Bulbophyllum is one of the largest genera and presents some of the most intricate taxonomic problems in the family Orchidaceae, including species of ornamental and medical importance. The lack of knowledge regarding the characterization of Bulbophyllum chloroplast (cp) genomes has imposed current limitations on our study. Here, we report the complete cp genomes of seven Bulbophyllum species, including B. ambrosia, B. crassipes, B. farreri, B. hamatum, B. shanicum, B. triste, and B. violaceolabellum, and compared with related taxa to provide a better understanding of their genomic information on taxonomy and phylogeny. A total of 28 Bulbophyllum cp genomes exhibit typical quadripartite structures with lengths ranging from 145,092 bp to 165,812 bp and a GC content of 36.60% to 38.04%. Each genome contained 125–132 genes, encompassing 74–86 protein-coding genes, 38 tRNA genes, and eight rRNA genes. The genome arrangements, gene contents, and length were similar, with differences observed in ndh gene composition. It is worth noting that there were exogenous fragment insertions in the IR regions of B. crassipes. A total of 18–49 long repeats and 38–80 simple sequence repeats (SSRs) were detected and the single nucleotide (A/T) was dominant in Bulbophyllum cp genomes, with an obvious A/T preference. An analysis of relative synonymous codon usage (RSCU) revealed that leucine (Leu) was the most frequently used codon, while cysteine (Cys) was the least used. Six highly variable regions (rpl32-trnL^UAG > trnT^UGU-trnL^UAA > trnF^GAA-ndhJ > rps15-ycf1 > rbcL-accD > psbI-trnS^GCU) and five coding sequences (ycf1 > rps12 > matK > psbK > rps15) were identified as potential DNA markers based on nucleotide diversity. Additionally, 31,641 molecular diagnostic characters (MDCs) were identified in complete cp genomes. A phylogenetic analysis based on the complete cp genome sequences and 68 protein-coding genes strongly supported that 28 Bulbophyllum species can be divided into four branches, sects. Brachyantha, Cirrhopetalum, and Leopardinae, defined by morphology, were non-monophyly. Our results enriched the genetic resources of Bulbophyllum, providing valuable information to illustrate the complicated taxonomy, phylogeny, and evolution process of the genus.

The TIFY gene family (formerly known as the zinc finger proteins expressed in inflorescence meristem (ZIM) family) not only functions in plant defense responses but also are widely involved in regulating plant growth and development. However, the identification and functional analysis of TIFY proteins remain unexplored in Orchidaceae. Here, we identified 19 putative TIFY genes in the Phalaenopsis aphrodite genome. The phylogenetic tree classified them into four subfamilies: 14 members from JAZ, 3 members from ZML, and 1 each from PPD and TIFY. Sequence analysis revealed that all Phalaenopsis TIFY proteins contained a TIFY domain. Exon–intron analysis showed that the intron number and length of Phalaenopsis TIFY genes varied, whereas the same subfamily and subgroup genes had similar exon or intron numbers and distributions. The most abundant cis-elements in the promoter regions of the 19 TIFY genes were associated with light responsiveness, followed by MeJA and ABA, indicating their potential regulation by light and phytohormones. The 13 candidate TIFY genes screened from the transcriptome data exhibited two types of expression trends, suggesting their different roles in cell proliferation and cell expansion of floral organ growth during Phalaenopsis flower opening. Overall, this study serves as a background for investigating the underlying roles of TIFY genes in floral organ growth in Phalaenopsis.

Fusarium wilt of Phalaenopsis spp., caused by Fusarium species, is a major soil-borne disease that severely threatens the yield and ornamental quality of Phalaenopsis. Endophytic fungi can serve as natural biocontrol agents against soil-borne pathogens and have potential applications in biological control. In this study, a pathogenic isolate, HT-1, was obtained from Fusarium-wilted Phalaenopsis plants, and its pathogenicity was confirmed through inoculation assays on detached leaves and intact plants. Morphological observations, together with internal transcribed spacer (ITS) and translation elongation factor 1-α (TEF-1α) sequence analyses, identified HT-1 as Fusarium proliferatum. Seven endophytic fungi with strong antagonism against F. proliferatum were screened using a dual-culture assay. All seven isolates consistently inhibited five representative plant pathogens in vitro, showing broad-spectrum and promising biocontrol potential. Detached leaf inoculation assays showed that both Trichoderma virens and Trichoderma asperellum significantly reduced lesion area and disease severity caused by F. proliferatum, indicating potential application in disease management. In summary, this study identified seven endophytic fungi from Phalaenopsis with broad-spectrum antagonistic potential against multiple plant pathogenic fungi. Meanwhile, the protective effects of T. virens and T. asperellum against F. proliferatum were preliminarily evaluated using detached leaves. These findings provide theoretical reference and fungal resources for screening candidate biocontrol strains against Phalaenopsis Fusarium wilt and related plant diseases, and lay a foundation for future studies on biocontrol mechanisms, whole-plant efficacy verification, and practical application.

Phalaenopsis is a widely cultivated ornamental plant of considerable economic value worldwide. However, traditional growing medium, sphagnum moss, is limited and non-renewable. It also decomposes slowly and is prone to environmental issues. Therefore, there is an urgent need to identify more environmentally friendly and efficient alternatives. Biochar, a sustainable material with excellent physical and chemical properties, has been recognized as an effective promoter of plant growth. In this study, we investigated the influence of biochar derived from three raw materials (corn straw, bamboo, and walnut) mixed1 with coconut shell at ratios of 1:2, 1:10, and 4:1, on the growth of Phalaenopsis ‘Big Chili’. Over a 150-day controlled experiment, we evaluated multiple growth parameters, including plant height, crown width, total root length, total projected area, total surface area, and root volume. Compared to the traditional growing medium, the optimal biochar-coconut shell mixture (maize straw biochar: coconut shell = 1:2) increased plant height and crown width by 7.55% and 6.68%, respectively. Root metrics improved substantially, with total root length increasing by 10.96%, total projected area by 22.82%, total surface area by 22.14%, and root volume by 38.49%. Root biomass in the optimal treatment group increased by 42.47%, while aboveground and belowground dry weights increased by 6.16% and 77.11%, respectively. These improvements were closely associated with favorable substrate characteristics, including low bulk density, high total and water-holding porosity, moderate aeration, and adequate nutrient availability. These findings demonstrate that substrate characteristics critically influence plant performance and that biochar–coconut shell mixtures, particularly at a 1:2 ratio, represent a viable and sustainable alternative to sphagnum moss for commercial cultivation of Phalaenopsis.

Urban streets are important heat-exposure environments, yet the relationships of two-dimensional (2D) planar plant morphology and three-dimensional (3D) vegetation structure with land surface temperature (LST) remain insufficiently integrated at a continuous street scale. We analyzed 42,603 street-scale study units in the central urban area of Wuhan, defined at 50 m sampling intervals with a 150 m radius. The 2D variables comprised green-space area (A), mean patch perimeter (P_mean), and perimeter–area ratio (P_A), while the 3D variables comprised green view index (GVI), mean 3D green volume (NV_mean), mean canopy height (CH_mean), and canopy-height variability (CH_sd). Anselin Local Moran’s I identified High–High (HH) and Low–Low (LL) zones; linear regression (LR), random forest (RF), and SHAP characterized linear, nonlinear, and model-based contributions; and buffered spatial cross-validation and spatial resampling evaluated robustness. Under the original random 80–20% train–test split, LR/RF R² values were 0.2153/0.4002 for the overall study area, 0.1940/0.3539 for the HH zone, and 0.0488/0.4853 for the LL zone. Under five-fold buffered spatial cross-validation, the corresponding pooled out-of-fold R² values were 0.1940/0.2237, 0.1427/0.0965, and −0.0942/−0.0231, showing that the RF advantage weakened after spatial separation and did not persist in the HH and LL zones. In the original fitted RF models, A, P_mean, and NV_mean had the largest mean absolute SHAP contributions overall; A, P_A, and P_mean ranked highest in the HH zone; and GVI, CH_sd, and CH_mean ranked highest in the LL zone. Repeated buffered spatial validation showed no stable overall 2D predominance because the median 2D share of 52.1% had a 46.5–58.8% percentile range, but it supported stable 2D relative predominance in the HH zone (61.0% [55.5–66.7%]) and stable grouped 3D relative predominance in the LL zone (61.9% [52.3–70.3%]), despite unstable LL variable-level rankings. SHAP relationships were nonlinear and zone-dependent: A and P_mean showed clearer directional transitions overall and in the HH zone, whereas the LL zone and most 3D variables exhibited multiple directional changes. Spatial-block bootstrap analysis examined 40 full-sample zero-crossing candidates, of which 39 met the predefined stability criteria; these ranges represent model-derived directional transitions rather than ecological thresholds or causal planning standards. The findings demonstrate thermal-context-dependent, model-based associations between 2D and 3D plant morphology and street-scale LST, while emphasizing that model performance and some importance rankings are spatially sensitive and require local validation.

Bulbophyllum is one of the largest genera and presents some of the most intricate taxonomic problems in the family Orchidaceae, including species of ornamental and medical importance. The lack of knowledge regarding the characterization of Bulbophyllum chloroplast (cp) genomes has imposed current limitations on our study. Here, we reported the complete cp genomes of seven Bulbophyllum species, including B. ambrosia, B. crassipes, B. farreri, B. hamatum, B. shanicum, B. triste and B. violaceolabellum, and compared with related taxa to provide a better understanding of their genomic information on taxonomy and phylogeny. A total of 28 Bulbophyllum cp genomes exhibit typical quadripartite structures with lengths ranging from 145,092 bp to 165,812 bp and GC content of 36.60% to 38.04%. Each genome contained 125–132 genes, encompassing 74–86 protein-coding genes, 38 tRNA genes, and eight rRNA genes. The genome arrangements, gene contents and length were similar with differences observed in ndh gene composition. A total of 18–49 long repeats and 38–80 simple sequence repeats (SSRs) were detected and the single-nucleotide (A/T) was dominant in Bulbophyllum cp genomes, with an obvious A/T preference. An analysis of relative synonymous codon usage (RSCU) revealed Leucine (Leu) was the most frequently used codon, while cysteine (Cys) was the least used. Six highly variable regions including ndhF-trnLUAG, trnTUGU-trnLUAA, trnFGAA-ndhJ, rps15-trnNGUU, rbcL-accD and psbI-trnSGCU were identified based on the ranking of the Pi values, had the potential to serve as DNA markers for species identification and phylogeny of the genus Bulbophyllum. Phylogenetic analysis based on the complete cp genome sequences and 68 protein-coding genes strongly supported 28 Bulbophyllum species can be divided into four branches and sects. Brachyantha, Cirrhopetalum, Leopardinae defined by morphology were non-monophyly. Our results enriched the genetic resources of Bulbophyllum species, providing valuable information to illustrate the complicated taxonomy, phylogeny and evolution process of the genus.