A simple method to evaluate the number of bradyrhizobia on soybean seeds and its implication on inoculant quality control

Soybean seeds are non-sterile and their bacterial population interferes with the enumeration of beneficial bacteria, making it difficult to assess survival under different conditions. Within this context, the principal aims of this work were: (1) to improve a selective media for the enumeration of B. japonicum recovered from inoculated soybean seeds; (2) to establish the most representative mathematical function for B. japonicum mortality on soybean seeds after inoculation; (3) to evaluate if environmental or physiological conditions modify B. japonicum mortality on soybean seeds; and (4) to create a new protocol for quality control of soybean inoculants. We successfully evaluated the combination of pentachloronitrobenzene and vancomycin added to the yeast-mannitol medium to inhibit most fungi and Gram-positive soybean microbiota, thus producing reliable counts of B. japonicum from inoculated soybean seeds. Percentages of recovery and survival factors were obtained and used to construct a two-phase exponential decay non-linear regression function. High temperature and desiccation decreased these parameters, while the optimization of temperature and the use of osmoprotective compounds with inoculants increased them. The use of this protocol minimized heterogeneity between experiments and may be considered more reliable than the simple expression of direct colony count of bacteria recovered from seeds

A simple method to evaluate the number of bradyrhizobia on soybean seeds and its implication on inoculant quality control

Soybean seeds are non-sterile and their bacterial population interferes with the enumeration of beneficial bacteria, making it difficult to assess survival under different conditions. Within this context, the principal aims of this work were: (1) to improve a selective media for the enumeration of B. japonicum recovered from inoculated soybean seeds; (2) to establish the most representative mathematical function for B. japonicum mortality on soybean seeds after inoculation; (3) to evaluate if environmental or physiological conditions modify B. japonicum mortality on soybean seeds; and (4) to create a new protocol for quality control of soybean inoculants. We successfully evaluated the combination of pentachloronitrobenzene and vancomycin added to the yeast-mannitol medium to inhibit most fungi and Gram-positive soybean microbiota, thus producing reliable counts of B. japonicum from inoculated soybean seeds. Percentages of recovery and survival factors were obtained and used to construct a two-phase exponential decay non-linear regression function. High temperature and desiccation decreased these parameters, while the optimization of temperature and the use of osmoprotective compounds with inoculants increased them. The use of this protocol minimized heterogeneity between experiments and may be considered more reliable than the simple expression of direct colony count of bacteria recovered from seeds

Indole-3-acetic acid attenuates the fungal lesions in infected potato tubers

In this report, we demonstrated that potato tubers pre-treated with 100 μM of indole-3-acetic acid (IAA) and then inoculated with Fusarium solani f. sp. eumartii (F. eumartii) showed a decrease in the fungal lesion compared with non-IAA pre-treated and inoculated tubers. Consequently, we tested the addition of different concentrations of IAA on the fungal growth and on the proteolytic activity when the fungus was grown in liquid culture medium. Fungal growth did not change under different phytohormone concentrations, but the activity of Fusarium extracellular serine protease (FESP) clearly decreased. The inhibition of FESP activity by IAA was dose dependent. Moreover, FESP as well as others extracellular pectinolytic activities detected in IAA pre-treated and inoculated tubers decreased compared with controls. In addition, the ability of IAA production by F. eumartii was tested, and the role of IAA on potato-F. eumartii interaction is discussed.Fil: Terrile, Maria Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Olivieri, Florencia Pia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Bottini, Ambrosio Ruben. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias. Departamento de Economía, Política y Administración Rural.; ArgentinaFil: Casalongue, Claudia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentin

Potassium phosphite increases tolerance to UV-B in potato

The use of biocompatible chemical compounds that enhance plant disease resistance through Induced Resistance (IR) is an innovative strategy to improve the yield and quality of crops. Phosphites (Phi), inorganic salts of phosphorous acid, are environment friendly, and have been described to induce disease control. Phi, similar to other plant inductors, are thought to be effective against different types of biotic and abiotic stress, and it is assumed that the underlying signaling pathways probably overlap and interact. The signaling pathways triggered by UV-B radiation, for instance, are known to crosstalk with other signaling routes that respond that biotic stress. In the present work, the effect of potassium phosphite (KPhi) pre-treatment on UV-B stress tolerance was evaluated in potato leaves. Plants were treated with KPhi and, after 3 days, exposed to 2 h/day of UV-B (1.5 Watt m(-2)) for 0, 3 and 6 days. KPhi pre-treatment had a beneficial effect on two photosynthetic parameters, specifically chlorophyll content and expression of the psbA gene. Oxidative stress caused by UV-B was also prevented by KPhi. A decrease in the accumulation of hydrogen peroxide (H2O2) in leaves and an increase in guaiacol peroxidase (POD) and superoxide dismutase (SOD) activities were also observed. In addition, the expression levels of a gene involved in flavonoid synthesis increased in UV-B-stressed plants only when pre-treated with KPhi. Finally, accumulation of glucanases and chitinases was induced by UV-B stress and markedly potentiated by KPhi pre-treatment. Altogether, this is the first report that shows a contribution of KPhi in UV-B stress tolerance in potato plants

First Insights into the Role of Polyamines in Biotic Stress Resistance Induced by Potassium Phosphite in Potato

Phosphites (Phi) are inorganic salts widely used to protect plants from pathogens, especially oomycetes, on a wide range of horticultural crops. They were studied at the molecular level in different pathosystems; however, their mode of action is not completely understood. In this work, we investigate the participation of polyamine (PA) metabolism in the defence mechanism induced by Phi. To this aim, potato plants were treated with potassium phosphite (KPhi), and 3 days later, leaves were inoculated with Phytophthora infestans (P. infestans). The content of free PAs was analysed and showed that KPhi induced the accumulation of spermine (Spm), spermidine (Spd) and putrescine (Put). However, when treated leaves were detached and inoculated with P. infestans, their accumulation was even higher. In addition, the expression of genes implicated in PA synthesis and degradation was measured. Most of the genes involved in PA biosynthesis showed no changes in their expression upon KPhi treatment, except for those involved in Spm synthesis: Spm synthase and S-adenosyl-L-methionine decarboxylase (SPMS and SAMDC, respectively). On the other hand, the catabolism genes: diamine oxidase (DAO), and polyamine oxidases PAO1, PAO2 and PAO5, but not PAO4, decreased their expression after inoculation with P. infestans, and this was reverted by KPhi pretreatment. To investigate this effect, assays were carried out using a PA oxidase inhibitor showing that defence responses induced by KPhi do not require products of PA catabolism. Additionally, we analysed in silico phytohormone and stress-responsive cis-elements in PAO gene promoters, which might explain, at least in part, the different behaviour of PAO4.Fil: Lobato, Maria Candela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Feldman, Mariana Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Machinandiarena, Milagros Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Olivieri, Florencia Pia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentin

Undaria pinnatifida (Harvey) Suringar aqueous extract activates potato defense responses against Phytophthora infestans

Undaria pinnatifida is an invasive macroalga on the Argentinean coast and its use as a biostimulant was rarely investigated. The present work aimed to evaluate the effect of U. pinnatifida aqueous extract (AE) as an inducer of potato defense responses. Our results showed that AE protected potato plants against Phytophthora infestans infection. Application of AE by foliar spray (10% dilution), resulted in a significant reduction in disease severity. However, when antimicrobial activity was tested in vitro, a higher concentration of AE was needed to reduce P. infestans growth, showing hyphae morphological alterations and a reduction in the number of sporangia. These results indicate that AE at 10% induced defense responses in treated plants. The analysis of PR1 and IPII genes (Salicylic Acid and Jasmonic acid pathways markers, respectively), showed that the SA pathway would participate in the AE-induced resistance. Additionally, the accumulation of β-1,3-glucanase, guaiacol peroxidase activity, and changes in free polyamines were analyzed. An accumulation or enhanced activity was stimulated by AE. Moreover, the effect of AE treatment might be positive for the plant performance, since all the dilutions of AE tested showed an increment of chlorophyll content. These preliminary results strongly encourage further studies of the potential use of AE as an alternative tool for potato disease management using locally available seaweed in environmentally friendly conditions.Fil: Lobato, Maria Candela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Olivieri, Florencia Pia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Machinandiarena, Milagros Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Becherucci, Maria Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; ArgentinaFil: Feldman, Mariana Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentin

The reinforcement of potato cell wall as part of the phosphite-induced tolerance to UV-B radiation

Phosphites (Phis), inorganic salts of phosphorous acid, have shown to be effective in protection of plants against biotic stress. Recently, we have described that potassium phosphite (KPhi) induces tolerance to UV-B radiation in potato. To counteract the harmful effect of UV radiation, plants accumulate UV-screening compounds, such as flavonoids, sinapate ester, and lignin. In previous work, we have shown an increase in guaiacol peroxidase (POD) activity in plants pretreated with KPhi and further exposed to UV-B radiation. In order to continue with this study, the expression of different enzymes and components involved in cell wall reinforcement were analyzed. An isoform of POD induced by KPhi was analyzed by isoelectric focusing and further identified as suberization-associated anionic peroxidase (POPA) by mass spectrometry. In addition, other enzymes participating in lignin biosynthesis, like caffeoyl-CoA O-methyltransferase (CCoAOMT), determined by accumulation of transcripts, and laccase activity, visualized in zymogrames, were increased by KPhi treatment previous to UV-B exposure. Further, the accumulations of extensin (EXT) transcripts and of conjugated polyamines (PAs) were increased by KPhi treatment previous to UV-B exposure. All these results suggest cell wall reinforcement in leaves due to KPhi pretreatment followed by UV-B exposure.Fil: Machinandiarena, Milagros Florencia. Universidad Nacional de Mar del Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Oyarburo, Natalia Soledad. Universidad Nacional de Mar del Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Daleo, Gustavo Raul. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina. Universidad Nacional de Mar del Plata; ArgentinaFil: Andreu, Adriana Balbina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina. Universidad Nacional de Mar del Plata; ArgentinaFil: Olivieri, Florencia Pia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina. Universidad Nacional de Mar del Plata; Argentin

Effect of β-aminobutyric acid (BABA) on protection against Phytophthora infestans throughout the potato crop cycle

The protective effect of â-aminobutyric acid (BABA) on two potato cultivars (Bintje and Pampeana) with different level of horizontal resistance against Phytophthora infestans was investigated during the crop cycle. Plants were treated with BABA 16, 23, 30, 38, 44, 51, 61 or 75 days after emergence. After each application, the percentage of protection and the content of glucanases, chitinases and phenolic compounds were determined in detached leaves. The foliar pretreatment with BABA up to 30 days after emergence showed a 60% protection percentage against P. infestans in cv. Pampeana, while cv. Bintje almost reached 20%. The results indicated that the time of application of BABA and the susceptibility of the cultivars affect both, the protection against late blight and the expression of defense molecules like glucanases, chitinases and phenolic compounds.Fil: Altamiranda, E.A.G.. Universidad Nacional de Mar del Plata; ArgentinaFil: Andreu, Adriana Balbina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Daleo, Gustavo Raul. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Olivieri, Florencia Pia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentin

Cell Wall Reinforcement in the Potato Tuber Periderm After Crop Treatment with Potassium Phosphite

The potato periderm is the outer tissue of potato tubers that protects the tuber from pathogen attack, dehydration, and wounding during harvest and storage. It is of secondary origin and replaces the epidermis early during tuber development. The outer layers of the periderm consist of cells with suberized walls. Phosphite compounds (Phi) are inorganic salts from phosphorous acid, and in previous works, we demonstrated that they are effective in protecting potato plants from biotic stress caused by different pathogens and also against UV-B radiation. The aim of the present work was to study the effect of Phi on the post-harvest potato periderm structure, cell wall components, and related enzymes. In three years of field experiments, potassium phosphite (KPhi) applications were compared with an untreated control. KPhi was applied to seed tubers before planting combined with foliage application of a conventional fungicide, to seed tubers and foliage without conventional fungicide, or to seed tubers combined with foliage application of KPhi plus a conventional fungicide. Observations of periderm sections showed increases in suberin and pectin depositions in post-harvest tubers from KPhi-treated plants compared to tubers from non-treated plants. In addition, peroxidase activity increased in the periderm tissue. The activity of laccases and superoxide dismutase was measured in the protein extract of the periderm, and an increase due to KPhi treatment was detected. The results presented here suggest that early KPhi treatment to seed tubers followed by foliage treatment leads to reinforcement of the cell wall of periderm cells, which represents a benefit for future response to stresses.Fil: Lobato, Maria Candela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Daleo, Gustavo Raul. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Andreu, Adriana Balbina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Olivieri, Florencia Pia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentin