Species in the Botryosphaeriaceae represent some of the most important fungal pathogens of woody plants. Although these fungi have been relatively well studied on economically important crops, hardly anything is known regarding their taxonomy or ecology on native or non-commercial tree species. The aim of this study was to compare the diversity and distribution of the Botryosphaeriaceae on Terminalia catappa, a tropical tree of Asian origin planted as an ornamental in Cameroon, Madagascar and South Africa. A total of 83 trees were sampled, yielding 79 Botryosphaeriaceae isolates. Isolates were initially grouped based on morphology of cultures and conidia. Representatives of the different morphological groups were then further characterised using sequence data for the ITS, tef 1-alpha, rpb2, BOTF15 and beta-tubulin gene regions. Five species of the Botryosphaeriaceae were identified, including Neofusicoccum parvum, N. batangarum sp. nov., Lasiodiplodia pseudotheobromae, L. theobromae and L. mahajangana sp. nov. Lasiodiplodia pseudotheobromae and L. theobromae, were the most commonly isolated species (62%), and were found at all the sites. Neofusicoccum parvum and N. batangarum were found in South Africa and Cameroon, respectively, whereas L. mahajangana was found only in Madagascar. Greenhouse inoculation trials performed on young T. catappa trees showed variation among isolates tested, with L. pseudotheobromae being the most pathogenic. The Botryosphaeriaceae infecting T. catappa appear to be dominated by generalist species that also occur on various other hosts in tropical and sub-tropical climates.

Phytophthora megakarya (Pmeg) and Phytophthora palmivora (Ppal) are closely related species causing cacao black pod rot. Although Ppal is a cosmopolitan pathogen, cacao is the only known host of economic importance for Pmeg. Pmeg is more virulent on cacao than Ppal. We sequenced and compared the Pmeg and Ppal genomes and identified virulence-related putative gene models (PGeneM) that may be responsible for their differences in host specificities and virulence. Pmeg and Ppal have estimated genome sizes of 126.88 and 151.23 Mb and PGeneM numbers of 42,036 and 44,327, respectively. The evolutionary histories of Pmeg and Ppal appear quite different. Postspeciation, Ppal underwent whole-genome duplication whereas Pmeg has undergone selective increases in PGeneM numbers, likely through accelerated transposable element-driven duplications. Many PGeneMs in both species failed to match transcripts and may represent pseudogenes or cryptic genetic reservoirs. Pmeg appears to have amplified specific gene families, some of which are virulence-related. Analysis of mycelium, zoospore, and in planta transcriptome expression profiles using neural network self-organizing map analysis generated 24 multivariate and nonlinear self-organizing map classes. Many members of the RxLR, necrosis-inducing phytophthora protein, and pectinase genes families were specifically induced in planta. Pmeg displays a diverse virulence-related gene complement similar in size to and potentially of greater diversity than Ppal but it remains likely that the specific functions of the genes determine each species’ unique characteristics as pathogens.

No abstract

Among the Phytophthora species that cause black pod of cacao, P. megakarya is the most virulent, posing a serious threat to cacao production in Africa. Correct identification of the species causing the black pod and understanding the virulence factors involved are important for developing sustainable disease management strategies. A simple PCR-based species identification method was developed using the species-specific sequences in the ITS regions of the rRNA gene. A phylogenetic tree generated for 119 Phytophthora isolates, based on the 60S ribosomal protein L10 gene and rDNA sequence, verified the PCR-based identification assay and showed high interspecific variation among the species causing black pod. Phytophthora megakarya isolates were uniformly virulent in an assay using susceptible cacao pod husks inoculated with zoospores, while the P. palmivora isolates showed greater divergence in virulence. The virulence of P. megakarya was associated with earlier production of sporangia and an accelerated induction of necrosis. While zoospore germ tubes of both species penetrated pods through stomata, only P. megakarya produced significant numbers of appressoria. A hypersensitive-like response was observed when attached SCA-6 pods were inoculated with P. palmivora. SCA-6 pods became vulnerable to P. palmivora when wounded prior to zoospore inoculation. Phytophthora megakarya was more aggressive than P. palmivora on attached SCA-6 pods, causing expanding necrotic lesions with or without wounding. Phytophthora megakarya is predominant in the Volta region of Ghana and it remains to be seen whether it can displace P. palmivora from cacao plantations of Ghana as it has in Nigeria and Cameroon.