Guvermectin, as a novel nucleoside-like biopesticide, could increase the rice yield excellently, but the potential environmental behaviors remain unclear, which pose potential health risks. Therefore, the uptake and biotransformation of guvermectin in three types of crops (rice, lettuce, and carrot) were first evaluated with a hydroponic system. Guvermectin could be rapidly absorbed and reached equilibrium in roots (12-36 h) and shoots (24-60 h) in three plants, and guvermectin was also vulnerable to dissipation in roots (t1/2 1.02-3.65 h) and shoots (t1/2 9.30-17.91 h). In addition, 8 phase I and 2 phase II metabolites, transformed from guvermectin degradation in vivo and in vitro exposure, were identified, and one was confirmed as psicofuranine, which had antibacterial and antitumor properties; other metabolites were nucleoside-like chemicals. Molecular simulation and quantitative polymerase chain reaction further demonstrated that guvermectin was metabolized by the catabolism pathway of an endogenous nucleotide. Guvermectin had similar metabolites in three plants, but the biotransformation ability had a strong species dependence. In addition, all the metabolites exhibit neglectable toxicities (bioconcentration factor <2000 L/kg b.w., LC50,rat > 5000 mg/kg b.w.) by prediction. The study provided valuable evidence for the application of guvermectin and a better understanding of the biological behavior of nucleoside-like pesticides.

Bacterial diseases caused substantial yield losses worldwide, with the rise of antibiotic resistance, there is a critical need for alternative antibacterial compounds. Natural products (NPs) from microorganisms have emerged as promising candidates due to their potential as cost-effective and environmentally friendly bactericides. However, the precise mechanisms underlying the antibacterial activity of many NPs, including Guvermectin (GV), remain poorly understood. Here, we sought to explore how GV interacts with Guanosine 5\'-monophosphate synthetase (GMPs), an enzyme crucial in bacterial guanine synthesis. We employed a combination of biochemical and genetic approaches, enzyme activity assays, site-directed mutagenesis, bio-layer interferometry, and molecular docking assays to assess GV\'s antibacterial activity and its mechanism targeting GMPs. The results showed that GV effectively inhibits GMPs, disrupting bacterial guanine synthesis. This was confirmed through drug-resistant assays and direct enzyme inhibition studies. Bio-layer interferometry assays demonstrated specific binding of GV to GMPs, with dependency on Xanthosine 5\'-monophosphate. Site-directed mutagenesis identified key residues crucial for the GV-GMP interaction. This study elucidates the antibacterial mechanism of GV, highlighting its potential as a biocontrol agent in agriculture. These findings contribute to the development of novel antibacterial agents and underscore the importance of exploring natural products for agricultural disease management.

Plant growth regulators (PGRs) play an important role in alleviating the detrimental effects of biotic and abiotic stress and improving crop yield and quality. As a novel PGR from Streptomyces registered in 2021, guvermectin (GV) has the potential to improve plant yield and defense, making its application in agriculture a subject of interest. Here, we describe the discovery process, functional activities, agricultural applications, toxicity, environmental safety, and biosynthetic mechanism of GV. This Perspective provides a guide for the development of novel PGRs from microorganisms.

Guvermectin is a novel biopesticide often used as seed soaking to promote the rice yield. However, its biotoxicity and degradation behavior in soils were still not disclosed, which posed a knowledge gap to guide its rational application. Therefore, the degradation behaviors of guvermectin in four typical soils under aerobic and anaerobic conditions were investigated in the laboratory. The results showed that guvermectin was degraded fast with DT50 ranging from 0.95 to 10.10 d, and the degradation rate was higher in aerobic condition than that in anaerobic condition. Eight transformation products were screened using UPLC-QTOF/MS. The acute toxicities tests of guvermectin to Coturnix coturnix japonica and Apis mellifera were measured by biological laboratory experiments, and the acute and chronic toxicities of transformation products to Danio rerio, Daphnia magna Straus and Green algae were predicted by ECOSAR software. The results showed that guvermectin has low toxic to quail and honeybee (LD50 2000 mg a.i./kg body weight, LD50 ˃ 100 μg a.i./bee), and its transformation products were also low toxic class to Danio rerio, Daphnia magna Straus and Green algae (LC50/EC50 > 100 mg a.i./L). However, the nucleoside-like metabolites may pose a potential risk due to their similarity to genetic material, which should be concerned. The findings provided important environmental risk assessment data for the rational use of guvermectin.

Flowering is a crucial stage for plant reproductive success; therefore, the regulation of plant flowering has been widely researched. Although multiple well-defined endogenous and exogenous flowering regulators have been reported, new ones are constantly being discovered. Here, we confirm that a novel plant growth regulator guvermectin (GV) induces early flowering in Arabidopsis. Interestingly, our genetic experiments newly demonstrated that WRKY41 and its homolog WRKY53 were involved in GV-accelerated flowering as positive flowering regulators. Overexpression of WRKY41 or WRKY53 resulted in an early flowering phenotype compared to the wild type (WT). In contrast, the w41/w53 double mutants showed a delay in GV-accelerated flowering. Gene expression analysis showed that flowering regulatory genes SOC1 and LFY were upregulated in GV-treated WT, 35S:WRKY41, and 35S:WRKY53 plants, but both declined in w41/w53 mutants with or without GV treatment. Meanwhile, biochemical assays confirmed that SOC1 and LFY were both direct targets of WRKY41 and WRKY53. Furthermore, the early flowering phenotype of 35S:WRKY41 lines was abolished in the soc1 or lfy background. Together, our results suggest that GV plays a function in promoting flowering, which was co-mediated by WRKY41 and WRKY53 acting as new flowering regulators by directly activating the transcription of SOC1 and LFY in Arabidopsis.

Rice direct seeding technology has been considered as a promising alternative to traditional transplanting because of its advantages in saving labor and water. However, the poor emergence and seedling growth caused by chill stress are the main bottlenecks in wide-scale adoption of direct-seeded rice in Heilongjiang Province, China. Here, we found that natural plant growth regulator guvermectin (GV) effectively improved rice seed germination and seedling growth under chilling stress. Results from 2 year field trials showed that seed-soaking with GV not only enhanced the emergence rate and seedling growth but also increased the panicle number per plant and grain number per panicle, resulting in 9.0 and 6.8% increase in the yield of direct-seeded rice, respectively. Integrative physiological, transcriptomic, and metabolomic assays revealed that GV promoted seed germination under chilling stress mainly by enhancing the activities of α-amylase and antioxidant enzymes (superoxide dismutase, peroxidase, and catalase), increasing the contents of soluble sugar and soluble protein, improving the biosynthesis of glutathione and flavonoids, as well as activating gibberellin-responsive transcription factors and inhibiting the abscisic acid signaling pathway. These findings indicate that seed-soaking with GV has good potential to improve seedling establishment and yield of direct-seeded rice even under chilling stress.

Food is a fundamental human right, and global food security is threatened by crop production. Plant growth regulators (PGRs) play an essential role in improving crop yield and quality, and this study reports on a novel PGR, termed guvermectin (GV), isolated from plant growth-promoting rhizobacteria, which can promote root and coleoptile growth, tillering, and early maturing in rice. GV is a nucleoside analogue like cytokinin (CK), but it was found that GV significantly promoted root and hypocotyl growth, which is different from the function of CK in Arabidopsis. The Arabidopsis CK receptor triple mutant ahk2-2 ahk3-3 cre1-12 still showed a GV response. Moreover, GV led different growth-promoting traits from auxin, gibberellin (GA), and brassinosteroid (BR) in Arabidopsis and rice. The results from a four-year field trial involving 28 rice varieties showed that seed-soaking treatment with GV increased the yields by 6.2 to 19.6%, outperforming the 4.0 to 10.8% for CK, 1.6 to 16.9% for BR, and 2.2 to 7.1% for GA-auxin-BR mixture. Transcriptome analysis demonstrated that GV induced different transcriptome patterns from CK, auxin, BR, and GA, and SAUR genes may regulate GV-mediated plant growth and development. This study suggests that GV represents a novel PGR with a unique signal perception and transduction pathway in plants.

Guvermectin is a biopesticide isolated from the secondary metabolites of Streptomyces sp. NEAU6, an endogenous actinomyces of a Chinese medicine named Paris polyphylla. However, the environmental degradation behavior and biotoxicity of guvermectin are still unclear, which may affect its rational application. Therefore, the degradation of guvermectin in water at different pH values (pH 4, pH 6, pH 7, and pH 9) and with or without light was investigated in the laboratory. The results showed that guvermectin could be degraded in pH 4 solution, and the presence of light irradiation enhanced the degradation process with a DT50 of 2.95 and 12 days for photolysis and hydrolysis, respectively. However, guvermectin was fairly stable in other conditions. Three products transformed from guvermectin degradation were identified by UPLC-QTOF/MS. Biotoxicity assessment was performed on Danio rerio and Daphnia magna Straus by ECOSAR prediction and in vivo biological tests. The test data showed that guvermectin and its transformation products exhibited low toxicities to D. rerio and D. magna Straus (LC50/EC50 > 100 mg a.i./L), and the transformation products had lower toxicity than their parent substance. The results provided a reference for elucidating the potential risk of guvermectin to nontarget organisms and promoting its rational use.

Guvermectin is a recently discovered microbial N9-glucoside cytokinin compound extracted from Streptomyces sanjiangensis NEAU6. Although some research has reported that N9-glucoside cytokinin compounds do not have the activity of cytokinin, it has been noted that guvermectin can promote growth and antifungal activity in Arabidopsis. Maize is an important food crop in the world and exploring the effect of guvermectin on this crop could help its cultivation in regions with adverse environmental conditions such as a high temperature. Here, we investigated the effects of guvermectin seed soaking treatment on the growth of maize at the seedlings stage and its yield attributes with different temperature stresses. The maize (cv. Zhengdan 958) with guvermectin seed soaking treatment were in two systems: paper roll culture and field conditions. Guvermectin seed soaking treated plants had increased plant height, root length, and mesocotyl length at the seedlings stage, and spike weight at maturity in the field. But only root length was increased at the paper roll culture by guvermectin seed soaking treatment. Guvermectin seed soaking treatment reduced the adverse effects on maize seedling when grow at a high temperature. Further experiments showed that, in high temperature conditions, guvermectin treatment promoted the accumulation of heat shock protein (HSP) 17.0, HSP 17.4 and HSP 17.9 in maize roots. Comparative transcriptomic profiling showed there were 33 common differentially expressed genes (DEGs) in guvermectin treated plants under high temperature and room temperature conditions. The DEGs suggested that guvermectin treatment led to the differential modulation of several transcripts mainly related with plant defense, stress response, and terpenoid biosynthesis. Taken together, these results suggested that the guvermectin treatment promoted the growth and tolerance of high temperature stresses, possibly by activation of related pathways. These results show that guvermectin is a novel plant growth regulator and could be developed as an application to maize seeds to promote growth in high temperature environments.