Orobanche crenata resistance and avoidance in pea (Pisum spp.) operate at different developmental stages of the parasite

Orobanche crenata (broomrape) is an important constraint to pea (Pisum sativum) cultivation in the Mediterranean area, because little resistance is available in commercial crop varieties. Field experiments have demonstrated that some resistance is present in a number of P. sativum and P. fulvum accessions. The goal of this work was to characterize such resistance. The Pisum–O. crenata interaction and the resistance symptoms were studied under controlled conditions by using Petri dish and polyethylene bag assays. The content of phenolics and peroxidase activity in host tissue from infected and non-infected plants were also measured. Resistance and avoidance mechanisms, acting at different developmental stages of the parasite, have been identified, including low stimulation of O. crenata seed germination, unsuccessful penetration of host roots, delay in post-attachment tubercle development and necrosis of the attached tubercles. Infection caused an increase in the content of total soluble phenolics in some Pisum genotypes. Peroxidase activity was higher in resistant than in susceptible accessions. Results obtained with different Pisum genotypes showed that resistance is the result of several mechanisms acting at different stages of the infection process. Resistance is also related to increased levels of peroxidase activity in host roots.     

AM真菌诱导的番茄信号物质及其对根结线虫的 抑制 效应

丛枝菌根(AM)真菌能诱导植物合成一些信号物质,如茉莉酸(JA)、水杨酸(SA)、一氧化氮(NO)、H2O2等,这些信号在AM真菌与植物识别、共生体建立和激活植物防御系统过程中发挥着重要作用[1].     

Revisiting strategies for reducing the seedbank of Orobanche and Phelipanche spp.

Weedy Orobanche and Phelipanche species are important in Southern and Eastern Europe, the Middle East and North Africa, and have recently been reported in the USA, Australia and some countries in South America. Rather than being controlled, the Orobanche and Phelipanche problem is increasing, both in intensity and in acreage. Large areas of new territory are at risk of invasion, if care is not immediately taken to limit the introduction of parasitic seeds and to educate farmers and others to be alert for new infestations. There is an urgent need to re-evaluate novel integrated Orobanche and Phelipanche management programmes that will allow a better control of the parasite species and limit their distribution. The main obstacle in the long-term management of Orobanche and Phelipanche infested fields is the durable seedbank, which may remain viable for decades in the field. Large quantities of long-lived seeds give the parasite genetic adaptability to changes in host resistance and cultural practices. As long as the seedbank is not controlled, the need to control the parasite will persist whenever a susceptible host is grown in the infested field. We discuss strategies for seedbank reduction, paying particular attention to cultural practices, whereas chemical and biological control methods are covered by other reviews in this issue.     

Pathogenic diversity within field populations of Orobanche cumana and different reactions on sunflower genotypes

Different races of the parasitic Orobanche cumana (sunflower broomrape) have been reported in Spain, race F being the most virulent. Full resistance in sunflower to races A–E is achieved with each of the single major genes Or₁ to Or₅ respectively. However, parasitised hybrids allegedly resistant to race F were observed in early 2002. The purpose of this study was to verify broomrape incidences (BI) on resistant sunflower genotypes, to assess the mixture of races within field populations and to test for partial resistance to race F in the sunflower hybrids showing a low degree of attack (DA) by the weed. Tests were conducted under field conditions in two locations of southern Spain. While no significant differences were found for yield and BI between locations, the DA on the cultivars depended on the location. With high infection levels and significantly lower yield in susceptible controls, marked differences in BI and DA were found within resistant cultivars, but all of them showed similar crop yield. When artificially inoculated with several populations of race F, line P96 and mainly line L86, were consistently slightly infected, suggesting they were inbred lines responsible for horizontal resistance in infested fields. L86 was extremely susceptible to race E populations, which is unusual as sunflower resistance to one race provided resistance to all the previously described races of O. cumana. No different virulences were detected within two groups of subpopulations (races E and F) inoculated onto resistant sunflower genotypes. However, race F subpopulations showed significant differences in aggressiveness, which seems to be related to horizontal (multigenic) resistance of the crop to the parasitic weed.     

Comparative proteomic analysis of Orobanche and Phelipanche species inferred from seed proteins

Orobanche and Phelipanche species are root parasitic plants some of which infect important crops, causing extensive damage. In an attempt to unravel molecular systematic in these genera using protein markers, we established the protein profile of twelve species by two-dimensional gel electrophoresis (2-DE). Proteins were detected by Sypro Ruby staining and the gel images were digitalised and analysed. The 2-DE maps contained from 30 to 140 protein spots for the analysed species. A virtual master gel was built with spots present in all species and revealing the presence of 260 protein spots. Only one protein was monomorphic, while 143 were unique to a single species. Proteins were scored for the presence or absence of homologous spots to create a binary matrix. Phylogenetic analysis was carried out using maximum parsimony and distance method. The trees obtained were rooted using Cistanche phelypaea as an out-group. Both analyses showed Orobanche as a monophyletic group. AMOVA analysis revealed phenotypic diversity between species within genera, as well as a considerable level of variation between genera. This is a first step to help in the phylogenetic classification of parasitic plants by means of proteomics.     

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.     

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。     

水分胁迫下AM真菌和根瘤菌对沙打旺生长及养分吸收的影响

利用温室盆栽试验研究水分胁迫下接种丛枝菌根（Arbuscular mycorrhizal,AM）真菌和根瘤菌（Rhizobium meliloti）对沙打旺(Astragalus adsurgens Pall.) 生长和养分吸收的影响。在土壤相对含水量65%和35%条件下,分别设不接种（对照）、单接根瘤菌、单接摩西球囊霉(Glomus mosseae)和双接根瘤菌与摩西球囊霉等4个处理。结果发现：干旱胁迫显著抑制沙打旺AM真菌侵染率（P<0.05）,而接种根瘤菌在两种水分条件下均显著促进摩西球囊霉对沙打旺根系的侵染（P<0.05）。接种AM真菌不仅显著提高沙打旺对P的吸收（P<0.05）,而且明显促进根瘤的生长。无论是在干旱条件下或是在正常供水条件下,双接根瘤菌与摩西球囊霉处理对沙打旺生长及养分吸收的效应显著高于单接菌处理,植株地上部、地下部生物量以及N、P、K等吸收量均为最大。结果表明：AM真菌与根瘤菌双接种技术在干旱、半干旱区受损生态系统的植被恢复中具有一定的应用潜力。     

Molecular and biochemical mechanisms of defence induced in pea by Rhizobium leguminosarum against Orobanche crenata

Orobanche species (broomrapes) are parasitic weeds which dramatically decrease the yields of many economically important dicotyledonous crops, including pea (Pisum sativum), in Mediterranean areas. Previously, we identified some Rhizobium leguminosarum strains, including P.SOM, which could both promote pea development and significantly reduce infection by Orobanche crenata, notably through induction of necrosis of attached parasites. In the present study, induced resistance against broomrape in the nodulated pea was shown to be associated with significant changes in rates of oxidative lipoxygenase (Lox) and phenylpropanoid/isoflavonoid pathways and in accumulation of derived toxins, including phenolics and pisatin (pea phytoalexin). Changes were followed for 5 weeks after inoculation and attack by Orobanche. In contrast to non‐inoculated plants or Orobanche only infected plants, polyphenoloxidase (PPO) activity and hydrogen peroxide content increased in response to bacteria inoculation indicating the involvement of oxidative processes. In parallel, the nodulated roots displayed high Lox activity related to the overexpression of the lox1 gene. Similarly, the expression of phenylalanine ammonia lyase (PAL) and 6a‐hydroxymaackiain 3‐O‐methyltransferase (Hmm6a) genes were induced early during nodule development, suggesting the central role of the phenylpropanoid/isoflavonoid pathways in the elicited defence. As a consequence, the derived products, phenolics and pisatin, accumulated in response to rhizobacteria and conferred mechanical and chemical barriers to the invading parasite. These results highlight the likely role of signalling pathways in induced resistance and suggest these mechanisms should be enhanced through integrated Orobanche management practices.     

Extent and pattern of genetic differentiation within and between European populations of Phelipanche ramosa revealed by amplified fragment length polymorphism analysis

The extent and pattern of genetic differentiation between Phelipanche ramosa populations colonising tobacco in different European regions were investigated by amplified fragment length polymorphism (AFLP) analysis, in order to determine levels of variation for tobacco resistance breeding and management programmes. Four different AFLP primer pairs amplified a total of 1050 clear and reproducible bands, of which 962 (91.62%) were polymorphic among the 35 individuals taken from four P. ramosa populations collected in Spain, Italy, Bulgaria and Germany. Cluster analysis based on the AFLP data categorised the plants into distinct groups, in line with their geographical origin, denoting clear genetic differentiation among the four populations. This differentiation was supported by both high bootstrap values and significant results of the analysis of molecular variance. The most divergent population was the one from Bulgaria. The majority of the genetic diversity was attributable to differences among populations (77.80%), as expected from the predominant autogamous behaviour of this species. Populations differed significantly in within-population diversity, as measured by Shannon's information index. The German population presented the lowest genetic diversity and the Italian population harboured the highest level of within-population genetic diversity. There are significant differences in genetic diversity level among the studied P. ramosa populations and clear population-specific genetic diversity structures. These need to be taken into account, together with the potential differences in parasite aggressiveness, when planning breeding and management strategies for P. ramosa control in tobacco cultivation.     

Resistance to Pseudomonas syringae in a collection of pea germplasm under field and controlled conditions

A total of 242 Pisum accessions were screened for resistance to Pseudomonas syringae pv. pisi under controlled conditions. Resistance was found to all races, including race 6 and the recently described race 8. Fifty‐eight accessions were further tested for resistance to P. syringae pv. syringae under controlled conditions, with some highly resistant accessions identified. Finally, a set of 41 accessions were evaluated for resistance to P. syringae pv. pisi and pv. syringae under spring‐ and winter‐sowing field conditions. R2, R3 and R4 race‐specific resistance genes to P. syringae pv. pisi protected pea plants in the field. Resistance sources to race 6 identified under controlled conditions were ineffective in the field. Frost effects were also evaluated in relation to disease response. Results strongly suggest that frost tolerance is effective in lowering the disease effects caused by P. syringae pv. pisi and pv. syringae under frost‐stress conditions, even in the absence of disease resistance genes, although the highest degree of this protection is reached when frost tolerance and disease‐resistance genes are combined in the same genetic background.     

Regulation of biosynthesis,perception, and functions of strigolactones for promoting arbuscular mycorrhizal symbiosis and managing root parasitic weeds

Strigolactones (SLs) are carotenoid‐derived plant secondary metabolites that play important roles in various aspects of plant growth and development as plant hormones, and in rhizosphere communications with symbiotic microbes and also root parasitic weeds. Therefore, sophisticated regulation of the biosynthesis, perception and functions of SLs is expected to promote symbiosis of beneficial microbes including arbuscular mycorrhizal (AM) fungi and also to retard parasitism by devastating root parasitic weeds. We have developed SL mimics with different skeletons, SL biosynthesis inhibitors acting at different biosynthetic steps, SL perception inhibitors that covalently bind to the SL receptor D14, and SL function inhibitors that bind to the serine residue at the catalytic site. In greenhouse pot tests, TIS108, an azole‐type SL biosynthesis inhibitor effectively reduced numbers of attached root parasites Orobanche minor and Striga hermonthica without affecting their host plants; tomato and rice, respectively. AM colonization resulted in weak but distinctly enhanced plant resistance to pathogens. SL mimics can be used to promote AM symbiosis and to reduce the application rate of systemic‐acquired resistance inducers which are generally phytotoxic to horticultural crops. © 2019 Society of Chemical Industry     

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。     

Role of seed infection by the Ascochyta blight pathogen of dried pea (Mycosphaerella pinodes) in seedling emergence, early disease development and transmission of the disease to aerial plant parts

The role of infected seed in the epidemiology of Ascochyta blight of pea, caused by Mycosphaerella pinodes, was studied both under growth chamber and field conditions, using healthy seeds, naturally infected seeds and artificially infected seeds. Results suggest that infected seeds caused serious losses, as a result of poor germination and high transmission of the disease, to parts of the plants under soil level. Foot rot symptoms often caused the death of young seedlings. Losses were increased by low temperatures during the early stage of crop development. M. pinodes progressed from seeds to aerial parts of the plants, but no Ascochyta blight symptoms occurred, the disease remaining near to the basal parts of the plants as a foot rot symptom. This suggests that seeds cannot be regarded as a source of contamination in the epidemiology of the disease.     

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.     

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.     

Effects of antagonistic fungi,plant growth-promoting rhizobacteria,and arbuscular mycorrhizal fungi alone and in combination on the reproduction of <Emphasis Type="Italic">Meloidogyne incognita</Emphasis> and growth of tomato

Antagonistic fungi (Aspergillus niger CA and Penicillium chrysogenum CA1), plant growth-promoting rhizobacteria (PGPR) (Burkholderia cepacia 4684 and Bacillus subtilis 7612) and AM fungi (Glomus intraradices KA and Gigaspora margarita AA) were assessed alone and in combination for their effects on the growth of tomato and on the reproduction of Meloidogyne incognita in glasshouse experiments. Application of antagonistic fungus, PGPR, or AM fungus alone or in combination significantly increased the length and shoot dry mass of plants both with and without nematodes. The increase in shoot dry mass caused by Gl. intraradices KA in plants without nematodes was greater than that caused by PGPR or antagonistic fungi. Similarly, the increase in shoot dry mass caused by Bu. cepacia 4684 in plants with nematodes was greater than that caused by P. chrysogenum CA1. Application of Bu. cepacia 4684 caused a 36.1% increase in shoot dry mass of nematode-inoculated plants followed by Ba. subtilis 7612 (32.4%), A. niger CA (31.7%), Gl. intraradices KA (30.9%), Gi. margarita AA (29.9%) and P. chrysogenum CA1 (28.8%). Use of Bu. cepacia 4684 with A. niger CA caused a highest (65.7%) increase in shoot dry mass in nematode-inoculated plants followed by Ba. subtilis 7612 plus A. niger CA (60.9%). Burkholderia cepacia 4684 greatly reduced (39%) galling and nematode multiplication, and the reduction was even greater (73%) when applied with A. niger CA. Antagonistic fungi had no significant effect on root colonization caused by AM fungi. Applying Bu. cepacia 4684 with A. niger CA may be useful in the biocontrol of M. incognita on tomato.     

不同真菌和植物种类对苜蓿滑刃线虫繁殖的影响

为探究美国进境苜蓿滑刃线虫对真菌的取食特性以及对植物的寄生性,通过接种试验观察该线虫在链格孢、灰葡萄孢、禾谷镰刀菌、拟盘多毛孢和核盘菌等5种真菌,以及在菊花、大豆、苜蓿和三叶草等4种植物8个品种上的繁殖情况.结果表明,苜蓿滑刃线虫能取食5种真菌的菌丝体,但是在各真菌上的繁殖系数(Rf)有明显差异,在链格孢和拟盘多毛孢上...