A semiaxenic phototrophic system to study interactions between arbuscular mycorrhizal and pathogenic fungi in woody plants

Among other benefits, arbuscular mycorrhizal (AM) fungi may increase plant tolerance to root diseases. The research on the underlying mechanisms requires growth conditions that are both controlled and realistic. To study these interactions, a semiaxenic phototrophic system was developed in which the roots grow in a controlled environment and can be inoculated with both pathogenic and symbiotic fungi. Micropropagated fig plantlets were grown in containers having shoots in the outside and roots in a growth medium without sugar, inoculated or not with the AM fungus Rhizophagus irregularis and the pathogenic fungus Armillaria mellea. Dual inoculated plants developed the mycorrhizal association and pathogen infection symptoms. Mycorrhizal inoculation lowered disease index and increased plant growth. Colonization of A. mellea in fig roots was quantified by real-time PCR, showing that R. irregularis did not significantly lower the quantity of Armillaria, suggesting that other mechanisms were involved in increased tolerance to the pathogen. The results show that the system proposed is suitable to study the triple interaction involving plant, AM and root pathogenic fungi.     

Arbuscular Mycorrhizal Fungi Reduce Development of Pea Root-rot caused by Aphanomyces euteiches using Oospores as Pathogen Inoculum

The effects of the arbuscular mycorrhizal (AM)-fungi Glomus intraradices and Glomus claroideum on pea root-rot development caused by the pathogen Aphanomyces euteiches were investigated in a greenhouse pot-experiment, over the course of three harvests, using oospores as pathogen inoculum. Signature whole cell fatty acids 16:15c and 14:19 were used to quantify AM-fungi and A. euteiches, respectively in both roots and soil. Disease incidence was reduced in AM plants, though this effect was more pronounced in plants with G. intraradices than plants with G. claroideum, and corresponded with a greater mycorrhiza development, both intra- and extra radical in plants with G. intraradices than with G. claroideum. At the final harvest, percentage of root length with oospores was similar in roots of mycorrhizal and non-mycorrhizal plants. Despite the fact that pea root-rot development was only slightly lower in mycorrhizal plants compared to that of non-mycorrhizal plants, in terms of shoot growth and disease severity, mycorrhizal plants suffered less. This suggests a possible mycorrhiza-induced tolerance against pea root-rot. Furthermore, the degree of tolerance induction differed between the two AM-fungi included in the present study.     

Management strategies and distribution of Aphanomyces root rot of alfalfa (Medicago sativa), a continuing threat to forage production in the United States

Alfalfa (Medicago sativa) is one of several legumes that is affected by Aphanomyces root rot (ARR) caused by Aphanomyces euteiches. Symptoms of ARR on alfalfa seedlings include a yellow-grey discolouration of roots, rotting and loss of lateral roots, stunted growth, chlorotic foliage and reduction of nitrogen-producing nodules on roots. Infection can also occur on adult plants leading to loss of lateral roots and nodules. At the seedling stage, ARR decreases alfalfa stand establishment, and field longevity is reduced when adult plants are infected. A. euteiches is an oomycete pathogen that has motile zoospores and thick-walled oospores that can survive for many years in soil. Two races are currently recognized by pathogenicity on differential alfalfa check cultivars. Most alfalfa cultivars contain race 1 resistance, but there is an increasing development of cultivars with resistance to race 2. Management strategies include planting resistant cultivars, avoiding planting in fields with poor drainage and rotating crops with nonhost plants.     

Effects of arbuscular mycorrhizal fungi on severity of root rot of bananas caused by Cylindrocladium spathiphylli

The interaction between four arbuscular mycorrhizal (AM) fungi, Glomus sp., G. proliferum , G. intraradices and G. versiforme , and the root-rot fungus Cylindrocladium spathiphylli , and subsequent effects on growth and phosphorus nutrition of banana ( Musa acuminata , AAA, cv. Grande Naine) were investigated under glasshouse conditions. Overall, root infection by C. spathiphylli reduced the growth of banana plants, but preinoculation with AM fungi significantly attenuated this detrimental effect. Lower disease severity, stimulation of growth and increase of shoot P content were observed for the plants inoculated with one of the four AM fungi. Glomus sp. and G. proliferum induced the largest increase in growth parameters and shoot P content as compared to G. intraradices and G. versiforme , in the presence as well as in the absence of C. spathiphylli . Root damage caused by C. spathiphylli was decreased in the presence of AM fungi, but the inoculation of mycorrhizal plants with C. spathiphylli also decreased the intensity of AM fungal root colonization, indicating a clear interaction between the two organisms.     

金龟子绿僵菌对苜蓿生长的影响及对苜蓿抗蚜性的诱导作用

为明确金龟子绿僵菌Metarhizium anisopliae在促进植物生长和提高植物抗性方面的作用,以金龟子绿僵菌拌土种植紫花苜蓿Medicago sativa,于室内测定植株的生长参数、苜蓿斑蚜Therioaphis trifolii的取食选择性及其发育历期和繁殖力。结果表明,金龟子绿僵菌处理后7 d,植株株高比对照显著增加了0.87 cm;处理后14 d,植株分枝数为3.88个,根长为54.67 mm,均显著高于对照。株龄14 d的植株接入苜蓿斑蚜,至株龄28 d时,金龟子绿僵菌处理的植株株高、分枝数和根长均较对照显著增加;同时生物量、叶绿素含量和类胡萝卜素含量也有不同程度的升高。选择性试验结果显示,24 h内选择并驻留在金龟子绿僵菌处理植株叶片上的苜蓿斑蚜数量较对照显著减少28.57%。以金龟子绿僵菌处理的植株叶片饲喂苜蓿斑蚜,1～4龄若虫的发育均有所减缓,各龄若虫的发育历期均较对照叶片饲喂的各龄若虫有所延长,而成虫寿命较以对照叶片饲喂时显著缩短了30.09%,总生命历期显著缩短了18.83%。表明金龟子绿僵菌促进了紫花苜蓿植株的生长,同时增强了植株对苜蓿斑蚜的耐害性、排趋性与...     

Mycorrhiza induced resistance in potato plantlets challenged by Phytophthora infestans

Biological control of soil-borne pathogens by arbuscular mycorrhizal (AM) fungi has been repeatedly demonstrated. However, their role in the control of above-ground hemibiotrophic pathogens is less conclusive. Here, we investigated in vitro the impact of an AM fungus on Phytophthora infestans in potato plants. The leaf infection index was decreased in mycorrhizal potato plants. Real-Time Quantitative PCR revealed the induction of two pathogenesis related genes (PR1 and PR2) in the leaves of mycorrhizal plants shortly after infection with P. infestans. These results suggested a systemic resistance in mycorrhizal plants, related to the priming of the two PR genes in potato.     

Role of methyl jasmonate in the expression of mycorrhizal induced resistance against Fusarium oxysporum in tomato plants

Arbuscular mycorrhiza (AM) colonization led to a decrease in the severity of fusarium wilt disease caused by Fusarium oxysporum f. sp. lycopersici in tomato plants. The involvement of two plant defense hormones, namely methyl jasmonate (MeJA) and salicylic acid (SA), in the expression of mycorrhiza induced resistance (MIR) against this vascular pathogen was studied in the AM colonized and non-colonized (controls) plants. Activity of lipoxygenase (LOX), which plays a role in jasmonic acid (JA) biosynthesis, as well as levels of methyl jasmonate (MeJA) increased in AM colonized plants as compared to controls, but did not show any further changes in response to F. oxysporum inoculation. On the other hand, activity of phenylalanine ammonia lyase (PAL), which is an enzyme from salicylic acid (SA) biosynthetic pathway, as well as SA levels, increased in both controls and AM colonized plants in response to application of F. oxysporum spores. Hence the JA and not the SA signalling pathway appeared to play a role in the expression of MIR against this vascular pathogen. The resistance observed in AM colonized plants was completely compromised when plants were treated with the JA biosynthesis inhibitor salicylhydroxamic acid (SHAM). This confirmed that the AM-induced increase in JA levels was involved in the expression of resistance toward F. oxysporum. The SA response gene pathogenesis-related 1 (PR1) showed an increased expression in response to F. oxysporum infection in SHAM treated AM colonized plants as compared to plants that were not treated with this JA inhibitor. This suggested the possibility that JA inhibited SA responses, at least in the roots. AM colonization therefore appeared to prime plants for improved tolerance against the vascular pathogen F. oxysporum, which was mediated through the JA signalling pathway.     

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。     

Guava decline: effect of root exudates from Meloidogyne enterolobii-parasitized plants on Fusarium solani in vitro and on growth and development of guava seedlings under controlled conditions

In guava decline, Fusarium solani-immune guava trees become susceptible to extensive root rot caused by this fungus after parasitism by Meloidogyne enterolobii. To understand the mechanisms involved in this disease, root exudates were collected from nematode-inoculated (NI) or uninoculated (UN) guava plants cultivated in sand. After filtration through a Millipore® membrane, NI and UN exudates were used: i) to prepare media to assess their effect on mycelial growth and production of propagules of F. solani isolate UENF/CF 163, and ii) to incubate macro- and microconidia to assess their effect on germination. NI exudates promoted (P?<?0.05) more mycelial growth and production of propagules than UN exudates or water. NI and UN exudates were used to water guava seedlings laid over seed germination paper inside plastic boxes. Half of the seedlings had an agar plug colonized by the fungus positioned in the collar region. Upon watering with NI exudates the fungus caused (P?<?0.05) extensive rotting of the seedlings’ rootlets. NI and UN exudates, either unlyophilized or lyophilized and re-suspended to the original concentration, were used to water guava seedlings grown in sterile sand before being inoculated (or left uninoculated) as described before. Solely upon watering with NI exudates, in its unlyophilized form or after lyophilization, the fungus caused a reduction (P?<?0.05) of shoot and root biomass associated with rotting of roots. These results suggest that M. enterolobii induces chemical changes in the root exudates of guava trees, which are necessary for root invasion causing root rotting by F. solani.     

Occurrence,genetic diversity and pathogenicity characteristics of Pseudomonas viridiflava inducing alfalfa bacterial wilt and crown root rot disease in Iran

Alfalfa (Medicago sativa), the world’s most influential forage crop, is infected by many diseases such as alfalfa bacterial wilt disease. The causal agent of bacterial crown and root rot and wilt disease is Pseudomonas viridiflava, which is a substantial pathogen of alfalfa worldwide. This pathogen spreads through the xylem and under field conditions, plants show growth stunting, chlorosis and wilting symptoms not previously reported. In this study- the first on Pseudomonas viridiflava on alfalfa from Iran, we have investigated the pathogenicity and genetic diversity of Pseudomonas viridiflava in some parts of Iran. To survey the causal agent of the disease, symptomatic plants were collected from the main alfalfa growing area. Pathogenicity of the collected strains was confirmed on alfalfa plants under green-house conditions using a completely randomized design. Determination of bacterial strains was done based on standard bacteriological methods and PCR assay using specific primers. Effects of bacterial strains on wet weight, dry weight, stem length and root length of infected plants were measured and the data were analyzed by SAS software and Duncan’s assessment. The diversity of liquid cellular proteins of bacterial strains was examined on Polyacrylamide gel. To delineate of genetic diversity the total DNA was drawn out. Fourteen random primers were used in a RAPD test. To sketch the dendrogram, RAPD fragments were used to calculate genetic diversity with NTSYS software. This data showed pathogenicity and genetic diversity of Pseudomonas viridiflava in Iran.     

Molecular characterization of a phytoplasma of the ash yellows group occurring in strawberry (Fragaria x ananassa Duch.) plants in Argentina

Strawberry (Fragaria x ananassa Duch. cv. Camino Real) plants showing phyllody symptoms were detected in production fields in Bella Vista (Corrientes province, Argentina). DNA from all symptomatic samples analyzed yielded fragments of the expected size in PCR reactions using phytoplasma universal primers. DNA from asymptomatic plants yielded no products. The associated phytoplasma was designated as Argentinean Strawberry Phyllody (ASP). The ASP sequence of the 16S rRNA gene showed 99?C98% homology with members of the 16SrVII ash yellows group. The putative RFLP profile was indistinguishable from the Argentinean alfalfa witches?? broom (ArAWB) phytoplasma. The phylogenetic analysis of nearly full-length 16S rDNA sequence and 16S-23S spacer region yielded a consensus tree wherein ASP clustered into the ash yellows group (16Sr VII) with high confidence values (95), generating a separated branch (100 bootstrap value) together with ArAWB (16Sr VII-C). In Argentina, the 16Sr VII group was also detected in alfalfa (Medicago sativa L.) crops, and weeds [Artemisa annua L. and Conyza bonariensis (L.) Cronquist]. To our knowledge, this is the first report of a phytoplasma from the 16Sr VII group affecting strawberries in Argentina and worldwide.     

丛枝菌根真菌和甜菜夜蛾的相互作用

为探索丛枝菌根(arbuscular mycorrhiza,AM)真菌与植食性昆虫的相互作用,将摩西管柄囊霉(Funneliformis mosseae BEG167,Fm)、根内球囊霉(Glomus intraradices BEG141,Gi)和地表球囊霉(Glomus versiforme Berch,Gv)3种AM真菌以及甜菜夜蛾Spodoptera exigua分别接种至花生和番茄上,通过测定AM真菌的侵染定殖和甜菜夜蛾的生长发育指标分析AM真菌与甜菜夜蛾的相互作用。结果显示,甜菜夜蛾取食菌根花生植株后,Fm、Gv和Gi的侵染率和泡囊数均有增加,侵染率分别是未取食的1.15、1.10和1.11倍,而Fm的泡囊数增加最显著,是未取食的1.27倍。在菌根番茄植株上,Fm和Gv的侵染率显著增加,分别是未取食的1.24倍和1.94倍,Fm、Gv和Gi的泡囊数均显著增加,分别是未取食的1.50、1.98和1.42倍。甜菜夜蛾取食菌根花生和菌根番茄植株后,幼虫存活率下降,幼虫历期和蛹期明显延长;Fm对其影响最显著,甜菜夜蛾在花生上的存活率、幼虫历期和蛹期分别为26.67%、26.78 d和10.67 d,在番茄上分别为35.52%、24.33 d和10.39 d,与取食非菌根植株处理差异显著。表明甜菜夜蛾能在一定程度上促进AM真菌的侵染和定殖,而AM真菌能抑制甜菜夜蛾的生长和发育,但其影响因AM真菌种类而不同。     

Biochemical factors involved in vetch resistance toOrobanche aegyptiaca

Experiments were performed to elucidate the biochemical nature of the differential response of resistantVicia atropurpureacv. Popany and susceptibleVicia sativacv. Yovel to the parasitic weedOrobanche aegyptiaca. Plant root material was obtained by growing vetch plants in association withOrobancheseeds on glass microfibre filter sheets inserted in polyethylene bags replenished by nutrient solution. Higher concentrations of bound phenolics, free phenolics and lignin, and higher activity of peroxidase were observed in cv. Popany roots as a result ofO. aegyptiacainfection than were found in cv. Yovel vetch. No differences in hydroxyproline-rich glycoproteins levels were detected between infected and non-infected roots of either vetch cultivar. These results suggest that the resistant vetch defense mechanism involves elevated induction of the phenylproponoid pathway uponOrobancheinfection, conferring mechanical and chemical barriers confronting the invading parasite.     

Effects of climatic factors on native arbuscular mycorrhizae and Meloidogyne exigua in a Brazilian rubber tree (Hevea brasiliensis) plantation

The root-knot nematode Meloidogyne exigua and arbuscular mycorrhizal (AM) fungi may both occur in the roots of Brazilian rubber trees ( Hevea brasiliensis ). AM fungi may stimulate plant growth whereas nematodes usually reduce it. Variations of native AM fungi and M. exigua populations in soil and roots of rubber trees were studied for one year in a Brazilian plantation. The number of AM spores in the soil was generally greater in the rainy season than in the dry season, although AM colonization of roots was unaffected by season. During the dry season, numbers of juveniles and eggs of M. exigua in roots were lower than in the rainy season. A site without nematodes in the soil or roots showed the greatest numbers of AM spores in soil and highest AM colonization of roots. A negative correlation was observed between the percentage of AM colonization and the number of second-stage juveniles in soil and second-stage juveniles and eggs in roots. Microscope observations revealed (i) tissue specificity for each of the microorganisms in the roots, with a cortical location of mycorrhizae and a mainly vascular cylinder location of nematodes, and (ii) that Gigaspora was the most abundant AM genus in the plantation soil.     

Glomus mosseae bioprotection against aster yellows phytoplasma (16srI-B) and Spiroplasma citri infection in Madagascar periwinkle

The bio-control potential of arbuscular mycorrhizal fungus Glomus mosseae against two pathogenic microorganisms aster yellows (AY) phytoplasma and Spiroplasma citri has been examined in Madagascar periwinkle (Catharanthus roseus). G. mosseae had a positive influence on healthy C. roseus plants and S. citri infection. It provided bioprotection against S. citri pathogen and induced significant degree of resistance to spiroplasma infection. Besides, symptom expression significantly reduced and shoot height, leaves number, root fresh and dry weight increased in spiroplasma-infected plants treated with mycorrhiza fungus. Although, G. mosseae had no positive effect on phytoplasma disease. The root architectures were affected by the phytoplasma pathogen, and the root surface area dramatically decreased in G. mosseae treated AY-infected periwinkles compared with the control. Nitrogen and Phosphorus concentrations notably increased in spiroplasma + G. mosseae compared with control plants. Potassium concentration did not differ significantly in all mycorrhizal treated and untreated infected plants except in G. mosseae treated healthy plants. The spore density and root colonization rate did not vary in both pathogen treatments G. mosseae + spiroplasma and G. mosseae + phytoplasma. To our knowledge, this is the first report showing the bioprotective effect of G. mosseae on S. citri. The possible mechanisms involved in complex interaction between plants, cell wall-less bacteria and arbuscular mycorrhizal fungi (AMF) are discussed and the underlying mechanisms for the functioning of AMF are hypothesized.     

几种除草剂和助剂对苜蓿Medicago sative出苗和生长的影响

在温室条件下研究了几种苗前除草剂单用或混用对苜蓿出苗和生长的影响,以及不同叶龄苜蓿对苗后除草剂的反应及添加助剂对药效的影响。结果表明:咪唑乙烟酸在60~120 g/hm2剂量下,于苗前施用,对苜蓿出苗和幼苗生长都有明显的抑制作用。咪唑乙烟酸(30 g/hm2)与二甲戊灵(495 g/hm2)混用,对苜蓿的药害显著降低。苗后施用除草剂对苜蓿的安全性与叶龄密切相关。咪唑乙烟酸以30~60 g/hm2剂量在苜蓿3叶期施用,对苜蓿安全;在120 g/hm2用量下,对5叶期的苜蓿无明显药害。乳氟禾草灵(130~260 g/hm2)和丙炔氟草胺(75~150 g/hm2) 无论是3叶期还是5叶期施用,对苜蓿幼苗都有严重的药害。与咪唑乙烟酸单用时比较,药液中添加1.0 mL/L 的平平加-15和AM-100及5.0 mL/L的SDP,对苜蓿幼苗生长无明显影响。     

Piriformospora indica reduces fusarium head blight disease severity and mycotoxin DON contamination in wheat under UK weather conditions

Piriformospora indica (Sebacinaceae) is a cultivable root endophytic fungus. It colonizes the roots of a wide range of host plants. In many settings colonization promotes host growth, increases yield and protects the host from fungal diseases. Evaluation was made of the effect of P. indica on fusarium head blight (FHB) disease of winter (cv. Battalion) and spring (cv. Paragon, Mulika, Zircon, Granary, KWS Willow and KWS Kilburn) wheat and consequent contamination by the mycotoxin deoxynivalenol (DON) under UK weather conditions. Interactions of P. indica with an arbuscular mycorrhizal fungus (Funneliformis mosseae), fungicide application (Aviator Xpro) and low and high fertilizer levels were considered. Piriformospora indica application reduced FHB disease severity and incidence by 70%. It decreased mycotoxin DON concentration of winter and spring wheat samples by 70 and 80%, respectively. Piriformospora indica also increased aboveground biomass, 1000‐grain weight and total grain weight. Piriformospora indica reduced disease severity and increased yield in both high and low fertilizer levels. The effect of P. indica was compatible with F. mosseae and foliar fungicide application. Piriformospora indica did not have any effects on plant tissue nutrients. These results suggest that P. indica might be useful in biological control of Fusarium diseases of wheat.     

Mycorrhizal colonization of chenopods and its influencing factors in different saline habitats,China

Chenopodiaceae is one of the most important families in arid and saline environments. Several studies have observed the mycorrhizal structure in Chenopodiaceae plants(i.e., chenopods), but the mycorrhizal colonization status of chenopods in saline habitats and the influencing factors are still not well understood. The mycorrhizal colonization of twenty chenopod species in three different saline habitats(a saline alkaline meadow in the Songnen Plain of northeastern China, a saline desert in the Junggar Basin of northwestern China, and a saline alpine meadow in the Tibetan Plateau of western China) and the chenopod-associated environmental factors(including soil moisture, soil available phosphorous(P) concentration, pH, and salt content) were analyzed. Our results showed that approximately 60% of the studied chenopods were colonized by arbuscular mycorrhizal(AM) fungi with a colonization percentage ranging from 5% to 33%. Structural analysis of mycorrhizal association indicated that vesicles were quite common, while arbuscules and hyphal coils were relatively rare. In addition, a positive correlation between mycorrhizal colonization rate and soil electrical conductivity(r=0.920, P0.01) and two negative correlations of mycorrhizal colonization rates with soil moisture(r= –0.818, P0.01) and the soil available P concentration(r= –0.876, P0.01) confirmed that mycorrhizal colonization rate in the roots of chenopods was environment-dependent.     

Arbuscular mycorrhizal fungi improved plant growth and nutrient acquisition of desert ephemeral Plantago minuta under variable soil water conditions

Desert ephemeral plants play an important role in desert ecosystem.Soil water availability is considered as the major restrictive factor limiting the growth of ephemeral plants.Moreover,arbuscular mycorrhizal fungi(AM fungi) are widely reported to improve the growth of desert ephemerals.The present study aimed to test the hypothesis of that AM fungi could alleviate drought stress of desert ephemeral Plantago minuta,and AM fungal functions reduced with the improvement of soil water content.A pot experiment was carried out with three levels of soil water contents(4.5%,9.0%,and 15.8%(w/w)),and three AM inoculation treatments(Glomus mosseae,Glomus etunicatum and non-inoculation).The results indicate that mycorrhizal colonization rate decreased with the increase of soil water availability.Inoculation improved plant growth and N,P and K acquisition in both shoots and roots regardless water treatments.When comparing the two fungi,plants inoculated with G.mosseae performed better than those inoculated with G.etunicatum in terms of plant growth and nutrient acquisition.These results showed that ameliorative soil water did not suppress arbuscular mycorrhizal fungal functions in improving growth and nutrient acquisition of desert ephemeral Plantago minuta.