Lasiodiplodia theobromae is a fungus of the Botryosphaeriaceae that causes grapevine vascular disease, especially in regions with hot climates. Fungi in this group often remain latent within their host and become virulent under abiotic stress. Transcriptional regulation analysis of L. theobromae exposed to heat stress (HS) was first carried out in vitro in the presence of grapevine wood (GW) to identify potential pathogenicity genes that were later evaluated for in planta expression. A total of 19,860 de novo assembled transcripts were obtained, forty-nine per cent of which showed homology to the Botryosphaeriaceae fungi, Neofusicoccum parvum or Macrophomina phaseolina. Three hundred ninety-nine have homology with genes involved in pathogenic processes and several belonged to expanded gene families in others fungal grapevine vascular pathogens. Gene expression analysis showed changes in fungal metabolism of phenolic compounds; where genes encoding for enzymes, with the ability to degrade salicylic acid (SA) and plant phenylpropanoid precursors, were up-regulated during in vitro HS response, in the presence of GW. These results suggest that the fungal L-tyrosine catabolism pathway could help the fungus to remove phenylpropanoid precursors thereby evading the host defense response. The in planta up-regulation of salicylate hydroxylase, intradiol ring cleavage dioxygenase and fumarylacetoacetase encoding genes, further supported this hypothesis. Those genes were even more up-regulated in HS-stressed plants, suggesting that fungus takes advantage of the increased phenylpropanoid precursors produced under stress. Pectate lyase was up-regulated while a putative amylase was down-regulated in planta, this could be associated with an intercellular growth strategy during the first stages of colonization. L. theobromae transcriptome was established and validated. Its usefulness was demonstrated through the identification of genes expressed during the infection process. Our results support the hypothesis that heat stress facilitates fungal colonization, because of the fungus ability to use the phenylpropanoid precursors and SA, both compounds known to control host defense.

Botryosphaeria dieback is one of the most prevalent grapevine trunk diseases (GTDs), and is caused by fungi in the Botryosphaeriaceae. Fungi invade grapevine vascular systems mainly through pruning wounds, and cause cankers and necrotic lesions, which lead to grapevine decline and death. Lasiodiplodia theobromae has been reported as a highly virulent pathogen of grapevine, and was previously reported in Mexican vineyards. The taxonomy of Lasiodiplodia was recently revised, adding new species, and some were reduced to synonymy. This study aimed to characterize Lasio-diplodia producing grapevine dieback symptoms in Sonora and Baja California, Mexico. Using the phylogenetic markers tef1-α and ITS regions, Lasiodiplodia brasiliensis, L. crassispora, L. exigua, and L. gilanensis were identified. Lasidiplodia exigua was the most prevalent species. Lasiodiplodia brasiliensis and L. gilanensis were very virulent to ‘Cabernet Sauvignon’ plants, while L. exigua and L. gilanensis were less virulent, and L. crassispora did not produce lesions at 2 months post-inoculation. The optimum temperature of the Lasiodiplodia spp. was 28°C, but all four species grew up to 37°C, and the isolates of L. exigua grew slowly at 40°C. This is the first report of the four of Lasio-diplodia species in vineyards of Mexico.