Effects of elevated atmospheric carbon dioxide on the biological control of coffee leaf rust under controlled conditions

The effect of elevated atmospheric CO2 concentration on biological control of coffee leaf rust, caused by Hemileia vastatrix, was evaluated by leaf disc assay, under controlled conditions. The biocontrol agents Bacillus subtilis, Bacillus pumilus and Lecanicillium longisporum were applied 24h before, 24h after, and simultaneously with the H. vastatrix on leaf discs (diameter of 1.5cm). The CO2 concentrations tested were: 380, 430, 700 and 1300ppm for B. subtilis and B. pumilus; and 380, 430, 670 and 1200ppm for L. longisporum. The antagonists were not affected by CO2 concentrations. B. subtilis was the most effective in controlling the disease when applied before and simultaneously with pathogen.

Performance of different rubber tree clones against South American leaf blight (Microcyclus ulei)

This study was aimed to evaluate the behaviour of eighteen clones of Hevea brasiliensis (rubber) against South American leaf blight and tostudy progress of the disease. The experiment was conducted in Cachoeiro de Itapemirim-ES, where 18 clones were evaluated. The experi-mental design was a randomized block with four replicates; each experimental unit consisted of three plants. Evaluations were performed at15-day intervals on 30 leaflets per tree. Disease incidence was quantified and infection classified according to the stages of development andtype of damage. Leaf blight occurred during the entire experimental period; however, disease intensity varied with the resistance level of theclones and the time of year. Clones FX 3864, RRIM 725, RRIM 711, IAC 300 and IAN 873 exhibited the highest resistance to leaf blight.

Effect of Fungicides and Plant Populations on Soybean Disease and Yield

Applications of foliar fungicides on soybeans have been shown to reduce disease pressure and protect yield under the right conditions, especially in environments that have very wet or humid conditions. In the past decade, fungicide use in Iowa has increased. Initially, growers were concerned with the potential threat of soybean rust, which is controlled effectively by foliar fungicides. In Iowa, however, there has not been any case of yield reduction due to soybean rust. New potential purposes for foliar fungicides include “plant health” benefits and the reduction of foliar diseases endemic in Iowa such as Septoria brown spot, Cercospora leaf blight, and frogeye leaf spot. Currently what is not known is how the efficacy of fungicides is affected when agricultural practices change. Our question: How does plant population affect the efficacy of fungicides?

Functional response of Ulmus minor Mill. to drought, flooding and dutch elm disease

Ulmus minor es una especie arbórea originaria de Europa cuyas poblaciones han sido diezmadas por el hongo patógeno causante de la enfermedad de la grafiosis. La conservación de los olmos exige plantearse su propagación a través de plantaciones y conocer mejor su ecología y biología. Ulmus minor es un árbol de ribera, pero frecuentemente se encuentra alejado del cauce de arroyos y ríos, donde la capa freática sufre fuertes oscilaciones. Por ello, nuestra hipótesis general es que esta especie es moderadamente resistente tanto a la inundación como a la sequía. El principal objetivo de esta tesis doctoral es entender desde un punto de vista funcional la respuesta de U. minor a la inundación, la sequía y la infección por O. novo-ulmi; los factores que posiblemente más influyen en la distribución actual de U. minor. Con este objetivo se persigue dar continuidad a los esfuerzos de conservación de esta especie que desde hace años se dedican en varios centros de investigación a nivel mundial, ya que, entender mejor los mecanismos que contribuyen a la resistencia de U. minor ante la inoculación con O. novo-ulmi y factores de estrés abiótico ayudará en la selección y propagación de genotipos resistentes a la grafiosis. Se han planteado tres experimentos en este sentido. Primero, se ha comparado la tolerancia de brinzales de U. minor y U. laevis – otro olmo ibérico – a una inmersión controlada con el fin de evaluar su tolerancia a la inundación y comprender los mecanismos de aclimatación. Segundo, se ha comparado la tolerancia de brinzales de U. minor y Quercus ilex – una especie típica de ambientes Mediterránea secos – a la falta de agua en el suelo con el fin de evaluar el grado de tolerancia y los mecanismos de aclimatación a la sequía. El hecho de comparar dos especies contrastadas responde al interés en entender mejor cuales son los procesos que conducen a la muerte de una planta en condiciones de sequía – asunto sobre el que hay una interesante discusión desde hace algunos años. En tercer lugar, con el fin de entender mejor la resistencia de algunos genotipos de U. minor a la grafiosis, se han estudiado las diferencias fisiológicas y químicas constitutivas e inducidas por O. novo-ulmi entre clones de U. minor seleccionados a priori por su variable grado de resistencia a esta enfermedad. En el primer experimento se observó que los brinzales de U. minor sobrevivieron 60 días inmersos en una piscina con agua no estancada hasta una altura de 2-3 cm por encima del cuello de la raíz. A los 60 días, los brinzales de U. laevis se sacaron de la piscina y, a lo largo de las siguientes semanas, fueron capaces de recuperar las funciones fisiológicas que habían sido alteradas anteriormente. La conductividad hidráulica de las raíces y la tasa de asimilación de CO2 neta disminuyeron en ambas especies. Por el contrario, la tasa de respiración de hojas, tallos y raíces aumentó en las primeras semanas de la inundación, posiblemente en relación al aumento de energía necesario para desarrollar mecanismos de aclimatación a la inundación, como la hipertrofia de las lenticelas que se observó en ambas especies. Por ello, el desequilibrio del balance de carbono de la planta podría ser un factor relevante en la mortalidad de las plantas ante inundaciones prolongadas. Las plantas de U. minor (cultivadas en envases de 16 litros a media sombra) sobrevivieron por un prolongado periodo de tiempo en verano sin riego; la mitad de las plantas murieron tras 90 días sin riego. El cierre de los estomas y la pérdida de hojas contribuyeron a ralentizar las pérdidas de agua y tolerar la sequía en U. minor. Las obvias diferencias en tolerancia a la sequía con respecto a Q. ilex se reflejaron en la distinta capacidad para ralentizar la aparición del estrés hídrico tras dejar de regar y para transportar agua en condiciones de elevada tensión en el xilema. Más relevante es que las plantas con evidentes síntomas de decaimiento previo a su muerte exhibieron pérdidas de conductividad hidráulica en las raíces del 80% en ambas especies, mientras que las reservas de carbohidratos apenas variaron y lo hicieron de forma desigual en ambas especies. Árboles de U. minor de 5 y 6 años de edad (plantados en eras con riego mantenido) exhibieron una respuesta a la inoculación con O. novo-ulmi consistente con ensayos previos de resistencia. La conductividad hidráulica del tallo, el potencial hídrico foliar y la tasa de asimilación de CO2 neta disminuyeron significativamente en relación a árboles inoculados con agua, pero solo en los clones susceptibles. Este hecho enlaza con el perfil químico “más defensivo” de los clones resistentes, es decir, con los mayores niveles de suberina, ácidos grasos y compuestos fenólicos en estos clones que en los susceptibles. Ello podría restringir la propagación del hongo en el árbol y preservar el comportamiento fisiológico de los clones resistentes al inocularlos con el patógeno. Los datos indican una respuesta fisiológica común de U. minor a la inundación, la sequía y la infección por O. novo-ulmi: pérdida de conductividad hidráulica, estrés hídrico y pérdida de ganancia neta de carbono. Pese a ello, U. minor desarrolla varios mecanismos que le confieren una capacidad moderada para vivir en suelos temporalmente anegados o secos. Por otro lado, el perfil químico es un factor relevante en la resistencia de ciertos genotipos a la grafiosis. Futuros estudios deberían examinar como este perfil químico y la resistencia a la grafiosis se ven alteradas por el estrés abiótico. ABSTRACT Ulmus minor is a native European elm species whose populations have been decimated by the Dutch elm disease (DED). An active conservation of this species requires large-scale plantations and a better understanding of its biology and ecology. U. minor generally grows close to water channels. However, of the Iberian riparian tree species, U. minor is the one that spread farther away from rivers and streams. For these reasons, we hypothesize that this species is moderately tolerant to both flooding and drought stresses. The main aim of the present PhD thesis is to better understand the functional response of U. minor to the abiotic stresses – flooding and drought – and the biotic stress – DED – that can be most influential on its distribution. The overarching goal is to aid in the conservation of this emblematic species through a better understanding of the mechanisms that contribute to resistance to abiotic and biotic stresses; an information that can help in the selection of resistant genotypes and their expansion in large-scale plantations. To this end, three experiments were set up. First, we compared the tolerance to experimental immersion between seedlings of U. minor and U. laevis – another European riparian elm species – in order to assess their degree of tolerance and understand the mechanisms of acclimation to this stress. Second, we investigated the tolerance to drought of U. minor seedlings in comparison with Quercus ilex (an oak species typical of dry Mediterranean habitats). Besides assessing and understanding U. minor tolerance to drought at the seedling stage, the aim was to shed light into the functional alterations that trigger drought-induced plant mortality – a matter of controversy in the last years. Third, we studied constitutive and induced physiological and biochemical differences among clones of variable DED resistance, before and following inoculation with Ophiostoma novo-ulmi. The goal is to shed light into the factors of DED resistance that is evident in some genotypes of U. minor, but not others. Potted seedlings of U. minor survived for 60 days immersed in a pool with running water to approximately 2-3 cm above the stem collar. By this time, U. minor seedlings died, whereas U. laevis seedlings moved out of the pool were able to recover most physiological functions that had been altered by flooding. For example, root hydraulic conductivity and leaf photosynthetic CO2 uptake decreased in both species; while respiration initially increased with flooding in leaves, stems and roots possibly to respond to energy demands associated to mechanisms of acclimation to soil oxygen deficiency; as example, a remarkable hypertrophy of lenticels was soon observed in flooded seedlings of both species. Therefore, the inability to maintain a positive carbon balance somehow compromises seedling survival under flooding, earlier in U. minor than U. laevis, partly explaining their differential habitats. Potted seedlings of U. minor survived for a remarkable long time without irrigation – half of plants dying only after 90 days of no irrigation in conditions of high vapour pressure deficit typical of summer. Some mechanisms that contributed to tolerate drought were leaf shedding and stomata closure, which reduced water loss and the risk of xylem cavitation. Obviously, U. minor was less tolerant to drought than Q. ilex, differences in drought tolerance resulting mostly from the distinct capacity to postpone water stress and conduct water under high xylem tension among species. More relevant was that plants of both species exhibited similar symptoms of root hydraulic failure (i.e. approximately 80% loss of hydraulic conductivity), but a slight and variable depletion of non-structural carbohydrate reserves preceding dieback. Five- and six-year-old trees of U. minor (planted in the field with supplementary watering) belonging to clones of contrasted susceptibility to DED exhibited a different physiological response to inoculation with O. novo-ulmi. Stem hydraulic conductivity, leaf water potential and photosynthetic CO2 uptake decreased significantly relative to control trees inoculated with water only in DED susceptible clones. This is consistent with the “more defensive” chemical profile observed in resistant clones, i.e. with higher levels of saturated hydrocarbons (suberin and fatty acids) and phenolic compounds than in susceptible clones. These compounds could restrict the spread of O. novo-ulmi and contribute to preserving the near-normal physiological function of resistant trees when exposed to the pathogen. These results evidence common physiological responses of U. minor to flooding, drought and pathogen infection leading to xylem water disruption, leaf water stress and reduced net carbon gain. Still, seedlings of U. minor develop various mechanisms of acclimation to abiotic stresses that can play a role in surviving moderate periods of flood and drought. The chemical profile appears to be an important factor for the resistance of some genotypes of U. minor to DED. How abiotic stresses such as flooding and drought affect the capacity of resistant U. minor clones to face O. novo-ulmi is a key question that must be contemplated in future research.

An evaluation of the training program: "The Alzheimer's disease afflicted: understanding the disease and the resident" /

On cover: New horizons in long term care.

Population differentiation in the banana leaf spot pathogen Mycosphaerella musicola, examined at a global scale

Single-copy restriction fragment length polymorphism (RFLP) markers were used to determine the genetic structure of the global population of Mycosphaerella musicola, the cause of Sigatoka (yellow Sigatoka) disease of banana. The isolates of M. musicola examined were grouped into four geographic populations representing Africa, Latin America and the Caribbean, Australia and Indonesia. Moderate levels of genetic diversity were observed for most of the populations (H = 0.22-0.44). The greatest genetic diversity was found in the Indonesian population (H = 0.44). Genotypic diversity was close to 50% in all populations. Population differentiation tests showed that the geographic populations of Africa, Latin America and the Caribbean, Australia and Indonesia were genetically different populations. Using F-ST tests, very high levels of genetic differentiation were detected between all the population pairs (F-ST > 0.40), with the exception of the Africa and Latin America-Caribbean population pair. These two populations differed by only 3% (F-ST = 0.03), and were significantly different (P < 0.05) from all other population pairs. The high level of genetic diversity detected in Indonesia in comparison to the other populations provides some support for the theory that M. musicola originated in South-east Asia and that M. musicola populations in other regions were founded by isolates from the South-east Asian region. The results also suggest the migration of M. musicola between Africa and the Latin America-Caribbean region.

Utility of cotyledon and detached leaf assays for assessing root reactions of lucerne to Phytophthora root rot caused by Phytophthora medicaginis

Phytophthora root rot, caused by Phytophthora medicaginis, is a major limitation to lucerne production but it can be managed through the use of resistant cultivars. Current resistance screening methods, using mature plants or post-emergence seedling assays, are costly and time consuming. The use of zoospore inoculum on detached leaves and intact cotyledons as an assay for plant resistance was assessed using genetically defined segregating populations. The detached leaf assay was a reproducible test, but this test could not be used for accurately predicting root ratings. The cotyledon tests using zoospores gave results at the population level that were indicative of the root responses of 19 cultivars and lines tested. The cotyledon reaction of individual plants also showed a strong association with root response. The cotyledon test, while not completely predictive of mature root responses, allowed the selection of Phytophthora resistant plants at a higher frequency than could be achieved by random selection.

Genetic uniformity of international isolates of Leifsonia xyli subsp xyli, causal agent of ratoon stunting disease of sugarcane

An international collection of the sugarcane ratoon stunting disease pathogen, Leifsonia xyli subsp. xyli, was analysed to assess genetic diversity. DNA fingerprinting using BOX primers was performed on 105 isolates, comprising 65 Australian isolates and an additional 40 isolates from Indonesia (n = 8), Japan (n = 1), USA (n = 3), Brazil (n = 2), Mali (n = 2), Zimbabwe (n = 13), South Africa (n = 9) and Reunion (n = 2). Sixty-two of these isolates were also screened using ERIC primers. No variation was found among any of the isolates. The intergenic spacer (IGS) region of the ribosomal RNA genes from 54 isolates was screened for sequence variation using single-stranded conformational polymorphism (SSCP), but none was observed. Direct sequencing of the IGS from a subset of nine isolates, representing all of the countries sampled in this study, confirmed the results of the SSCP analysis. Likewise, no sequence variation was found in the 16S ribosomal RNA genes of the same subset. Four Colombian isolates from sugarcane, morphologically similar to L. xyli subsp. xyli, were putatively shown to be an undescribed Agrococcus species of unknown pathogenicity. The lack of genetic variation among L. xyli subsp. xyli isolates, independent of time of sampling, cultivar of isolation, or country of origin, suggests the worldwide spread of a single pathogenic clone, and further suggests that sugarcane cultivars resistant to ratoon stunting disease in one area should retain this property in other regions.

Is the distribution of Fiji leaf gall in Australian sugarcane explained by variation in the vector Perkinsiella saccharicida?

Fiji leaf gall (FLG) is an important virally induced disease in Australian sugarcane. It is confined to southern canegrowing areas, despite its vector, the delphacid planthopper Perkinsiella saccharicida, occurring in all canegrowing areas of Queensland and New South Wales. This disparity between distributions could be a result of successful containment of the disease through quarantine and/or geographical barriers, or because northern Queensland populations of Perkinsiella may be poorer vectors of the disease. These hypotheses were first tested by investigating variation in the ITS2 region of the rDNA fragment among eastern Australian and overseas populations of Perkinsiella. The ITS2 sequences of the Western Australian P. thompsoni and the Fijian P. vitiensis were distinguishable from those of P. saccharicida and there was no significant variation among the 26P. saccharicida populations. Reciprocal crosses of a northern Queensland and a southern Queensland population of P. saccharicida were fertile, so they may well be conspecific. Single vector transmission experiments showed that a population of P. saccharicida from northern Queensland had a higher vector competency than either of two southern Queensland populations. The frequency of virus acquisition in the vector populations was demonstrated to be important in the vector competency of the planthopper. The proportion of infected vectors that transmitted the virus to plants was not significantly different among the populations tested. This study shows that the absence of FLG from northern Queensland is not due to a lack of vector competency of the northern population of P. saccharicida.

Susceptibility of source plants to Sugarcane Fiji disease virus influences the acquisition and transmission of the virus by the planthopper vector Perkinsiella saccharicida

Fiji leaf gall (FLG) caused by Sugarcane Fiji disease virus (SCFDV) is transmitted by the planthopper Perkinsiella saccharicida. FLG is managed through the identification and exploitation of plant resistance. The glasshouse-based resistance screening produced inconsistent transmission results and the factors responsible for that are not known. A series of glasshouse trials conducted over a 2-year period was compared to identify the factors responsible for the erratic transmission results. SCFDV transmission was greater when the virus was acquired by the vector from a cultivar that was susceptible to the virus than when the virus was acquired from a resistant cultivar. Virus acquisition by the vector was also greater when the vector was exposed to the susceptible cultivars than when exposed to the resistant cultivar. Results suggest that the variation in transmission levels is due to variation in susceptibility of sugarcane cultivars to SCFDV used for virus acquisition by the vector.

Production locality influences postharvest disease development and quality in mangoes

Postharvest disease management is one of the key challenges in commercial mango supply chains. Comprehensive investigations were made regarding the impact of geographic locality on postharvest disease development and other quality parameters in 'Sindhri' and 'Samar Bahisht (S.B.) Chaunsa' mangoes under ambient (33±1°C; 55-60% RH) and low temperature storage/simulated shipping (12±1°C; 80- 85% RH) conditions (28 or 35 days storage for 'Sindhri' and 21 or 28 days for 'S.B. Chaunsa'). Physiologically mature (days from fruit set were 95-100 and 110-115 for 'Sindhri' and 'S.B Chaunsa', respectively) 'Sindhri' and 'S.B. Chaunsa' fruits were harvested from five geographic localities and subjected to ambient and simulated shipping conditions. Under ambient conditions, no disease incidence was observed till fruit eating stage in 'Sindhri'. However, in 'S.B. Chaunsa', significant variation in different localities was observed with respect to disease incidence. Maximum and at par disease was exhibited by the fruit collected from district Vehari and Khanewal in 'S.B. Chaunsa'. Under simulated shipping conditions, disease development varied significantly with respect to different locations and storage durations. In 'Sindhri', fruit of M. Garh, while, 'S.B. Chaunsa' fruit of districts R.Y. Khan, M. Garh and Khanewal showed higher disease incidence. Fruit peel colour development was significantly reduced as storage days increased. Fruit firmness, skin shriveling, fresh weight loss, dry matter, biochemical and organoleptic attributes also varied significantly among the fruit sourced from different orchards of different localities. Analysis of N contents in leaves and fruit peel revealed that N contents of leaf and peel were positively correlated with disease severity in mango. Botryodiplodia spp., Phomopsis mangiferae, Alternaria alternata, Colletotrichum gloeosporioides were the pathogens isolated from fruits of all locations; however, the prevalence frequency varied with the geographic localities. In conclusion, the production locality, cultivar and nutrition (nitrogen content of fruit peel) had significant effect on fruit quality out-turn at ripe stage in terms of disease development so area specific disease management system needs to be implemented for better quality at retail.

Potential of Trichoderma harzianum for control of banana leaf fungal pathogens when applied with a food source and an organic adjuvant

Trichoderma isolates were obtained from diseased leaves and fruit collected from plantations in the main banana production area in Northern Queensland. Phylogenetic analyses identified the Trichoderma isolates as T. harzianum and T. virens. The Trichoderma spp. were found to be antagonistic against the banana leaf pathogens Mycosphaerella musicola, Cordana musae, and Deight-oniella torulosa in vitro. Several products used by the banana industry to increase production, including molasses, Fishoil and Seasol, were tested as food source for the Trichoderma isolates. The optimal food substrate was found to be molasses at a concentration of 5 %, which when used in combination with a di-1-p-menthene spreader-sticker enhanced the survivability of Trichoderma populations under natural conditions. This formulation suppressed D. torulosa development under glasshouse conditions. Furthermore, high sensitivity was observed towards the protectant fungicide Mancozeb but Biopest oil (R), a paraffinic oil, only marginally suppressed the growth of Trichoderma isolates in vitro. Thus, this protocol represents a potential to manage banana leaf pathogens as a part of an integrated disease approach.

Assessing the relationship between seagrass health and habitat quality with wasting disease prevalence in the Florida Keys

Marine pathogens of the genus Labyrinthula have been identified as the cause of wasting disease in seagrass systems in both temperate and subtropical regions. An understanding of the association between environmental factors and the prevalence of wasting disease in seagrass meadows is important for elucidating plant-pathogen interactions in coastal environments. We conducted a survey of 7 turtle grass-dominated beds within the Florida Keys National Marine Sanctuary to assess the relationship between environmental and biological parameters on seagrass health. All sites contained Labyrinthula spp.; the most pathogenic strain was obtained from an anthropogenically impacted site. Leaf and total biomass, in addition to root/rhizome carbon content, canopy light and % light transmitted, all displayed strong negative correlations with a wasting index (WI). It was noted that many of the same environmental measurements that showed negative correlations with WI also displayed strong positive correlations with canopy light levels. These data suggest that light availability may be an important factor that has previously been understated in the seagrass disease literature yet warrants more attention with respect to turtle grass susceptibility to infection. Studies such as this are important because they identify gaps in our understanding of plant-pathogen interactions in subtropical marine ecosystems. Furthermore, the relationships identified in this study may offer insight into which factors are most useful in identifying "at-risk" meadows prior to the onset of larger scale die-off events.

Development of Glomerella leaf spot is enhanced in virus-infected Maxi Gala apples.

Apples are commercially grown in Brazil in a subtropical environment that favors the development of fungal diseases such as Glomerella leaf spot (GLS) caused mainly by Glomerella cingulata (anamorph Colletotrichum gloeosporioides). The main objective of this work was to evaluate the effect of mixed infections by Apple stem grooving virus (ASGV) and Apple stem pitting virus (ASPV) on the infection and the colonization processes of C. gloeosporiodes in cv. Maxi Gala plants. Leaves of 16-month-old potted plants were spray-inoculated and both the disease incidence and lesion count were monitored over time and leaf severity was assessed in the final evaluation using an image analysis tool. Results showed that initial infection estimated from a monomolecular model fitted to progress of lesion count was higher and the incubation period (time to reach 50% incidence) was on average 10 h shorter in virus-infected plants compared to non-infected plants. It is hypothesized that initial events such as conidial germination and fungal penetration into plant cells were facilitated by the presence of viral infection. Also, final GLS severity was significantly higher in the virus-infected plants. Mixed infections by ASGV/ASPV seemed to make apple leaves more susceptible to the initial infection and colonization by C. gloeosporioides.