Infusions prepared from raw pu-erh tea (RAPT) and ripened pu-erh tea (RIPT) showed remarkable aroma differences. Predominant odorants in RAPT and RIPT infusions were identified and compared by the combined use of gas chromatography–olfactometry, aroma extract dilution analysis, odor activity values (OAVs), and multivariate analysis. A total of 35 and 19 odorants (OAV > 1) were detected in RIPT and RAPT, respectively. Odorants in RAPT and RIPT are significantly different in both odor properties and aroma compound intensities. Overall, RAPT contained a complex variety of chemical classes with diverse odors and moderate odor intensities, while RIPT is dominated by structurally and organoleptically similar compounds with high potency. Specifically, stale and musty smelling methoxybenzenes contributed the most to RIPT, while floral-, sweet-, and woody-smelling terpene alcohols, terpene ketones, and phenolic compounds were the predominant odorants in RAPT. Orthogonal partial least squares discriminant analysis revealed that linalool, α-ionone, 1,2,4-trimethoxybenzene, 1,2,3-trimethoxy-5-methylbenzene, 1,2,3,4-tetramethoxybenzene, and 1,2,3-trimethoxybenzene underwent remarkable changes during pile fermentation and could be used as potential odor-active markers for RIPT and RAPT discrimination. The comprehensive aroma characterization of pu-erh tea and determination of the effect of pile fermentation on odorant alteration herein will provide guidance for pu-erh tea flavor quality control and evaluation.

Ralstonia solanacearum species complex (RSSC) is a diverse group of plant pathogens that attack a wide range of hosts and cause devastating losses worldwide. In this study, we conducted a comprehensive analysis of 131 RSSC strains to detect their genetic diversity, pathogenicity, and evolution dynamics. Average nucleotide identity analysis was performed to explore the genomic relatedness among these strains, and finally obtained an open pangenome with 32,961 gene families. To better understand the diverse evolution and pathogenicity, we also conducted a series of analyses of virulence factors (VFs) and horizontal gene transfer (HGT) in the pangenome and at the single genome level. The distribution of VFs and mobile genetic elements (MGEs) showed significant differences among different groups and strains, which were consistent with the new nomenclatures of the RSSC with three distinct species. Further functional analysis showed that most HGT events conferred from Burkholderiales and played a great role in shaping the genomic plasticity and genetic diversity of RSSC genomes. Our work provides insights into the genetic polymorphism, evolution dynamics, and pathogenetic variety of RSSC and provides strong supports for the new taxonomic classification, as well as abundant resources for studying host specificity and pathogen emergence.

Ralstonia pseudosolanacearum, previously known as R. solanacearum species complex (RSSC) phylotypes I and III, is a plant pathogenic bacterium causing significant yield losses in economical crops. In the May of 2020 and 2021, cigar tobacco bacterial wilt was first observed in fields in Danzhou, Hainan Province, China. Eight bacterial isolates were isolated and identified as R. pseudosolanacearum with race 1, biovar III by 16S rRNA gene sequencing, Biolog and host identification. The amino acid sequence showed that Hainan strains and 15 R. pseudosolanacearum reference strains from flue-cured tobacco in Shandong and Guizhou Provinces, all belonged to RS1000 type containing the avrA gene, only Guizhou strains also had the popP1 gene. On the basis of phylotype-specific multiplex PCR amplification, mismatch repair gene and endoglucanase gene-base tree, Hainan strains were identified as phylotype I sequevar 70, and showed stronger pathogenic capabilities on three different varieties than those reference strains. This is the first report of cigar tobacco bacterial wilt caused by R. pseudosolanacearum sequevar 70. The results revealed the diversity of RSSC in Nicotiana tabacum in China and provided useful information regarding the epidemiology of cigar tobacco wilt disease, as well as the breeding for disease resistance in local cigar tobacco.