Ineffectiveness of pruning to control citrus huanglongbing caused by <Emphasis Type="Italic">Candidatus</Emphasis> Liberibacter americanus

The huanglongbing (HLB) disease of citrus trees, caused by Candidatus Liberibacter asiaticus and Ca. Liberibacter americanus, was first reported in Brazil in March, 2004. The presence of the disease has caused serious concerns among growers. Pruning experiments were conducted to determine if removal of symptomatic branches or the entire canopy (decapitation) would eliminate infected tissues and save HLB-affected trees. Pruning was done in five blocks on a total of 592 3- to 16 year-old ‘Valência’, ‘Hamlin’ or ‘Pêra’ sweet orange trees showing no symptoms or with two levels of symptom severity. Ten decapitated trees per block were caged and all trees were treated with insecticides to control the psyllid vector, Diaphorina citri. Mottled leaves reappeared on most symptomatic (69.2%) as well on some asymptomatic (7.6%) pruned trees, regardless of age, variety, and pruning procedure. Presence of the pathogen (Ca. Liberibacter americanus) in all symptomatic trees was confirmed by PCR. In general, the greater the symptom severity before pruning the lower the percentage of trees that remained asymptomatic after pruning.     

Calcium ion promotes successful penetration of powdery mildew fungi into barley cells

To determine whether Ca²⁺ promotes powdery mildew penetration, Ca²⁺-treated barley coleoptiles were inoculated with conidia of pathogenic and nonpathogenic fungi. Penetration efficiency of the pathogenic powdery mildew Blumeria graminis was enhanced by Ca²⁺ treatment, but that of the necrotrophic pathogen Helminthosporium sp. remained unaffected. Similarly, when actin-dependent penetration resistance is suppressed with cytochalasin A, Ca²⁺ treatment specifically enhanced penetration of the nonpathogenic powdery mildew Erysiphe pisi but not that of other nonpathogens. Calmodulin inhibitors suppressed the promotive effect of Ca²⁺ on B. graminis penetration. These results suggest that barley powdery mildew specifically requires Ca²⁺ and calmodulin for penetration.     

Molecular characterization of <Emphasis Type="Italic">Phoma tracheiphila</Emphasis>, causal agent of Mal secco disease of citrus,in Israel

Mal secco disease of citrus caused by Phoma tracheiphila is a devastating disease in the Mediterranean basin. Susceptible citrus species include lemon, citron, lime and others. Trees attacked by the fungus show characteristic symptoms; the smallest twigs die first, followed by the larger branches. Eventually, the whole tree is killed. The symptoms are clear in the orchards but by the time they are visible the disease is already well established. The need for a sensitive, reliable and rapid diagnostic method for the early identification of the fungus in trees and fruit exists. We have developed a PCR-based method for the identification of P. tracheiphila from plant tissues including fruit. Any such method must take into account the genetic variability in the pathogen population. Molecular methods were used to compare different isolates of P. tracheiphila. This study found no significant differences between different isolates from different citrus species from different parts of Israel.     

Cloning of DNA fragments specific to the pathotype and race of <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

Japanese isolates of Verticillium dahliae, a causal agent of wilt disease in many plants, are classifiable into pathotypes based on their pathogenicity. Because these pathotypes are morphologically indistinguishable, establishing a rapid identification method is very important for the control of this pathogen in Japan. For cloning DNA fragments that are useful for identification and specific detection of V. dahliae pathotypes, we performed random amplified polymorphic DNA (RAPD) analyses using various isolates. One polymerase chain reaction (PCR) product, E10-U48, was specific to isolates pathogenic to sweet pepper. The other product, B68-TV, was specific to race 1 of isolates pathogenic to tomato. The specificity of these sequences was confirmed by genomic Southern hybridization. Further analyses revealed that the region peripheral to B68-TV obtained from the genomic DNA library includes the sequence specific to all isolates pathogenic to tomato (races 1 and 2). Moreover, sequence tagged site (STS) primers designed from B68-TV and its peripheral region showed race-specific and pathotype-specific amplification in a PCR assay. The probes and primers obtained in this study are likely to be useful tools for the identification and specific detection of pathotypes and races of V. dahliae. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession number AB095266.     

Management approach using simple indices of deer density and status of understory vegetation for conserving deciduous hardwood forests on a regional scale

We investigated the validity and efficiency of a survey using sight per unit effort (SPUE) of sika deer and shrub-layer decline rank (SDR), which is an index of decline in the physical structure of a whole stand caused by sika deer, based on data collected on a broad scale. This survey was to be used to manage a deer population in order to conserve a forest ecosystem. First, we evaluated the spatial and temporal scales of deer density that are most appropriate for predicting decline in the status of understory vegetation. The model with SPUE calculated in a buffer with a radius of 4.5 km using data for the past 4 years was found to be the best. We showed that our knowledge of the relationship between deer density and status of shrub-layer vegetation is improved by identifying the most suitable spatial and temporal scales of SPUE for predicting SDR. Next, we quantified the effects of SPUE and environmental components on SDR in stands. We found that SPUE had the greatest effect on SDR among all explanatory variables. Moreover, the area under the curve (AUC) was large in a model that only used SPUE (AUC = 0.718). This result suggests that the variation in SDR among stands was explained well by SPUE regardless of differences in the forest environment. Furthermore, we identified the effective values of SPUE for preventing shrub-layer vegetation from declining through deer density control. We conclude that a management system based on SPUE and SDR is a simple and valid method for managing deer populations in order to conserve forest ecosystems.     

Effects of carbon and nitrogen amendment on soil carbon and nitrogen mineralization in volcanic immature soil in southern Kyushu,Japan

Soil N mineralization is affected by microbial biomass and respiration, which are limited by available C and N. To examine the relationship between C and N for soil microbial dynamics and N dynamics, we conducted long-term laboratory incubation (150 days) after C and N amendment and measured changes in C and N mineralization, microbial biomass C, and dissolved C and N throughout the incubation period. The study soil was volcanic immature soil from the southern part of Japan, which contains lower C and N compared with other Japanese forest soils. Despite this, the area is covered by well-developed natural and plantation forests. Carbon amendment resulted in an increase in both microbial biomass and respiration, and net N mineralization decreased, probably due to increasing microbial immobilization. In contrast, N amendment resulted in a decrease in microbial respiration and an increase in net N mineralization, possibly due to decreased immobilization by microbes. Amendment of both C and N simultaneously did not affect microbial biomass and respiration, although net N mineralization was slightly increased. The results suggested that inhibitory effect on microbial respiration by N amendment should be reduced if carbon availability is higher. Thus, soil available C may limit microbial biomass and respiration in this volcanic immature soil. Even in immature soil where C and N substrate is low, soil C, such as plant root exudates and materials from above- and belowground dead organisms, might help to maintain microbial activity and N mineralization in this study site.