Light intensity strongly influences the morphological development and photoprotective responses of in vitro plantlets, yet the optimal conditions for hydrangea remain undefined. This study investigated the effects of five light intensity gradients (TrA: 80–120 lux, TrB: 380–480 lux, TrC: 1500–1800 lux, TrD: 3800–4000 lux, TrE: 6000–6400 lux) on Hydrangea macrophylla ‘Qingtian’ plantlets. Plantlets exhibited optimal growth at TrB, showing maximal biomass, leaf expansion, chlorophyll content, and root activity, accompanied by low antioxidant enzyme activities and soluble sugar levels. Nutrient accumulation was greater under low light than under high light conditions. Transcriptome analysis of treatments (TrB and TrE) with marked phenotypic differences revealed 7119 differentially expressed genes (DEGs). Of these, 4582 genes were up-regulated and 2537 were down-regulated. The up-regulated genes were significantly enriched in pathways related to cell walls, the microtubule cytoskeleton, and developmental processes, which are involved in the plant growth and development process, such as photosynthesis, nutrient ion transport and regulation, as well as plant hormone responses and transport; whereas the down-regulated genes were significantly enriched in pathways related to carbohydrate metabolism, oxidoreductase activity, and glutathione metabolism, suggesting that high light stress impairs growth by disrupting carbon and antioxidant processes. These results demonstrated that 380–480 lux is the optimal light intensity for ‘Qingtian’ Hydrangea macrophylla in vitro plantlets. This study provides a foundation for optimizing culture conditions and offers new insights into the molecular regulation of light-responsive genes.

The floricultural value of Hydrangea macrophylla is significantly influenced by its distinctive blue flower coloration, which results from aluminum uptake and vacuolar complexation. However, commercial cultivation faces challenges in achieving consistent bluing while avoiding Al toxicity. This study investigated the effects of Al concentration (0–971.62 mg/L) and application period on six blue-modifiable cultivars. Optimal results were achieved with an Al₂(SO₄)₃·18H₂O concentration of 6 g/L applied from two weeks after pinching until bloom. This regimen successfully induced vibrant sepal bluing without impairing chlorophyll content or plant growth. Furthermore, it enhanced tissue concentration of essential nutrients, including N, K, Mg, Fe, Zn, Mn, Cu, B, and Co. This study proposes a robust technical and theoretical framework for optimizing Al application to ensure consistent sepal color, which provides technical support for commercial production.

Background: Upland cotton (Gossypium hirsutum) is a major natural fiber crop and an important model for studying genome evolution and gene function in polyploid plants. However, its large and highly redundant genome presents substantial challenges for efficient and coordinated multiplex genome editing. Methods: Here, we developed a high-efficiency CRISPR/Cas12a-based multiplex genome editing system in cotton by integrating a tRNA–crRNA polycistronic expression strategy with a Bean yellow dwarf virus (BeYDV)-derived replicon. Results: This platform enabled coordinated expression of multiple crRNAs and simultaneous targeting of 16 loci within a centromere-proximal region of chromosome D03 (18.65–24.47 Mb). In individual transgenic lines, up to 10 target sites were edited concurrently, with nine targets exhibiting editing efficiencies above 56% and the highest efficiency reaching 96.46%. High-density multiplex editing predominantly induced small insertions and deletions at target loci. Notably, edited plants exhibited reduced growth and pronounced cytological abnormalities, including chromosome bridges, lagging chromosomes, and abnormal meiotic products. Transcriptome analysis revealed widespread dysregulation of genes involved in chromosome segregation and cell cycle regulation. Despite these functional perturbations, HiFi long-read sequencing detected no large-scale chromosomal rearrangements, indicating that genome instability arises from cumulative local perturbations rather than global structural alterations. Conclusions: Together, our results establish an efficient multiplex genome editing platform in cotton and highlight potential constraints of high-density editing on genome stability in complex plant genomes.