分以获得理想图谱的最佳反应浓度方法，能够显著减少反应组分优化次数，大丽轮枝菌（Verticillium dahliae)RAPD扩增最优反应组分浓度为：MgCl2 3.5mmol/ ,dNTP0.25mmol/L ,引物15pmol/L,Taq酶1.5U,DNA5n，实验表明，MgCl2与dNTP浓度之比是影响RAPD的增效果的最主要因素。根据优化的反应组分，用9个随机引物对来自江苏南通，射阳，泗阳，盐都4个群体53个大丽轮枝菌菌株进行RAPD扩增，对各个群体的遗传结构分析表明，各地群体遗传相似性较高。     

诱导模式对真菌激发子的影响

选择抗（耐）黄萎病的豫8号和感黄萎病的豫棉6两个棉花（Gossypium hirsutum L.)品种作为细胞系，选择高毒的V44和低毒的V64两种大丽轮枝菌9Verticillium dahlium Kle.b)为黄萎病原菌。设计不同毒力的病原菌分别直接诱导不同抗性的棉花细胞，不同毒力的病原菌和不同抗性的棉花细胞互相诱导的几种互作模式。发现棉花过氧化物酶（POD），超氧化物歧化酶（SOD）和多酚氧化酶（PPO）的比活经病原菌激发子直接诱导后有不同程度的提高；经交互诱导3种酶比活有进一步的变化，豫棉6号细胞交互诱导后3种酶的比活有了进一步的升高，而豫棉8号细胞交互诱导后3种酶的比活基本保持不变，并探讨了这些变化与棉花抗病性之间的关系。     

Potential of wild species for genetic enhancement of some semi-arid food crops

Discovery and incorporation of genes from wild species provide means to sustain crop improvement, particularly when levels of resistance in the cultigens are low and virulent strains of pests and pathogens overcome the host plant resistance. The extent of utilization and the potential of the wild genepool for genetic enhancement were reviewed in five important food crops viz. sorghum, pearl millet, chickpea, pigeonpea and groundnut grown in the semi-arid tropics. Introgression from compatible wild germplasm in the primary gene pool resulted in transfer of new cytoplasmic male sterility systems in pearl millet and pigeonpea, development of high protein, cleistogamous flower and dwarf pigeonpea lines and foliar disease resistant groundnut cultivars. Utilization of wild species in secondary and tertiary gene pools has been generally limited due to sterility, restricted recombination or cross incompatibility. Nevertheless, these species are extremely important as they contain high levels of resistance to several important biotic and abiotic stresses. Several of them, like those belonging to the Parasorghum section and the rhizomatous Arachis species are sources of multiple resistances and hold great promise to sustain crop productivity.     

“绿色革命”以来大气CO2浓度升高对C3作物营养品质的影响

自20世纪60年代"绿色革命"以来,育种技术和农耕技术的发展促进了农作物产量的大幅提升,然而作物的营养品质出现下降趋势。在相似的遗传背景下,大气CO_2浓度升高会使单位体积农作物产品的营养元素含量下降,因此"绿色革命"至今,农作物产品的营养元素下降可能受大气CO_2浓度升高影响。通过植物生长箱模拟"绿色革命"初期和目前的大气CO_2浓度水平(310μmol/mol和400μmol/mol),针对主要C_3作物水稻、小麦和大豆,研究"绿色革命"以来大气CO_2浓度升高对其籽粒的C、N、Fe、Zn元素含量的影响,结果表明:CO_2浓度升高对3种作物籽粒的C元素含量几乎没有影响,变化幅度在±1.5%之间;籽粒的N、Fe、Zn元素含量普遍呈现下降趋势,但均未达到显著水平。     

Faculty positions:Center for Agricultural Resources Research,Chinese Academy of Sciences

正The Center for Agricultural Resources Research(CARR),the Institute of Genetics and Developmental Biology(IGDB),Chinese Academy of Sciences,invites applicants for several research group leader positions.CARR is one of the research organizations in Chinese Academy of Sciences(CAS).We seek nominations and applications     

Collection and preliminary evaluation of native turfgrass accessions in Italy

Despite a raising interest on turfs in Italy, all theavailable varieties of this kind in the Country are of foreign origin, and areoften poorly adapted to the prevailing climatic conditions. This prompted tobegin a collection activity of indigenous turfgrass species, with the ultimategoal of identifying promising materials for future breeding based on localgenetic resources. The collection was carried out in three areas of Italy, viz.the northern Po Plain, the coastal region of Liguria, and the island of Sardiniathat are characterised, respectively, by subcontinental, warm temperate, andtypical Mediterranean climate. Altogether, 141 sites were visited, yielding 226accessions belonging to eight species of potential interest for turfs:Poa pratensis, Poa trivialis,Festuca rubra, Festuca arundinacea,Lolium perenne, Agrostis stolonifera,Agrostis tenuis, and Cynodon dactylon,this last being a warm-season grass. Poa pratensis andCynodon dactylon were mostly collected in northern Italyand Sardinia, respectively, whereas Festuca arundinacea andLolium perenne were rather ubiquitous. The collection sitesranged from 0 to 1040 m asl, but sites over 750 m wereonly visited in the inner part of Sardinia. All the accessions, collected aswhole plants, were transplanted at Lodi, northern Italy, where they are beingevaluated. Their preliminary evaluation for traits of importance for turf use,such as sward colour and overall quality, highlighted the great variation andthe occurrence of interesting accessions in all species. Other characters wererecorded, bearing specific importance in individual species, and in all casespromising accessions were identified. The germplasm of Festucarubra, Festuca arundinacea, and Loliumperenne proved highly infected by endophytic (symbiotic) fungi of thegenus Neotyphodium.     

Leaf position and genotype differences for mineral element concentrations in sorghum grown on tropical acid soil 1

Mineral element deficiencies and toxicities are common problems associated with sorghum [Sorghum bicolor (L.) Moench] production on acid soils. To better understand some of the mineral element problems and the analysis of plant tissue of sorghum plants grown on acid soils, four sorghum genotypes were grown on an acid Oxisol at Carimagua, Colombia limed with dolomite at 2 and 6 Mg ha^‐1.

Samples for mineral element analyses were obtained from leaves at different positions on the four genotypes. Concentrations of P and Mg were highest in the flag leaf (Leaf No. 1) and decreased as the position on the plant declined from the top of the plant for plants grown at 2 Mg lime ha^‐1. Similar decreases in P, Mg, K, and Zn concentrations occurred in plants grown with 6 Mg lime ha^‐1. Concentrations of Ca, S, Si, Mn, Fe, Cu, and Al increased as leaf position declined from the flag leaf for plants grown at 2 and 6 Mg lime ha^‐1. The higher lime supply enhanced Ca and reduced Mn and Fe concentrations in leaves. Differences in mineral element concentrations for the four genotypes used were fairly extensive. The elements to show the greatest range among genotypes were Al and Si and the elements to show the least range among genotypes were P, K, and S. Care should be used in collecting leaf samples for plant analysis and genotypic differences for accumulation of mineral elements should be considered in interpretation of results.     

Short-term variation in the concentration of selected ions within individual tropical rainstorms

Rainwater was collected at the campus of the University of Brunei Darussalam in Bandar Seri Begawan, Brunei Darussalam, using a funnel-in-bottle sampler. Polypropylene bottles were changed at intervals during rainstorm events. The pH and conductivity were determined immediately after collection on aliquots of the sample. Samples were refrigerated at 5°C for subsequent chemical analysis. Analyses for Na, Mg, Ca, Zn and Fe were carried out by means of inductively coupled plasma atomic emission spectroscopy (ICP-AES); Cu and Mn were analysed by graphite furnace atomic absorption spectroscopy (GFAAS); K was analysed using flame atomic emission spectroscopy (FAES); and Cl^–, NO₃ ^– and SO₄ ^2– were analysed by ion chromatography (IC). Concentration versus time profiles are reported for three rainstorm events. All ions exhibited a decrease in concentration during the rainstorm. The first sample contained the highest concentration of ions, consistent with a first-flush effect. The contribution of the initial stages of the shower to the total quantity of ion deposited during the entire rainstorm is quite overwhelming; in many cases 20 to 30% of the mass was deposited in less than 5% of rainstorm duration. On the other hand, the pH and conductivity variation during rainstorms did not exhibit a consistent pattern.     

Leaf litter selection by detritivore and geophagous earthworms

Summary Litterbag experiments with 10 different kinds of leaf litter showed that detritivore (Lumbricus species) and geophagous (Aporrectodea species) earthworms prefer certain litter types over others, since different numbers of worms were found below the litter after 50–52 days of exposure in a pasture. The detritivores preferred Fraxinus, Tilia, and predecomposed Ulmus and Fagus litter to Fagus litter and paper, while geophages preferred Tilia litter to Alnus and Ulmus litter, so that the two groups of earthworms showed different preferences. The detritivores seemed to be more selective than the geophages. The palatability of the litter was examined in relation to the C: N ratio, the lignin concentration and the initial and final polyphenol concentration. The numbers of detritivores were significantly correlated with the C: N ratio and the final polyphenol concentration, so that selection of litter seems to be related to palatability. The numbers of geophages were not significantly correlated with any of the parameters for palatability. The disappearance of litter after 50–52 days appeared to be due to detritivore activity, since the numbers found below the litter were positively and significantly correlated with the litter disappearance. There was no significant correlation with geophage activity. This indicates that detritivores use litter as food, and therefore influence the composition of the litter layer.     

The 5S rRNA gene diversity in the genus <Emphasis Type="Italic">Secale</Emphasis> and determination of its closest haplomes

We analyzed 127 rDNA sequences (5S DNA units) obtained from 23 seed accession samples from more or less 10 taxa in wild and cultivated rye, genus Secale L. The sequences fell into two known groups, here assigned to two unit classes, viz. long R1 and short R1 (designations to reflect on R haplome of rye). The different taxa could not be fully differentiated based on the 5S DNA units. We searched for 5S DNA sequences from known unit classes most closely similar to the long R1 and the short R1. One set with the long R1 unit class contained sequences of the long P1 unit class from Agropyron (P haplome) and from Kengyilia (StYP haplome), long J1 from Thinopyrum (J haplome), whereas the set with the short R1 included the long S1 from Pseudoroegneria (St haplome) and Kengyilia (StYP haplome), the short J1 from Thinopyrum (J haplome) and the short V1 from Dasypyrum (V haplome). Each of the two sets was analyzed separately by maximum likelihood (ML) phylogenetic analysis from which we were able to infer that the 5S DNA units of Secale differentiated in a non-clock fashion and followed the HKY substitution model in the gene tree with the long R1 unit class and the HKY + G in the gene tree with the short R1 unit class. A complementary Bayesian analysis yielded identical tree topologies to the ML ones for each of the two sequence sets. In the tree with the long R1 units the long P1 and long J1 unit classes were closest to the long R1 unit class, whereas in the tree with the short R1 units the long S1 and short J1 unit classes were closest to the short R1 unit class, indicating possibly a close relationship between the St, J and R haplomes.     

Nematode succession and microfauna-microorganism interactions during root residue decomposition

The quality of plant material affects the vigor of the decomposition process and composition of the decomposer biota. Root residues from hairy vetch (Vicia villosa Roth), rye (Secale cereale L.) and vetch+rye, packed in litterbags were placed in pots of soil at 15 C and the content of the bags was analyzed after 2, 4, 8 and 12 weeks. Bacterial biomass did not differ between residues with contrasting composition. Among bacterivores groups of nematodes that require high bacterial production dominated in fast decomposing resources whereas flagellates with smaller requirements prevail in slower decomposing resources. Biomass of bacterial feeding nematodes correlated positively with early phase (0-2 wk) decomposition that increased in the order: rye< vetch+rye<vetch. Bacterial biomass therefore seems to be under top-down (predation) control during early decomposition. In contrast, the fungal biomass differed between resources with highest values for rye. Moreover, this increase in fungal biomass occurred later during succession and was correlated with decomposition activity for rye in that period. Fungal biomass therefore seems to be under bottom-up (resource) control. The composition of the nematode assemblages (composed of 25 taxa) showed a clear relationship to initial plant resource quality as well as decomposition phase. Early successional microbivorous nematodes vary according to resource quality with demanding bacterivores+predators (Neodiplogasteridae) dominating in vetch and less demanding bacterivores (Rhabditidae) and fungivores (Aphelenchus) being equally common in vetch and rye. Later in the succession (2-4 wk) bacterivorous Cephalobidae and fungivorous Aphelenchoides prevailed similarly on the different root materials whereas bacterivorous protozoa and the amoebal fraction thereof dominated in rye. At week 12 no species dominated the nematode assemblages that were similar between the resources. The differences between nematode assemblages among plant resources at 2 week were similar to the results of a field study sampled after 6 weeks with the same soil and plant resources. This lends support to the relevance of the successional patterns observed in this incubation study.     

Cycling of extracellular DNA in the soil environment

Upon entering the soil environment, extracellular DNA is subjected to dynamic biological, physical, and chemical factors that determine its fate. This review concerns the fate of both recombinant and non-recombinant sources of DNA. A schematic of DNA cycling coupled with genetic transformation is presented to understand its behavior in soil. Extracellular DNA may persist through cation bridging onto soil minerals and humic substances, be enzymatically degraded and restricted by DNases of microbial origin, and/or enter the microbial DNA cycle through natural transformation of competent bacteria. Lateral gene transfer may disseminate DNA through the microbial community. An understanding of DNA cycling is fundamental to elucidating the fate of extracellular DNA in the soil environment.