No abstract

Entomopathogenic fungi from the genus Beauveria (Vuillemin) play an important role in controlling insect populations and have been increasingly utilized for the biological control of insect pests. Various studies have reported that Beauveria bassiana (Bals.), Vuill. also has the ability to colonize a broad range of plant hosts as endophytes without causing disease but while still maintaining the capacity to infect insects. Beauveria is often applied as an inundative spore application, but little research has considered how plant colonization may alter the ability to persist in the environment. The aim of this study was to investigate potential interactions between B. bassiana and Zea mays L. (maize) in the rhizosphere following inoculation, in order to understand the factors that may affect environmental persistence of the fungi. The hypothesis was that different isolates of B. bassiana have the ability to colonize maize roots and/or rhizosphere soil, resulting in effects to the plant microbiome. To test this hypothesis, a two-step nested PCR protocol was developed to find and amplify Beauveria in planta or in soil; based on the translation elongation factor 1-alpha (ef1α) gene. The nested protocol was also designed to enable Beauveria species differentiation by sequence analysis. The impact of three selected B. bassiana isolates applied topically to roots on the rhizosphere soil community structure and function were consequently assessed using denaturing gradient gel electrophoresis (DGGE) and MicroRespTM techniques. The microbial community structure and function were not significantly affected by the presence of the isolates, however, retention of the inocula in the rhizosphere at 30 days after inoculation was enhanced when plants were subjected to intensive wounding of foliage to crudely simulate herbivory. The plant defense response likely changed under wound stress resulting in the apparent recruitment of Beauveria in the rhizosphere, which may be an indirect defensive strategy against herbivory and/or the result of induced systemic susceptibility in maize enabling plant colonization.

Species in the fungal genus Beauveria are pathogens of invertebrates and have been commonly used as the active agent in biopesticides. After many decades with few species described, recent molecular approaches to classification have led to over 25 species now delimited.

Background. The New Zealand forest industry would greatly benefit from a successful way of controlling insect pests. The entomopathogenic fungus, Beauveria bassiana could hold such potential and has previously been shown to be capable of endophytic colonisation of the Monterey pine Pinus radiata. Nevertheless clarifications on its mode of transmission, persistence and action in this plant are required. In this study we investigated B. bassiana transmission and persitence in P. radiata and whether this fungus is beneficial to P. radiata by testing its effect as a plant endophyte on the fitness performance of above and belowground insect feeders. Methods. Both culturing and molecular approaches were used to detect the occurrence B. bassiana in pines. Transmission electron microscopy of positive germinating seeds was also used to locate the fungus. Bioassays were conducted on root and needle feeding insects using Beauveria positive and endophyte free pine seedlings. Results. Beauveria bassiana was detected in seedlings which had not previously been exposed to the fungus, indicating a vertical mode of transmission. The fungus could colonise all parts of the pines, but did not always persist. We found that the presence of the fungus negatively affects the fitness of the below-ground insect feeding on the plant by reducing their survival by over 10% and their weight by 5%. This study also showed that the mode of action of endophytic B. bassiana in pine is likely to be by feeding deterrence of insects induced locally by fungal metabolites, rather than by direct fungal infection of the insects. Discussion. A vertically transmitted beneficial endophyte of pine could be used as a cost effective approach to control insect pest in these commercially grown trees.

The identification of fungal endophytes often relies on culturing isolates from surface-sterilized plant tissue. However, molecular techniques have enabled the rapid detection and identification of targeted endophyte species, and next-generation sequencing technology provides an opportunity to obtain comprehensive information on endophytic communities, directly from plant tissue. In order to achieve accurate detection from internal tissues, surface microbes and associated deoxyribonucleic acid (DNA) must be eliminated, with particular consideration for the type of plant tissue and the efficacy of the surface sterilization procedure used. The methodology described later was developed specifically for detection of DNA from the entomopathogenic fungal endophyte Beauveria bassiana (Vuillemin) (Ascomycota: Hypocreales) in various tissues of Zea mays (L.). However, the protocol may be easily applied to other fungi and bacterial endophytes. Included is a stringent sodium hypochlorite-based surface sterilization protocol for plant material in preparation for polymerase chain reaction (PCR) to detect target DNA within plant tissue. Included are a treatment for dealing with surface DNA contamination and a novel procedure for assessing the efficacy of surface sterilization using PCR.

Background. The New Zealand forest industry would greatly benefit from a successful way of controlling insect pests. The entomopathogenic fungus, Beauveria bassiana could hold such potential and has previously been shown to be capable of endophytic colonisation of the Monterey pine Pinus radiata. Nevertheless clarifications on its mode of transmission, persistence and action in this plant are required. In this study we investigated B. bassiana transmission and persitence in P. radiata and whether this fungus is beneficial to P. radiata by testing its effect as a plant endophyte on the fitness performance of above and belowground insect feeders. Methods. Both culturing and molecular approaches were used to detect the occurrence B. bassiana in pines. Transmission electron microscopy of positive germinating seeds was also used to locate the fungus. Bioassays were conducted on root and needle feeding insects using Beauveria positive and endophyte free pine seedlings. Results. Beauveria bassiana was detected in seedlings which had not previously been exposed to the fungus, indicating a vertical mode of transmission. The fungus could colonise all parts of the pines, but did not always persist. We found that the presence of the fungus negatively affects the fitness of the below-ground insect feeding on the plant by reducing their survival by over 10% and their weight by 5%. This study also showed that the mode of action of endophytic B. bassiana in pine is likely to be by feeding deterrence of insects induced locally by fungal metabolites, rather than by direct fungal infection of the insects. Discussion. A vertically transmitted beneficial endophyte of pine could be used as a cost effective approach to control insect pest in these commercially grown trees.

No abstract

Certain crops depend upon pollination services for fruit set, and, of these, almonds are of high value for Australia. Stressors, such as diseases, parasites, pesticides, and nutrition, can contribute to honey bee Apis mellifera L. colony decline, thereby reducing bee activity and pollination efficiency. In Australia, field studies are required to monitor honey bee health and to ascertain whether factors associated with colony decline are impacting hives. We monitored honey bee colonies during and after pollination services of almond. Video surveillance technology was used to quantify bee activity, and bee-collected pollen was periodically tested for pesticide residues. Plant species diversity was also assessed using DNA metabarcoding of the pollen. Results showed that bee activity increased in almond but not in bushland. Residues detected included four fungicides, although the quantities were of low risk of oral toxicity to bees. Floral diversity was lower in the pollen collected by bees from almonds compared to bushland. However, diversity was higher at the onset and conclusion of the almond bloom, suggesting that bees foraged more widely when availability was low. Our findings suggest that commercial almond orchards may sustain healthier bee colonies compared to bushland in early spring, although the magnitude of the benefit is likely landscape-dependent.