Genome-Wide Identification, Expression Analysis and Functional Study of DELLA Genes in Chinese Cabbage (Brassica rapa L. ssp. pekinensis).

BACKGROUND: DELLA protein is a crucial factor which played pivotal roles in regulating numerous intriguing biological processes in plant development and abiotic stress responses. However, little is known about the function and information of DELLA protein in Chinese cabbage. METHODS: Using 5 DELLA gene sequences in Arabidopsis Thaliana as probes, 5 DELLA genes in Chinese cabbage were identified by Blast search in Chinese cabbage database (Brassica database (BRAD)). The National Center for Biotechnology Information (NCBI), ExPaSy, SWISS-MODEL, DNAMAN, MEGA 11, PlantCARE were used to identify and analyze the DELLA gene family of Chinese cabbage. Gene expression was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). The function of BraA10gRGL3 was verified by overexpression and phenotypic analysis of BraA10gRGL3 and yeast hybrid. RESULTS: In this study, 5 BraDELLAs homologous to Arabidopsis thaliana were identified and cloned based on the Brassica database, namely, BraA02gRGL1, BraA05gRGL2, BraA10gRGL3, BraA06gRGA and BraA09gRGA. All BraDELLAs contain the DELLA, TVHYNP, and GRAS conserved domains. Cis-element analysis revealed that the promoter regions of these 5 DELLA genes all contain light-responsive elements, TCT motif, I-box, G-box, and box 4, which are associated with GA signaling. Transcriptome analysis results proved that the expression of BraA02gRGL1, BraA05gRGL2, and BraA10gRGL3 in Y2 at different growth stages were lower than them in Y7, which is consistent with the phenotype that Y7 exhibited stronger stress tolerance than Y2. It is worth emphasizing that even through the overexpression of BraA10gRGL3-Y7 in Arabidopsis resulted in smaller leaf size and lower fresh weight compared to the wild type (WT) Arabidopsis: Columbia, a stronger response to abiotic stresses was observed in BraA10gRGL3-Y7. It indicated that BraA10gRGL3-Y7 can improve the stress resistance of plants by inhibiting their growth. Moreover, the yeast two-hybrid experiment confirmed that BraA10gRGL3-Y7 can interacted with BraA05gGID1a-Y7, BraA04gGID1b1, BraA09gGID1b3-Y2, and BraA06gGID1c, whereas BraA10gRGL3-Y2 cannot interact with any BraGID1. CONCLUSIONS: Collectively, BraDELLAs play important role in plant development and response to abiotic stress. The differences in amino acid sequences between BraA10gRGL3-Y2 and BraA10gRGL3-Y7 may result in variations in their protein binding sites, thus affecting their interaction with the BraGID1 family proteins. This systematic analysis lays the foundation for further study of the functional characteristics of DELLA genes of Chinese cabbage.

Promoting effect of low concentration strontium on photosynthetic performance of Chinese cabbage seedlings: Combined leaf characteristics, photosynthetic carbon assimilation and chlorophyll fluorescence.

Low concentration strontium (LC-Sr) can promote the growth of plants. In order to explore its promoting mechanism from the aspect of photosynthesis, the leaf characteristics, CO2 assimilation and chlorophyll (Chl) a fluorescence kinetics were investigated with hydroponically LC-Sr-treated Chinese cabbage seedlings. After a 28-d treatment to SrCl2 at different concentrations (0.1, 0.2, 0.5, and 1.0 mmol L-1), we observed an increase in the specific leaf weight (SLW) of Chinese cabbage compared with the control group. Notably, as the strontium concentration increased, a more pronounced improvement trend in the contents of Chl and protein in the leaves was observed, contributing to the enhancement of photosynthesis. However, the statistical differences in Pn among various LC-Sr treatments were not significant. Nevertheless, the leaf starch content exhibited a significant increase after LC-Sr treatments. Additionally, Chl a fluorescence transient has been used as a sensitive indicator of the promotional effect of LC-Sr on photosynthesis. The results of fluorescence parameters showed that LC-Sr treatments accelerated the light reaction speed of leaves (Tfm, dV/dto, dVG/dto), improved the energy utilization efficiency of photosystem (PSI and PSII) (ETo/CSo, ψET,ψRE, δRo, φRo), and ultimately enhanced the photosynthetic performance of leaves (PIabs, SFIabs, DFabs). The increased RCs/CSo and Sm contributed to the enhancement of the light reaction activity of strontium-treated leaves. The LC-Sr treatments had no interference with the calcium absorption, and notably enhanced the photosynthetic capacity of Chinese cabbage, shedding light on potential benefits of LC-Sr for crop cultivation.

Promotion of osteogenesis by bioactive glass-ceramic coating: Possible involvement of the Hedgehog signaling pathway.

PURPOSE: Bioactive glass-ceramic (BGC) coatings have been extensively studied and clinically used as bone substitute materials because of their osteogenesis, osteoinduction, and osteoconduction characteristics. Although the Hedgehog (Hh) signaling pathway plays an important role in skeletal development, the relationship between BGC coatings and the Hh signaling pathway is unknown. METHODS: In this study, a BGC coating is fabricated by furnace sintering, and its surface is investigated by a scanning electron microscope (SEM) and a transmission electron microscope (TEM). Furthermore, the expression of Ki67 is evaluated using immunofluorescence, and osteogenesis-related factors and Hh signaling pathway molecules on the BGC coating are examined by real-time reverse transcription polymerase chain reaction (real-time RT-PCR) and Western blotting in bone marrow mesenchymal stem cells (BMSCs). RESULTS: The SEM and the TEM show that the BGC coating surface is smooth, without cracks, and composed of particles with mesoporous structure. The expression of Ki67 positive BMSCs of the BGC group is higher than that of the control group. Real-time RT-PCR and Western blotting assay reveal that the expression levels of osteoblast-related genes (BMP2, Osteocalcin, ALP, Runx2) and Hh signaling pathway molecules (Gli1, Smo) are much higher for the BGC coating group than those for the control group. Furthermore, after treating with Smo inhibitor cyclopamine, the Smo and Gli1 expressions in BMSCs are dramatically down-regulation for the BGC coating compared to those for the control group. Both mRNA and protein expression levels of osteogenesis-related factors was downregulated after treating Smo inhibitor cyclopamine in BMSCs with the BGC coating. CONCLUSIONS: The BGC coatings promote osteogenesis probably via the Hh signaling pathway, which provides a theory reference for future clinical application of bone formation.