Identification of valid reference genes for quantitative RT-PCR in Caragana microphylla under salt and drought stresses.

Caragana microphylla is a leguminosae plant and grows mainly in semi-arid areas of northwest China and Mongolia. However, the lack of studies on C. microphylla reference genes limits the accurate understanding of the molecular biology mechanisms in this crop under abiotic stresses. In this study, we selected nine candidate genes from salt-treated C. microphylla transcriptome data and evaluated their stability by using geNorm, NormFinder, BestKeeper, and RefFinder in salt and drought conditions. In addition, the relative expressions of Delta 1-pyrroline-5-carboxylate synthase 2 (P5CS2) and Catalase 2 (CAT2) were examined to confirm the stability of the candidate reference genes. As a results, glyceraldehyde-3-phosphate dehydrogenase C2 (GAPC2) and 26S proteasome regulatory subunit (RPN5) were the most stable in both salt and drought treatments. The relative expression of P5CS2 and CAT2 also showed more stable levels in normalization by GAPC2 and RPN5 than the most unstable gene, Ubiquitin 4 (UBQ4). Therefore, it is believed that these candidate reference genes selected and validated in our study could be used to study the molecular biological study of response to salt and drought stress in C. microphylla.

Seedling growth and photosynthetic response of Pterocarpus indicus L. to shading stress.

In tropical forests, the shade provided by tree canopies and extreme climate causes inhibition of plant seedling growth due to the lack of light. However, the plants can acclimate to such environmental stress by generating specific responses. The present study aimed to investigate the effects of shading conditions on ecophysiological performance of Narra seedlings (Pterocarpus indicus L.) via a mesocosm experiment. A pot experiment was conducted for 20 weeks in a greenhouse with different shading treatments, 75% (control), 25%, and 4% of full sunlight (FS). As a result, the photosynthetic rate (PN), Rubisco enzyme activity, maximum carboxylation rate (VCmax), and maximum electron transport rate (Jmax) in 25% FS treatment were higher or similar to those in control after three weeks of the beginning of shade treatment, whereas the highest values after ten weeks were observed in control. In contrast, the photosynthetic pigments were highest in control after three weeks, while the values were highest in 25% FS treatment after ten weeks. The growth parameters, such as biomass and leaf area, were highest in 75% FS treatment. The expression of Rubisco, phosphoglycerate kinase, glyceraldehyde-3-phosphate dehydrogenase, and fructose-1,6-bisphosphatase were up-regulated in 4% FS treatment compared to control after ten weeks, contributing to tolerating the shade stress. Our findings indicated the capacity of P. indicus seedlings to tolerate and acclimate low light conditions causing shade stress by generating specific physiological and morphological responses, especially Rubisco enzyme activity as well as gene expression related to photosynthetic activity. The present study will improve our understanding of the tolerance mechanism of Narra plant under light-deficient conditions, thereby providing a better strategy for efficiently growing seedlings of this species in tropical rainforests.

Saponarin, a Di-glycosyl Flavone from Barley (Hordeum vulgare L.): An Effective Compound for Plant Defense and Therapeutic Application.

Saponarin (SA) is a major di-C-glycosyl-O-glycosyl flavone, which is predominantly accumulated in the young green leaves of barley (Hordeum vulgare L.), with numerous biological functions in plants, such as protection against environmental stresses. Generally, SA synthesis and its localization in the mesophyll vacuole or leaf epidermis are largely stimulated in response to biotic and abiotic stresses to participate in a plant's defense response. In addition, SA is also credited for its pharmacological properties, such as the regulation of signaling pathways associated with antioxidant and anti-inflammatory responses. In recent years, many researchers have shown the potential of SA to treat oxidative and inflammatory disorders, such as in protection against liver diseases, and reducing blood glucose, along with antiobesity effects. This review aims to highlight natural variations of SA in plants, biosynthesis pathway, and SA's role in response to environmental stress and implications in various therapeutic applications. In addition, we also discuss the challenges and knowledge gaps concerning SA use and commercialization.

Variation of Saponarin Content in Barley Sprouts (Hordeum vulgare L.) by Natural Light Shielding: Implication of the Importance of Light Intensity.

Saponarin is a functional metabolite produced by barley sprouts, and the mass production of saponarin by this crop is attractive for dietary supplement manufacturing. Light is the most important environmental factor determining plant growth, survival, and the production of secondary metabolites including flavonoids. This study was conducted to investigate the importance of light intensity for saponarin production in barley sprouts using a hydroponic growth system. Light intensity was manipulated by using shielding treatments to 100, 80, 70, and 50% natural sunlight (NS), and crop cultivation was performed on a monthly cycle. We found that the growth rate and biomass of barley sprouts did not differ in response to the shield treatments, whereas the saponarin content did. The highest saponarin content (i.e., from 1329 to 1673 mg 100 g-1) was observed in the 100% NS treatment, and it gradually decreased as light intensity also decreased. Statistical analysis revealed a significant polynomial relationship of saponarin content with cumulative PPFD (R2 = 76%), implying that the absolute total amount of light exposure over the growth period has a large effect on saponarin productivity in a hydroponic facility. Taken together, our results showed that shielding conditions, which are often unintentionally created by the design of cultivation facilities, can adversely affect saponarin production in barley sprouts. In addition, it was confirmed through our findings that light conditions with at least 70% NS in the cultivation facility enable the production of an amount corresponding to the saponarin content of the sprouts (>1000 mg 100 g-1) produced in the open field. Further studies are needed to investigate the underlying physiological and molecular mechanisms responsible for the relationship of saponarin content with light quantity and quality in barley sprouts.

Biostimulatory Effects of Chlorella fusca CHK0059 on Plant Growth and Fruit Quality of Strawberry.

Green algae have been receiving widespread attention for their use as biofertilizers for agricultural production, but more studies are required to increase the efficiency of their use. This study aimed to investigate the effects of different levels of Chlorella fusca CHK0059 application on strawberry plant growth and fruit quality. A total of 800 strawberry seedlings were planted in a greenhouse and were grown for seven months under different Chlorella application rates: 0 (control), 0.1, 0.2, and 0.4% of the optimal cell density (OCD; 1.0 × 107 cells mL-1). The Chlorella application was conducted weekly via an irrigation system, and the characteristics of fruit samples were monitored monthly over a period of five months. The growth (e.g., phenotype, dry weight, and nutrition) and physiological (e.g., Fv/Fm and chlorophylls) parameters of strawberry plants appeared to be enhanced by Chlorella application over time, an enhancement which became greater as the application rate increased. Likewise, the hardness and P content of strawberry fruits had a similar trend. Meanwhile, 0.2% OCD treatment induced the highest values of soluble solid content (9.3-12 °Brix) and sucrose content (2.06-2.97 g 100 g-1) in the fruits as well as fruit flavor quality indices (e.g., sugars:acids ratio and sweetness index) during the monitoring, whilst control treatment represented the lowest values. In addition, the highest anthocyanin content in fruits was observed in 0.4% OCD treatment, which induced the lowest incidence of grey mold disease (Botrytis cinerea) on postharvest fruits for 45 days. Moreover, a high correlation between plants' nutrients and photosynthetic variables and fruits' sucrose and anthocyanin contents was identified through the results of principal component analysis. Overall, C. fusca CHK0059 application was found to promote the overall growth and performance of strawberry plants, contributing to the improvement of strawberry quality and yield, especially in 0.2% OCD treatment.

Synergetic effect of complex soil amendments to improve soil quality and alleviate toxicity of heavy metal(loid)s in contaminated arable soil: toward securing crop food safety and productivity.

Globally, various types of soil amendments have been used to improve the fertility and quality of soils in agricultural lands. In heavy metal(loid) (HM)-contaminated land, the soil amendments can also act as an immobilizing agent, thereby detoxifying HMs. A pot experiment was conducted to investigate the effects of three different complex amendments, including T1 (gypsum + peat moss + steel slag; GPMSS), T2 (GPMSS + lime), and T3 (GPMSS + lime + sulfate), on biogeochemical properties of the HM-contaminated arable soils, including Soil A and Soil B, and the magnitude of HM uptake by Chinese cabbage (Brassica rapa L.) for 6 weeks. All the examined complex amendments improved soils' physical and biological properties by increasing the water-stable aggregate (WSA) ratio by 18-54% and dehydrogenase activity (DHA) by 300-1333 mg triphenyl formazan (TPF) kg-1 24 h-1 in comparison to control soils. The concentrations of HMs accumulated in B. rapa appeared to decrease tremendously, attributed to effectively immobilizing the HMs in soils by incorporating complex amendments mediated by soil pH, dissolved organic carbon (DOC), and complexation with the components of amendments. All these positive changes in soil properties resulted in the elevation of B. rapa productivity. For instance, T1 treatment induced an increase of plant dry weight (DW) by 3.7-3.9 times compared to the controls. Suppose there are no typical differences in the efficiency among the treatments. In that case, our findings still suggest that using complex amendments for the HM-contaminated arable soils would be beneficial by bringing a synergetic effect on improving soil biogeochemical properties and alleviating HM toxicity, which eventually can enhance plant growth performance.

Auxin-Glucose Conjugation Protects the Rice (Oryza sativa L.) Seedlings Against Hydroxyurea-Induced Phytotoxicity by Activating UDP-Glucosyltransferase Enzyme.

Hydroxyurea (HU) is the replication stress known to carry out cell cycle arrest by inhibiting ribonucleotide reductase (RNR) enzyme upon generating excess hydrogen peroxide (H2O2) in plants. Phytohormones undergo synergistic and antagonistic interactions with reactive oxygen species (ROS) and redox signaling to protect plants against biotic and abiotic stress. Therefore, in this study, we investigated the protective role of Indole-3-acetic acid (IAA) in mitigating HU-induced toxicity in rice seedlings. The results showed that IAA augmentation improved the growth of the seedlings and biomass production by maintaining photosynthesis metabolism under HU stress. This was associated with reduced H2O2 and malondialdehyde (MDA) contents and improved antioxidant enzyme [superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), and peroxidase (POD)] activity that was significantly affected under HU stress. Furthermore, we showed that the HU stress-induced DNA damage leads to the activation of uridine 5'-diphosphate-glucosyltransferase (UGT), which mediates auxin homeostasis by catalyzing IAA-glucose conjugation in rice. This IAA-glucose conjugation upregulates the RNR, transcription factor 2 (E2F2), cyclin-dependent kinase (CDK), and cyclin (CYC) genes that are vital for DNA replication and cell division. As a result, perturbed IAA homeostasis significantly enhanced the key phytohormones, such as abscisic acid (ABA), salicylic acid (SA), cytokinin (CTK), and gibberellic acid (GA), that alter plant architecture by improving growth and development. Collectively, our results contribute to a better understanding of the physiological and molecular mechanisms underpinning improved growth following the HU + IAA combination, activated by phytohormone and ROS crosstalk upon hormone conjugation via UGT.