Determination of the effects of nanoparticles synthesis by different methods on germination, glycomannan content and associated microbiome of anacamptis laxiflora seeds

This study investigated the potential effects of nanoparticles on Anacamptis laxiflora (Lam.) R.M.Bateman, Pridgeon & M.W.Chase seed germination, seed development, glucomannan content, and the accompanying seed microbiome, which is an under-researched area. For this purpose, comprehensive evaluations were made by adding Anacamptis laxiflora seeds to three concentrations (10, 50, and 150 mg) of chemically and biologically synthesized nanoparticles to different growth media (solid, liquid, and soil). Four of the endophytic fungi isolated from Anacamptis laxiflora seeds were determined to be capable of producing nanoparticles, and these isolates were identified as Dactylonectria torresensis, Fusarium tricinctum, Alternaria alternate, and Alternaria tenuissima. Seed germination studies have revealed interesting and diverse responses among treatments. While chemical nanoparticles exhibited anti-germination effects in the solid environment, their negative effects in the soil environment were negligible. In contrast, 50 mg application of Fe BNp1 nanoparticles, biologically synthesized (green synthesis) through Dactylonectria torresensis, showed a positive effect on germination in solid media, and Fe BNp6 nanoparticles biologically synthesized through Fusarium tricinctum showed promising results in soil environment. Interestingly, the effect of a liquid medium on the germination of Anacamptis laxiflora seeds was unfavorable. Regarding the assessment of glucomannan content, significant differences emerged in the response to nanoparticle exposure. Glucomannan content was increased, especially in solid and soil media treated with FeBNp6 nanoparticles(50 mg). Analysis of the seed-associated microbial community performed through high-throughput sequencing of 16S rRNA and ITS rRNA genes provided valuable information on the role of FeBNp6 nanoparticles (50 mg) selected from treatments and its impact on the seed microbiome. Within the bacterial community, an increased abundance of certain genera, including Pseudomonas, Cellulomonas, and Chthoniobacter, was observed in the presence of FeBNp6 nanoparticles (50 mg). These findings also shed light on specific metabolic pathways that are enhanced in the presence of these nanoparticles via the PICRUSt tool. Among fungi, genera such as Barnettozyma, Aspergillus, Didymella, Albifimbria, and Malassezia were dominant and relatively abundant, especially in response to specific pathways. This comprehensive, pioneering study contributes to understanding the complex interactions between nanoparticles, Anacamptis laxiflora seeds, and their associated microbial communities.