Salinity Effects on Gene Expression, Morphological, and Physio-Biochemical Responses of Stevia rebaudiana Bertoni In Vitro

Stevia rebaudiana Bertoni is a little bush, which is cultivated on a large scale in many countries for medicinal purposes and used as a natural sweetener in food products. The present work aims to conduct a protocol for stevia propagation in vitro to produce and introduce Stevia rebaudiana plants as a new sweetener crop to Egyptian agriculture. To efficiently maximize its propagation, it is important to study the influence of stress factors on the growth and development of Stevia rebaudiana grown in vitro. Two stevia varieties were investigated (Sugar High A3 and Spanti) against salt stress. Leaves were used as the source of explants for callus initiation, regeneration, multiplication and rooting. Some stress-related traits, i.e., photosynthetic pigments, proline contents, and enzyme activity for peroxidase (POD), polyphenol oxidase (PPO), and malate dehydrogenase (MDH) were studied. Murashig and Skoog (MS) medium was supplemented with four NaCl concentrations: 500, 1000, 2000, and 3000 mgL−1, while a salt-free medium was used as the control. The data revealed that salinity negatively affected all studied characters: the number of surviving calli, regeneration%, shoot length, the number of multiple shoots, number of leaf plantlets−1, number of root plantlets−1, and root length. The data also revealed that Sugar High A3 is more tolerant than Spanti. The total chlorophyll content decreased gradually with increasing NaCl concentration. However, the opposite was true for proline content. Isozyme’s fractionation exhibited high levels of variability among the two varieties. Various biochemical parameters associated with salt tolerance were detected in POD. Namely, POD4, POD6, POD 9 at an Rf of 0.34, 0.57, and 0.91 in the Sugar High A3 variety under high salt concentration conditions, as well as POD 10 at an Rf of 0.98 in both varieties under high salt concentrations. In addition, the overexpression of POD 5 and POD 10 at Rf 0.52 and 0.83 was found in both varieties at high NaCl concentrations. Biochemical parameters associated with salt tolerance were detected in PPO (PPO1, PPO2 and PPO4 at an Rf of 0.38, 0.42 and 0.62 in the Sugar High A3 variety under high salt concentrations) and MDH (MDH 3 at an Rf of 0.40 in both varieties at high salt concentrations). Therefore, these could be considered as important biochemical markers associated with salt tolerance and could be applied in stevia breeding programs (marker-assisted selection). This investigation recommends stevia variety Sugar High A3 to be cultivated under salt conditions

Natural Biostimulant Attenuates Salinity Stress Effects in Chili Pepper by Remodeling Antioxidant, Ion, and Phytohormone Balances, and Augments Gene Expression

A biostimulant is any microorganism or substance used to enhance the efficiency of nutrition, tolerance to abiotic stress and/or quality traits of crops, depending on its contents from nutrients. Plant biostimulants like honey bee (HB) and silymarin (Sm) are a strategic trend for managing stressed crops by promoting nutritional and hormonal balance, regulating osmotic protectors, antioxidants, and genetic potential, reflecting plant growth and productivity. We applied diluted honey bee (HB) and silymarin-enriched honey bee (HB- Sm) as foliar nourishment to investigate their improving influences on growth, yield, nutritional and hormonal balance, various osmoprotectant levels, different components of antioxidant system, and genetic potential of chili pepper plants grown under NaCl-salinity stress (10 dS m‒1). HB significantly promoted the examined attributes and HB-Sm conferred optimal values, including growth, productivity, K+/Na+ ratio, capsaicin, and Sm contents. The antioxidative defense components were significantly better than those obtained with HB alone. Conversely, levels of oxidative stress markers (superoxide ions and hydrogen peroxide) and parameters related to membrane damage (malondialdehyde level, stability index, ionic leakage, Na+, and Cl− contents) were significantly reduced. HB-Sm significantly affects inactive gene expression, as a natural biostimulator silencing active gene expression. SCoT primers were used as proof in salt-treated or untreated chili pepper plants. There were 41 cDNA amplicons selected by SCoT-primers. Twenty of them were EcDNA amplicons (cDNA-amplicons that enhanced their genes by one or more treatments) representing 49% of all cDNA amplicons, whereas 7 amplicons for ScDNA (whose genes were silenced in one or more treatments) represented 17%, and 14 McDNA (monomorphic cDNA-amplicons with control) amplicons were represented by 34% from all cDNA amplicons. This indicates the high effect of BH-Sm treatments in expression enhancement of some inactive genes and their silenced effect for expression of some active genes, also confirming that cDNA-SCoT markers succeeded in detection of variable gene expression patterns between the untreated and treated plants. In conclusion, HB-Sm as a natural multi-biostimulator can attenuate salt stress effects in chili pepper plants by remodeling the antioxidant defense system and ameliorating plant productivity

Natural Biostimulant Attenuates Salinity Stress Effects in Chili Pepper by Remodeling Antioxidant, Ion, and Phytohormone Balances, and Augments Gene Expression

A biostimulant is any microorganism or substance used to enhance the efficiency of nutrition, tolerance to abiotic stress and/or quality traits of crops, depending on its contents from nutrients. Plant biostimulants like honey bee (HB) and silymarin (Sm) are a strategic trend for managing stressed crops by promoting nutritional and hormonal balance, regulating osmotic protectors, antioxidants, and genetic potential, reflecting plant growth and productivity. We applied diluted honey bee (HB) and silymarin-enriched honey bee (HB- Sm) as foliar nourishment to investigate their improving influences on growth, yield, nutritional and hormonal balance, various osmoprotectant levels, different components of antioxidant system, and genetic potential of chili pepper plants grown under NaCl-salinity stress (10 dS m‒1). HB significantly promoted the examined attributes and HB-Sm conferred optimal values, including growth, productivity, K+/Na+ ratio, capsaicin, and Sm contents. The antioxidative defense components were significantly better than those obtained with HB alone. Conversely, levels of oxidative stress markers (superoxide ions and hydrogen peroxide) and parameters related to membrane damage (malondialdehyde level, stability index, ionic leakage, Na+, and Cl− contents) were significantly reduced. HB-Sm significantly affects inactive gene expression, as a natural biostimulator silencing active gene expression. SCoT primers were used as proof in salt-treated or untreated chili pepper plants. There were 41 cDNA amplicons selected by SCoT-primers. Twenty of them were EcDNA amplicons (cDNA-amplicons that enhanced their genes by one or more treatments) representing 49% of all cDNA amplicons, whereas 7 amplicons for ScDNA (whose genes were silenced in one or more treatments) represented 17%, and 14 McDNA (monomorphic cDNA-amplicons with control) amplicons were represented by 34% from all cDNA amplicons. This indicates the high effect of BH-Sm treatments in expression enhancement of some inactive genes and their silenced effect for expression of some active genes, also confirming that cDNA-SCoT markers succeeded in detection of variable gene expression patterns between the untreated and treated plants. In conclusion, HB-Sm as a natural multi-biostimulator can attenuate salt stress effects in chili pepper plants by remodeling the antioxidant defense system and ameliorating plant productivity

Integrative Seed and Leaf Treatment with Ascorbic Acid Extends the Planting Period by Improving Tolerance to Late Sowing Influences in Parsley

Abnormal production of reactive oxygen species (ROS) is an undesirable event which occurs in plants due to stress. To meet this event, plants synthesize ROS-neutralizing compounds, including the non-enzymatic oxidant scavenger known as vitamin C: ascorbic acid (AsA). In addition to scavenging ROS, AsA modulates many vital functions in stressed or non-stressed plants. Thus, two-season (2018/2019 and 2019/2020) trials were conducted to study the effect of integrative treatment (seed soaking + foliar spray) using 1.0 or 2.0 mM AsA vs. distilled water (control) on the growth, seed yield, and oil yield of parsley plants under three sowing dates (SDs; November, December, and January, which represent adverse conditions of late sowing) vs. October as the optimal SD (control). The ion balance, osmotic-modifying compounds, and different antioxidants were also studied. The experimental layout was a split plot in a completely randomized block design. Late sowing (December and January) noticeably reduced growth traits, seed and oil yield components, and chlorophyll and nutrient contents. However, soluble sugar, proline, and AsA contents were significantly increased along with the activities of catalase (CAT) and superoxide dismutase (SOD). Under late sowing conditions, the use of AsA significantly increased growth, different yields, essential oil fractions, CAT and SOD activities, and contents of chlorophylls, nutrients, soluble sugars, free proline, and AsA. The interaction treatments of SDs and AsA concentrations indicated that AsA at a concentration of 2 mM was more efficient in conferring greater tolerance to adverse conditions of late sowing in parsley plants. Therefore, this study recommends 2.0 mM AsA for integrative (seed soaking + foliar spraying) treatment to prolong the sowing period of parsley seeds (from October up to December) and avoid damage caused by adverse conditions of late sowing

Integrative Seed and Leaf Treatment with Ascorbic Acid Extends the Planting Period by Improving Tolerance to Late Sowing Influences in Parsley

Abnormal production of reactive oxygen species (ROS) is an undesirable event which occurs in plants due to stress. To meet this event, plants synthesize ROS-neutralizing compounds, including the non-enzymatic oxidant scavenger known as vitamin C: ascorbic acid (AsA). In addition to scavenging ROS, AsA modulates many vital functions in stressed or non-stressed plants. Thus, two-season (2018/2019 and 2019/2020) trials were conducted to study the effect of integrative treatment (seed soaking + foliar spray) using 1.0 or 2.0 mM AsA vs. distilled water (control) on the growth, seed yield, and oil yield of parsley plants under three sowing dates (SDs; November, December, and January, which represent adverse conditions of late sowing) vs. October as the optimal SD (control). The ion balance, osmotic-modifying compounds, and different antioxidants were also studied. The experimental layout was a split plot in a completely randomized block design. Late sowing (December and January) noticeably reduced growth traits, seed and oil yield components, and chlorophyll and nutrient contents. However, soluble sugar, proline, and AsA contents were significantly increased along with the activities of catalase (CAT) and superoxide dismutase (SOD). Under late sowing conditions, the use of AsA significantly increased growth, different yields, essential oil fractions, CAT and SOD activities, and contents of chlorophylls, nutrients, soluble sugars, free proline, and AsA. The interaction treatments of SDs and AsA concentrations indicated that AsA at a concentration of 2 mM was more efficient in conferring greater tolerance to adverse conditions of late sowing in parsley plants. Therefore, this study recommends 2.0 mM AsA for integrative (seed soaking + foliar spraying) treatment to prolong the sowing period of parsley seeds (from October up to December) and avoid damage caused by adverse conditions of late sowing