The samples were stored in cold rooms (−20 ☌), and then subdivided into two portions: one portion for the isolation of spores and identification of AMF through microscopy, and another portion for DNA extraction and sequencing of the ITS region. The zones were characterized into agricultural areas of cocoa, orange, and green cultivation, among other plants associated with this agroecosystem. The sample was collected at a depth of 20 cm, in the upper humic layer of each tree. Semi-conservative practice involves the low use of synthetic fertilizers and herbicides (NPK and glufosinate-ammonium, frequency of no more than 4 times per year), supplemented by organic fertilizers and cultural practices. ![]() From each canton, five cocoa trees cultivated using conservative practices and five cultivated using semi-conservative practices were selected. Soil and root samples were collected between October and December 2020 in three cantons in Ecuador (Milagro, Vinces, and Calceta) ( Figure 1 and Table A2). AMF hyphae can extend beyond depletion zones, increasing the area of mineral uptake and mineral content in the plant, exploring a larger volume of soil than could be achieved by root growth alone. Once an interaction is established, AMF modifies the morphology of the root, intracellularly colonizing its cortex using hyphae that generate specialized structures that act as organs of nutrient exchange between the plant cell and host. These fungi are associated with 90% of the plant families on earth and sustain terrestrial ecosystems through mutualistic symbiotic associations. ![]() In recent years, new genera continue to be added to the phylum Glomeromycota, and others already classified have either been reclassified or kept their original classification following much discussion ( Table A1). In this regard, the importance of arbuscular mycorrhizal fungi (AMF) has increased in the last decade, with numerous reports of beneficial effects on crops. Ĭountries such as Spain, Mexico, Canada, the United States, and China, among others, have been using mycorrhizal microorganisms to develop biological products applied to commercial crops. The main root of fine aroma cocoa extends up to 2 m deep, and the secondary roots develop in the upper humic layer (between the first 20 cm of depth) and extend horizontally up to 6 m around the main stem, where a wide diversity of mycorrhizal fungi cohabit. The average size of a cocoa crop is 3 ha, and each cocoa tree reaches a height of 4 to 8 m, except for fine aroma cocoa cultivars, which can reach up to 12 m in height. According to the Ministry of Production, Foreign Trade, Investment, and Fisheries, the Ecuadorian cocoa sector exported USD 815.5 million in 2020, and reached USD 266.4 million between January and May 2021 the main destination countries for Ecuadorian cocoa in 2020 included the United States, Indonesia, Malaysia, and the Netherlands. Ecuador is considered the world’s leading producer of fine and aromatic cocoa ( Theobroma cacao L.), with a contribution of approximately 63%, followed by Indonesia with 10%. ![]() bernensis.Įcuador is an agricultural country by tradition, with cocoa being one of the main exportation products. Seven genera were observed for the first time in cocoa crop agroforestry systems, including P. An abundance of 348 and 114 taxa were observed, corresponding to the conservative and semi-conservative practices, respectively. Although the metabarcoding analysis showed that diversity indices showed a higher increase in the roots than in the cocoa soil, independently of the cultivation practice. Morphological analysis showed that the density of Glomeromycota spores exhibited significant differences between production systems. In this context, our work aims to discover the diversity of arbuscular mycorrhizae associated with two cocoa cultivation practices (conservative and semi-conservative) through the isolation of spores using microscopy and metabarcoding of the internal transcribed spacer region (ITS). To the best of our knowledge, there are no known studies of the fungal communities in the fine aroma cocoa cultivars. Studies report different short- and long-term cultivation practices that influence the abundance and diversity of Glomeromycota. Arbuscular mycorrhizal fungi are key to plant nutrition and agriculture. ![]() In recent years, new data on the diversity of genera and species in the phylum Glomeromycota continue to be added and rearranged.
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