Cota tinctoria and Orosius albicinctus: A new plant host and potential insect vector of ‘Candidatus Phytoplasma trifolii’ Chamran Hemmati 1,2 & Mehrnoosh Nikooei 1 & Hossein Pasalari 1 Received: 3 March 2018 /Accepted: 26 April 2018 /Published online: 2 May 2018 # Australasian Plant Pathology Society Inc. 2018 Abstract Samples of Cota tinctoria with phytoplasma symptoms including witches’ broom, stunting, twisting of the shoots and little leaf were observed in Bandar Abbas, Iran, in 2017. Phylogenetic analysis revealed that the phytoplasma associated with Cota tinctoria witches’ broom (CtWB) is a strain of ‘Candidatus Phytoplasma trifolii’. Furthermore, the phytoplasma was successfully detected in the leafhopper, Orosius albicinctus collected from the symptom-bearing plants. This work therefore reports a new phytoplasma-associated disease on Cota tinctoria and its putative insect vector. Keywords Golden marguerite . Phytoplasma . Potential vector . Witches’ broom Phytoplasmas (wall-less and phloem-restricted plant pathogen- ic bacteria) are recognized as a serious constraint for the culti- vation of many ornamental plants around the world. They can reduce the quality and yield of ornamental plants because of their unspecific symptoms and can cause severe losses (Bertaccini and Duduk 2010). Epidemics of these diseases have limited the cultivation of many ornamental plant species such as gladiolus, lily, chrysanthemum and rose. Phytoplasmas affect more than 600 plant species from tropical, subtropical to tem- perate climates all over the world (Jones 2002). The general symptoms include flower malformation, growth abnormalities, yellowing of leaves, elongation and etiolation of internodes, witches ’ broom, stunting, little leaf and virescence (Chaturvedi et al. 2010). In plant disease epidemics, insect vec- tors play a major role. Furthermore, polyphagous vectors have the potential to inoculate a wide range of plant species, which may differ in susceptibility (Weintraub and Beanland 2006). Cota tinctoria, also known as golden marguerite, yellow chamomile, is a species of perennial flowering plant in the Asteraceae family. It is a short-living plant often treated as biennial, native to Europe, the Mediterranean and Western Asia and naturalized in scattered locations in North America (Franke 2005). It produces excellent yellow, buff and golden- orange dyes, used in the past for fabrics. This species is grown in gardens for its bright attractive flowers and fine lacy foliage, and in Iran, especially in southern regions, this species is wide- ly planted as an ornamental plant in green landscapes. So far, no diseases and pests have been reported on Cota tinctoria. In January 2017, during a survey, typical symptoms of phy- toplasma disease, including witches’ broom, stunting, twisting of the shoots and little leaf were observed in several Cota tinctoria flowers planted in a green landscape in Bandar Abbas, Hormozgan province, Iran (Fig. 1). A preliminary study to determine whether a phytoplasma was associated with the symptom-bearing plant was performed. Simultaneously, insect samples, mainly planthoppers and leafhoppers (Cicadellidae), were collected using yellow sticky cards. The yellow sticky cards were replaced at weekly intervals and spe- cies were captured and checked for phytoplasma presence. Insects were preserved in absolute acetone until DNA extrac- tion (Fukatsu 1999). Ten samples of both symptom-bearing and three samples of asymptomatic Cota tinctoria were collected from green land- scapes of Bandar Abbas (N27°22′36″; E56°35′25″). Total DNA was extracted from leaves of all collected symptom- bearing and symptomless samples by using the cetyltrimethylammoniumbromide (CTAB) extraction proce- dure described by Sahu et al. (2012). Total DNA was extracted from ten individual leafhoppers using the CTAB protocol de- scribed by Reineke et al. (1998). To determine the presence of phytoplasmas in Cota tinctoria and insect samples, a nested polymerase chain * Chamran Hemmati Chamran.hemmati@hormozgan.ac.ir 1 Department of Agriculture, Faculty of Agriculture and Natural Resources, University of Hormozgan, Bandar Abbas, Iran 2 Plant Protection Research Group, University of Hormozgan, Bandar Abbas, Iran Australasian Plant Disease Notes (2018) 13: 13 https://doi.org/10.1007/s13314-018-0298-1