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The production of inoculum from arbuscular mycorrhizal fungi (AMF) at a large scale and low cost is essential for establishing methods to assist in producing pineapple plantlets with high nutritional and phytosanitary quality. However, this objective is difficult to accomplish because of the biotrophic nature of these fungi. The on-farm multiplication method for AMF inoculum presents a good alternative to supply the demand for the production of glomerospores. This study aimed to multiply and evaluate AMF inoculum originating from isolated species (including Rhizophagus clarus, Claroideoglomus etunicatum) versus native AMF from pineapple and coffee plantations multiplied by the on-farm method on the colonization in pineapple plantlets. Initially, inocula of R. clarus, C. etunicatum, and native AMF (pineapple and coffee) were multiplied by the on-farm method in Sorghum bicolor. After four months, the number of AMF spores and the percentage of viable spores at the layers of 0.00-0.05 and 0.05-0.10 m were evaluated. There were no differences in spore numbers in relation to the source of the inoculum (R. clarus, C. etunicatum, pineapple, and coffee) and evaluated layers, with an average number of 605 spores per 100 cm of soil. The percentage of viable spores was greater at the layer of 0.00-0.05 m (76.32 %) compared to the layer of 0.05-0.10 m (72.05 %), regardless of the inoculum source. The viability of the inoculum obtained from C. etunicatum was higher than that from the coffee crop (77.93 and 68.06 %, respectively). Subsequently, the spores were inoculated in pineapple plantlets to assess the rate of colonization. Pineapple plantlets inoculated with AMF had an average of colonization Division – Soil Processes and Properties | Commission – Soil Biology Moreira et al. Effect of inoculation of pineapple plantlets with arbuscular mycorrhizal... 2 Rev Bras Cienc Solo 2019;43:e0180148 INTRODUCTION Arbuscular mycorrhizal fungi (AMF) are important components of soil that can be exploited to enhance the development of crops and contribute to the establishment of more sustainable agriculture. This approach would reduce, or even eliminate, the need for chemical fertilizers and pesticides in organic farming (Douds Jr et al., 2008, 2010). This is possible because plants colonized by AMF exploit higher volumes of soil (Smith and Read, 1997) and have higher nutrient uptake (Smith et al., 2011), as well as increased tolerance to drought, saline stress (Augé, 2001), and heavy metals (Rozpadek et al., 2014). Thus, they provide many additional benefits to acting as plant growth promoting agents (Azcón-Aguilar et al., 1997) and as a biological control against phytopathogens (Pozo et al., 2002; Moreira et al., 2016). Arbuscular mycorrhizal fungi are obligate biotrophic (Douds Jr et al., 2006, 2008), and complete their life cycles solely associated with the roots of living plants. Consequently, they cannot be multiplied separately in a defined culture medium (Douds Jr et al., 2006). Because of this, it is very difficult to develop low-cost methods for high-quality inocula production at large scales (IJdo et al., 2011). Thus, these fungi are usually multiplied in host plants in culture pots, aeroponic cultivation, hydroponics, or in vitro culture with genetically transformed roots (IJdo et al., 2011). These processes are carried out under controlled or semi-controlled conditions, such as greenhouses or growth chambers (IJdo et al., 2011). To stimulate inocula production, studies have been developed to test the multiplication of spores under field conditions, called the on-farm method. These studies explore mycorrhizal colonization with fungal isolates that are environmentally adapted to local conditions, potentially representing a low-cost alternative for farmers (Douds Jr et al., 2006, 2008, 2010; Schlemper and Stürmer, 2014). This method allows farmers and nursery workers to access inoculums with the most effective strains for their culture and their soil and climate conditions; furthermore, they can produce seedlings already mycorrhizal, with this benefit enhancing the establishment of seedlings in the field (Douds Jr et al., 2008). Schlemper and Stürmer (2014) showed that multiplication on-farm with Rhizophagus clarus and Claroideoglomus etunicatum grown in agro-industrial residues, such as sugarcane bagasse, is a good strategy for the multiplication of AMF, leading to excellent inoculum potential and large numbers of spores. The production of pineapple plantlets [Ananas comosus (L.) Merril] inoculated with AMF at the acclimatization stage improves the growth and nutrient uptake by the plants, especially under conditions with low P levels (Moreira et al., 2015). This approach also enhances the activity of various antioxidant enzymes responsible for ensuring greater resistance to pathogens (Moreira et al., 2016). The inoculum production in large-scale, easy to apply and low cost would then be the next step to establishing a methodology that would help in the production of pineapple plantlets with nutrition and phytosanitary of 18 and 67.73 % after 50 and 180 days cultivation, respectively, with no significant difference being detected between treatments. Therefore, on-farm inoculum production was effective at multiplying the AMF of both isolates of R. clarus and C. etunicatum, as well as for commercial crops (pineapple and coffee), with spores having high viability. Arbuscular mycorrhizal fungi colonized pineapple plantlets independently of the inoculum utilized and favored its growth.

revista brasileira de ciência do solo

Effect of Inoculation of Pineapple Plantlets with Arbuscular Mycorrhizal Fungi Obtained from Different Inoculum Sources Multiplied by the On-Farm Method