Heat Stress Alleviation by Exogenous Calcium in the Orchid Dendrobium nobile Lindl: A Biochemical and Transcriptomic Analysis.

The growth of Dendrobium nobile is sensitive to heat stress. To find an effective method for enhancing heat tolerance, this study investigated the relieving effect of exogenous calcium at different concentrations (0 mmol/L, 5 mmol/L, 10 mmol/L, 15 mmol/L, 20 mmol/L CaCl2) on heat stress in D. nobile. Principal component analysis was used to screen the optimal exogenous calcium concentration, and transcriptome analysis was used to reveal its possible heat tolerance mechanism. The results showed that compared with the T0, a 10 mmol/L calcium treatment: increased the average leaf length, leaf width, plant height, and fresh matter accumulation of D. nobile by 76%, 103.39%, 12.97%, and 12.24%, respectively (p < 0.05); significantly increased chlorophyll a (Chla), chlorophyll b (Chlb), carotenoids(Car), ascorbic acid (ASA), glutathione (GSH), and flavonoids by 15.72%, 8.54%, 11.88%, 52.17%, 31.54%, and 36.12%, respectively; and effectively enhanced the enzyme activity of the antioxidant system, increasing superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) by 1.38, 1.61, and 2.16 times, respectively (p < 0.05); At the same time, the treatment can effectively reduce the yellow leaf rate and defoliation rate of D. nobile under heat stress. The principal component analysis method and membership function were used to calculate the D value to rank the relief effects of each calcium treatment group, and the results also showed that 10 mmol/L CaCl2 had the best relief effect. Transcriptomics testing identified 7013 differentially expressed genes, of which 2719 were upregulated, and 294 were downregulated. Among them, genes such as HSPA1s, HSP90A, HSPBP1, ATG8, COMT, REF1, E1.11.1.7, along with transcription factors such as MYB, bHLH, WRKY, and NAC, formed the network of tolerance to heat stress in D. nobile. This study provides new insights for improving the cultivation techniques of D. nobile.

Transcriptional and Physiological Analysis Reveal New Insights into the Regulation of Fertilization (N, P, K) on the Growth and Synthesis of Medicinal Components of Dendrobium denneanum.

Dendrobium denneanum is an important medicinal and ornamental plant. Its ornamental and medicinal values are affected by its vegetative growth conditions and chemical composition accumulation. This study adopted an orthogonal experimental design to treat D. denneanum with nine different levels of nitrogen (N), potassium (K), and phosphorus (P). The morphological indicators of the plant were positively correlated with the nitrogen concentration. The polysaccharide content was the highest at 1500 mg·L-1 nitrogen and 3000 mg·L-1 phosphorous and was 26.84% greater than the control. The flavonoid content increased by 36.2% at 500 mg·L-1 nitrogen, 2000 mg·L-1 phosphorous, and 300 mg·L-1 potassium. Principal component score analysis showed that nitrogen had the most significant impact on the various indicators of D. denneanum, followed by phosphorus and potassium. The comprehensive score showed that the T9 treatment (N: 1500 mg·L-1, P: 3000 mg·L-1, K: 500 mg·L-1) had the strongest effect on D. denneanum. Transcriptional analysis showed that compared with the control, the T9 treatment led to 2277 differentially expressed genes (1230 upregulated and 1047 downregulated). This includes fifteen genes enriched in the MAPK signaling pathway, five genes in phenylpropanoid biosynthesis, and two genes in flavonoid biosynthesis. These genes may be involved in regulating plant growth and the biosynthesis of polysaccharides and flavonoids. This study provides guidance for the optimal use of N, P, and K in the cultivation of D. denneanum.

Isolation, Structural Characteristics Analysis of a Vigna unguiculata Polysaccharide VUP80-3 and Its Protective Effect on GES-1 Cells In Vitro.

Cowpea (Vigna unguiculata) is one of the main edible legume vegetables in China, and it can improve spleen and stomach function. A polysaccharide component (VUP80-3) has been isolated from V. unguiculata in this study. The average molecular weight of VUP80-3 is 6.43 × 104 Da, and the main monosaccharide group is glucose. The mass ratio of monosaccharide groups in the polysaccharide was glucose:galactose:arabinose:rhamnose:xylose:mannose:fucose = 152.36:24.50:16.53:8.13:1.26:0.97:0.82. NMR analysis showed that VUP80-3 has →4)-α-D-Galp (1→ and →4)-α-D-Glcp(1→ main chain and →3,4)-ß-D-Glcp(1→, →4,6)-α-D-Glcp(1→ branch chains, and the terminal sugar is α-D-Glcp(1→. Biological activity test results showed that VUP80-3 at 1000 µg·mL-1 significantly increased the activity of ethanol injured GES-1 cells (p < 0.01) and significantly reduced reactive oxygen species (ROS) in ethanol injured GES-1 cells and inflammatory factors (IL-8, IL-1ß and TNF-α,) in GES-1 cells. This compound also reduced the apoptosis rate (p < 0.05), thereby significantly reducing the oxidative damage caused by ethanol in GES-1 cells. Therefore, VUP80-3 is a potential drug to protect the gastric mucosa from damage.

Transcriptomics combined with physiological analysis provided new insights into the Zn enrichment capacity and tolerance mechanism of Dendrobium denneanum Kerr.

In this study, we investigated the tolerance and accumulation capacity of Dendrobium denneanum Kerr (D.denneanum) by analyzing the growth and physiological changes of D.denneanum under different levels of Zn treatments, and further transcriptome sequencing of D.denneanum leaves to screen and analyze the differentially expressed genes. The results showed that Zn400 treatment (400 mg·kg-1) promoted the growth of D.denneanum while both Zn800 (800 mg·kg-1) and Zn1600 treatment (1600 mg·kg-1) caused stress to D.denneanum. Under Zn800 treatment (800 mg·kg-1), the resistance contribution of physiological indexes was the most obvious: antioxidant system, photosynthetic pigment, osmoregulation, phytochelatins, and ASA-GSH cycle (Ascorbic acid-Glutathione cycle). D.denneanum leaves stored the most Zn, followed by stems and roots. The BCF(Bioconcentration Factor) of the D.denneanum for Zn were all more than 1.0 under different Zn treatments, with the largest BCF (1.73) for Zn400. The transcriptome revealed that there were 1500 differentially expressed genes between Zn800 treatment and group CK, of which 842 genes were up-regulated and 658 genes were down-regulated. The genes such as C4H, PAL, JAZ, MYC2, PP2A, GS, and GST were significantly induced under the Zn treatments. The differentially expressed genes were associated with phenylpropane biosynthesis, phytohormone signaling, and glutathione metabolism. There were three main pathways of response to Zn stress in Dendrobium: antioxidant action, compartmentalization, and cellular chelation. This study provides new insights into the response mechanisms of D.denneanum to Zn stress and helps to evaluate the phytoremediation potential of D.denneanum in Zn-contaminated soils.

Transcriptome analysis reveals the role of polysaccharide biosynthesis in the detoxification of Dendrobium nobile under zinc stress.

Plants can bind excessive heavy metals by synthesizing compounds to alleviate the harm caused by heavy metals. To reveal the mechanism by which Dendrobium nobile alleviates zinc stress, metabolome combined transcriptome analysis was used in this research. The results showed that zinc was mainly enriched in the roots and leaves and the biomass of the roots and leaves of D. nobile decreased significantly by 18.21 % and 49.22 % (P < 0.05) compared to the control (CK), respectively. Meanwhile, the contents of nonprotein thiol(NPT), glutathione(GSH), and phytochelatins (PCs) in the roots were significantly increased by 48.8 %, 78.3 %, and 45.4 % compared to CK, respectively. Through TEM testing, it was found that D. nobile exhibited toxic symptoms. Metabolome analysis showed that the metabolites of D. nobile under zinc stress were mainly enriched in biosynthesis of other secondary metabolites and carbohydrate metabolism. Nova-seq results identified 1202 differentially expressed genes(DEGs), of which 603 were upregulated and 599 were downregulated. Through GO and KEGG annotation analysis of these DEGs, it was found that PMR6 and PECS-2.1, SS1 and GLU3 genes were significantly upregulated, leading to an increase in the biosynthesis of xylan, pectin, starch and other polysaccharides in D. nobile. These polysaccharides can form a "Polysaccharide-Zn" with excess zinc. Meanwhile, the GSTs in glutathione metabolism were significantly upregulated, leading to a significant increase in the content of NPT, GSH, and PCs. These zinc complexes were transported to vacuoles through ABC transporters for compartmentalization, effectively alleviating the damage of zinc. The results can provide new insights for phytoremediation and quality assurance of medicinal D. nobile.

Cerium improves the physiology and medicinal components of Dendrobium nobile Lindl. under copper stress.

Heavy metal stress affects the quality of medicinal plants, and rare earth elements can effectively alleviate heavy metal stress. In this paper, we investigated the effects of rare earth element cerium (0, 5, 10, 20, 40, 80, and 160 mg/L) on the physiological and medicinal components of Dendrobium nobile Lindl. under copper (200 mg/L) stress. The results revealed that cerium (Ce) had a good alleviating effect on copper (Cu) stress, low concentrations of Ce (10-20 mg/L) significantly improved the resistance and medicinal qualities of the plant such as polysaccharide, polyphenol and flavonoid, it also increased the content of photosynthetic pigment, proline, soluble sugar and soluble protein of D. nobile Lindl., effectively balance the osmotic pressure and the generation and removal of reactive oxygen species in the plant, thereby the toxic effect of copper on D. nobile Lindl. is alleviated. From the point of view of the treatment time when the optimal relieving concentration appeared, the optimal concentration for relieving antioxidant enzyme activity all appeared at the treatment time of 10 d, the optimum concentrations of other indicators all appeared at the treatment time of 15 d. Overall, this study suggests that the optimum level of Ce (10-20 mg/L) might be promising for alleviating the adverse impacts of copper stress and promoting the accumulation of medicinal components in D. nobile Lindl.