Background: TheLycorisgenus includes many ornamentally and medicinally important species. Polyploidization and hybridization are considered modes of speciation in this genus, implying great genetic diversity. However, the lack of effective molecular markers has limited the genetic analysis of this genus.Results: In this study, mining of EST-SSR markers was performed using transcriptome sequences ofL. aurea, and 839 primer pairs for non-redundant EST-SSRs were successfully designed. A subset of 60 pairs was randomly selected for validation, of which 44 pairs could amplify products of the expected size. Cross-species transferability of the 60 primer pairs amongLycorisspecies were assessed inL. radiataHreb,L. sprengeriComes ex Baker,L. chinensisTraub andL. anhuiensis, of which between 38 to 77% of the primers were able to amplify products in theseLycorisspecies. Furthermore, 20 and 10 amplification products were selected for sequencing verification inL. aureaandL. radiatarespectively. All products were validated as expected SSRs. In addition, 15 SSRs, including 10 sequence-verified and 5 unverified SSRs were selected and used to evaluate the genetic diversity of sevenL. radiatalines. Among these, there were three sterile lines, three fertile lines and one line represented by the offspring of one fertile line. Unweighted pair group method with arithmetic mean analysis (UPGMA) demonstrated that the outgroup,L. aureawas separated fromL. radiatalines and that the sevenL. radiatalines were clustered into two groups, consistent with their fertility. Interestingly, even a dendrogram with 34 individuals representing the sevenL. radiatalines was almost consistent with fertility.Conclusions: This study supplies a pool of potential 839 non-redundant SSR markers for genetic analysis ofLycorisgenus, that present high amplification rate, transferability and efficiency, which will facilitate genetic analysis and breeding program inLycoris.

Corydalis yanhusuo, a precious herb of the Papaveraceae family, is widely used in multiple traditional Chinese medicines for the treatment of many painful conditions, and its medicinal part is the dried tuber. Yet how to improve this plant’s medicinal yield as well as its economic efficiency remains a key problem in its cultivation. The planting of C. yanhusuo in rotation with peanut (Arachis hypogaea L.) aims to improve land utilization efficiency, but the total production of tubers is severely reduced relative to fields without rotation. However, an increased yield was observed in C. yanhusuo plants grown in previously flooded fields (HR field) compared to the ones grown in the fields that had been used to cultivate peanut (PL field) or in fields without rotation or flooding (N field). Based on these phenomena, in this study, we explored the potential factors responsible for the altered growth/yield of C. yanhusuo under different field conditions. Soil physicochemical properties and the diversity and community of rhizobacteriome of C. yanhusuo were both analyzed. By testing several soil physicochemical properties, we found that the cation exchange capacity (CEC), soil organic matter (SOM), total nitrogen (TN), and pH value differed significantly among these three types of fields. 16S rRNA amplicon sequencing revealed stark differences in the composition, diversity, and potential functions of the bacterial community in the rhizosphere of C. yanhusuo plants grown in field with the peanut rotation or flooding. Notably, the Acidobacteria were enriched in the HR field, while Actinobacteria were enriched in the PL field. More importantly, further analysis showed that changed soil physicochemical properties could be one reason for why the rhizospheric bacterial community has changed; hence, soil physicochemical properties might also be affecting plant performance indirectly by regulating the rhizospheric bacterial community. The RDA analysis distinguished CEC as the most important soil physicochemical property influencing the structure and composition of the C. yanhusuo rhizobacteriome. In summary, our results suggest peanut rotation- and flooding-induced soil physicochemical properties changes would further impact the rhizobacteriome of C. yanhusuo albeit differentially, culminating in opposite effects upon the plant growth and medicinal yield of C. yanhusuo.

As a kind of Amaryllidaceae alkaloid which is accumulated in the species of Lycoris plants, lycorine has a range of physiological effects. The biosynthesis pathway of lycorine has been partly revealed, but the transport and accumulation mechanisms of lycorine have rarely been studied. In this study, an ATP-binding cassette (ABC) transporter from Lycoris aurea (L’Hér) Herb., namely LaABCB11, was cloned and functionally characterized. Heterologous expression showed that LaABCB11 transported lycorine in an outward direction, increased the tolerance of yeast cells to lycorine, and caused a lower lycorine accumulation in transformants than control or mutant in yeast. LaABCB11 is associated with the plasma membrane, and in situ hybridization indicated that LaABCB11 was mainly expressed in the phloem of leaves and bulbs, as well as in the cortical cells of roots. These findings suggest that LaABCB11 functions as a lycorine transport and it might be related to the translocation and accumulation of lycorine from the leaves and bulbs to the roots.

Introduction: Changnienia amoena is an endangered orchid endemic to China, with notable medicinal and ornamental significance. Like other orchids, it relies heavily on microbial symbionts for germination, growth, and survival; however, its associated microbiomes remain poorly understood. Methods: This study employed high-throughput amplicon sequencing targeting bacterial 16S rRNA and fungal ITS regions and provided an in-depth characterization of the root-associated microbiomes of C. amoena across its native range. Results: Both bacterial and fungal communities exhibited significant biogeographic patterns, but only fungal Shannon-Wiener diversity showed a decreasing trend with increasing latitude. The neutral model revealed predominantly stochastic assembly of bacterial communities. In contrast, fungal communities exhibited a lower estimated migration rate, indicating a relatively stronger influence of non-neutral processes on their assembly. Soil nutrients emerged as key environmental drivers shaping microbial communities. Functional predictions highlighted multiple bacterial functions related to nitrogen cycling and the enrichment of mycorrhizal fungi, suggesting the functional potential of root-associated microbes in promoting plant nutrient acquisition, growth, and environmental adaptation. Co-occurrence network analysis further revealed positive associations between mycorrhizal fungi and potentially beneficial bacteria based on taxonomic inference. Discussion: This study reveals distinct biogeographic patterns, assembly processes, and functional potentials of root-associated microbiomes of C. amoena , which not only expands the understanding of the plant–soil-microbe interactions, but also provides crucial insights for manipulating them for the conservation of this endangered orchid.

Introduction: Parrotia subaequalis , an endangered Tertiary relict species endemic to eastern China, holds significant values for both evolutionary ecology and ornamental horticulture. However, it is increasingly threatened by leaf spot disease. The causal pathogens and the potential role of bacterial endophytes in disease suppression remain uncharacterized. Methods: In this study, fungal pathogens were isolated from diseased leaves collected from native habitats and their pathogenicity was confirmed by Koch’s postulates. Simultaneously, bacterial endophytes were isolated from the non-lesioned portions of diseased leaves and screened for their antagonistic activity. Additionally, the antifungal potential of bacterial secondary metabolites was evaluated and analyzed using untargeted metabolomic profiling. Results: Four fungal strains, Alternaria sp. S1, Schizophyllum sp. S2, Diaporthe sp. S5, and Botryosphaeria sp. S6, were confirmed as causal agents of leaf spot disease, with S5 causing the most severe symptoms. A total of 206 bacterial endophytes were isolated and 25 strains exhibited strong inhibition against fungal pathogens. Among them, Bacillus species showed the strongest antagonistic effects. Additionally, there were 103 bacterial metabolites identified. Seven abundant metabolites were tested for their antifungal activity, revealing that dihydrojasmone (DJ) and δ-tridecalactone (DT) significantly inhibited fungal growth. DJ displayed broad-spectrum inhibition, suppressed fungal melanin synthesis, and induced hyphal deformation, while DT exerted weaker but significant effects against S2 and S6. Discussion: This study provides the first identification of fungal pathogens responsible for leaf spot disease in P. subaequalis . The strong antifungal activities of endophytic Bacillus species and their metabolites were demonstrated, highlighting their promise as eco-friendly biocontrol agents.