Influence of Fusarium solani on citrus root rot caused by Phytophthora parasitica and Phytophthora citrophthora

Interactions between Fusarium solani and Phytophthora parasitica or F. solani and P. citrophthora influenced the development of root rot of citrus but depended on the temporal order of inoculation with F. solani or the two Phytophthora spp. Inoculation of citrus with either Fusarium solani and Phytophthora parasitica or Phytophthora citrophthora increased root rot compared to inoculation with P. parasitica or P. citrophthora alone when plants were inoculated with Phytophthora by dipping their roots in zoospore suspensions and subsequently transplanted into soil infested with F. solani. However, root rot was not increased by simultaneous co-inoculation of P. parasitica and F. solani or when plants were inoculated with F. solani first. Root rot was not increased when heat-stressed or non-stressed plants were inoculated with P. parasitica 30 days after transplanting into soil infested with F. solani. In most but not all experiments, F. solani alone reduced growth of tops or roots a small but significant amount.Co-inoculation of citrus by root-dipping into zoospore suspensions of P. parasitica and transplanting into soil infested with F. solani reduced feeder root length by 62% and root weight by 61% but did not significantly reduce the percentage of living roots when compared to inoculation with P. parasitica alone. When citrus roots were immersed in zoospore suspensions of P. citrophthora and transplanted into soil infested with F. solani, feeder root length was reduced by 68%, but feeder root weight and the percentage of living roots were not significantly reduced when compared to plants inoculated with P. citrophthora alone.Propagule densities of both P. parasitica and P. citrophthora in the rhizosphere of plants inoculated by root-immersion and then transplanting into soil infested with F. solani were not significantly different than propagule densities from plants transplanted into non-infested soil. Propagule densities of P. parasitica were suppressed an average of 41% when citrus was inoculated with P. parasitica 30 days after transplanting into soil infested with F. solani and by 41% when citrus was co-inoculated by transplanting into soil infested with both F. solani and P. parasitica.     

Behaviour of Phytophthora citrophthora and P. nicotianae var. in soil, and Differences in Their Tolerance to Antimicrobial Components of Selective Media Used for Isolation of Phytophthora spp.

Phytophthora citrophthora was inhibited to a greater extent than P. nicotianac var. parasitica by chloramphenicol, hymexazol, PCNB and pimaricin at concentrations used in selective media for the isolation of Phytophthora spp. Reduced concentrations of the antimicrobial components of the selective media to tolerant levels for P. citrophthora markedly increased the recovery of the two brown rot pathogens from soil. Mycelium of both Phytophthora spp. survived in air-dried soil for at least 5 months while mycelium of most Phytophthora spp. do not survive in dry soil. In moist soil, P. nicotianae var. parasitica produced hyphal swellings, sporangia and chlamydospores. P. citrophthora produced hyphal swellings and sporangia, but no chlamydospores. No oospores were produced, even in pairing cultures on agar plates with isolates obtained from several locations of citrus groves andfruits by both species. Sporania were obtained in both species in citrus groves on mycelium mats, in the rhizosphere, in infected leaves and fruits buried at soil depths of 5–35 cm. Numbers of propagules declined during the incubation period, but conside, rable numbers survived throughout the experimental period (6 months). Although P. nicotianae var. parasitica produced chlamydospores while P. citrophthora did not, numbers of surviving propagules recovered from soil after 6 months were comparable with both species. The brown rot pathogens survived in soil both as colonizers of detached leaves and fruits and as parasites in living root tissues.     

Distribution of Phytophthora citrophthora R.E. Sm. and E.H. Sm. and P. parasitica Dastur within citrus orchards in Kerman province,Iran

In this research, distribution of Phytophthora species were determined in Kerman Province (Bam, Shahdad and Arzuiyeh) during 2004–2007. The Phytophthora species were isolated from infected root, crown and soil. Root and crown pieces were washed and cultured on a CMA-PARPH medium. The isolation from infected soil was performed by bating method using citrus leaves. It was identified by morphological and several physiological characteristics. Distribution of species was determined by recording the number of isolates recovered from samples from each city. In this study, from 220 soil samples collected from 52 citrus orchards, 80 isolates of Phytophthora were recovered. Among of all isolates of Phytophthora, P. parasitica and P. citrophthora were the most important species of causal agent of gummosis on Citrus sp. Distribution of P. citrophthora was highest in Arzuiyeh and lowest in Bam and Shahdad cities, while distribution of P. parasitica was highest in Bam and Shahdad cities. The majority of soil samples contained only P. parasitica, but a few of percentage samples containing P. citrophthora alone and both of fungi in cites samples.     

Increased tolerance to Phytophthora citrophthora in transgenic orange plants constitutively expressing a tomato pathogenesis related protein PR-5

Phytophthora citrophthora is the most widely spread oomycete plant pathogen over all the citrus growing areas and represents one of the major causes of crop losses. Constitutive over-expression of genes encoding proteins involved in plant defence mechanisms to disease is one of the strategies proposed to increase plant tolerance to oomycete and fungal pathogens. P23 (PR-5), a 23-kDa pathogenesis-related protein similar to osmotins, is induced in tomato (Lycopersicon esculentum Mill. cv. Rutgers) plants when they are infected with citrus exocortis viroid, and its antifungal activity has been demonstrated in in vitro assays. We have successfully produced transgenic orange (Citrus sinensis L. Obs. cv. Pineapple) plants bearing a chimeric gene construct consisting of the cauliflower mosaic virus 35S promoter and the coding region of the tomato pathogenesis-related PR-5. Nine regenerated transgenic lines constitutively expressed the PR protein. They were challenged with Phytophthora citrophthora using a detached bark assay. A significant reduction in lesion development was consistently observed in one transgenic line in comparison to the control plants. This same line achieved plant survival rates higher than control plants when transgenic trees were inoculated with oomycete cultures. These results provide evidence for the in vivo activity of the tomato PR-5 protein against Phytophthora citrophthora, and suggest that this may be employed as a strategy aimed at engineering Phytophthora disease resistance in citrus.     

Identification of Phytophthora citrophthora with Cloned DNA Probes

Two different DNA fragments, one of 2.9 kilobases and the other of 5.1 kilobases, were cloned from Phytophthora citrophthora and showed no homology with DNA from plants and other related fungi. These DNA probes hybridized with DNA from 12 different P. citrophthora isolates obtained from a variety of hosts but did not hybridize with DNA from 6 P. citrophthora isolates obtained from cacao. Southern blot analysis revealed that the probes contained repetitive DNA, and restriction fragment length polymorphisms were identified among several P. citrophthora isolates. Of the isolates tested, two major groups were observed whose genetic similarity correlated with geographical distribution. One of the DNA probes was used to detect P. citrophthora growing from infected citrus roots incubated on semiselective medium. P. citrophthora was not detected by a hybridization assay of total DNA extracted directly from infected roots.     

The Influence of Glomus mosseae on Tylenchulus semipenetrans-Infected and Uninfected Citrus limon Seedlings

Greenhouse studies have shown that when rough lemon (Citrus limon) seedlings infected with TyIenchulus semipenetrans were transplanted into soil infested with Glomus mosseae, the mycorrhizal fungus infection increased seedling growth compared to nonntycorrhizal seedlings. Tylenchulus semipenetrans significantly suppressed seedling growth below that of mycorrhizal seedlings. Histological observations of nematode-free mycorrhizal roots showed that hyphae penetrated the epidermis and invaded the cortex, giving rise to arbuscules and vesicles. Nematode infection sites in T. semipenetrans-infected roots grown in soil infested with G. mosseae did not show evidence of vesicle development in the cortex but did show arbuscule development.     

Citrus replant problem in Iraq I. Possible role of soil fungi and nematodes

The role of soil fungi and nematodes in citrus replant problems was investigated. Several pathogenic and non-pathogenic fungi and the nematodeTylenchulus semipenetrans were isolated in about equal abundance from soils which displayed or did not display replant problems. However, in almost all trees showing evidence of decline, infection by the fungusPhytophthora citrophthora had advanced through the roots until it had reached the tree crown. Fumigation of old citrus soil with methyl bromide greatly increased the growth of sour organe. The application of Nemacure, Benomyl and Ridomil 5G confirmed the important role of pathogenic and non-pathogenic fungi and nematodes in the citrus replant problem. Inoculation of methyl bromide-treated soil with the pathogenic and non-pathogenic fungi separately or in combination further confirmed the previous experiments and also revealed an important role ofP. citrophthora andFusarium solani when replanting citrus in old citrus orchards.     

The infection of potatoes by Phoma exigua

The pathogenicities of Phoma exigua var. foveata and P. exigua var. exigua were compared by determining the relationship between incidence of infection of whole tubers or tuber pieces and various inoculum levels in soil. In natural soil var. foveata was more pathogenic than var. exigua, but in sterilized soil there were no clear differences between the two. Both varieties were isolated from some lesions derived from inoculum to which only one variety had been added indicating either natural contamination of soil or latent infection in tubers; the proportion of such lesions increased at the lower inoculum concentrations. In studies where potato plants were grown in infested soils, the incidence of stem disease and tuber disease increased at increased inoculum concentrations of var. foveata; var. exigua was not added to the inoculum but was isolated from some lesions, the proportions of such lesions again increasing at lower inoculum concentrations. The infectivity of the infested soil, assessed by a tuber-baiting method, declined soon after planting but increased prior to harvest. Laboratory studies indicated that in natural soil the infectivity was highest after 2–3 months and lowest after 10 months incubation, but in sterilized soil infectivity was highest after 5 months and lowest after 7 months incubation.     

The response of seedlings of two dipterocarp species to nutrient additions and ectomycorrhizal infection

An experiment was conducted to investigate the effect of ectomycorrhizal infection on growth and nutrient uptake, especially of P and K of dipterocarp seedlings.Hopea helferi (Dyer) Blanco andHopea odorata Roxb. seedlings were grown in a sandy loam soil given a basal dressing. Nutrient treatments were unamended soil (NIL), amended soil with the addition of P and K (F), amended soil without P but with K (-P), amended soil without K but with P (-K), amended soil without P or K addition (-PK). Seedlings grown in the amended soil treatments showed foliar symptoms suggestive of calcium deficiency. Ectomycorrhizal infection appeared to improve shoot Ca concentration and relieved the foliar symptoms. Ectomycorrhizal infection inH. odorata plants increased shoot P concentration and increased shoot and total dry weight to the same or greater extent than those of uninfected plants growing on P amended soil.H. helferi showed a positive response to ectomycorrhizal infection in shoot, root and total dry weight in all nutrient treatments but no response to the nutrient treatments themselves.     

Potential of endophytic Streptomyces spp. for biocontrol of Fusarium root rot disease and growth promotion of tomato seedlings

Sixteen endophytic actinobacteria isolated from roots of native plants were evaluated for their antagonistic potential against soil-borne phytopathogenic fungi. Among them, three strong antagonistic isolates were selected and characterised for in vitro plant-growth-promoting and biocontrol traits, including production of hydrogen cyanide, indole-3-acetic acid and siderophores, chitinase and β-1,3-glucanase activities, and inorganic phosphate solubilisation. In all trials, the strain Streptomyces sp. SNL2 revealed promising features. The selected actinobacteria were investigated for the biocontrol of Fusarium oxysporum f. sp. radicis lycopersici and for growth promotion of tomato (Solanum lycopersicum L. cv. Aïcha) seedlings in autoclaved and non-autoclaved soils. All seed-bacterisation treatments significantly reduced the root rot incidence compared to a positive control (with infested soil), and the isolate SNL2 exhibiting the highest protective activity. It reduced the disease incidence from 88.5% to 13.2%, whereas chemical seed treatment with Thiram® provided 14.6% disease incidence. Furthermore, isolate SNL2 resulted in significant increases in the dry weight, shoot and root length of seedlings. 16S rDNA sequence analysis showed that isolate SNL2 was related to Streptomyces asterosporus NRRL B-24328^T (99.52% of similarity). Its interesting biocontrol potential and growth enhancement of tomato seedlings open up attractive uses of the strain SNL2 in crop improvement.     

Phytophthora root-rot of container-grown citrus as affected by foliar sprays and soil drenches of phosphorous and acetyl salicylic acids

Mycelial growth ofPhytophthora nicotianae var.parasitica in vitro was inhibited by phosphorous and acetyl salicylic acids at 10 ppm; mycelial growth ofP. citrophthora was inhibited by phosphorous acid at 10 ppm and acetyl salicylic acid at 100 ppm. Foliar sprays and soil drenches with these chemicals were compared with soil applications of metalaxyl as treatments for Phytophthora root-rot of container-grown citrus (var. Cleopatra mandarin) caused by the aforementioned spp. Foliar sprays with 0.1% phosphorous acid were as effective as the metalaxyl treatment in promoting root growth and phosphorous acid offers considerable potential as an alternative treatment for Phytophthora root-rot of citrus.     

The effect of temperature on the infection of onions by Sclerotium cepivorum

Studies on Sclerotium cepivorum infection of salad onions in artificially infested soil showed that under field conditions infection was greatest in late spring and summer and declined to a low level in the late autumn and winter months. Change of infection levels was found to be correlated with the effect of soil temperature in the field on the germination of sclerotia. This was substantiated in laboratory studies which demonstrated that sclerotium germination, mycelial growth and seedling infection were all markedly influenced by temperature.     

Efficiency of different application methods of biocontrol agents and biocides in control of Fusarium root rot on some citrus rootstocks

Abstract

The efficiency of two biocontrol agents (Trichoderma harzianum NB and Bacillus subtilis NB) and two commercial biocides (Plant Guard and Rhizo-N) in controlling Fusarium root rot disease on some citrus rootstocks was evaluated under artificially infested soil in green house.

Fusrium root rot on citrus rootstocks seedlings i.e. sour orange (SO), volkamer lime (VL), rangpur lime (RP) and cleopatra mandarin (CL) was successfully controlled by dipping the root system of such seedlings in water suspensions of each biological treatment i.e. Trichoderma harzianum (spore suspension 5×10⁶ spore/ml), Bacillus subtilis (cell suspension 8×10⁷ cell/ml), Plant Guard (3 g/l) and Rhizo-N (4 g/l), then transplanted into artificially infested soil with Fusarium solani and drenched with enough water suspension of such biological treatments. Plant Guard (3 g/l) and Rhizo-N (4 g/l) were highly effective treatments in decreasing infection and severity of the disease, Fusarium density in rhizosphere soil and colonization of Fusarium solani in the roots of all tested seedlings.

Meanwhile, root dipping or soil drenching with the same treatments individually gave the least effect in reducing root rot incidenceon all tested rootstocks compared with application of the two methods together.

It should be noted that using biocontrol agents and commercial biocides could be successfully used in controlling root rot pathogens on citrus in commercial greenhouses or under field conditions before transplanting in new reclaimed lands in the desert.     

Species-mediated soil moisture availability and patchy establishment of Pseudotsuga menziesii in chaparral

The occurrence of mature individuals of Pseudotsuga menziesii in stands of Arctostaphylos species mark the initial stages of mixed evergreen forest invasion into chaparral in central coastal California. We planted two cohorts of P. menziesii seeds at three sites under stands of two Arctostaphylos species and Adenostoma fasciculatum in order to determine whether first-year seedling emergence and survival, particularly during the regular summer drought, underlie the spatial distribution of mature trees observed in chaparral. Regardless of the chaparral species they were planted under, P. menziesii seeds that were not protected from vertebrate predation displayed very little emergence and no survival. In contrast, emergence of P. menziesii that were protected from vertebrate predators was much higher but still did not significantly differ among the three chaparral species. However, survival of protected seedlings under Arctostaphylos glandulosa was much greater than under A. fasciculatum, with intermediate survival under Arctostaphylos montana. While mortality of protected seedlings due to insect herbivory, fungal infection, and disturbance displayed no consistent patterns, summer drought mortality appeared to drive the patterns of survival of P. menziesii under the different chaparral species. These emergence, mortality, and survival data suggest that spatial patterns of P. menziesii recruitment in chaparral are driven by first-year summer drought seedling mortality, but only in years when seeds and seedlings are released from vertebrate predation pressure. Because the first-year drought mortality and survival patterns of P. menziesii seedlings differed strongly depending on the chaparral species, we examined the additional hypothesis that these patterns are associated with differences in the availability of soil moisture under different chaparral species. Both higher survival and lower drought mortality of P. menziesii seedlings were associated with higher soil water potential under Arctostaphylos stands during the summer drought, especially in the subsurface soil. The data suggest that Arctostaphylos stands, particularly stands of A. glandulosa, ameliorate xeric summer conditions to a degree that facilitates first-year establishment of P. menziesii and strongly influences spatial distribution of mature trees. Received: 18 September 1998 / Accepted: 23 December 1998     

Interaction of Rotylenchulus reniformis with Seedling Disease Pathogens of Cotton

The impact of 10 Fusarium species in concomitant association with Rotylenchulus reniformis on cotton seedling disease was examined under greenhouse conditions. In experiment 1, fungal treatments consisted of Fusarium chlamydosporum, F. equiseti, F. lateritium, F. moniliforme, F. oxysporum, F. oxysporum f.sp. vasinfectum, F. proliferatum, F. semitectum, F. solani, and F. sporotrichioides; Rhizoctonia solani; and Thielaviopsis basicola. The experimental design was a 2 × 14 factorial consisting of the presence or absence of R. reniformis and the 12 fungal treatments plus two controls in autoclaved field soil. In experiment 2, the same fungal and nematode treatments were examined in autoclaved or non-autoclaved soil. This experimental design was a 2 × 2 × 14 factorial consisting of field or autoclaved soil, presence or absence of R. reniformis, and the 12 fungal treatments plus two controls. In both tests, Fusarium oxysporum f. sp. vasinfectum, F. solani, R. solani, and T. basicola consistently displayed extensive root and hypocotyl necrosis that was more severe (P ≤ 0.05) in the presence of R. reniformis. Soil treatment (autoclaved vs. non-autoclaved) influenced the impact of the Fusarium species on cotton seedling disease, with disease being more severe in the autoclaved soil. Rotylenchulus reniformis reproduction on cotton seedlings was greater in field soil compared to autoclaved soil (P ≤ 0.05). This study suggests the importance of Fusarium species and R. reniformis in cotton seedling disease.     

Taxon-specific multiplex-PCR for quick,easy, and accurate identification of encyrtid and aphelinid parasitoid species attacking soft scale insects in California citrus groves

Citricola scale, Coccus pseudomagnoliarum Kuwana (Hemiptera: Coccidae), is a serious pest of citrus in California’s San Joaquin Valley, but not in southern California where a complex of Metaphycus spp. Mercet (Hymenoptera: Encyrtidae) suppress it. This has created interest in using these (and other Metaphycus) species for biological control in the San Joaquin Valley. A critical step in assessing an organism’s potential for biological control is the ability to accurately identify it. For Metaphycus spp., this currently requires slide mounted adult specimens and expert taxonomic knowledge. We present a simple, quick and accurate method to identify any life stage of the ten major parasitoids of soft scales in California citrus, based on amplification of ribosomal DNA, using the polymerase chain reaction (PCR). Three multiplex-PCR protocols amplify products of taxon-specific sizes, allowing direct diagnosis of taxa accommodated by the PCR, and reducing identification time to a fraction of that of existing methods.     

Chemical,physical and biological control of carrot seedling diseases

In naturally infested soil containingPythium ultimum, P. acanthicum andPhytophthora megasperma, onlyP. ultimum was associated with root rot and damped-off seedlings. Damping-off was promoted by low soil temperatures and by flooding. Seedling stands were markedly reduced when seed was pre-incubated in soil at 12°C but not at 25°C or 35°C. Dusting carrot seed with metalaxyl significantly increased seedling stands in the field at rates from 1.5–6 g kg⁻¹ seed and in both flooded and unflooded, naturally infested soil at 3.15 g kg⁻¹. In greenhouse experiments using artifically infested soil,P. ultimum andP. paroecandrum caused damping-off of carrot seedlings andRhizoctonia solani reduced root and shoot weights.R. solani caused damping-off in nutrient-enriched soil.P. acanthicum andP. megasperma were not pathogenic to seedlings, although both fungi colonized roots. Soil populations of allPythium spp., particularlyP. ultimum, increased during growth of seedlings and population growth ofP. megasperma was promoted by periodic flooding. Infestation of soil withP. acanthicum did not reduce damping-off of carrot seedlings byP. ultimum orP. paroecandrum, but significantly increased root and shoot weights and decreased root colonization byR. solani P. acanthicum has potential as a biocontrol agent againstR. solani.