An Evaluation of the Effectivity of the Green Leaves Biostimulant on Lettuce Growth, Nutritional Quality, and Mineral Element Efficiencies under Optimal Growth Conditions.

The use of biostimulants is becoming a useful tool for increasing crop productivity while enhancing nutritional quality. However, new studies are necessary to confirm that the joint application of different types of biostimulants, together with bioactive compounds, is effective and not harmful to plants. This study examined the impact of applying the biostimulant Green Leaves, comprising Macrocystis algae extract and containing a mixture of amino acids, corn steep liquor extract, calcium, and the bioactive compound glycine betaine. The effect of applying two different doses (3 and 5 mL L-1) of this biostimulant was evaluated on lettuce plants, and growth and quality parameters were analyzed along with photosynthetic efficiency, nutritional status, and nutrient efficiency parameters. The application of Green Leaves improved plant weight (25%) and leaf area and enhanced the photosynthetic rate, the accumulation of soluble sugars and proteins, and the agronomic efficiency of all essential nutrients. The 3 mL L-1 dose improved the nutritional quality of lettuce plants, improving the concentration of phenolic compounds and ascorbate and the antioxidant capacity and reducing NO3- accumulation. The 5 mL L-1 dose improved the absorption of most nutrients, especially N, which reduced the need for fertilizers, thus reducing costs and environmental impact. In short, the Green Leaves product has been identified as a useful product for obtaining higher yield and better quality.

Evaluation of the alkalinity stress tolerance of three Brassica rapa CAX1 TILLING mutants.

Alkalinity is an important environmental factor that affects crop production and will be exacerbated in the current climate change scenario. Thus, the presence of carbonates and high pH in soils negatively impacts nutrient assimilation and photosynthesis and causes oxidative stress. A potential strategy to improve tolerance to alkalinity could be the modification of cation exchanger (CAX) activity, given that these transporters are involved in calcium (Ca2+) signaling under stresses. In this study, we used three Brassica rapa mutants (BraA.cax1a-4, BraA.cax1a-7, and BraA.cax1a-12) from the parental line 'R-o-18' that were generated by Targeting Induced Local Lesions in Genomes (TILLING) and grown under control and alkaline conditions. The objective was to assess the tolerance of these mutants to alkalinity stress. Biomass, nutrient accumulation, oxidative stress, and photosynthesis parameters were analyzed. The results showed that BraA.cax1a-7 mutation was negative for alkalinity tolerance because it reduced plant biomass, increased oxidative stress, partially inhibited antioxidant response, and lowered photosynthesis performance. Conversely, the BraA.cax1a-12 mutation increased plant biomass and Ca2+ accumulation, reduced oxidative stress, and improved antioxidant response and photosynthesis performance. Hence, this study identifies BraA.cax1a-12 as a useful CAX1 mutation to enhance the tolerance of plants grown under alkaline conditions.

Application of an Enzymatic Hydrolysed L-α-Amino Acid Based Biostimulant to Improve Sunflower Tolerance to Imazamox.

Herbicides, commonly used in agriculture to control weeds, often cause negative effects on crops. Safeners are applied to reduce the damage to crops without affecting the effectiveness of herbicides against weeds. Plant biostimulants have the potential to increase tolerance to a series of abiotic stresses, but very limited information exists about their effects on herbicide-stressed plants. This study aims to verify whether the application of a potential safener such as Terra-Sorb®, an L-α-amino acid-based biostimulant, reduces the phytotoxicity of an Imazamox-based herbicide and to elucidate which tolerance mechanisms are induced. Sunflower plants were treated with Pulsar® 40 (4% Imazamox) both alone and in combination with Terra-Sorb®. Plants treated with the herbicide in combination with Terra-Sorb® showed higher growth, increased acetolactate synthase (ALS) activity, and amino acid concentration with respect to the plants treated with Imazamox alone. Moreover, the biostimulant protected photosynthetic activity and reduced oxidative stress. This protective effect could be due to the glutathione S-transferase (GST) induction and antioxidant systems dependent on glutathione (GSH). However, no effect of the biostimulant application was observed regarding phenolic compound phenylalanine ammonium-lyase (PAL) activity. Therefore, this study opens the perspective of using Terra-Sorb® in protecting sunflower plants against an imazamox-based herbicide effect.

Effect of l-amino acid-based biostimulants on nitrogen use efficiency (NUE) in lettuce plants.

BACKGROUND: Biostimulants are increasingly integrated into production systems with the goal of modifying physiological processes in plants to optimize productivity. Specifically, l-α-amino acid-based biostimulants enhance plant productivity through improved photosynthesis and increased assimilation of essential nutrients such as nitrogen (N). This element is a major component of fertilizers, which usually are applied in excess. Thus, the inefficient use of N fertilizers has generated a serious environmental pollution issue. The use of biostimulants has the potential to address problems related to N fertilization. Therefore, the objective of this study is to analyze whether two biostimulants based on l-α-amino acid (Terra Sorb® radicular and Terramin® Pro) designed by Bioiberica, S.A.U company can compensate deficient N fertilization and test its effect on lettuce plants. Growth, photosynthetic, N accumulation, and N use efficiency (NUE) parameters were analyzed on lettuce leaves. RESULTS: Results showed that regardless of N fertilization, the use of both biostimulants, especially Terramin® Pro, increased biomass production. Moreover, both biostimulants enhanced photosynthetic, NO3 - and total N accumulations as well as NUE parameters. CONCLUSION: Therefore, Terra Sorb® radicular and Terramin® Pro constitute a useful tool for crops development in N-limiting areas, and in intensive agricultural areas without N deficiency allowing the reduction of N inputs without impairing crop yields and reducing environmental impact. © 2022 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Improvement of the physiological response of barley plants to both Zinc deficiency and toxicity by the application of calcium silicate.

An adequate availability of Zinc (Zn) is crucial for plant growth and development given the essentiality of this element. Thus, both Zn deficiency and Zn toxicity can limit crop yields. In plants, the responses to Zn imbalances involve important physiological aspects such as reactive oxygen species (ROS) accumulation, phytohormone balance, tricarboxylic acid cycle (TCA) metabolism, and organic acids (OAs) accumulation. However, a way to improve tolerance to stresses such as those produced by nutritional imbalances is the application of beneficial elements such as silicon (Si). In this study, we grew barley plants in hydroponics under Zn deficiency and toxicity conditions, applying Si in the form of CaSiO3 in order to assess its effectiveness against Zn imbalances. Parameters related to plant growth, oxidative stress, TCA enzyme activities, phytohormones and OAs accumulation were analyzed. Both Zn deficiency and toxicity reduced leaf biomass, increased ROS accumulation, and affected phytohormone and OAs concentrations and TCA enzyme activities. CaSiO3 treatment was effective in counteracting these effects enhancing Zn accumulation under Zn deficient conditions and limiting its accumulation under toxic conditions. In addition, this treatment decreased ROS levels, and improved ascorbate/glutathione and phytohormonal responses, citrate synthase activity, and malate/oxalate ratio. Therefore, this study enhanced the notion of the efficacy of CaSiO3 in improving tolerance to Zn imbalances.

Effect of CAX1a TILLING mutations on photosynthesis performance in salt-stressed Brassica rapa plants.

Salinity is an important environmental factor that reduces plant productivity in many world regions. It affects negatively photosynthesis causing a growth reduction. Likewise, calcium (Ca2+) is crucial in plant stress response. Therefore, the modification of Ca2+ cation exchangers (CAX) transporters could be a potential strategy to increase plant tolerance to salinity. Using Targeting Induced Local Lesions in Genomes (TILLING), researchers generated three mutants of Brassica rapa CAX1a transporter: BraA.cax1a-7, BraA.cax1a-4, and BraA.cax1a-12. The aim of this study was to test the effect of those mutations on salt tolerance focusing on the response to the photosynthesis process. Thus, the three BraA.cax1a mutants and the parental line (R-o-18) were grown under salinity conditions, and parameters related to biomass, photosynthesis performance, glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49), and soluble carbohydrates were measured. BraA.cax1a-4 provided higher biomass and a better photosynthetic performance manifested by higher water use efficiency (WUE), Fv/Fm, electron fluxes, and Rubisco (EC 4.1.1.39) values. In addition, BraA.cax1a-4 presented increased osmotic protection through myo-inositol accumulation. On the other hand, BraA.cax1a-7 produced some negative effects on photosynthesis performance and lower G6PDH and Rubisco accumulations. Therefore, this study points out BraA.cax1a-4 as a useful mutation to improve photosynthetic performance in plants grown under saline conditions.

Assaying the use of sodium thiosulphate as a biostimulant and its effect on cadmium accumulation and tolerance in Brassica oleracea plants.

An optimal uptake of mineral elements is crucial to ensure both crop yield and quality. The use of biostimulants is taking relevance to improve the nutrition of crops. Sulphur (S) is one of the elements with great potential within biostimulants. Furthermore, soil contamination by heavy metals such as cadmium (Cd) has become a serious environmental problem. Different studies have suggested the use of thiosulphate (TS) as a biostimulant and to increase the phytoremediation capacity of plants. Therefore, in the present study, we use a crop plant with high S requirements such as Brassica oleracea, to test whether TS serves as a biostimulant and whether affects Cd accumulation and tolerance. B. oleracea plants were grown with two different TS doses (2 mM and 4 mM), under Cd toxicity, and with the combination of Cd toxicity and both TS doses. Parameters of biomass, mineral elements accumulation, and stress tolerance were analyzed. The results showed that TS reduced biomass of B. oleracea plants. The application of 2 mM TS increased Cd accumulation whereas the 4 mM dose reduced it. On the other hand, TS incremented micronutrient accumulation on plants subjected to Cd toxicity and increased Zn contents. Besides, the application of 2 mM to Cd-stressed plants enhanced photosynthesis performance and reduced oxidative stress. Finally, TS increased the antioxidant capacity of B. oleracea plants. Briefly, although TS can not be used as a biostimulant it could be used for Cd phytoremediation purposes and to enhance Zn accumulation in B. oleracea plants.

Tolerance to cadmium toxicity and phytoremediation potential of three Brassica rapa CAX1a TILLING mutants.

Cadmium (Cd) is one of the most toxic heavy metals that reduces crop productivity and is a threat to all the food chain including human health. Phytoremediation is an environmentally friendly strategy to clean up soil contaminated with heavy metals. Researchers are selecting new varieties with an enhanced capacity for phytoremediation purposes. Three Brassica rapa mutants for CAX1 transporter were obtained through TILLING. The objective of this work is to evaluate the tolerance of these mutants to Cd toxicity and its potential for Cd phytoremediation. For this, the mutants and the parental R-o-18 were grown under control and Cd toxicity conditions (100 µM CdCl2) and growth, Cd accumulation and physiological parameters were analyzed. The results show that BraA.cax1a mutation provides greater Cd uptake capacity although only BraA.cax1a-12 would be useful for phytoremediation because it registered more than three-fold the Cd content of R-o-18 and presented greater Cd tolerance. This tolerance could be due to the higher Ca and Mg accumulations, the maintaining of photosynthesis performance, the enhanced ROS detoxification and AsA/GSH and TCA cycles, the higher malate, and GA4 concentrations and the lower ethylene levels. Briefly, this study identifies BraA.cax1a-12 as a potential mutant for phytoremediation of Cd contaminated soil and identifies possible physiological elements that contribute to this capacity.

Study of Zn accumulation and tolerance of HMA4 TILLING mutants of Brassica rapa grown under Zn deficiency and Zn toxicity.

Nowadays, Zinc (Zn) deficiency is the most widespread micronutrient deficiency but simultaneously Zn toxicity is produced due to environmental pollution. A potential method to alleviate Zn deficiency and to reduce Zn concentration in soils is through the generation of plants with enhanced capacity for Zn accumulation and higher tolerance. This could be achieved through the modification of HMA4 transporter. BraA.hma4a-3 is a TILLING mutant plant that presents one modification in HMA4 transporter. Thus, in this study we analyzed the potential of BraA.hma4a-3 for Zn accumulation and Zn deficiency and toxicity tolerance. BraA.hma4a-3 and parental R-o-18 plants were grown with different Zn doses: 1 µM ZnSO4 (Control), 0.01 µM ZnSO4 (Zn deficiency) and 100 µM ZnSO4 (Zn toxicity). Parameters of biomass, Zn concentration, photosynthesis, oxidative stress, N metabolism and amino acids (AAs) were measured. BraA.hma4a-3 did not affect plant biomass but did increase Zn accumulation in leaves under an adequate Zn supply and Fe under control and Zn deficiency doses. Regarding stress tolerance parameters and N metabolism, BraA.hma4a did not produce alterations under control conditions. In addition, under Zn toxicity, parameters suggest a greater tolerance. Briefly, the obtained results point to BraA.hma4a-3 as a useful mutant to increase Zn accumulation.

Possible role of HMA4a TILLING mutants of Brassica rapa in cadmium phytoremediation programs.

Cadmium (Cd) is a dangerous transition element that causes environmental and health problems due to its high mobility in the soil-plant system. In plants, Cd causes serious alterations in physiological processes, affecting different vital functions such as photosynthesis. Species such as Brassica juncea and Brassica rapa have been selected as suitable plants for phytoremediation purposes due to their ability to tolerate the toxic effect of heavy metals. In order to improve this strategy, techniques of plant mutagenesis such as TILLING (Targeting Induced Local Lessions in Genomes) have been employed. In the present work we studied the role of the HMA4 gene in the tolerance to Cd toxicity (100 µM CdCl2) using a TILLING mutant of B. rapa (BraA.hma4a-3). These mutant plants presented a lower biomass reduction and a higher Cd concentration in leaves. An increase in the GSH/GSSG ratio, in the content of photosynthetic pigments and a reduction of oxidative stress was observed, as well as a better photosynthetic index, confirming that BraA.hma4a-3 plants showed a higher tolerance to Cd. In conclusion, according to the results obtained in this work, BraA.hma4a-3 plants could be used for phytoremediation purposes of Cd contaminated soils.

Effect of CAX1a TILLING mutations and calcium concentration on some primary metabolism processes in Brassica rapa plants.

Cation/H+ exchanger transporters (CAXs) are crucial in Ca2+ homeostasis and in the generation of Ca2+ profiles involved in signalling processes. Given the crucial role of CAX1 in Ca2+ homeostasis, CAX1 modifications could have effects on plant metabolism. Three Brassica rapa mutants for CAX1 were obtained through TILLING. The aim of this work is to assess the effect of the different mutations and different Ca2+ doses on plant metabolism. For this, the mutants and the parental line were grown under low, control and high Ca2+ doses and parameters related to nitrogen (N) and tricarboxylic acid (TCA) metabolisms, and amino acid (AAs) and phytohormone profiles were measured. The results show that BraA.cax1a mutations affect metabolism especially under high Ca2+ dose. Thus, BraA.cax1a-7 inhibited some N metabolism enzymes and activated photorespiration activity. On the opposite side, BraA.cax1a-12 mutation provides a better tolerance to high Ca2+ dose. This tolerance could be provided by an improved N and TCA metabolisms enzymes, and a higher glutamate, malate, indole-3-acetic acid and abscisic acid concentrations. Therefore, BraA.cax1a-12 mutation could be used for B. rapa improving; the metabolomics changes observed in this mutant could be responsible for a better tolerance to high Ca2+.

Physiological profile of CAX1a TILLING mutants of Brassica rapa exposed to different calcium doses.

Calcium (Ca) is an essential macronutrient for plants and its homeostasis is basic for many processes in plants. Therefore, both Ca deficiency and toxicity constitute potential issues for crops. CAX1 transporter is a potential target to obtain plants with better Ca homeostasis and higher Ca concentration in edible parts. Three Brassica rapa mutants for CAX1 were obtained through TILLING. The objective of this work is to evaluate the growth, physiological state and nutrients concentration of these mutants grown with different Ca doses. The mutants and the parental line were grown under low, control and high Ca doses and parameters related to their oxidative stress, photosynthetic performance and nutrients concentration were determined. BraA.cax1a-4 and BraA.cax1a-7 mutants presented lower total Chl, an altered photosynthesis performance and higher ROS levels. BraA.cax1a-12 mutant grew better under high Ca conditions. All mutants accumulated more Ca and Mg in leaves under control and high Ca doses and accumulated more Fe regardless the Ca dose. The results obtained point to BraA.cax1a-12 as a potential candidate for biofortification with Fe, Ca and Mg since it accumulate higher concentrations of these elements, do not present an altered growth and is able to tolerate higher Ca doses.

Study of phytohormone profile and oxidative metabolism as key process to identification of salinity response in tomato commercial genotypes.

Climatic change, intensive agriculture, and worsening water quality induce abiotic stress conditions for plants. Among these factors, salinity stress is a limit factor for plant growth. Therefore, the purpose of this study was to analyze the phytohormones role and oxidative metabolism in response to salt stress of two genotypes of tomato cv. Grand Brix and cv. Marmande RAF, the crops were carried out in a growth chamber. Salinity stress reduces biomass and relative growth rate (RGR) in both genotypes, this effect being greater in cv. Marmande RAF. These results, together with main stress indicator response, the O2.-, indicate that cv. Marmande RAF is more sensitive to Saline stress. Grand Brix showed less oxidative stress, because it presented greater detoxification of the O2-, due to SOD enzyme activity induction and greater antioxidant capacity. Furthermore, Grand Brix has a better hormonal profile adapted to salt stress resistance, the accumulation of IAA, GA4 and CKs and their beneficial role against oxidative stress could make the difference between resistance and sensitivity to salt stress. On the other hand, a lower ACC concentration, ethylene precursor, combined with a greater O2.- detoxification in the cv. Grand Brix could play a fundamental role in tolerance to saline stress. Besides, an increase in ABA levels promotes better stomatal closure, better photosynthesis control and a lower rate of water loss. This data could be essential to select plants with greater resistance to saline stress.

Phytohormone profile in Lactuca sativa and Brassica oleracea plants grown under Zn deficiency.

Phytohormones, structurally diverse compounds, are involved in multiple processes within plants, such as controlling plant growth and stress response. Zn is an essential micronutrient for plants and its deficiency causes large economic losses in crops. Therefore, the purpose of this study was to analyse the role of phytohormones in the Zn-deficiency response of two economically important species, i.e. Lactuca sativa and Brassica oleracea. For this, these two species were grown hydroponically with different Zn-application rates: 10 µM Zn as control and 0.1 µM Zn as deficiency treatment and phytohormone concentration was determined by U-HPLC-MS. Zn deficiency resulted in a substantial loss of biomass in L. sativa plants that was correlated with a decline in growth-promoting hormones such as indole-3-acetic acid (IAA), cytokinins (CKs), and gibberellins (GAs). However these hormones increased or stabilized their concentrations in B. oleracea and could help to maintain the biomass in this species. A lower concentration of stress-signaling hormones such as ethylene precursor aminocyclopropane-1-carboxylic acid (ACC), abscisic acid (ABA), salicylic acid (SA) and jasmonic acid (JA) and also CKs might be involved in Zn uptake in L. sativa while a rise in GA4, isopentenyl adenine (iP), and ACC and a fall in JA and SA might contribute to a better Zn-utilization efficiency (ZnUtE), as observed in B. oleracea plants.

Comparative study of Zn deficiency in L. sativa and B. oleracea plants: NH4(+) assimilation and nitrogen derived protective compounds.

Zinc (Zn) deficiency is a major problem in agricultural crops of many world regions. N metabolism plays an essential role in plants and changes in their availability and their metabolism could seriously affect crop productivity. The main objective of the present work was to perform a comparative analysis of different strategies against Zn deficiency between two plant species of great agronomic interest such as Lactuca sativa cv. Phillipus and Brassica oleracea cv. Bronco. For this, both species were grown in hydroponic culture with different Zn doses: 10µM Zn as control and 0.01µM Zn as deficiency treatment. Zn deficiency treatment decreased foliar Zn concentration, although in greater extent in B. oleracea plants, and caused similar biomass reduction in both species. Zn deficiency negatively affected NO3(-) reduction and NH4(+) assimilation and enhanced photorespiration in both species. Pro and GB concentrations were reduced in L. sativa but they were increased in B. oleracea. Finally, the AAs profile changed in both species, highlighting a great increase in glycine (Gly) concentration in L. sativa plants. We conclude that L. sativa would be more suitable than B. oleracea for growing in soils with low availability of Zn since it is able to accumulate a higher Zn concentration in leaves with similar biomass reduction. However, B. oleracea is able to accumulate N derived protective compounds to cope with Zn deficiency stress.