The Begomovirus genus belongs to the Geminiviridae family, which includes pathogens that have circular genomes of single-stranded DNA with one or two components of 2700-3000 pb within incomplete icosahedral particles (geminated). They are responsible for several diseases that affect economically important crops in tropical and subtropical regions worldwide (^{Moffat, 1999}). Based on its host range, insect vector, genomic composition and sequence similarity, the Geminiviridae family is divided into seven genera (^{Varsani et al., 2009}; ^{Varsani et al., 2014}). One of them is the Begomovirus genus that includes over 60 species exclusively transmitted by a Bemisia tabaci species complex (^{Markham et al., 1994}). In Mexico, roselle (Hibiscus sabdariffa L.) is a crop of great economic importance, and Ayutla and Tecoanapa, Guerrero, are the municipalities with the greatest cultivated area at the national level (^{SIAP, 2015}). Worldwide, four viruses associated with this crop have been reported: Cotton leaf curl virus (CLCuV), Malva vein clearing virus (MVCV) (^{Brunt et al., 1996}), Okra mosaic virus (OkMV) (^{Stephan et al., 2008}) and Mesta yellow vein mosaic virus (MYVMV) (^{Chatterjee et al., 2008}). In Mexico, a begomovirus complex was found associated with roselle yellowing, including Okra yellow mosaic Mexico virus (OYMMV) (^{Velázquez et al., 2016}). In the 2015 cycle, the disease was detected in two plots in Tecoanapa with 100% incidence. Whitefly populations are very low on crops in that production area. Based on this, the objective of the present study was to find out if there are other insects that carry OYMMV or other begomoviruses.

In 2016, two samplings of the municipalities of Tecoanapa and Ayutla, Guerrero, were done (Table 1). The first sampling took place in the 2015 cycle, from August 3 to 5, when the crop was in its vegetative stage, in locations where there was a high incidence of plants showing yellowing. The second sampling took place during the 2016 cycle, from November 26 to 28, when calyces were being harvested, in plots where the incidence of yellowing was high. In all cases, insects were collected with a sweep net from roselle plants and surrounding weeds showing yellowing, vein clearing and mosaic, and placed in plastic containers containing 96% ethanol. In the laboratory, the insects were separated according to their morphological similarities and kept at -20 °C. Total DNA was extracted from 2 or 3 individuals in each insect group using CTAB (^{Sambrook and Rusell, 2001}). The remaining insects in each group were kept in ethanol to be identified later, in case they tested positive for begomoviruses through PCR using AV494/AC1048 universal primers and under the amplification conditions reported by ^{Wyatt and Brown (1996)}, which amplify a 550 pb fragment. The amplified products were sequenced and compared to those in the GenBank database. The insects that tested positive for begomoviruses were mounted, identified using taxonomic keys (Table 1) and photographed using an optical microscope.

During the first sampling, insects of the Thysanoptera (207 individuals), Coleoptera (76 individuals) and Hemiptera orders were found. Four families of the Hemiptera order were identified: Membracidae (17 individuals), Pyrrhocoridae (62 individuals from the Dysdercus genus) and Aleyrodidae and Cicadellidae. ^{Pérez et al. (2009)} studied the entomofauna associated with roselle in Chiautla de Tapia, Puebla, and reported 17 species belonging to six orders, 11 families and 19 genera. The authors reported Atta mexicana, Sphenarium purpurascens, Melanoplus spp. and Aphis gossypii as pests that cause considerable damage to the roselle crop, but they were not found in this study. The density of leafhoppers observed was higher than that of whiteflies.

From the first batch of insects collected, 45 groups of insects were analyzed by PCR, 8 of which were found to have the expected 550-pb fragment of begomovirus. On insects obtained from the first sampling, Okra yellow mosaic Mexico virus (OYMMV) was detected in Trypanalebra maculata, Agallia sp., Kunzeana scimetara and A. excavata, while in A. modesta we found Melon chlorotic leaf curl virus (MCLCuV) (Figure1, Table 2). MCLCuV was reported by ^{Brown et al. (2001)} in Guatemala, and they suggested that it is a new species derived from the Squash leaf curl virus (SLCV) group that includes bipartite begomoviruses native to Central America and Mexico. MCLCuV had not been detected before in roselle or in weeds associated with it in the study area. In the second sampling, OYMMV was found in T. maculata collected in San Miguel and Cortijo. Sida golden mosaic Buckup virus (SiGMBuV) was found in Agallia sp. from Cortijo. ^{Stewart et al. (2014)} point out that plants of the Sida genus are SiGMBuV hosts. However, ^{Ortega et al. (2017)} detected OYMMV in Sida collina, S. aggregata, S. acuta, S. hankeana and Malacra fasiata plants that were associated with roselle crops in the study area, but no SiGMBuV. This may be due to the fact that OYMMV has a higher transmission efficiency or is better able than SiGMBuV to infect the diversity of Sida species in this region. The four insect species that tested positive for begomoviruses belong to the Cicadellidae family. T. maculata and K. scimetara belong to the Typhlocybinae subfamily, which includes the Empoasca genus, including E. papayae (^{Acosta et al., 2017}) and E. devastans (^{Hague and Parasram, 1973}), known to be the vector of 16SrII phytoplasma that causes papaya bunchy top (PBT) disease (^{Acosta et al., 2017}). Another species from this subfamily known to be a phytoplasma vector is Alebroides nigroscutellatus, which transmits the phytoplasma Potato purple top roll (16SrIII-B) (^{Rojas, 2009}). To date, no viruses are reported to be transmitted by species of this subfamily. ^{Dietrich (2013)} mentions that the members of the Typhlocybinae subfamily prefer to eat parenchyma cells, which suggests that there is low or no probability that they acquired begomovirus that are limited to the phloem. On the other hand, Agallia excavata and A. modesta belong to the Deltocephalinae subfamily, whose members prefer phloem (^{Zahniser and Dietrich, 2008}) and, given this habit, would be able to transmit viruses. Such is the case of Dalbulus maidis, vector of Maize rayado fino virus (MRFV) that causes one of the most important diseases affecting maize in Latin America (^{Nault et al., 1980}). Within the Agallia genus there are species such as A. constricta and A. cuadripundata, which are confirmed vectors of the New Jersey variant of Potato yellow dwarf virus (Rhabdoviridae) and Wound tumor virus (Reoviridae) in eastern United States (^{Belatra et al., 2017}).

Figure 1 Leafhoppers associated with roselle that were collected at different locations in Ayutla and Tecoanapa, Guerrero, and tested positive for begomoviruses. a) Trypanalebra maculata ; b) Kunzeana scimetara; c) Agallia excavata; d) A. modesta. 

Although little is known about the interactions that cause geminivirus-vector specificity, several studies indicate that the coat protein is responsible. ^{Briddon et al. (1990)} demonstrated that the exchange of the African cassava mosaic virus (ACMV) coat protein gene (transmitted by whiteflies) along with the Beet curly top virus (BCTV) (transmitted by leafhoppers) altered the vector’s specificity and resulted in the transmission of this ACMV chimerical isolate by leafhoppers. On the other hand, ^{Roumagnac et al. (2015)} recently demonstrated that Alfalfa leaf curl virus is transmitted by Aphis craccivora and suggested that this virus be considered a new genus within the Geminiviridae family (and proposed it should be named Capulavirus), which would include different recently discovered geminiviruses that have no known vector.

In this study no begomoviruses were detected in the B. tabaci individuals analyzed (Access No. MG675920). However, this species is made up of multiple “biotypes” that differ in their level of competence as a vector, the number and type of endosymbionts, and their genetic composition (^{Brown et al., 1995}). Several studies indicate that at least two different mechanisms can explain the fact that begomoviruses are not transmitted by whiteflies: (1) the particles lose their capacity to penetrate the insect’s intestinal epithelium; (2) the virions can reach the insect’s hemolymph, but they cannot become correctly associated with the GroEL protein (^{Rosell et al., 1999}; ^{Morin et al., 2000}). Also, it has been demonstrated that certain Abutilon mosaic virus (AbMV) isolates are not transmitted by B. tabaci (^{Wu et al., 1996}; ^{Höfer et al., 1997}). In this case, the epithelial cells in the intestine of B. tabaci are the first barrier that begomovirus must cross in order to be transmitted, and it is probable that those AbMV isolates have lost their capacity to join the receptors within the whitefly’s digestive tract (^{Morin et al., 2000}).

It was demonstrated that Trypanalebra maculata, Kunzeana scimetara, Agallia excavata and Agallia modesta collected from roselle plants and weeds associated with yellowing, vein clearing and mosaic are carriers of three begomoviruses.