Foliar Spraying of Glycine Betaine Alleviated Growth Inhibition, Photoinhibition, and Oxidative Stress in Pepper (Capsicum annuum L.) Seedlings under Low Temperatures Combined with Low Light.

Low temperature combined with low light (LL stress) is a typical environmental stress that limits peppers' productivity, yield, and quality in northwestern China. Glycine betaine (GB), an osmoregulatory substance, has increasingly valuable effects on plant stress resistance. In this study, pepper seedlings were treated with different concentrations of GB under LL stress, and 20 mM of GB was the best treatment. To further explore the mechanism of GB in response to LL stress, four treatments, including CK (normal temperature and light, 28/18 °C, 300 µmol m-2 s-1), CB (normal temperature and light + 20 mM GB), LL (10/5 °C, 100 µmol m-2 s-1), and LB (10/5 °C, 100 µmol m-2 s-1 + 20 mM GB), were investigated in terms of pepper growth, biomass accumulation, photosynthetic capacity, expression levels of encoded proteins Capsb, cell membrane permeability, antioxidant enzyme gene expression and activity, and subcellular localization. The results showed that the pre-spraying of GB under LL stress significantly alleviated the growth inhibition of pepper seedlings; increased plant height by 4.64%; increased root activity by 63.53%; and decreased photoinhibition by increasing the chlorophyll content; upregulating the expression levels of encoded proteins Capsb A, Capsb B, Capsb C, Capsb D, Capsb S, Capsb P1, and Capsb P2 by 30.29%, 36.69%, 18.81%, 30.05%, 9.01%, 6.21%, and 16.45%, respectively; enhancing the fluorescence intensity (OJIP curves), the photochemical efficiency (Fv/Fm, Fv'/Fm'), qP, and NPQ; improving the light energy distribution of PSΠ (Y(II), Y(NPQ), and Y(NO)); and increasing the photochemical reaction fraction and reduced heat dissipation, thereby increasing plant height by 4.64% and shoot bioaccumulation by 13.55%. The pre-spraying of GB under LL stress also upregulated the gene expression of CaSOD, CaPOD, and CaCAT; increased the activity of the ROS-scavenging ability in the pepper leaves; and coordinately increased the SOD activity in the mitochondria, the POD activity in the mitochondria, chloroplasts, and cytosol, and the CAT activity in the cytosol, which improved the LL resistance of the pepper plants by reducing excess H2O2, O2-, MDA, and soluble protein levels in the leaf cells, leading to reduced biological membrane damage. Overall, pre-spraying with GB effectively alleviated the negative effects of LL stress in pepper seedlings.

Effects of exogenous glycine betaine and cycloleucine on photosynthetic capacity, amino acid composition, and hormone metabolism in Solanum melongena L.

Although exogenous glycine betaine (GB) and cycloleucine (Cyc) have been reported to affect animal cell metabolism, their effects on plant growth and development have not been studied extensively. Different concentrations of exogenous glycine betaine (20, 40, and 60 mmol L-1) and cycloleucine (10, 20, and 40 mmol L-1), with 0 mmol L-1 as control, were used to investigate the effects of foliar spraying of betaine and cycloleucine on growth, photosynthesis, chlorophyll fluorescence, Calvin cycle pathway, abaxial leaf burr morphology, endogenous hormones, and amino acid content in eggplant. We found that 40 mmol L-1 glycine betaine had the best effect on plant growth and development; it increased the fresh and dry weight of plants, increased the density of abaxial leaf hairs, increased the net photosynthetic rate and Calvin cycle key enzyme activity of leaves, had an elevating effect on chlorophyll fluorescence parameters, increased endogenous indoleacetic acid (IAA) content and decreased abscisic acid (ABA) content, and increased glutamate, serine, aspartate, and phenylalanine contents. However, cycloleucine significantly inhibited plant growth; plant apical dominance disappeared, plant height and dry and fresh weights decreased significantly, the development of abaxial leaf hairs was hindered, the net photosynthetic rate and Calvin cycle key enzyme activities were inhibited, the endogenous hormones IAA and ABA content decreased, and the conversion and utilization of glutamate, arginine, threonine, and glycine were affected. Combined with the experimental results and plant growth phenotypes, 20 mmol L-1 cycloleucine significantly inhibited plant growth. In conclusion, 40 mmol L-1 glycine betaine and 20 mmol L-1 cycloleucine had different regulatory effects on plant growth and development.

Exogenous Proline Enhances Systemic Defense against Salt Stress in Celery by Regulating Photosystem, Phenolic Compounds, and Antioxidant System.

This study aimed to explore how exogenous proline induces salinity tolerance in celery. We analyzed the effects of foliar spraying with 0.3 mM proline on celery growth, photosystem, phenolic compounds, and antioxidant system under salt stress (100 mM NaCl), using no salt stress and no proline spraying as control. The results showed that proline-treated plants exhibited a significant increase in plant biomass due to improved growth physiology, supported by gas exchange parameters, chlorophyll fluorescence, and Calvin cycle enzyme activity (Ketosasaccharide-1,5-diphosphate carboxylase and Fructose-1,6-diphosphate aldolase) results. Also, proline spraying significantly suppressed the increase in relative conductivity and malondialdehyde content caused by salt stress, suggesting a reduction in biological membrane damage. Moreover, salt stress resulted in hydrogen peroxide, superoxide anions and 4-coumaric acid accumulation in celery, and their contents were reduced after foliar spraying of proline. Furthermore, proline increased the activity of antioxidant enzymes (superoxide dismutase, peroxidase, and catalase) and the content of non-enzymatic antioxidants (reduced ascorbic acid, glutathione, caffeic acid, chlorogenic acid, total phenolic acids, and total flavonoids). Additionally, proline increased the activity of key enzymes (ascorbate oxidase, ascorbate peroxidase, glutathione reductase, and dehydroascorbate reductase) in the ascorbic acid-glutathione cycle, activating it to counteract salt stress. In summary, exogenous proline promoted celery growth under salt stress, enhanced photosynthesis, increased total phenolic acid and flavonoid contents, and improved antioxidant capacity, thereby improving salt tolerance in celery.

Response of rhizosphere microbial community of Chinese chives under different fertilization treatments.

Soil microorganisms play an irreplaceable role in agricultural production, however, an understanding of response of soil microorganisms to slow-release and common fertilizer applications is limited. In this study, different amounts of slow- release fertilizer were used to overwintering Chinese chives growing area in a plastic greenhouse to investigate the effects of on rhizosphere soil physicochemical properties and soil microbial communities (bacteria and fungi) of Chinese chives. The result displayed that application of slow-release fertilizer significantly improved soil nutrients, soil enzyme activity, and soil microbial community structure and diversity compared to conventional fertilizer application. Compared with T1 treatment, the content of total nitrogen (TN) and available phosphorus (AP), and the SU-E activity in the soil of T2 (NPK: 62.8 kg · 667 m-2) increased by 42.58%, 16.67%, and 9.70%, respectively, showing the best effects. In addition, soil bacterial diversity index and soil microbial community structure were improved as indicated by increased relative abundance of each species, such as Byssovorax, Sandaracinus, and Cellvibrio. Oppositely, the both soil fungal diversity and the number of species decreased after fertilizationthe relative abundance of Ascomycota increased in each fertilization treatment detected by ITS sequencing. Further, the relative abundance of pathogenic fungi such as Pezizomycetes, Cantharellales, and Pleosporales decreased in the T2 treatment. Principal Coordinates Analysis (PCoA) showed that both the amount of fertilizer applied and the type of fertilizer applied affected the soil microbial community structure. RDA evidenced that soil bacteria, Proteobacteria and Gemmatimonadetes, were closely correlated with soil AN, SOM, and AK. Acidobacteria were closely correlated with soil pH, TN, and AP. Ascomycota was closely correlated with soil pH and TN. In conclusion, the application of slow-release fertilizers and reduced fertilizer applicationcould improve soil physical and chemical properties as well as soil microbial community structure and diversity, contributing to sustainable soil development. The recommended fertilization rate for overwintering Chinese chives is NPK: 62.8 kg · 667 m-2.

Zinc oxide nanoparticles improve lettuce (Lactuca sativa L.) plant tolerance to cadmium by stimulating antioxidant defense, enhancing lignin content and reducing the metal accumulation and translocation.

Cadmium (Cd) contamination is a serious global concern that warrants constant attention. Therefore, a hydroponic study was conducted to evaluate the effect of different concentrations (0, 1, 2.5, 5, 10, 15 mg/l) of zinc oxide nanoparticles (ZnONPs) on the Cd content in lettuce (Lactuca sativa L.) under Cd stress conditions. The results showed that Cd stress triggered a decrease in plant biomass, an increase in relative electrolyte conductivity (REC), a decrease in root activity, accumulation of reactive oxygen species (ROS) accumulation, and nutrient imbalance. The application of ZnONPs reduced the toxicity symptoms of lettuce seedlings under Cd stress, with the most pronounced effect being observed 2.5 mg/l. ZnONPs promoted the growth of lettuce under Cd stress, mainly in terms of increase in biomass, chlorophyll content, antioxidant enzyme activity, and proline content, as well as reduction in Cd content, malondialdehyde, and reactive oxygen species (ROS) in plant tissues. ZnONPs also enhanced the uptake of ions associated with photosynthesis, such as iron, manganese, magnesium, and zinc. In addition, ZnONPs increase the amount of lignin in the roots, which blocks or reduces the entry of Cd into plant tissues.

Melatonin enhanced low-temperature combined with low-light tolerance of pepper (Capsicum annuum L.) seedlings by regulating root growth, antioxidant defense system, and osmotic adjustment.

Melatonin (MT) is an important biologically active hormone that plays a vital role in plant growth and development. In particular, it has been investigated for its roles in abiotic stress management. In this study, pepper seedlings were subjected to low-temperature combined with low-light stress (LL) (15/5°C, 100 µmol m-2s-1) prior to a foliar spray of 200mM MT for 168h to investigate the protective role of MT in pepper seedlings. Our results demonstrated that LL stress negatively affected root growth, and accelerated the accumulation of reactive oxygen species (ROS), including H2O2 and O 2 - , changed the osmolytes contents, and antioxidative system. However, these were reversed by exogenous MT application. MT effectively promoted the root growth as indicated by significant increase in root length, surface area, root volume, tips, forks, and crossings. In addition, MT reduced the burst of ROS and MDA contents induced by LL, enhanced the activities of SOD, CAT, POD, APX, DHAR, and MDHAR resulted by upregulated expressions of CaSOD, CaPOD, CaCAT, CaAPX, CaDHAR, and CaMDHAR, and elevated the contents of AsA and GSH, declined DHA and GSSH contents, which prevented membrane lipid peroxidation and protected plants from oxidative damages under LL stress. Furthermore, seedlings treated with MT released high contents of soluble sugar and soluble protein in leave, which might enhance LL tolerance by maintaining substance biosynthesis and maintaining cellular homeostasis resulted by high levels of osmolytes and carbohydrate in the cytosol. Our current findings confirmed the mitigating potential of MT application for LL stress by promoting pepper root growth, improving antioxidative defense system, ascorbate-glutathione cycle, and osmotic adjustment.

Trehalose alleviates salt tolerance by improving photosynthetic performance and maintaining mineral ion homeostasis in tomato plants.

Trehalose (Tre), which was an osmoprotective or stabilizing molecule, played a protective role against different abiotic stresses in plants and showed remarkable perspectives in salt stress. In this study, the potential role of Tre in improving the resistance to salt stress in tomato plants was investigated. Tomato plants (Micro Tom) were treated with Hoagland nutrient solution (CK), 10 mM Tre (T), 150 mM sodium chloride (NaCl, S), and 10 mM Tre+150 mM NaCl (S+T) for 5 days. Our results showed that foliar application of Tre alleviated the inhibition of tomato plant growth under salt stress. In addition, salt stress decreased the values of net photosynthetic rate (Pn, 85.99%), stomata conductance (gs, 57.3%), and transpiration rate (Tr, 47.97%), but increased that of intercellular carbon dioxide concentration (Ci, 26.25%). However, exogenous application of Tre significantly increased photosynthetic efficiency, increased the activity of Calvin cycle enzymes [ribulose diphosphate carboxylase/oxygenase (Rubisco), fructose-1,6-bisphosphate aldolase (FBA), fructose-1, 6-bisphosphatase (FBPase), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and transketolase (TK)], up-regulated the expression of genes encoding enzymes, induced stomatal opening, and alleviated salt-induced damage to the chloroplast membrane and structure. In the saline environment, photosynthetic electron transport was restricted, resulting the J-I-P phase to decrease. At the same time, the absorption, capture, and transport energies per excited cross-section and per active reaction center decreased, and the dissipation energy increased. Conversely, Tre reversed these values and enhanced the photosystem response to salt stress by protecting the photosynthetic electron transport system. In addition, foliage application with Tre significantly increased the potassium to sodium transport selectivity ratio (S K-Na ) by 16.08%, and increased the levels of other ions to varying degrees. Principal component analysis (PCA) analysis showed that exogenous Tre could change the distribution of elements in different organs and affect the expressions of SlSOS1, SlNHX, SlHKT1.1, SlVHA, and SlHA-A at the transcriptional level under salt stress, thereby maintaining ion homeostasis. This study demonstrated that Tre was involved in the process of mitigating salt stress toxicity in tomato plants and provided specific insights into the effectiveness of Tre in mediating salt tolerance.

Effect of Methyl Jasmonate Treatment on Primary and Secondary Metabolites and Antioxidant Capacity of the Substrate and Hydroponically Grown Chinese Chives.

Hydroponic culture has become a commercial planting model for leafy vegetables, herbs, and other plants with medicinal value. Methyl jasmonate (MeJA) is involved in primary and secondary plant metabolism; moreover, it regulates plant bioactive compounds and enhances the nutritional and medicinal value of plants. We performed targeted metabolomic analysis of the primary and secondary metabolites in substrate-grown and hydroponic Chinese chive leaves sprayed with MeJA (0, 300, 500, and 800 µM). Using ultra-performance liquid chromatography (UPLC), UPLC tandem mass spectrometry, and chemometric tools, and analyzed the antioxidant activity of these plants. We identified the biomarkers of amino acids (serine, proline, lysine, and arginine) and phenolic compounds (4-coumaric acid and protocatechuic acid) using chemometric tools to distinguish between substrate-grown and hydroponic Chinese chives treated with MeJA. MeJA (500 µM) treatment significantly increased the total sugar and amino acid (essential and non-essential amino acids and sulfur-containing amino acids) contents of hydroponically grown Chinese chives. However, the changes in total sugar and amino acid contents in Chinese chive grown in substrates showed the opposite trend. The organic acid content of hydroponically grown Chinese chives treated with MeJA decreased significantly, whereas that of substrate-grown plants treated with 300 µM MeJA increased significantly. Further, MeJA treatment significantly increased the phenolic content of substrate-grown Chinese chives. Treatment with 800 µM MeJA significantly increased the carotenoid content of substrate-grown Chinese chives and the phenolic content of hydroponic Chinese chives. In addition, the 500 µM MeJA treatment significantly increased the antioxidant activity of Chinese chives in both substrate-grown and hydroponic cultures, and promoted the accumulation of nutrients and bioactive substances. This treatment also improved the flavor quality of these plants and their nutritional and medicinal value. Thus, the results suggested that MeJA-treated plants could be used as value-added horticultural products.

Effect of slow-release fertilizer on soil fertility and growth and quality of wintering Chinese chives (Allium tuberm Rottler ex Spreng.) in greenhouses.

To avoid the negative impact of excessive fertilization on vegetable production, a decreased fertilization experiment was conducted in a multi-layer covered plastic greenhouse in 2017 to 2018. Treatments included no fertilizer (CK), traditional fertilization (TF), slow-release fertilizers (SRF), and decreased fertilization with slow-release fertilizers (DSRF). Results showed that the SRF and DSRF increased leaf length (13% and 8.3%) and chlorophyll content (7.1% and 8.2%) of Chinese chives compared to TF. Similarly, DSRF was found to increase the accumulation of dry matter accumulation of roots (22%) and the dry matter accumulation of shoots (36%) of Chinese chives. Flavonoid, soluble sugar, and soluble protein content were enhanced by 18%, 8.5%, and 4.6%, respectively, in DSRF compared to TF. Nitrate content of the SRF and SRFR decreased significantly by 26% and 35%, respectively. In addition, there was a significant increase in soil nutrient and enzyme activity in the middle and late harvest of Chinese chives under DSRF compared to TF, and there was a high correlation between soil nutrients and the quality of Chinese chives. The available P and total P content significantly differed among the different greenhouse soil samples, and this significantly affected the quality of Chinese chives. The content of available P and total P in greenhouse soil was 125.07 g kg-1 and 1.26 mg kg-1, respectively. Optimal quality was obtained. Hence, the application of DSRF promoted the growth of Chinese chives and improved soil fertility, thereby enhancing the productivity and quality of Chinese chives.

Effects of Preharvest Methyl Jasmonate and Salicylic Acid Treatments on Growth, Quality, Volatile Components, and Antioxidant Systems of Chinese Chives.

Methyl jasmonate (MeJA) and salicylic acid (SA) regulate the production of biologically active compounds in plants and stimulate the accumulation of plant aromatic substances. However, the underlying mechanisms of how MeJA and SA influence characteristic flavor compounds and the antioxidant activity of vegetables are poorly understood. Five MeJA and SA concentrations were used to investigate the dose-dependent effects of these phytohormones on the dry and fresh weight; chlorophyll abundance; the contents of vitamin C, soluble protein, and sugar, nitrate, total phenols, flavonoids, volatile components, and enzymatically produced pyruvic acid; and antioxidant activity in Chinese chive. We found that MeJA and SA at concentrations of 500 and 150 µM, respectively, significantly increased the levels of total chlorophyll, phenols and flavonoids, vitamin C, and volatile components and significantly reduced the accumulation of nitrate. In addition, compared with the control, 500 µM of MeJA significantly increased the soluble sugar and protein content, and 150 µM SA significantly increased the dry and fresh weight of Chinese chive. Furthermore, these concentrations of MeJA and SA significantly increased the enzymatic pyruvate content and the amount of sulfide and aromatic volatile compounds and improved the characteristic flavor compounds. The 2,2-diphenyl-1-picrylhydrazyl radical scavenging capacity, Trolox-equivalent antioxidant capacity, and ferric-reducing antioxidant capacity were significantly improved after a preharvest treatment with 500 µM MeJA and 150 µM SA, which could improve the antioxidant activity, thus improving the postharvest quality and preservation characteristics of Chinese chives. Taken together, a preharvest treatment with 500 µM MeJA and 150 µM SA is optimal to improve the growth, quality, antioxidant activity, and flavor of Chinese chive, thereby enhancing its commercial value.

Corrigendum to: Promoting pepper (Capsicum annuum) photosynthesis via chloroplast ultrastructure and enzyme activities by optimising the ammonium to nitrate ratio.

Optimal plant growth in many species is achieved when the two major forms of N are supplied at a particular ratio. In this pot experiment, the effects of five different ammonium:nitrate ratios (ANRs) (0:100, 12.5:87.5, 25:75, 37.5:62.5, and 50:50) on photosynthesis efficiency in chilli pepper (Capsicum annuum L.) plants were evaluated. The results showed that an ANR of 25:75 increased the contents of chl a, leaf area and dry matter, whereas chl b content was not affected by the ANRs. Regarding chlorophyll fluorescence, an ANR of 25:75 also enhanced the actual photochemical efficiency, photochemical quenching and maximum photosynthetic rate. However, the 0:100 and 50:50 ANRs resulted in higher values for nonphotochemical quenching. An inhibition of maximal photochemical efficiency was found when 50% NH4+ was supplied at the later stage of plant growth. The addition of 25% or 37.5% NH4+ was beneficial for gas exchange parameters and the 25% NH4+ optimised the thylakoid of chloroplasts. Compared with nitrate alone, 12.5-50% NH4+ upregulated glutamate dehydrogenase (GDH), the large subunit and the small subunit of Rubisco. It can be concluded that the 25:75 ANR accelerated N assimilation through active GDH, which provides a material basis for chloroplast and Rubisco formation, resulting in the increased photosynthetic rate and enhanced growth in chilli pepper.

Promoting pepper (Capsicum annuum) photosynthesis via chloroplast ultrastructure and enzyme activities by optimising the ammonium to nitrate ratio.

Optimal plant growth in many species is achieved when the two major forms of N are supplied at a particular ratio. In this pot experiment, the effects of five different ammonium:nitrate ratios (ANRs) (0:100, 12.5:87.5, 25:75, 37.5:62.5, and 50:50) on photosynthesis efficiency in chilli pepper (Capsicum annuum L.) plants were evaluated. The results showed that an ANR of 25:75 increased the contents of chl a, leaf area and dry matter, whereas chl b content was not affected by the ANRs. Regarding chlorophyll fluorescence, an ANR of 25:75 also enhanced the actual photochemical efficiency, photochemical quenching and maximum photosynthetic rate. However, the 0:100 and 50:50 ANRs resulted in higher values for nonphotochemical quenching. An inhibition of maximal photochemical efficiency was found when 50% NH4+ was supplied at the later stage of plant growth. The addition of 25% or 37.5% NH4+ was beneficial for gas exchange parameters and the 25% NH4+ optimised the thylakoid of chloroplasts. Compared with nitrate alone, 12.5­50% NH4+ upregulated glutamate dehydrogenase (GDH), the large subunit and the small subunit of Rubisco. It can be concluded that the 25:75 ANR accelerated N assimilation through active GDH, which provides a material basis for chloroplast and Rubisco formation, resulting in the increased photosynthetic rate and enhanced growth in chilli pepper.