Polyhydroxyalkanoates (PHAs) from Endophytic Bacterial Strains as Potential Biocontrol Agents against Postharvest Diseases of Apples.

Due to the increasing use and accumulation of petrochemical plastics in the environment and the rapid depletion of natural resources, microbial polyhydroxyalkanoates have great potential to replace them. This study provides new insights in the field of obtaining of polyhydroxyalkanoates (PHAs) from endophytic bacterial strains and applying them as potential biocontrol agents against postharvest diseases of apples. Two strains-Pseudomonas flavescens D5 and Bacillus aerophilus A2-accumulated PHAs in amounts ranging from 2.77 to 5.9 g L-1. The potential to use low-cost substrates such as beet molasses and soapstock for PHA accumulation was shown. The PHAs produced by the Ps. flavescens D5 strain had pronounced antagonistic activity against Penicillium expansum (antifungal property = 62.98-73.08%). The use of PHAs as biocontrol agents significantly reduced the severity of apple blue mold, especially in the preventive treatment option.

Ultrastructural observation and transcriptome analysis provide insights into mechanisms of Penicillium expansum invading apple wounds.

Penicillium expansum is a pathogen causing enormous postharvest losses of fruits, especially apples. In this study, we first investigated the morphological changes of P. expansum within apple wounds during infectious process by microscopic observation. We found that conidia swelled and secreted potential hydrophobin in 4 h, germinated in 8 h, and finally formed conidiophores in 36 h, a critical control time point to prevent the second contamination of spores. We then compared the transcript accumulation of P. expansum in apple tissues and liquid culture at 12 h. In total, 3168 and 1318 up-regulated and down-regulated genes were identified. Among them, genes regarding the biosynthesis of substances such as ergosterol, organic acid, cell wall degrading enzymes, and patulin were induced in expression. Pathways were activated, including autophagy, the mitogen-activated protein kinase, and pectin degradation. Our findings provide insights into the lifestyle and the mechanisms of P. expansum invading apple fruits.

Elaborated regulation of griseofulvin biosynthesis in Penicillium griseofulvum and its role on conidiation and virulence.

Penicilium griseofulvum, the causal agent of apple blue mold, is able to produce in vitro and on apple a broad spectrum of secondary metabolites (SM), including patulin, roquefortine C and griseofulvin. Among them, griseofulvin is known for its antifungal and antiproliferative activity, and has received interest in many sectors, from medicine to agriculture. The biosynthesis of SM is finely regulated by filamentous fungi and can involve global regulators and pathway specific regulators, which are usually encoded by genes present in the same gene cluster as the backbone gene and tailoring enzymes. In the griseofulvin gene cluster, two putative transcription factors were previously identified, encoded by genes gsfR1 and gsfR2, and their role has been investigated in the present work. Analysis of P. griseofulvum knockout mutants lacking either gene suggest that gsfR2 forms part of a different pathway and gsfR1 exhibits many spectra of action, acting as regulator of griseofulvin and patulin biosynthesis and influencing conidia production and virulence on apple. The analysis of gsfR1 promoter revealed that the regulation of griseofulvin biosynthesis is also controlled by global regulators in response to many environmental stimuli, such as carbon and nitrogen. The influence of carbon and nitrogen on griseofulvin production was further investigated and verified, revealing a complex network of response and confirming the central role of gsfR1 in many processes in P. griseofulvum.