Epiphytic microflora of poplar clones susceptible and resistant to infection by Dothichiza populea

One-year-old poplar shoots (nodes, internodes and lenticels) of clones susceptible to infection by the pathogenic fungus Dothichiza populea, viz. Populus nigra Italica and P. Robusta, resistant ones, viz. P. Grandis and P. Hybrida 275, as well as a hybrid of a susceptible and a resistant clone, viz. P. maximowiczii x P. nigra (P. Kórnik 42), were used. The plate method was employed to determine: 1. the abundance of the epiphytic microflora on a mineral medium with glucose; 2. the quantitative composition of epiphytic communities by determining the numbers of typical bacteria (including rod-shaped, spherical and sporulating forms), actinomycetes and yeasts in microscopic preparations from epiphyte colonies; 3. the abundance and level of activity of epiphytes antagonistic towards Dothichiza populea. In all poplar clones the epiphytic microflora was most abundant on nodes and least abundant on lenticels. In the resistant clones epiphytes were 7 (P. Grandis) to as many as 84 times (P. Hybrida 275) less numerous than in the susceptible ones. In the microflora communities of the susceptible poplars, rod-shaped bacteria were the most abundant, and in the resistant ones and the hybrid, yeasts, which made up from 60% to 70% of the strains tested. Spherical and sporulating bacteria as well as actinomycetes were found in numbers not exceeding 4% of the total number of epiphytes. The proportion of antagonistic microflora in whole epiphytic communities was higher in the resistant clones and the hybrid than in the susceptible clones, with the microflora having a more restrictive effect on the development of the pathogen.     

Effect of epiphytes on the extent of necrotic injuries of resistant and susceptible poplar clones infected with Dothichiza populea.

Poplar cuttings of a resistant clone, Populus ‘Grandis’, and susceptible clones, Populus nigra ‘Italica’ and Populus ‘Robusta’, were infected with the pathogenic fungus Dothichiza populea alone, or with the pathogen and one of five strains of epiphytes antagonistic towards it (in vitro), isolated from poplar bark. The extent of injury was examined for 28 days after infection by determining the length of necrotic patches and their area as expressed in per cent of the total area of a cutting or the area of necrotic injuries caused by the pathogen alone.

All the poplar cuttings of both the resistant and susceptible clones became diseased when infected with the pathogen alone. Surprisingly enough, however, the least affected clone was the susceptible P. ‘Robusta’, in which necrotic injuries covered 28% of the total area, as against 40% and 70% in the resistant P. ‘Grandis’ and the susceptible P. nigra ‘Italica’, respectively.

When the cuttings were infected simultaneously with Dothichiza populea and its antagonistic epiphytes, the diseased area in the resistant clone diminished by as much as two-thirds, and in the susceptible P. nigra ‘Italica’, by one-third in comparison with the area affected by the pathogen alone. In turn, in the susceptible P. ‘Robusta’ the introduction of three out of five epiphytes stimulated the growth of the pathogenic fungus producing on average a double increase in the necrotic area. The differences in the response of the pathogen to the presence of epiphytes recorded in the susceptible clones indicate a marked influence of the plant on the nature of interactions between its epiphytic microflora and the pathogen.     

Candidatus Monilibacter spp., common bulking filaments in activated sludge, are members of Cluster III Defluviicoccus

Two alphaproteobacterial Neisser negative ‘Nostocoida limicola’ morphotypes differing slightly in their trichome diameter and filament regularity were dominant populations in the Bendigo, Victoria, Australia activated sludge community removing phosphorus (P). Neither responded to the FISH probes available for any of the other alphaproteobacterial ‘N. limicola’ morphotypes. Instead both fluoresced with the DF988 FISH probe designed originally to target alphaproteobacterial cluster II Defluviicoccus tetrad forming organisms. A 16S rRNA based clone library from this biomass revealed that the alphaproteobacterial clones grouped closely with Candidatus ‘Monilibacter batavus’ and Defluviicoccus clones in a cluster separate from the existing cluster I and II Defluviicoccus. When a FISH probe was designed against these, it only hybridized to the thinner and less abundant ‘N. limicola’ morphotype. Micromanipulation–RT-PCR was used to selectively recover the main ‘N. limicola’ morphotype and a FISH probe designed against the 16S rRNA clones generated from it showed only this filament fluoresced. From FISH based surveys, both ‘N. limicola’ variants occurred frequently in phosphorus removal activated sludge systems in Australia treating domestic waste. The data suggest that they represent two new strains of Candidatus ‘Monilibacter’, which on this evidence are filamentous members of the genus Defluviicoccus, a potential competitor for the polyphosphate accumulating organisms in these communities.     

Can elevated CO(2) improve salt tolerance in olive trees?

We compared growth, leaf gas exchange characteristics, water relations, chlorophyll fluorescence, and Na⁺ and Cl⁻ concentration of two cultivars (‘Koroneiki’ and ‘Picual’) of olive (Olea europaea L.) trees in response to high salinity (NaCl 100 mM) and elevated CO₂ (eCO₂) concentration (700 μL L⁻¹). The cultivar ‘Koroneiki’ is considered to be more salt sensitive than the relatively salt-tolerant ‘Picual’. After 3 months of treatment, the 9-month-old cuttings of ‘Koroneiki’ had significantly greater shoot growth, and net CO₂ assimilation (A_CO2) at eCO₂ than at ambient CO₂, but this difference disappeared under salt stress. Growth and A_CO2 of ‘Picual’ did not respond to eCO₂ regardless of salinity treatment. Stomatal conductance (g_s) and leaf transpiration were decreased at eCO₂ such that leaf water use efficiency (WUE) increased in both cultivars regardless of saline treatment. Salt stress increased leaf Na⁺ and Cl⁻ concentration, reduced growth and leaf osmotic potential, but increased leaf turgor compared with non-salinized control plants of both cultivars. Salinity decreased A_CO2, g_s, and WUE, but internal CO₂ concentrations in the mesophyll were not affected. eCO₂ increased the sensitivity of PSII and chlorophyll concentration to salinity. eCO₂ did not affect leaf or root Na⁺ or Cl⁻ concentrations in salt-tolerant ‘Picual’, but eCO₂ decreased leaf and root Na⁺ concentration and root Cl⁻ concentration in the more salt-sensitive ‘Koroneiki’. Na⁺ and Cl⁻ accumulation was associated with the lower water use in ‘Koroneiki’ but not in ‘Picual’. Although eCO₂ increased WUE in salinized leaves and decreased salt ion uptake in the relatively salt-tolerant ‘Koroneiki’, growth of these young olive trees was not affected by eCO₂.     

Effects of Commercial and Indigenous Microorganisms on Fusarium Wilt Development in Chickpea

The purpose of this research was to determine whetherBacillus subtilis,nonpathogenicFusarium oxysporum,and/orTrichoderma harzianum,applied alone or in combination to chickpea (Cicer arietinumL.) cultivars ‘ICCV 4’ and ‘PV 61’ differing in their levels of resistance to Fusarium wilt, could effectively suppress disease caused by the highly virulent race 5 ofFusarium oxysporumf. sp.ciceris.Seeds of both cultivars were sown in soil amended with the three microbial antagonists, alone or in combination, and 7 days later seedlings were transplanted into soil infested with the pathogen. All three antagonistic microorganisms effectively colonized the roots of both chickpea cultivars, whether alone or in combination, and significantly suppressed Fusarium wilt development. In comparison with the control, the incubation period for the disease was delayed on average about 3 days and the final disease severity index and standardized area under the disease progress curve were reduced significantly between 14 and 33% and 16 and 42%, respectively, by all three microbial antagonists. Final disease incidence only was reduced byB. subtilis(18–25%) or nonpathogenicF. oxysporum(18%). The extent of disease suppression was higher and more consistent in ‘PV 61’ than in ‘ICCV 4’ whether colonized byB. subtilis,nonpathogenicF. oxysporum,orT. harzianum.The combination ofB. subtilis+T. harzianumwas effective in suppressing Fusarium wilt development but it did not differ significantly from treatments with either of these antagonists alone. In contrast, the combination ofB. subtilis+ nonpathogenicF. oxysporumtreatment was not effective but either antagonist alone significantly reduced disease development.     

The elevated anthocyanin level in the shaded peel of ‘Anjou’ pear enhances its tolerance to high temperature under high light

Pigments, chlorophyll fluorescence, dark respiration, and the antioxidant system in the shaded peel of green ‘Anjou’ pear (Pyrus communis L.) and its bud mutation, red ‘Anjou’, were compared in response to high peel temperature, high light alone or in combination to determine the protective role of anthocyanins under high temperature with or without light. Under high temperature treatment alone, no difference in the maximum quantum yield of PSII (F_V/F_M) was detected between red ‘Anjou’ and green ‘Anjou’; the superoxide dismutase activity and the glutathione pool were up-regulated in green ‘Anjou’ peel but remained unchanged in red ‘Anjou’ peel. Under high temperature coupled with high light, the F_V/F_M of green ‘Anjou’ peel was decreased to a lower value than that of red ‘Anjou’, and significant interaction was detected between temperature and light for both cultivars. Furthermore, the difference in F_V/F_M between red ‘Anjou’ and green ‘Anjou’ under high temperature coupled with high light was significantly larger than that under high light alone, indicating that this larger difference was caused by the interaction between high temperature and high light as no significant difference was detected in F_V/F_M between the two cultivars under high temperature treatment alone at any sampling point. It is concluded that the elevated anthocyanin level in the shaded peel of red ‘Anjou’ does not alter its thermotolerance in the dark, but makes it more tolerant of high temperature under high light.     

Impact of intercropping aphid-resistant wheat cultivars withoilseed rape on wheat aphid (Sitobion avenae) and its natural enemies

The effects of intercropping of wheat cultivars and oilseed rape on the densities of wheat aphid, Sitobion avenae, and their arthropod natural enemies were evaluated. Three winter wheat cultivars with different resistant levels to S. avenae were used: ‘KOK’ (high resistance), ‘Xiaobaidongmai’ (low resistance) and ‘Hongmanghong’ (susceptible). The results showed that the densities of S. avenae were significantly higher on the monoculture pattern than on either the 8-2 intercropping pattern (eight rows of wheat with two rows of oilseed rape) or the 8-4 intercropping pattern (eight rows of wheat with four rows of oilseed rape). The mean number of predators and the mummy rates of S. avenae were significantly higher in two intercropping patterns than those in the monoculture pattern. The densities of S. avenae, ladybeetles, and mummy rate of S. avenae were significantly different among different wheat cultivars. The highest densities of S. avenae and ladybeetles were found on wheat cultivar Hongmanghong. The lowest densities of S. avenae associated with high mummy rate of S. avenae were found on wheat cultivar Xiaobaidongmai. The results showed that wheat-oilseed rape intercropping conserved more predators and parasitoids than in wheat monoculture fields, and partial resistance of wheat cultivar Xiaobaidongmai had complementary or even synergistic effects on parasitoid of S. avenae.     

Identification of PaPKS1, a polyketide synthase involved in melanin formation and its use as a genetic tool in Podospora anserina

In the filamentous fungus Podospora anserina, many pigmentation mutations map to the median region of the complex locus ‘14’, called segment ‘29’. The data presented in this paper show that segment 29 corresponds to a gene encoding a polyketide synthase, designated PaPKS1, and identifies two mutations that completely or partially abolish the activity of the PaPKS1 polypeptide. We present evidence that the P. anserina green pigment is a (DHN)-melanin. Using the powerful genetic system of PaPKS1 cloning, we demonstrate that in P. anserina trans-duplicated sequences are subject to the RIP process as previously demonstrated for the cis-duplicated regions.     

The distribution and conservation of threatened Sphaeriidae on British grazing marshland

Freshwater sphaeriid bivalves are poorly known ecologically, particularly uncommon species such as Pisidium pseudosphaerium (UK RDB (Red Data Book) staus = ‘rare’). In the UK, this species occupies grazing marsh where conservation opportunities might be shared with other threatened molluscs. We surveyed sphaeriids including P. pseudosphaerium and snails in 106 drainage ditches in SE England in 1999. P. pseudosphaerium occupied over half the ditches surveyed, at slightly elevated BOD (6 ± 7 mg l⁻¹ sd) but reduced calcium (64 ± 31 mg l⁻¹) and nitrate (0.5 ± 1.2 mg l⁻¹). As part of a sphaeriid assemblage comprising Sphaerium corneum, Musculium lacustre and Pisidium obtusale (=Assemblage 1), P. pseudosphaerium occurred in ditches with floating vegetation and abundant snails of conservation importance, co-occurring significantly with either Valvata macrostoma (RDB ‘vulnerable’) or Anisus vorticulus (RDB: ‘vulnerable’). A more diverse sphaeriid assemblage (=Assemblage 2) included species common in wider, deeper ditches with open water. We suggest that traditional ditch management can support both rare and representative snails and bivalves on grazing marsh. Quasi-traditional and rotational ditch clearance will favour common sphaeriids and pioneer snails during early succession; P. pseudosphaerium, V. macrostoma, A. vorticulus during mid-succession; and the snail Segmentina nitida (RDB: ‘Endangered’) in late successional ditches. In common with threatened snails, P. pseudosphaerium will also benefit from reduced eutrophication. We recommend that P. pseudosphaerium retain RDB status to ensure protection and to emphasise the need for improved ecological information.     

Ribulose-1, 5-bisphosphate carboxylase activity,chlorophyll and protein concentrations in differentPopulus clones

Ribulose-1,5-bisphosphate carboxylase activity (RuBPC), chlorophyll (chl) and protein (prot) concentrations and chlorophyll/protein (chl/prot) ratios were determined in five differentPopulus clones together with their maximal net CO₂ uptake rates (P_max). A classic reference clone (Populus ×euramericana “Robusta” (Dode) Guinier) was compared with four recently selected euramerican and interamerican crossings. Chl/prot ratio and RuBPC activity varied among the different clones, while chl a/chlb ratio showed only a very low coefficient of variation (1.7%) for the five clones. Poplar clone “Robusta” could be distinguished from the recent faster growing clones based on the different biochemical characteristics. A significant correlation was found between both total chl concentration and chl/prot ratio with P_max for the five clones.     

Fusarium head blight biological control with Lysobacter enzymogenes strain C3

Fusarium head blight (FHB), caused by Fusarium graminearum (= Gibberella zeae), is a destructive disease of wheat for which biological controls are needed. Lysobacter enzymogenes strain C3, a bacterial antagonist of fungal pathogens via lytic enzymes and induced resistance, was evaluated in this study for control of FHB. In greenhouse experiments, chitin broth cultures of C3 reduced FHB severity to <10% infected spikelets as compared to >80% severity in the controls in some experiments. C3 broth cultures heated to inactivate cells and lytic enzymes, but retaining the elicitor factor for induced resistance, also were effective in reducing FHB severity, suggesting induced resistance is one mechanism of action. C3 broth cultures also were effective when applied in highly diluted form and when applied 1 week prior to pathogen inoculation. When applied to 8 cultivars of hard red spring wheat in the greenhouse, C3 treatments reduced FHB in 5 cultivars but not in the others. These findings also are consistent with induced resistance. Protection offered by C3 treatments, however, was not systemic and required that C3 be applied uniformly to all susceptible florets. Field tests were conducted in South Dakota and Nebraska to evaluate the efficacy of C3 chitin broth cultures in spring and winter wheat, respectively. In experiments involving two hard red spring wheat cultivars, treatment with C3 reduced FHB severity in ‘Russ’ but not in ‘Ingot’. In three other field experiments comparing C3, the fungicide tebuconazole, and the combination of C3 and tebuconazole, treatments with the bacterial culture alone and the fungicide alone were inconsistent across experiments, each treatment being ineffective in controlling FHB in one experiment. The biocontrol agent–fungicide combination was more consistently effective, reducing FHB incidence or severity in all three experiments. Thus, the potential for using L. enzymogenes C3 as a biological control agent for FHB was demonstrated along with a number of factors that might affect control efficacy in the field.     

Isoenzyme diversity and cryptic speciation in Juniperus excelsa (Cupressaceae) complex in Iran

An isoenzyme analysis was performed on 109 individuals from 11 populations of Juniperus excelsa complex collected from Iran. A total of 61 allozymes from five enzyme systems were scored. The mean numbers of bands per presumed isoenzyme, ranged from 2.07 to 3.23. The value of Euclidean distance ranged from 1.920 to 4.411. The average value of Shannon diversity index (H′) estimated for each population ranged from 0.317 in the population sampled from ‘Fasa’ to 1.053 in the population of ‘Golestan’, and the average H′ value for each enzyme system ranged from 0.031 (SOD-3) to 1.670 (PPO-1). To reveal the relationships among these populations, a cluster analysis based on Euclidean distances was performed. The results of the analysis suggest recognizing three taxa in the complex in Iran: Juniperus polycarpos subsp. polycarpos, Juniperus polycarpos subsp. turcomanica, and one cryptic species not recorded from Iran until now. Moreover, the analysis presented, does not confirm the occurrence of J. excelsa in Iran.     

Epiphytic microflora of poplar clones susceptible and resistant to infection by Dothichiza populea

One-year-old poplar shoots (nodes, internodes and lenticels) of clones susceptible to infection by the pathogenic fungus Dothichiza populea, viz. Populus nigra Italica and P. Robusta, resistant ones, viz. P. Grandis and P. Hybrida 275, as well as a hybrid of a susceptible and a resistant clone, viz. P. maximowiczii x P. nigra (P. Kórnik 42), were used. The plate method was employed to determine: 1. the abundance of the epiphytic microflora on a mineral medium with glucose; 2. the quantitative composition of epiphytic communities by determining the numbers of typical bacteria (including rod-shaped, spherical and sporulating forms), actinomycetes and yeasts in microscopic preparations from epiphyte colonies; 3. the abundance and level of activity of epiphytes antagonistic towards Dothichiza populea. In all poplar clones the epiphytic microflora was most abundant on nodes and least abundant on lenticels. In the resistant clones epiphytes were 7 (P. Grandis) to as many as 84 times (P. Hybrida 275) less numerous than in the susceptible ones. In the microflora communities of the susceptible poplars, rod-shaped bacteria were the most abundant, and in the resistant ones and the hybrid, yeasts, which made up from 60% to 70% of the strains tested. Spherical and sporulating bacteria as well as actinomycetes were found in numbers not exceeding 4% of the total number of epiphytes. The proportion of antagonistic microflora in whole epiphytic communities was higher in the resistant clones and the hybrid than in the susceptible clones, with the microflora having a more restrictive effect on the development of the pathogen.     

Influence of free air CO2 enrichment (EUROFACE) and nitrogen fertilisation on the anatomy of juvenile wood of three poplar species after coppicing

Populus × euramericana, P. alba, and P. nigra clones were exposed to ambient or elevated (about 550 ppm) CO₂ concentrations under field conditions (FACE) in central Italy. After three growing seasons, the plantation was coppiced. FACE was continued and in addition, one-half of each experimental plot was fertilised with nitrogen. Growth and anatomical wood properties were analysed in secondary sprouts. In the three poplar clones, most of the growth and anatomical traits showed no uniform response pattern to elevated [CO₂] or N-fertilisation. In cross-sections of young poplar stems, tension wood amounted to 2–10% of the total area and was not affected by elevated CO₂. In P. nigra, N-fertilisation caused an about twofold increase in tension wood, but not in the other clones. The formation of tension wood was not related to diameter or height growth of the shoots. In P. × euramericana N-fertilisation resulted in significant reductions in fibre lengths. In all three genotypes, N-fertilisation caused significant decreases in cell wall thickness. In P. × euramericana and P. alba elevated [CO₂] also caused decreases in wall thickness, but less pronounced than nitrogen. In P. nigra and P. × euramericana elevated [CO₂] induced increases in vessel diameters. These results show that elevated [CO₂] and N-fertilisation affect wood structural development in a clone specific manner. However, the combination of these environmental factors resulted in overall losses in cell wall area of 5–12% in all three clones suggesting that in future climate scenarios negative effects on wood quality are to be anticipated if increases in atmospheric CO₂ concentration were accompanied by increased N availability.     

Interaction dynamics between saprobic lignicolous fungi and Armillaria in controlled environments: Exploring the potential for competitive exclusion of Armillaria on peach

Armillaria root rot, caused by Armillaria tabescens and Armillaria mellea, is a major cause of premature tree death in peach orchards in the southeastern United States. The root systems of infected trees can become entirely colonized by Armillaria, serving as an inoculum source for adjacent trees and providing massive inoculum levels in replant situations. If dead or dying trees could be colonized by an effective competitor of Armillaria before their removal, the extent of root colonization by the pathogen could be reduced, thus decreasing the threat to adjacent trees and/or subsequent plantings. Interactions between five species of saprobic lignicolous fungi (Ganoderma lucidum, Hypholoma fasciculare, Phanerochaete velutina, Schizophyllum commune, and Xylaria hypoxylon) and the two Armillaria species were examined in controlled conditions to provide proof of concept for competitive exclusion of Armillaria from peach roots. On agar-coated glass slides, all five potential antagonists induced detrimental reactions in >58% of the Armillaria hyphae observed, with the majority resulting in hyphal swelling or granulation. On poplar wood blocks, all antagonists consistently either overgrew Armillaria colonies or—in the case of S. commune—engaged in deadlock reactions; in all cases, the viability of Armillaria colonies was reduced to <30% of that of unchallenged controls. When inoculated simultaneously onto opposite ends of peach root segments, all antagonists consistently reduced growth and viability of Armillaria on and under the bark, whereby reduction of pathogen growth underneath the bark, Armillaria’s primary ecological niche, was most pronounced for G. lucidum, S. commune, and X. hypoxylon. When root segments were allowed to be colonized entirely by Armillaria before being inoculated with the antagonists, the latter were able to overgrow the pathogen on the root surface but unable to pre-empt it from underneath the bark. In summary,G. lucidum, S. commune, and X. hypoxylon caused strong hyphal and mycelial interference reactions and the most pronounced reductions in growth of Armillaria above and below the bark, indicating that they would be the most promising candidates for field-scale evaluations to restrict colonization of dead or dying peach trees by Armillaria in the orchard.     

Suppression of Botrytis cinerea sporulation by Clonostachys rosea on rose debris: a valuable component in Botrytis blight management in commercial greenhouses

Botrytis blight, caused by Botrytis cinerea (Bc), is an important disease on roses grown in plastic greenhouses in Brazil. Biocontrol with Clonostachys rosea (Cr) applied to leaves and crop debris to reduce pathogen sporulation can complement other control measures for disease management. Two experiments, each with a rose cultivar, were conducted in a plastic greenhouse. For ‘Red Success,’ four treatments were compared: (1) control; (2) fortnightly sprays of Cr; (3) weekly sprays of mancozeb; and (4) weekly sprays of either Cr or mancozeb to the lower third of the plants and the debris. For ‘Sonia,’ treatment 4 was not included. Samples were taken from debris (leaves and petals) at ten 15-day intervals and plated on PCA medium. Sporulation of fungi and incidence of Botrytis blight on buds were assessed. For both cultivars, C treatments significantly (P=0.05) reduced Bc sporulation. However, disease incidence was not consistently reduced, probably because the applications of C. rosea started when Botrytis blight epidemic was advanced and no sanitation practices were performed on nontreated plots. From the present and previous studies, continuous application of Cr on debris, associated with sanitation practices, has the potential to reduce Bc sporulation and disease incidence in the buds.     

Induced defence responses and protective effects on tomato against Xanthomonas vesicatoria by an aqueous extract from Solanum lycocarpum infected with Crinipellis perniciosa

This study investigated the induced defence responses and protective effects on susceptible tomato (Lycopersicon esculentum Mill.) against Xanthomonas vesicatoria (Doidge) by a heat-treated aqueous extract (VLA) from dry necrotic tissue of ‘Lobeira’ (Solanum lycocarpum St. Hil.) branches infected with the fungus Crinipellis perniciosa (Stahel) compared with acibenzolar-S-methyl (ASM), a commercial inducer of resistance. Plantlets were sprayed with VLA and ASM and challenged 4 days later with a virulent strain of X. vesicatoria, under greenhouse conditions. The disease severity, fresh weight of shoots, the activities of phenol peroxidase (POX), polyphenol oxidase (PPO), chitinase (CHI), phenylalanine ammonia-lyase (PAL), lignin deposition, and soluble phenolic contents were evaluated in the leaf tissues. Reduction of the bacterial spot severity was observed in plantlets treated with VLA which conferred 63% of the ASM protection. This protective effect and lesion reduction promoted by VLA were probably associated particularly with POX and PAL activities, lignin deposition on leaf tissues and, to a less extent, CHI activity.     

Biosynthesis of scopoletin in Hevea brasiliensis leaves inoculated with Phytophthora palmivora

Inoculation of resistant (R) and susceptible (S) Hevea brasiliensisleaves with Phytophthora palmivorainduced foliar necrosis and biosynthesis of scopoletin (Scp), considered as a Heveaphytoalexin. The degree of resistance of four clones, as classified by the necrotic lesions, was related to the rapidity and intensity of Scp production. The resistant BPM-24, and marked partially resistant clone PB-235, displayed an early secretion of scopoletin that intensified and lasted longer than the weak partially resistant RRIT251, and susceptible clone RRIM600. The lesion size and amount of Scp after infection were positively correlated to the concentration of spores applied to Hevealeaves. In addition, in leaflets inoculated with high spore concentration, Scp reached the highest level earlier than those with low spore concentration. A fungitoxic effect of Scp on mycelium growth was shown in bioassays; the I₅₀ value tested on Phytophthora palmivora was relatively the same as on Phytophthora botryosa, but much lower than those found on other leaf pathogens of rubber tree, Corynespora cassiicolaand Colletotrichum gloeosporioides. The lesion size and level of Scp upon spore inoculation may be appropriate for classifying Heveaclones according to their R/S with respect to P. palmivora     

N,N′-bis-(2,3-Methylenedioxyphenyl)urea and N,N′-bis-(3,4-methylenedioxyphenyl)urea enhance adventitious rooting in Pinus radiata and affect expression of genes induced during adventitious rooting in the presence of exogenous auxin

We have analyzed the effect of N,N′-bis-(2,3-methylenedioxyphenyl)urea (2,3-MDPU) and N,N′-bis-(3,4-methylenedioxyphenyl)urea (3,4-MDPU), two symmetrically substituted diphenylurea derivatives with no auxin or cytokinin-like activity, on the rooting capacity of Pinus radiata stem cuttings. Results indicate that both diphenylurea derivatives enhance adventitious rooting in the presence of exogenous auxin (indole-3-butyric acid, IBA), even at low auxin concentration, in rooting-competent cuttings, but have no effect on the adventitious rooting of low or null competent-to-root cuttings. Histological analyses show that, in the simultaneous presence of MDPUs and low concentration of exogenous auxin, adventitious root formation is induced in the cell types that retain intrinsic competence to form adventitious roots in response to auxin. The time course of cellular events leading to root formation and the time of root emergence are closely similar to that observed in cuttings treated only with higher auxin concentration. In addition, the mRNA level of a P. radiata SCARECROW-LIKE gene, which is significantly induced in the presence of the optimal concentration (10 μM) of exogenous auxin needed for cuttings to root, is increased in the presence of MDPUs and low concentration of exogenous auxin (1 μM). The expression of a P. radiata SHORT-ROOT gene in rooting-competent cuttings during adventitious rooting is also affected by the presence of MDPUs when combined with auxin. As MDPUs do not affect the expression of either gene in the absence of exogenous auxin, but only in its presence, we suggest that MDPUs could interact, directly or indirectly, with the auxin-signalling pathways in rooting-competent cuttings during adventitious rooting.