Yellowing of part or the whole canopy of chestnut trees was observed during the summer of 2014 in the areas of Tenno, Pranzo and Drena in Trentino, where chestnut cultivation is traditional on the lateral moraine deposit in a context of a prevalent limestone lithological substrate. Symptoms were observed on chestnut trees of different ages, either grafted or not, scattered or in stripes along the maximum slope. Investigations were carried out in the form of field surveys, chemical and molecular analyses of soil and leaves and greenhouse trials, to assess the damage evolution and its causes. No known pathogens emerged during the field surveys, and laboratory tests excluded the presence of phytoplasma infections. A comparison of yellow and asymptomatic leaves evidenced significant deficiencies of manganese (Mn) and iron (Fe) in the yellow leaves, though the level of the two nutrients was the same in the soil below green and yellowed trees. Experimental runoff trials in the greenhouse reproduced the foliage damage with a continuous basic water regime; open-field fertilizations with Mn and Fe led to a slightly recovery on the part of some treated leaves, thus confirming the hypothesis of a probable, sudden deficiency of the mentioned microelements. A possible explanation of this phenomenon could be, on one side, the high precipitation level of 2014, and, on the other side, the previous outbreaks of the Asian chestnut gall wasp, that had reduced foliage volume and thus the organic substance in the soil. The natural recovery from symptoms was clearly visible in trees after four years in a context of normal rainfall regime and increasing organic acid in the soil, after effective biological control of wasp. Even if related to a particular and rare geological condition, this is the first description of Mn deficiency in Castanea sativa.