苇状羊茅与多年生黑麦草内生真菌菌丝基因组DNA提取的优化及PCR初步检测

感染内生真菌的禾草在牧草和草坪业上具有重要的生态和经济意义，家畜采食感染Neotyphodium coenophialum和N.lolii的苇状羊茅和多年生黑麦草会发生中毒。本研究收集天津口岸1998年以来进境的部分苇状羊茅和多年生黑麦草种子，对经镜检确认带有内生真菌的种子进行分离培养，对疑似菌株的菌丝用改进的Moiler等方法进行基因组DNA抽提，测定浓度及纯度，对照原方法，DNA的纯度有较大提高，浓度略有上升。Tubulin2基因的引物IS1-IS3扩增结果显示为单一的条带，结合形态学和序列比对，分离培养得到的菌株可以基本确定为N．coenophialum和N。lolii。根据Genbank中N.coenophialum和N.lolii的NC25基因序列设计出引物F1-R1，扩增得到能区分开N.coenophialum和N．lolii的单一条带（相差160bp），建立了N．coenophialum和N．lolii的PCR检测方法，结果准确可靠。     

大田条件下丛枝菌根真菌对西瓜生长和枯萎病的影响

 在大田条件下研究丛枝菌根(AM)真菌对西瓜(Citrullus lanatus)生长和枯萎病的影响。接种AM真菌能显著降低西瓜枯萎病的发病率、病情指数、根内和根围土壤中镰刀菌繁殖体的数量,促进植株对N、P、B和Zn等矿物质的吸收,提高西瓜叶片中叶绿素的含量、叶片净光合速率、气孔导度和水分利用效率,促进植株生长,增加西瓜产量。     

进境苇状羊茅和多年生黑麦草种子携带内生真菌带菌率的研究

苇状羊茅内生真菌和多年生黑麦草内生真菌曾给一些国家的畜牧业造成巨大损失.为防止内生真菌随进境草籽传入我国并在中国扩散危害,我们对该2种进境草籽内生真菌的带菌率进行了研究.首先通过分离培养,得到了苇状羊茅内生真菌和黑麦草内生真菌,然后采用染色法对1998年以来收集到的从天津口岸进境的部分苇状羊茅和黑麦草种子进行了初步检验,经检验,37份苇状羊茅样品中有18个品种带菌,34份多年生黑麦草样品中14个品种带菌,带菌率最高可达86.7%.早熟禾、剪股颖等其它禾本科草籽目前尚未发现带有内生真菌.     

接种丛枝菌根对玉米生长与抗旱性的影响

采用盆栽试验,研究了水分胁迫下接种丛枝菌根真菌对玉米生长及其抗旱性的影响。结果表明:玉米与丛枝菌根可形成良好的共生关系,接种丛枝菌根显著提高了玉米生物量、株高、地径。与未接菌相比,接菌提高了植株水分利用效率,水分胁迫和正常供水条件下,较对照分别提高117%和24.6%。接菌后玉米叶片SPAD值和可溶性蛋白含量增加,叶片过氧化氢酶活性增强。同时,接种菌根显著降低了植株叶片脯氨酸、丙二醛含量,干旱胁迫和正常供水条件下,较未接菌处理分别降低了14.1%、18.9%和59%、69%。由此可见,接种丛枝菌根真菌能够促进玉米的生长,缓解干旱胁迫对玉米生长造成的不利影响,提高了玉米的抗旱性。     

荒漠生态系统中丛枝菌根真菌多样性

到目前为止,在全球荒漠生态系统中,共报道了69个科389种丛枝菌根真菌的寄主植物,其中,Compositae,Gramineae和Leguminosae 3个科的植物为荒漠生态系统中丛枝菌根真菌寄主植物的优势类群;共发表了7个属的89种丛枝菌根真菌,其中,Acaulospora和Glomus 属的真菌为荒漠生态系统中的优势类群.在此基础上,分析了荒漠生态系统中AM真菌多样性研究中存在的问题,并探讨了荒漠生态系统中AM真菌的研究方向.     

利用丛枝菌根真菌提高植物抗病性研究进展

本文就近年来国内外有关丛枝菌根真菌对植物土传病害的防效及其抗性机制进行了阐述,同时展望了丛枝菌根真菌资源在设施蔬菜生产中的广阔应用前景。     

丛枝菌根真菌与小麦孢囊线虫的相互作用

 近年来, 小麦孢囊线虫(cereal cyst nematodes, CCN:主要病原为燕麦孢囊线虫Heterodera avenae)对小麦(Triticum aestivum)的危害日益严重, 亟待探索新的生防途径。丛枝菌根真菌(arbuscular mycorrhizal fungi, AMF)作为环境功能生物, 对寄主植物具有生物药肥双重作用, 不仅能促进植物吸收利用养分, 而且能拮抗土传病原物、提高植物抗病性。本研究以AMF与CCN相互作用为切入点, 试图明确AMF与CCN相互作用关系, 并科学评价不同AMF抑制CCN、降低病害的效应。试验于温室盆栽条件下进行, 设接种AMF Gigaspora margarita(Gi.m)、Glomus mosseae(G.m)、Glomus intraradices(G.i)、Glomus versiforme(G.v)、Gi.m +G.m+G.i+G.v、CCN、CCN+Gi.m、CCN+G.m、CCN+G.i、CCN+G.v、CCN+Gi.m+G.m+ G.i+G.v和不接种对照(CK)共12个处理。结果表明, 接种AMF各处理均能降低小麦孢囊线虫侵染率、土壤中孢囊数和根内J2数量, 其中Gi.m处理抑制效果最大;CCN不同程度减少AMF侵入点数和产孢数量。Gi.m 和CCN+Gi.m处理的根内丛枝着生数量最多, 而后者根内的超氧化物歧化酶、苯丙氨酸解氨酶和过氧化氢酶活性显著高于其他处理、丙二醛含量则显著低于其他CCN+AM真菌处理。Gi.m和G.i处理的小麦株高、地上部干重高于其他处理;Gi.m 和CCN+Gi.m的处理的单穗重和单株产量均高于其他处理。表明AMF能不同程度拮抗CCN、促进小麦生长和提高产量, 其中, Gi.m是高效菌种。结论认为, AMF与CCN之间存在一定相互抑制作用, AMF能通过诱导植株防御反应拮抗CCN。     

茉莉酸介导的丛枝菌根真菌诱导植物抗病性研究进展

丛枝菌根（arbuscular mycorrhizal, AM）真菌能与植物根系建立共生体提高植物抗病性。植物激素茉莉酸（jasmonic acid,JA）参与AM的形成,介导AM真菌诱导的植物抗病性。目前,关于AM真菌诱导的植物抗病性内容较为丰富,但是关于JA介导的AM真菌诱导的植物抗病性却有待进一步研究。本文首先总结了JA对AM形成和AM对植物内源JA积累的影响;其次介绍了AM真菌对植物抗病性影响及诱导机制;最后从诱导形成防御结构、提高酶活性及合成防御蛋白、诱导次生代谢物质合成、诱导调控信号分子、JA与其他激素相互作用五个方面探讨了JA在AM真菌诱导抗病性中的作用,并对本领域的研究方向进行展望,以期为植物病害防治和菌根研究提供可借鉴的思路。     

丛枝菌根真菌Glomus mosseae单寄主培养体系的建立

 By using transformed Ri T-DNA carrot roots as host, the monoxenic culture of Glomus mosseae was successfully established. Some morphological processes of germination, colonization and spore formation, as well as bi-directional movement of hyphal cytoplasm flow were observed in this system.     

AM真菌对加拿大一枝黄花与菌根植物和非菌根植物种间作用的影响

为明确丛枝菌根(arbuscular mycorrhizal,AM)真菌对加拿大一枝黄花Solidago canadensis与本地菌根植物和非菌根植物种间竞争格局的调控作用,采用温室盆栽试验,通过接种摩西球囊霉Glomus mosseae(GM)、根内球囊霉G. intraradices(GI)及其混合菌种(GM+GI)3种处理,分析AM真菌对加拿大一枝黄花与本地菌根植物玉米Zea mays和非菌根植物油菜Brassica campestris种间作用的影响。结果表明:与对照相比,接种AM真菌均显著提高了加拿大一枝黄花和玉米的菌根侵染率,菌根侵染率为13.720%～50.015%,且前者的菌根侵染率明显高于后者。单独种植时,与对照相比,接种AM真菌尤其是接种混合菌种显著提高了加拿大一枝黄花的株高、叶片数和总干重。在加拿大一枝黄花与玉米混合种植时,与单独种植相比,加拿大一枝黄花的株高、叶片数、根长和总干重均较低;同时,与对照相比,接种AM真菌显著提高了玉米的相对竞争强度而对加拿大一枝黄花的相对竞争强度没有显著影响。在加拿大一枝黄花和油菜混合种植时,与对照相比,接种AM真菌则显著提高了加拿大一枝黄花的株高、叶片数、净光合速率和总干重;同时,接种AM真菌促进了入侵种的竞争优势而抑制了非菌根植物油菜的生长。说明加拿大一枝黄花与本地种的竞争格局受到与之混生物种的菌根依赖性强度以及AM真菌的种类差异影响。     

Dynamics of arbuscular mycorrhizal fungi associated with desert ephemeral plants in Gurbantunggut Desert

Previous studies documented that most desert plants can be colonized by arbuscular mycorrhizal (AM) fungi,however,little is known about how the dynamics of AM fungi are related to ephemerals in desert ecosystems.The dynamics of AM fungi with desert ephemerals were examined to determine the effects of host plant life stages on the development of AM fungi.Mean colonization of ephemeral annual plants was 45% lower than that of ephemeral perennial plants.The colonizations were much higher in the early part of the growing season than in later parts,peaking at flowering times.The phenology of AM fungi in root systems varied among different ephemerals.The density of AM fungal spores increased with the development of ephemeral annual plants,reached its maximum at flowering times,and then plateaued about 20 days after the aboveground senescence.A significant positive correlation was found between AM fungi spore density and biomass of ephemeral annual plants.The life cycles of AM fungi associated with desert ephemerals were very short,being about 60-70 days.Soil temperature and water content had no direct influence on the development of AM fungal spores.We concluded that the development of AM fungi was in response to desert ephemeral phenology and life history strategy.     

丛枝菌根菌丝桥传递作用对烟草抗病性相关酶活性的影响

为明确供体与受体植株间菌丝桥传递抗病信号对受体植株生长以及抗病性相关酶活性的影响,利用丛枝菌根真菌在供体与受体烟苗植株间建立菌根菌丝桥,对供体植株接种青枯菌的方法进行研究。结果表明:在供体烟苗接种丛枝菌根真菌条件下,再接种青枯菌,比只有菌丝连接的受体烟苗叶片内的过氧化物酶(peroxidase,POD)、多酚氧化酶(polyphenol oxidase,PPO)、苯丙氨酸解氨酶(phenylalanine ammonialyas,PAL)的活性分别提高了21%、29%和14%,地上部干重和植株磷含量也有相同的趋势;而在不接种丛枝菌根真菌条件下,供体烟苗接种青枯菌与否对酶的活性、植株干重及氮、磷含量均无显著影响。表明供体烟苗产生的抗病信号可以通过菌丝桥传递给受体烟苗,增强受体烟苗的抗病性。     

Isolation and identification of desert habituated arbuscular mycorrhizal fungi newly reported from the Arabian Peninsula

Arbuscular mycorrhizal fungi（AMF） are known to facilitate the growth and vigour of many plants, particularly in arid ecosystems. In a survey of AMF in a date palm plantation and two natural sites of a desert in Oman, we generated many single spore-derived cultures of AMF. We identified a number of these isolates based on spore morphotyping and molecular phylogenetic analysis using the sequence of the LSU-rDNA. Here, we presented the characteristics of four species of AMF recovered, namely Claroideoglomus drummondii, Diversispora aurantia, Diversispora spurca and Funneliformis africanum. The four species have been described previously, but for the Arabian Peninsula they are reported here for the first time. Our endeavor of isolation and characterization of some AMF habituated to arid sites of Arabia represents a first step towards application for environmental conservation and sustainable agriculture in this region.     

AMF+PGPR组合提高黄瓜抗枯萎病的作用机制

对黄瓜(Cucumis sativus)品种香翠16接种尖孢镰刀菌黄瓜专化型(Fusarium oxysporum f.sp.cucumerinum,Foc)、丛枝菌根真菌(AMF)Funneliformis mosseae(Fm)+Foc、丛枝菌根真菌Glomus versiforme(Gv)+Foc、根围促生细菌(PGPR)PS2-6+Foc、PS3-2+Foc、Fm+PS3-2+Foc以及Gv+PS2-6+Foc,以不接种处理作为对照。结果表明,接种Foc条件下,Fm+PS3-2和Gv+PS2-6组合处理的抗病效果显著优于单接种AMF或PGPR处理;镜检观测到靠近AMF菌丝的Foc菌丝纤细,生长不良。AMF菌丝能缠绕Foc菌丝,并阻挡其侵入根系;AMF侵染率与其防效均呈正相关(P0.05);Fm+PS3-2+Foc和Gv+PS2-6+Foc处理的根系活力是单接种Foc的4.6倍;与不接种对照相比,Fm+PS3-2+Foc和Gv+PS2-6+Foc处理的黄瓜叶片中吲哚乙酸(IAA)含量分别提高了205%和166%,赤霉素(GA3)提高了33%和22%,玉米素核甘(ZR)提高了100%和80%,JA平均提高了65%,ABA降低23%和17%;而与单接种Foc处理相比,IAA提高了667%和568%,GA3提高了223%和196%,ZR提高了254%和219%,茉莉酸(JA)含量平均提高了215%,而脱落酸(ABA)含量只有单接种Foc处理的53%;Fm+PS3-2+Foc和Gv+PS2-6+Foc处理显著提高了黄瓜叶内苯丙氨酸解氨酶(PAL)防御酶活性和PAL基因表达量。综上推断AMF+PGPR组合是通过拮抗病原物、促进合成抗病信号物质、上调防御基因表达、提高防御酶活性和降低植株体内有毒物质等机制降低病害和改善植物抗病性。     

Effect of native and allochthonous arbuscular mycorrhizal fungi on Casuarina equisetifolia growth and its root bacterial community

Exotic trees are often planted to recover degraded lands. Inoculation with mycorrhizal fungi can improve their survival. Plant growth is partly dependent on the strain used, but little attention has been paid to the selection of mycorrhizal fungi. The aim of this study was to determine whether the growth of Casuarina equisetifolia L. (Johnson) is affected by two different mycorrhizal inocula generated using fungal spores retrieved from an Australian site (allochthonous soil) and a Senegalese site (native soil) under C. equisetifolia trees. Comparative experiments were conducted with plants in a Senegalese soil, previously sterilized or not, and grown in a greenhouse. At harvest, parameters related to plant growth and mycorrhization were evaluated and soil bacterial communities were compared. Tree growth was significantly influenced by both types of inoculants. In unsterilized soil, plants inoculated with the native inoculant were taller than plants inoculated with the allochthonous inoculant and control plants. The frequency of mycorrhization with both inoculants was higher in unsterilized soil. The strongest effects of the mycorhizosphere on the soil microbiome were obtained with the allochthonous inoculum, and analysis of the taxonomic composition revealed mycorrhizal communities specific to each inoculum. These results suggest that the development of C. equisetifolia and its root bacterial community are dependent on the composition of the mycorrhizal inoculum. The functional consequences of this rhizosphere effect in terms of soil fertility should be further studied to better guide reforestation operations.     

AM真菌和镰刀菌对西瓜根系膜脂过氧化作用和膜透性的影响

 在温室盆栽条件下研究了丛枝菌根(arbuscular mycorrhiza,AM)真菌Glomus versiforme (Karsten) Berch和西瓜枯萎镰刀菌(Fusarium oxysporum f. sp. niveum)对西瓜根系中膜脂过氧化产物丙二醛(MDA)含量和细胞膜透性的影响。结果表明,接种AM真菌的西瓜根系中MDA含量、组织自动氧化速率和膜透性均低于对照,先接种G. versiforme,后接种F. oxysporum f. sp. niveum处理的MDA含量、组织自动氧化速率和细胞膜透性均低于只接种F. oxysporum f. sp. niveum的处理。接种G. versiforme感枯萎病西瓜品种"郑杂5号"MDA含量、组织自动氧化速率和膜透性降低幅度大于抗病品种"京欣1号"的接种处理,说明G. versiforme可降低感病西瓜品种的膜透性和MDA的产生,从而有效地保护细胞膜系统,减轻F. oxysporum f. sp. niveum对西瓜的为害程度。     

抗黄瓜枯萎病丛枝菌根真菌与根围促生细菌组合菌剂的筛选

 丛枝菌根真菌(AMF)和植物根围促生细菌(PGPR)同时接种具有促生防病的作用。本试验旨在评价AMF与PGPR组合菌剂防治黄瓜枯萎病(Fusarium oxysporum f. sp. cucumerinum )的效果,以获得高效生防菌剂。于温室盆栽条件下将AMF Funneliformis mosseae(Fm)、Glomus intraradices(Gi)和 Glomus versiforme(Gv)与PGPR菌株(PR2-1、PS1-3、PS1-5、PS2-6和PS3-2)不同组合菌剂处理黄瓜(Cucumis sativus, 品种:香翠16号),测定各处理对黄瓜植株生长量、枯萎病防治效果和黄瓜产量的影响。供试AMF+PGPR各组合菌剂处理能不同程度的增加黄瓜根系AMF侵染率和根围细菌的定殖数量、促进黄瓜生长发育和降低枯萎病危害程度。其中,以Fm+PS3-2和 Gv+PS2-6组合菌剂促生和增产效果最好, 以Fm+PS1-5、Fm+PS3-2和Gv+PS2-6组合菌剂防治枯萎病效果最好。综合评价筛选获得高效组合为Fm+PS3-2和Gv+PS2-6。这两个处理可使黄瓜枯萎病病情指数分别下降50.0%和58.4%,防效达到54.5%和63.7%,具有一定的田间应用潜力。     

西部矿区接菌对风化煤与黄土配比的土壤改良效应

针对西部矿区土壤贫瘠、有效养分缺乏和作物难以生长的现状,采用盆栽试验方法,风化煤与黄土按不同配比混合为供试基质,玉米为供试植物,研究接种丛枝菌根真菌对玉米生长的影响及其对不同混合基质的改良效应,寻求风化煤与黄土最佳配比。结果表明:接种丛枝菌根真菌促进了玉米的生长和对矿质元素的吸收,在风化煤与黄土质量比(1∶1)时菌根生态效应达到最大,玉米干质量、叶色值、地上部分氮磷钾累积量分别达到4.61 g·株~(-1),41.17,53.01 mg·株~(-1),7.15 mg·株~(-1),79.42 mg·株~(-1);风化煤与黄土(1∶1)处理菌丝密度达到4.91 m·g~(~(-1)),且玉米根系侵染率达到最大。随着风化煤比例的增加,根际土壤中球囊霉素相关蛋白和酸性磷酸酶活性逐渐递增。丛枝菌根与风化煤的协同作用促进了作物的生长,改善了根际土壤微环境,实现了对沟壑区土壤的改良和培肥。