Screening of entomopathogenic fungi against citrus mealybug, Plannococcus citri (Hemiptera: Pseudococcidae)

Planococcus citri (citrus mealybug) is a common and damaging citrus crop pest which has proven difficult to control using conventional methods, such as chemical pesticides and insect growth regulators, particularly late in the citrus growing season. The virulence of two entomopathogenic fungal species was studied in laboratory bioassays against the crawlers and adults of P. citri. Isolates of Metarhizium anisopliae and Beauveria bassiana, collected from citrus orchards in the Eastern Cape Province in South Africa, were verified using and molecular techniques. Mealybug bioassays were performed in 24-well plates. Beauveria bassiana (GAR 17 B3) and M. anisopliae (FCM AR 23 B3) isolates both resulted in 67.5 % mortality of mealybug crawlers and B. bassiana (GB AR 23 13 3) resulted in 64 % crawler mortality with concentrations of 1 x 107 conidia/ml. These three isolates were further tested in multipledose bioassays to determine the median lethal concentration (LC50), which were 5.29 x 105conidia/ml for the M. anisopliae isolate (FCM AR 23 B3), 4.25 x 106 conidia/ml for B. bassiana (GAR 17 B3), and 6.65 x 107 conidia/ml B. bassiana (GB AR 23 13 3) for crawlers, respectively. The results of this study suggested that two isolates (M. anisopliae FCM AR 23 B3 and B. bassiana GAR 17 B3) showed potential for further development as biological control agents against citrus mealybug. Further research would be required to determine their ability to perform under field conditions

Biological control potential of ectomycorrhizal fungi against Fusarium circinatum on Pinus patula seedlings

The South Africa forestry industry, covering ∼1.3 million hectares, is dependent on exotic pine and Eucalyptus species. Nursery seedlings are not inoculated with ectomycorrhizal (ECM) fungi or other beneficial microbes. Fusarium circinatum is an economically important pathogen affecting seedling survival. The purpose of this investigation was to assess the effects of ectomycorrhizal fungal inoculation on Pinus patula seedling growth and resistance to the fungal pathogen F. circinatum. Explants from ECM basidiocarps, collected from Pinus stands, were plated onto MMN medium to obtain isolates that were then verified via DNA extraction and PCR amplification and sequencing of the ITS rDNA region. These isolates were identified as Boletus edulis f. reticulatus, Lactarius quieticolor, Suillus granulatus and an unknown Suillus species. P. patula growth in the presence of the pathogen F. circinatum was significantly increased and promoted by the L. quieticolor and Suillus isolates. Preventative inoculation of seedlings in the nursery using these isolates would ensure the production of stronger, healthier plants that would be more resistant to F. circinatum infection, increasing survival in the plantation

Screening of ectomycorrhizal and other associated fungi in South African forest nurseries

The South African forestry industry covers approximately 1.3 million hectares and is dependent on exotic pine and eucalypt species. Nursery seedlings are not inoculated with ectomycorrhizal (ECM) fungi. This investigation assessed levels of naturally occurring ECM colonisation of Pinus patula seedlings from 10 different South African forestry nurseries using a grid line intersect method. Fungi from colonised roots were identified using morphological characteristics and Illumina sequencing. Colonisation of seedlings in production nurseries was low (2-21%). Morphologically, the ECM fungi Thelophora terrestris, Suillus sibiricus, and the genera Russula and Pseudotomentella were identified. Molecularly, the ECM fungi T. terrestris, Inocybe jacobi and the genus Sphaerosporella, as well as several other ECM-containing families were identified, along with many saprotrophic/ endophytic fungi belonging to genera such as Penicillium, Ramasonia and Talaromyces. As can be seen, a combination of both molecular and morphological identification techniques are needed as neither is able to give a full picture of the species present in isolation. This study reveals an initial insight into the root microbiome community associated with Pinus patula seedlings, which should be taken into account when inoculation with beneficial microbes is considered. It determined that natural ECM fungal root colonisation levels are very low throughout the South African nurseries investigated, indicating the need for ECM fungal inoculation, which can increase seedling growth, viability and resistance to pathogens