Isolation and identification of indigenous Arbuscular Mycorrhizal Fungi (AMF) of forest clove rhizosphere from Maluku, Indonesia

Mahulette AS, Alfian A, Kilkoda KA, Lawalata IJ, Marasabessy DA, Tanasale VL, Makaruku MH. 2021. Isolation and identification of indigenous Arbuscular Mycorrhizal Fungi (AMF) of forest clove rhizosphere from Maluku, Indonesia. Biodiversitas 22: 3613-3619. Forest clove is classified as wild-type and endemic to the Maluku (Moluccas) Islands, Indonesia. The different condition of growing areas causes various types of Arbuscular Mycorrhizal Fungi (AMF) associated with forest clove. The study aimed to identify and obtain indigenous AMF inoculums from the forest clove rhizosphere from two distribution areas in Maluku. The results of AMF identification found two types of spores from the genus Glomus in the rhizosphere of forest cloves from Ambon Island with a spore density of 35/50 g of soil. In comparison, three spores were found in Seram Island, two from the genus Scutellospora and one from the Acaulospora. With an overall spore density of 5/50 g of soil. After culture trapping, there was a change in type and an increase in spore density in soil samples from the rhizosphere of the two forest clove distribution areas. Soil samples from Ambon after trapping culture obtained two new types of spores from the genus Acaulospora with a total spore number of 57/50 g soil while in soil samples from Seram found three new types of spores from the genus Glomus with a total spore count of 104/50 g of soil.

Yield and Rhizosphere - microflora of Cowpea in Response to Magnesium Fertilization in Lateritic Soils of Kerala

Background: Magnesium deficiency has become a major nutritional disorder in lateritic soils of Kerala. Appropriate magnesium fertilization is the best strategy to combat deficiency issues. Apart from correcting nutritional deficiency, magnesium fertilization has an influence on the growth of beneficial microbes such as nitrogen fixing bacterias and arbuscular mycorrhizal fungi. The experiment aimed to investigate the effect of magnesium fertilization on crop yield and population rhizosphere micoflora of cowpea in lateritic soils of Kerala.Methods: A pot culture experiment was conducted with a gradient of magnesium additions ranging from 5 mg kg-1 to 80 mg kg-1 of soil along with recommended dose of fertilizers. Population of rhizobium, free living nitrogen fixing bacteria, spore count of arbuscular mycorrhizal fungi and per cent root colonization of arbuscular mycorrhizal fungi were studied during flowering. The available magnesium and magnesium uptake were also worked out during harvest. Yield and yield contributing characteristics of cowpea were measured during harvest stage.Result: Magnesium addition produced significant variations in population of rhizobium and free- living nitrogen fixing bacteria whereas spore count of AMF and per cent root colonization of AMF did not vary according to the added doses of magnesium. A higher population of rhizobium, free living nitrogen fixers, root nodules, magnesium uptake, plant height and yield were obtained in the treatment where magnesium was applied @ 10 mg kg-1 soil.

Shelf Life Studies of Seed Coat Formulation of Rhizobium and Arbuscular Mycorrhizal Fungus (AMF) for Pulses - A New Perspective in Biofertilizer Technology

Background: Arbuscular Mycorrhizal Fungi (AMF) and Rhizobium are beneficial plant partners exhibiting mutual association with crop plants. Conventional carrier based formulation has lesser population and limited shelf life. Present study was aimed to evaluate and to assess the shelf life in new formulation as well as in coated seeds under storage.Methods: Three experiments are conducted in this study. Population of Rhizobium and AM fungi spore count were assessed in newer seed coat formulation under room temperature storage. Secondly survival of these organisms on the coated seeds of blackgram and greengram were evaluated. In third experiment the coated seeds stored for different duration were evaluated for germination, growth and vigour index.Result: Rhizobial population was maintained at 1011 g-1 and AM spore load was 900-1000 spores g-1 up to one year of storage. AM spore count was also found sufficient up to 9 months of storage in coated seeds of both crops. Growth and vigour index were significantly enhanced in inoculated plants over uninoculated control. From the study, it was evident that these formulations can be stored upto one year at room temperature without any loss in viable count and can be used for seed coating.

Soil P, Arbuscular mycorrhizal spore count and root colonization of cowpea in biochar amended soils under maize/cowpea cropping systems

<p><strong>Abstract</strong></p><p>Cowpea, a food and nutrition security crop is being threatened by decline in soil fertility especially in small holder farmstead. The natural arbuscular mycorrhizal fungi in the soil could improve its ability to acquire and retain nutrients thereby leading to higher yield. This irrigated field research was conducted to determine the effect of biochar rates and cropping systems on selected soil chemical properties, soil microbial biomass carbon (SMBC), nitrogen (SMBN), phosphorus (SMBP), and vesicular-arbuscular mycorrhizal (VAM) spore count and mycorrhizal fungi colonization (AMF) of cowpea. Experimental design was 3 x 3 factorial in randomized complete block design (RCBD). Factor A was three cropping systems; sole cowpea, intercropping and intra-cropping, while factor B was three biochar rates; control (biochar at 0 t ha<sup>-1</sup> (B<sub>0</sub>)), biochar at 2.5 t ha<sup>-1 </sup>(B<sub>1</sub>) and biochar at 5 t ha<sup>-1</sup> (B<sub>2</sub>). These were replicated in three blocks to constitute 27 plots. The entire plot was cleared, ploughed and demarcated into beds with hoes and diggers. Cowpea sole or inter- or intra- cropped with maize were planted in a spacing distance of 25cm by 75cm, with intercropped cowpea being in-between the interrow spacing (75 cm), while the intracropped cowpeas was planted between the intrarow spacing (25 cm). Biochar soil amendment were applied two weeks after planting by making a groove in-between the rows in the soil and covering them with soil. The result showed that biochar soil amendment and interaction of biochar with cropping system significantly (p<0.05) affected SMBN, SMBC, total  VAM spore count and AMF colonization by cowpea, whereas cropping system significantly affected only total VAM spore count and AMF colonization by cowpea. B<sub>2</sub> amended soil had the highest SMBC content (0.028 mg kg<sup>-1</sup>) while the least was from control plot (0.021 mg kg <sup>-1</sup>), SMBN was highest in B<sub>1 </sub>amended soil (0.004 mg kg<sup>-1</sup>), followed by control plot (0.002 mg kg<sup>-1</sup>). Control had higher AMF and total VAM spore count while biochar amended soil had higher soil microbial properties. Considering the cropping systems, inter and intra-cropping had higher microbial biomass and total VAM spore count than sole cowpea whereas sole cowpea had higher AMF infection of cowpea than the intercropped cowpea. Biochar at 5 tha<sup>-1</sup> had the highest available P. Generally, this study showed superiority of the interaction of biochar with cropping systems over sole cropping in the improvement of soil properties in degraded soils of North-West province of South Africa.</p><p>Key words: Cropping systems; Chromic Luvisol; Microbial properties; Soil fertility; Soil amendment</p>

Arbuscular Mycorrhizal Fungi Colonization in the Rhizosphere of Aspilia pruliseta Schweif. ext Schweif in the Semiarid Eastern Kenya

The use of arbuscular mycorrhizal fungi (AMF) to enhance soil phosphate uptake is a biological technique considered to cure phosphate deficiency in soils. This study investigated association of Aspilia pruliseta Schweif shrub with AMF in Kenya. The study aims at profiling a tropical shrub with multiple ecological benefits that could reduce addition of chemical phosphatic fertilizer into the soil and reverse negative consequences of eutrophication. Sampling was purposive to have areas with or without Aspilia pruliseta vegetation growing. A small amount (10 g) of the soil from 27 composite samples was used for spore count determination and a similar amount for next generation sequencing. Spore counts varied significantly among soil textural types, sample locations, and soil depth. Sandy loam had the highest spore counts with a mean average of 404 spores. The spore count decreased significantly ( P < .05) with the depth of soil from a mean of 514 spores to 185 along the rhizosphere. The intensity of spore morphotypes was significantly higher at P < .05 for soils whose vegetation was covered with Aspilia pruliseta than those without. Aspilia pruliseta vegetation used together with sandy loam soil could culture commercial mycorrhiza fungi production for use in agrisystems.

An aero mycological analysis of Mucormycetes in indoor and outdoor environments of northern India

Mucormycosis is an angio-invasive infection, predominantly acquired by inhalation of sporangiospores from the environment. However, the burden of Mucormycetes sporangiospores in the air is not well studied. We aimed to estimate the burden of Mucormycetes spores in the outdoor and indoor (hospital) environment across different seasons in north India. A total of 380 air samples from outdoor (n = 180) and indoor (n = 200) environment were included in the study. Air samples were suctioned using air sampler (100 l/min) and cultured on Dichloran Rose Bengal Chloramphenicol (DRBC) with benomyl for selective isolation of Mucormycetes. The isolates were identified by phenotypic and genotypic methods. The mean spore count (±SD) of Mucormycetes (cfu/m3) in outdoor samples varied from 0.73 (±0.96) to 8.60 (±5.70) across different seasons. In hospital, the mean spore count varied from 0.68 (±1.07) to 1.12 (±1.07) and 0.88 (±1.01) to 1.72 (±2.17) for air-conditioned wards and non-air-conditioned wards, respectively. Rhizopus arrhizus was the predominant agent isolated from both indoor and outdoor environment followed by Cunninghamella species. We also report a single isolate of the rare mucormycete agent, Apophysomyces variabilis from outdoor environment. The present study highlights the presence of low spore burden of Mucormycetes in outdoor and hospital settings in north India. This study also reports the first isolation of A. variabilis from air samples in the Indian subcontinent.

Arbuscular Mycorrhizal and endophytic (DSE) association in the dominant grasses of Melghat forest (Phase -I), India

Investigations on arbuscular mycorrhizal Fungi (AMF) and dark septate endophytic (DSE) association in some of the dominant grasses from Melghat forest (phase I) of Satpura terrain India, was carried out to examine their existence and symbiotic relationships with the host plants. This forest area was not surveyed earlier by anybody to evaluate the AMF status diversity hence it was decided to survey the buffer and core area. Rhizosphere soil of each sampled grass was analyzed for the AM fungal structures in the roots to study percent root colonization by AMF and AMF spore density in rhizospheric soils of respective samples. The composite soil sample was prepared for each site and used for physicochemical analysis by standard methods. In the first phase of project, forty-eight dominant grass species from twenty-one different sites were collected along with roots and rhizospheric soil to find out AMF and DSE status of grasses. Both type of fungal associations was found in almost all the grass species collected during studies. All of them were found colonized by AMF hyphae along with moderate to poor development of mycorrhizal structures in roots. DSE colonization was also found in maximum forty-three grasses. Physico- chemical characterization of all the soil samples were performed to find out its correlation with AM percent colonization and spore count. Mean AMF percent colonization were in between 1.33 to 52.85 and DSE in with 0.00 to 18.97. Viable AM spore count were in between 0 to 98 per 100g of soil. Altogether four AMF genera with its thirty-nine-different species were isolated and identified. A data base of indigenous AM species richness for Melghat forest has been generated to plan and design the future management practices for grasses establishment and development especially in burnt and over grazed areas.

Soil organic matter and its active pools as affected by legume/cereal cropping systems under some agronomic practices in the derived savannah zone of Nigeria

Introduction of grain legumes facilitates farmers’ acceptance in adopting legume cropping system. This study evaluated effect of four cropping systems on soil organic carbon (SOC), total vesicular arbuscular mycorrhiza (VAM) spore count, soil microbial biomass carbon (SMBC), soil microbial biomass nitrogen (SMBN) and soil microbial biomass phosphorus (SMBP) within two locations (Nsukka and Moniya) of the derived savannah zone of Nigeria. The experiment was set up in a 4 x 2 x 2 factorial in randomized complete block design (RCBD), represent- ing 4 cropping systems, 2 fertilizer application rates (0 and 60 kg N ha-1) and 2 residue management methods (incorporated and not-incorporated). This set-up was replicated three times and the experiment was repeated the following year (2008 and 2009). At the end of the experiment, soil samples for laboratory analy- ses were collected from 0 – 20 cm soil depth using auger. The result showed that SOC was not significantly (p≤0.05) affected by all factors and their interaction at both locations. In Nsukka, cropping systems and their interaction with residue management significantly (p≤0.05) affected SMBP and total VAM spore count. Continuous maize had the highest total VAM spore count (280), which was sta- tistically the same with cowpea/maize rotation (277). Soybean/maize cropping system had the least value (259). Velvet bean/maize cropping system had the highest SMBP (0.0007 mg g-1). In Moniya, SMBC, SMBN and SMBP were sig- nificantly (p≤0.05) affected by the cropping systems with velvet bean/maize plots having highest values (0.053, 0.006, 0.0007 mg g-1). Residue incorporation and fertilizer application (60 kg ha-1) significantly (p≤0.05) increased SMBC and SMBN at Moniya. Interaction between cropping systems and residue manage- ments at this location also significantly affected SMBC, SMBP and SMBC. In this study, SMBP distinguished the effect of the factors on soil quality better than the other measured parameters. Velvet bean-based cropping system improved soil microbial properties better than other cropping systems.

Aerobiological studies with special reference to airborne basidiospores of Ganoderma Karst. at Pune, Maharashtra, India.

The basidiospores of Ganoderma were trapped in abundance during the aerobiological studies at Pune, during the year 2012. The sampling was conducted during the rainy months from 18th June to 10th September. Volumetric continuous Tilak Air sampler was used for sampling. Maximum number of spores was observed in the month of August (25760/m3 of air), followed by September (16548/m3 of air). The total spore number was 47460/m3 of air. The highest spore catch (3472/m3 of air) was recorded on 1st September. The relationship between the airborne concentration of Ganoderma spore count and meteorological factors are discussed in the paper. This information could be important for the sensitive individuals as Ganoderma spores are known to be allergic.