Short Communication Escola Superior de Agricultura ‘Luiz de Queiroz’- ESALQ USP Broccoli Stunt, a New Disease in Broccoli Plants Associated with Three Distinct Phytoplasma Groups in Brazil B arbara Eckstein 1 ,J ulio C esar Barbosa 2 , Patr  ıcia Fabretti Kreyci 2 , Maria Cristina Canale 2 ,K atia Regiane Brunelli 3 and Ivan Paulo Bedendo 2 Authors’ addresses: 1 Empresa Brasileira de Pesquisa Agropecu aria – Embrapa Florestas, Estrada da Ribeira, Colombo, Paran a, CEP 83411-000, Brazil; 2 Departamento de Fitopatologia e Nematologia, Universidade de S~ ao Paulo, CP 09, CEP 13418-900, Piracicaba, S~ ao Paulo, SP, Brazil; 3 Sakata Seed Sudamerica Ltda, CP 427, CEP 12906-840, Braganc ßa Paulista, S~ ao Paulo, SP, Brazil (correspondence to B. Eckstein. E-mail: barbaraeckstein@gmail.com) Received November 3, 2012; accepted January 13, 2013 Keywords: Brassica oleracea var. italica, broccoli stunt, phytoplasmas, Brazil Abstract Since 2007, a new disease in broccoli (Brassica oleracea var. italica Plenck) has been observed in the S~ ao Paulo state, Brazil. The characteristic symptoms of the disease are plant stunting, inflorescence malformation, redden- ing of the leaves and phloem necrosis. Nested polymer- ase chain reaction with P1/Tint and F2n/R2 primer pairs revealed the presence of phytoplasmas in diseased broccoli plants. Restriction fragment length polymor- phism and phylogenetic analysis of the 16S rDNA gene showed that phytoplasmas belonging to 16SrI, III and XIII groups were associated with the plants. To the best of our knowledge, this is the first report of phytoplas- mas in this Brassica species in Brazil, as well the first time phytoplasmas of 16SrIII and XIII groups have been associated with broccoli plants. Introduction Broccoli (Brassica oleracea var. italica Plenck; Brass- icaceae) is considered to be an important vegetable worldwide. Recently, a disease in broccoli named broccoli stunt has been observed in the S~ ao Paulo state, Brazil, causing losses of c. 3 to 10% of the crop. Disease symptoms include plant stunting with shortened internodes, inflorescence malformation, reddening leaves and vessel necrosis (Fig. 1). The affected plants are unmarketable; therefore, even low incidences cause economic damage for growers. In Brazil, phytoplasmas have been associated with diseases in brassica crops (Canale et al. 2012), includ- ing cauliflower stunt disease in Brassica oleracea L. subsp. botrytis (L.) Meztg (Rappussi et al. 2012), the symptoms of which are similar to those observed in broccoli plants (Fig. 1). Considering this symptom similarity, we investigated the association of phytoplas- mas with diseased broccoli plants in Brazil. Materials and Methods Plant samples and DNA extraction Diseased broccoli plants were collected between 2007 and 2009 from three municipalities (Braganc ßa Paulista, Morungaba and Sorocaba) of the S~ ao Paulo state. Total DNA from 51 symptomatic plants (Table 1) and (a) (b) Fig. 1 Broccoli plants infected by phytoplasmas. (a) stunted plants, reddening leaves of the plant; (b) necrosis of phloem vessels J Phytopathol doi: 10.1111/jph.12087 © 2013 Blackwell Verlag GmbH