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Soil Application of Effective Microorganisms (EM)

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Soil Application of Effective Microorganisms (EM) International Journal of Molecular Sciences Article Soil Application of Effective Microorganisms (EM) Maintains Leaf Photosynthetic Efficiency, Increases Seed Yield and Quality Traits of Bean (Phaseolus vulgaris L.) Plants Grown on Different Substrates Marcello Iriti 1,* , Alessio Scarafoni 2 , Simon Pierce 1 , Giulia Castorina 1 and Sara Vitalini 1,* 1 Department of Agricultural and Environmental Sciences, Milan State University, 20133 Milan, Italy; [email protected] (S.P.); [email protected] (G.C.) 2 Department of Food, Environmental and Nutritional Sciences, Milan State University, 20133 Milan, Italy; [email protected] * Correspondence: [email protected] (M.I.); [email protected] (S.V.); Tel.: +39-025-0316766 (M.I.) Received: 18 March 2019; Accepted: 9 May 2019; Published: 10 May 2019 Abstract: EM (effective microorganisms) is a biofertilizer consisting of a mixed culture of potentially beneficial microorganisms. In this study, we investigated the effects of EM treatment on leaf in vivo chlorophyll a fluorescence of photosystem II (PSII), yield, and macronutrient content of bean plants grown on different substrates (nutrient rich substrate vs. nutrient poor sandy soil) in controlled environmental conditions (pot experiment in greenhouse). EM-treated plants maintained optimum leaf photosynthetic efficiency two weeks longer than the control plants, and increased yield independent of substrate. The levels of seed nutritionally-relevant molecules (proteins, lipids, and starch) were only slightly modified, apart from the protein content, which increased in plants grown in sandy soil. Although EM can be considered a promising and environmentally friendly technology for sustainable agriculture, more studies are needed to elucidate the mechanism(s) of action of EM, as well as its efficacy under open field conditions. Keywords: EM technology; food security; sustainable crop production; pulses; biofertilizer; biocontrol agents 1. Introduction The global food security challenge is straightforward: by 2050, the world must feed around 9 billion people, and, consequently, the demand for food will increase by 60%. Therefore, progress towards food security requires that food is available, accessible, and of sufficient quantity and quality to ensure good nutritional outcomes, particularly in protracted socioeconomic crises [1]. In this alarming scenario, new approaches for crop production are more than ever of paramount importance. Biostimulants, including plant-growth promoting microorganisms, have been shown to increase plant nutrient uptake, growth, and yield via different underlying mechanisms such as changes in soil structure, nutrient solubility, root growth and morphology, plant physiology, and symbiotic relationships. In addition, they can improve the plant tolerance to abiotic stresses, as well as the resistance to pathogens [2,3]. EM (effective microorganisms) is an environmentally friendly technology consisting of a fermented mixed culture of coexisting and mutually compatible microorganisms in an acidic medium. This biofertilizer contains up to 80 different species belonging to five main groups of microorganisms, including photosynthetic bacteria (Rhodopseudomonas palustris, Rhodobacter sphaeroides), Int. J. Mol. Sci. 2019, 20, 2327; doi:10.3390/ijms20092327 www.mdpi.com/journal/ijms Int. J. Mol. Sci. 2019, 20, 2327 2 of 9 lactic acid bacteria (Lactobacillus plantarum, L. casei, Streptococcus lactis), yeasts (Saccharomyces cerevisiae, Candida utilis), actinomycetes (Streptomyces albus, S. griseus), and fermenting fungi (Aspergillus oryzae, Penicillium spp., Mucor hiemalis)[4,5]. Similarly to other biostimulants, EM can positively affect plant nutrition, modify root morphology, and selectively change the rhizosphere–microbial community structure. The use of EM as an amendment has been reported for different crops by some authors to enhance soil fertility, increase crop yield, and control plant diseases [5–12]. However, in cotton plants, application of EM alone did not increase yield significantly over control, though in combination with organic matter it resulted in a 44% increase in yield over control [13]. Similarly, in green manure Int. J. Mol. Sci. 2019, 20, x 2 of 9 amendment, EM application resulted in a significant decline of 23% in grain yield of mung bean, while it significantlybiostimulants, increased EM can positively grain yieldaffect byplant 24% nutrition, and 46%modify in farmyardroot morphology, manure and andselectively NPK fertilizer amendments,change respectively the rhizosphere–microbial [7]. In an open community field experiment structure. The over use twoof EM years, as an EMamendment alone orhas in been combination reported for different crops by some authors to enhance soil fertility, increase crop yield, and control decreased tomato yield by 27–49% [14]. plant diseases [5–12]. However, in cotton plants, application of EM alone did not increase yield The esignificantlyffects of EM over oncontrol, photosynthesis though in combination have alsowith organic been investigated.matter it resulted in EM a 44% treatment increase in increased the photosynthesisyield over control rate in [13]. cabbage Similarly, plants, in green as wellmanure as amendment, stomatal conductance EM application and resulted intracellular in a CO2 concentrationsignificant [15]. decline Similarly, of 23% in photosynthetic grain yield of mung effi bean,ciency while increased it significantly in increased periwinkle grain plantsyield by after EM 24% and 46% in farmyard manure and NPK fertilizer amendments, respectively [7]. In an open field application [11]. Nonetheless, little is known about the possible variations of the nutritional experiment over two years, EM alone or in combination decreased tomato yield by 27–49% [14]. value of grainsThe fromeffects plantsof EM on treated photosynthesis with thishave liquidalso been microbial investigated. inoculant, EM treatment nor increased the physiological the mechanismsphotosynthesis underlying rate any in variation.cabbage plants, We as hypothesize well as stomatal that increasedconductance yieldand intracellular and nutritional CO2 quality of bean (Phaseolusconcentration vulgaris [15]. L.)Similarly,occur phot andosynthetic are associated efficiency with increased maintenance in periwinkle of photosynthetic plants after EM efficiency. application [11]. Nonetheless, little is known about the possible variations of the nutritional value of Additionally, we hypothesize that yield increases are evident when EM is applied even on nutrient grains from plants treated with this liquid microbial inoculant, nor the physiological mechanisms poor substrates.underlying Thus, any wevariation. investigated We hypothesize the eff thatects increased of EM onyield leaf andin nutritional vivo chlorophyll quality of abeanfluorescence of photosystem(Phaseolus II vulgaris (PSII), L.) seed occur yield, and are and associated macronutrient with maintenance content of ofphotosynthetic bean plants efficiency. grown in two different substrates.Additionally, we hypothesize that yield increases are evident when EM is applied even on nutrient poor substrates. Thus, we investigated the effects of EM on leaf in vivo chlorophyll a fluorescence of 2. Resultsphotosystem II (PSII), seed yield, and macronutrient content of bean plants grown in two different substrates. The ratio of variable to maximal fluorescence (Fv/Fm), which reflects the maximal photochemical 2. Results yield of PSII centers, is highly correlated with the quantum yield of net photosynthesis of treated and untreatedThe leaves. ratio of Forty variable days to maximal after sowing,fluorescence di (Ffferencesv/Fm), which were reflects significant the maximal among photochemical treatments and yield of PSII centers, is highly correlated with the quantum yield of net photosynthesis of treated and substrates.untreated In particular, leaves. Forty Fv/F mdaysremained after sowing, at optimal differences levels were (~0.83) significant for atamong least treatments two weeks and longer for EM-treatedsubstrates. plants, In regardless particular, F ofv/F soilm remained (Figure at 1optimal). By thelevels time (~0.83) of for plant at least senescence two weeks (atlonger 53 for DAS, days after sowing),EM-treated differences plants, regardless in photochemical of soil (Figure yield 1). By of th PSIIe time were of plant determined senescence (at mainly 53 DAS, by days substrate, after being particularlysowing), low on differences sandy soil in photochemical compared to yield the of richer PSII were substrate. determined mainly by substrate, being particularly low on sandy soil compared to the richer substrate. Figure 1. EFigureffects 1. of Effects EM (eofff EMective (effective microorganisms) microorganisms) treatments treatments on on maximum maximum efficiency efficiency of photosystem of photosystem II (Fv/Fm) ofII bean (Fv/F plantsm) of bean grown plants on grown greenhouse on greenhouse substrate substr andate and sandy sandy soil. soil. DAS, DAS, daysdays after sowing. sowing. Results are expressedResults as are mean expressed (n = 15) as mean and ( errorn = 15) barsand error indicate bars indicate the standard the standard deviation deviation (SD). (SD). Int. J. Mol. Sci. 2019, 20, 2327 3 of 9 Int.Int. J. J.Mol. Mol. Sci. Sci. 2019 2019, 20, 20, x, x 3 of3 of 9 9 EM application significantly increased all the seed yield properties, i.e., seed number per plant, EMEM application application significantly significantly increased increased all all the the seed seed yield yield
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