Patients with grass pollen allergy, commonly called pollinosis, often present reactivity to pollen allergens from a number of grass species due to cross-reactivity of IgE antibodies to pollen proteins present in pollen grasses. In this context, Italian rye grass (Lolium multiflorum) pollen of the Poaceae family cultivated in Southern Brazil has been considered a major sensitizing agent in patients with pollinosis. In this region, Italian rye grass is capable of producing a great amount of pollen. In addition to L. multiflorum, other Poaceae grasses are naturally grown as weed in Southern Brazil, but with no clinical relevance. Pollen extracts derived from homologous or heterologous grasses are often used for both diagnosis and treatment of seasonal allergy. However, no standardized L. multiflorum pollen extract is commercially available in Brazil and mixed grass extracts are commonly used for diagnosis and immunotherapy of grass pollen allergy. Further studies are required to better characterize the cross-reactivity between L. multiflorum and other grass pollen allergens for improving the diagnosis and immunotherapy to L. multiflorum pollen allergy.