Microbial isolation The Degree of Mycorrhizal Infection

commit to user “biofertilizer” Andrés et al. 2009; Fedderman 2010. Recent studies have focused on the application of AM fungi to crops Tan et al. 2011; Golubski 2011; Hart and Forsythe 2012. The effects of AM fungi on leguminous crops have been confirmed with sterilized pot experiments and by studying plant growth under ecologically damaged conditions Meghvansi et al. 2008; Miransari 2011. Studies have shown that AM fungi can enhance the ability of soybean to absorb nutritional elements while improving both the nitrogen-fixing ability of Rhizobium and the colonization structure in the rhizosphere niche, thus increasing yields and economic efficiency of soybean Tian et al. 2013. Furthermore, the results of research by Jie 2013 showed that AM fungi community structures in the rhizosphere soils were significantly different among different soybean cultivars at the branching stage. This research aims to study the effect of the use of sorts manure cow, goat, quail and mycorrhiza on soil microbial biodiversity, as well as its influence on soybean yields in marginal lands. Materials and Methods The study was conducted by the method of isolation of therhizosphere soil directly from perakaran area rhizosphere of some soybean plants. The identification was conducted atSoil Biology Laboratory, Faculty of Agriculture, University of Sebelas Maret. Research used Complete Randomized Block Design consists of two factors, namely the treatment of mycorrhiza treatment with two levels M0: without inoculation mycorrhiza and M1: inoculation mycorrhiza and the type of manure treatment with five levels P0: without manure, P1: 10 ton cowshed fertilizer Ha -1 , P2: 10 ton goat manure Ha -1 , P3: 10 ton quail manure Ha -1 , P4: 5 ton straw Ha -1 to obtain 10 combinations of treatments, each of which was repeated 3 times. Implementation of the study include: tillage, manure application, the application of mycorrhiza and soybean planting, maintenance, harvesting.

A. Microbial isolation

Soil sampling carried out for the isolation and identification of soil microbes. Soil samples were taken for each treatment plot near soybean roots, soil sampling was repeated 3 times, each soil sample was taken ± 200 g, sample was put into the soil in a plastic bag and given code. Put in10 g sample of soil into 90 ml physiological saline, then shaken until homogeneous 10-1 dilution. Take 1 ml solution of 10-1, enter into 9 ml physiological saline, then shaken until it became homogeneous dilution 10-2, did the same thing until the dilution became 10-5. Took 0.1 ml solution of 10-5 and poured into NA medium for bacteria and PDA media for mushrooms, then smooth over the media, did the same thing to all of dilution. Incubating the isolates at room temperature with petridish in inverted position, stored at room temperature, isolated after purification performed 4-7 days old. Isolates were identified morphologically after the age of 3 days, then count colonies of bacteria and fungi that grow.

B. The Degree of Mycorrhizal Infection

The degree of mycorrhiza infectionwas done using root staining technique Phillip and Hayman 1970. Roots were washed and cut into pieces, and then put in a film canister, after that added 2.5 KOH, closed the tubes, left for a day. After that, the roots were washed and fed with a solution of 2 HCl, soaked for 24 hours. Then replaced with a staining solution, left for 24 hours and kept on film canisters. To calculate the root infection, take root pieces with a length of ± 1 cm of 5 pieces per treatment, then placed in a preparation glass and closed with cover glass. Calculation of the percentage of the root infection was seen using microscope. Then gave positive sign when there was arbuscular mycorrhizal structure hyphae, vesicles, arbuscular every visibility distance. Every 1 cm root gave visibility 7- 10. When the infected roots couldn’t have been calculated yet, it could be stored in the refrigerator. The percentage of infected root is calculated with the following formula:

C. Diversity Index