Diversity and Pathogenicity of Botryosphaeriaceae Associated with Macadamia Branch Dieback in Australia

Abstract

Botryosphaeria branch dieback is a serious disease of macadamia in Australia, but its etiology has not been clearly defined, which limits effective disease control. Therefore, this study examined whether the causal agents of branch dieback in commercial macadamia orchards in five agroecological regions in Australia are similar in prevalence and aggressiveness. The identity of the causal agents was determined using conventional culturing techniques and DNA sequencing that targets the internal transcribed spacer (ITS), translation elongation factor 1-alpha (tef1α), β-tubulin (tub2), and DNA-directed RNA polymerase II second largest subunit (rpb2) gene loci. The pathogenic variation of the isolates, relative to the source (region and host plant part), was examined using in vivo and in planta assays. Lasiodiplodia and Neofusicoccum were the dominant fungal genera obtained from surveys of 59 macadamia orchards across the agroecological regions. Phylogenetic analysis of 52 representative isolates identified four putative novel Lasiodiplodia clades, with three other Lasiodiplodia spp. (Lasiodiplodia iraniensis, L. pseudotheobromae, and L. theobromae) and three Neofusicoccum spp. (Neofusicoccum luteum, N. mangroviorum, and N. parvum) from macadamia. L. pseudotheobromae that constituted 40% of the isolates from symptomatic tissues was the most prevalent in all the regions. Both the in vivo and in planta pathogenicity assays revealed that all isolates of the Botryosphaeriaceae, except N. mangroviorum, were pathogenic to macadamia. L. theobromae, N. luteum, and L. iraniensis were the most aggressive species causing dieback symptoms in macadamia.