Transcriptome analysis of gene expression in Lasiodiplodia theobromae

Botryosphaeriaceae is an important fungal family that can colonize a wide range of woody hosts including economically important plants and cause serious diseases worldwide. To uncover the virulence variation in Lasiodiplodia theobromae, we sequenced the transcriptome of Las1 in L. theobromae inoculated with the stems of V.vinifera using dual-RNAseq method. We found a lot of expanded genes were induced after inoculated with grapevine compared with the transcriptome of Las1 that was maintained in PDA under the same condition. The L. theobromae strain Las1 was selected for inoculation tests on detached Vitis vinifera cv. ‘Summer Black’ branches. Briefly, wounds of green grapevine shoots were inoculated with mycelium plugs (5 mm in diameter) cut with a cork borer and then maintained under an alternating dark/light cycle inside a growth chamber with constant humidity and a temperature of 25 °C. All the inoculated grapevine tissues were collected at 24 h post-inoculation, and fungal mycelia collected from PDA at 24 h post-culturing at 25 °C were used for subsequent RNA-seq analyses. Total RNA was extracted and processed following Novogene Company methodologies for preparing Illumina RNA-seq libraries. Paired-end sequencing was performed on an Illumina HiSeq 4000 platform to generate 150-bp paired-end reads. RNA-seq was performed on three independent biological replicates.