Co-Inoculation Effects of Thiobacillus thiooxidans Bacteria and Mycorrhiza (Glomus spp.) on Maize Nutrition at Different Levels of Sulfur

Sulfur is the key element for higher crops and plays an important role in the formation of proteins, vitamins, and enzymes. It is a constituent of amino acids such as cysteine and methionine, which act for the synthesis of other compounds containing reduced sulfur, such as chlorophyll and utilization of phosphorus and otheressential nutrients.Deficiency of this nutrient in soil is usually compensated by using chemical fertilizers. However, these fertilizers have harmful effects on the environment and decrease the quality of the agriculture products. Therefore, biological fertilizers are more useful for using in agricultural ecosystems.Sulfurshould be addedto the soil, usually in a reduced form such as elemental sulfur. Use of S oxidizers enhances the rate of natural oxidation of S and speeds up the production of sulfates and makes them available to plants consequently resulting in an increased plant yield. The role of chemolithotrophic bacteria of the genus Thiobacillus through oxidation process in the soil is usually emphasized. Sulfur oxidation is the most important step of sulfur cycle, which improves soil fertility. The result is formation of sulfate, which can be used by the plants, while the acidity produced by oxidation helps to solubilize nutrients in alkaline soils. These bacteria can solubilise the soil minerals through the production of H2SO4 that reacts with these non-soluble minerals and oxidised them to be available nutrients to the cultivated plants. Arbuscular MycorrhizalFungi isan important component ofthe microbiota, mutualistic symbioticsoilfungithatcolonizesthe rootsofmost cropplants.The AM symbiosis involves an about 80% of land plant species and 92% of plant families. They have theability to enhance host uptake of relativelyimmobile nutrientsparticularly phosphorus (P) andzinc (Zn),Manganese (Mn) andiron(Fe).Arbuscular mycorrhizal fungi increased plant uptake of phosphorus, nitrogen and water absorption. Inoculation withthesefungihas increased the yield of numerous field-grown crops. This study was aimed to evaluate the effects of thiobacillus bacteria and sulfur application on soil pH, and also their interactions with mycorrhizal fungi in order to improve nutrients uptake and grain yield of maize under alkaline soil condition. Treatments arranged as factorial experiment were based on RCBD with three replications. Treatments consisted of mycorrhizal inoculation: inoculated (m1) and non-inoculated (m0), thiobacillus in two levels of inoculated (t1) and non-inoculated (t0) and three levels of sulfur (S0: 0 kg.ha-1, S1: 250 kg.ha-1 and S2: 500 kg.ha-1). Four-row plots were prepared with row width and intra-row space of 60 and 20 cm, respectively. Seeds of maize (Zea Mays, Sc:647) were surface sterilized in a 10% (v/v) solution of hydrogen peroxide for 10 min, were rinsed with sterile distilled water. Before sowing, 300 kg of urea per hectare were applied according to the results of soil analysis. In order to facilitate oxidation of sulfur to sulfate form,, S was applied and thoroughly mixed into top 30 cm of soil 30 days before sowing. One week before sowing, thiobacillus (Thiobacillus thiooxidans) was inoculated. Inoculum of AM fungus Glomus intraradices, were added to soil just before planting at about 2 centimeters below seed sowing dept. To measure Arbuscular Mycorrhizal colonization, root plants collected one week before harvesting, cleared in 10% KOH at 80˚C for 2 h, and then acidified in 1% HCL for 60 min. Then the cleared roots were stained in a solution of Trypan blue. For nutrient analysis, the following procedure was applied. Zn, Fe, S, and P were determined by Inductively Coupled Plasma-atomic emission spectrometry apparatus. For this purpose, ash of seed samples was prepared at 500-550 degree of Celsius and then 5 ml of HCl 37% was added and with dionized water to reach to 50 ml. Kjeldahl method was used to determine nitrogen. Analysis of variance was performed on all experimental data and means were compared using the least Significant Differences (LSD) test with SAS software. The significance level was p>0.05 unless stated otherwise. Results showed sulfur application increased significantly the amount of S, P, N, Fe, Zn, shoot dry weight and leaf chlorophyll of maize. With increasing Sulfur, sulfur concentration in plant shoot increased with linear trend. The highest S concentration was obtained with 200 mg.kg-1 S and the lowest amount was obtained from control plots. Applications of 50, 100, 150 and 200 mg.kg-1 S increased P content about 0.45, 3.91, 4.74 and 5.56 %, respectively. The highest N contentwas obtained with 100 mg.kg-1 S. The thiobacillus significantly increased P, Fe, Zn anddecreased root colonization and soil pH compared to control. Thiobacillus bacteria increased shoot P only with application of 100 mg.kg-1 S. Mycorrhizal inoculation increased the amount of N, P, S, Fe, Zn, shoot dry weight and root colonization. Inoculation with G.intra and G.mosseae increased shoot P content about 4.18 and 3.34% in comparison with the control plots. Single or combination of sulfur and thiobacillus had a negative impact on the root colonization. Based on the results it seems that sulfur, thiobacillus and mycorrhiza in alkaline soils improved crops nutrition and growth. S application and thiobacillus interaction on S concentration of maize shoot were significant. In condition of 0 or 50 mg.kg-1 S application, inoculation of thiobacillus is recommended. Also, the effects of mycorrhiza on P shoot was significant with no application of S.