First report of the cherry borer Grapholita packardi (Zeller) (Lepidoptera: Tortricidae) attacking hawthorn fruits (Crataegus mexicana ) in Veracruz, Mexico

. Mexican hawthorn Crataegus mexicana (Moc. Sessé) is a Mexican fruit consumed as part of the Mexican culinary traditions. We conducted a study to identify and describe the incidence of a tortricid (Lepidoptera: Tortricidae) in C. mexicana from two areas where the fruit tree is present on its wild form. From the total of the collected fruits, 10% was visibly damaged during the first year, increasing to 30% the following year. Adult tortricids were monitored and captured using pheromone traps. Identification was carried out using larvae chaetotaxy in the larva and analysis of genitalia in the adults, as well as the amplification and sequencing of the gene 5.8 S rDNA. The presence of the species Grapholita packardi (Zeller) was determined in the fruits. Since there are no sequences of the gene 5.8 S rDNA reported in the GenBank database for this species, this is the first study in providing the amplification and sequence of this molecular marker for this insect. It is important to follow-up the evolution and distribution of G. packardi and the damages it causes. In this work, we suggest that this species should be considered as a potential pest in Mexico.

and 2016 with pheromone traps (TRECE) coated with Pherecon® pheromone. A total of 120 traps were positioned as follows: 60 in Jalacingo and 60 in Las Vigas de Ramirez. Traps were placed in creole apple, plum and wild hawthorn trees presenting healthy fruits and foliage in a quadrant of 200 m 2 per trap in family homegardens.

Taxonomic identification of adults and larvae
Adults and larvae in fruits samples were processed at the Laboratory of High Technology Xalapa, S.C. of the Veracruzana University, for the preparation of samples and identification according to morphological characteristics. Identification was carried out using taxonomic keys, setal maps (MacKay, 1959;Weisman, 1986;Stehr, 1987), and extraction of genitalia, using a stereomicroscope (Carl Zeiss, Stemi 1000) and a compound microscope (Carl Zeiss bright field, Axiostar).

Molecular analysis
DNA extraction was performed using a modified version of the method proposed by Yu et al. (2011). Briefly, tissue was extracted from the abdominal larval segments 4 to 8 and PCR performed according to the protocol (SAGARPA-CNRF, 2014). The TW81 (GTTTCCGTAGGTGAACCTGC) and AB28 (ATATGCTTAAGTTCAGCGGGT) molecular markers were used. The 5.8S rDNA amplified gene fragment was purified with Wizard® SV Gel kit and PCR Clean-Up System (Promega, WI, USA), and sequenced in the Institute of Biotechnology of the UNAM (sequencer Applied Bio systems). BLAST sequence homology search was performed within GenBank database NCBI (National Center for Biotechnology Information) on the nucleotide sequences obtained.
Out of the 210 adult specimens, 95 were intercepted on hawthorn trees and 115 on plum and apple trees. However, in the hawthorn fruits, 40% of visual damage observed was caused by larvae of Conotrachelus crataegi Walsh (Coleptera: Curculionidae) as Muñiz-Merino et al. (2012) have previously reported as a usual plague in hawthorn in México. Only 10% was due to G. packardi larvae in 2015. However, in 2016, the damage by G. packardi increased to 30%.
Both larval and male genitalia characters were coincident with those reported in this species (MacKay, 1959;Weisman, 1986;Stehr, 1987) (Figs. 1, 2). BLAST analysis indicated 94% of similarity in the 900 bp fragment sequence when compared with Eucosma siskiyouana (accession number KC430350.1). The proportion of sequenced gene is greater than 1:1. No similarity to other species of Grapholita was obtained. Since no gene sequences for 5.8 s rDNA have been The Mexican tejocote Crataegus mexicana (Moc. Sessé) (Rosales: Rosaceae) is commonly found in the states of San Luis Potosi, Jalisco, Veracruz, Puebla and Chiapas, although its distribution also includes Central America and Ecuador (CONABIO, 2015). In Mexico, tejocote is consumed as part of the culinary traditions during the Christmas festivities called "posadas". More than 940 hectares of tejocote are cultivated in Mexico, with an estimated production of 8,877 tons per year (SIAP, 2016).
Among the pests of economic importance affecting fruit trees cultivation worldwide, species of the genus Grapholita are one of the most reported. In Mexico, there are 57 pests reported under epidemiological phytosanitary surveillance in the 31 states and in Mexico city (SENASICA, 2012). Grapholita packardi (Zeller) (Lepidoptera: Tortricidae), known as cherry moth, is one of the major pests in fruit trees from the Ericaceae and Rosaceae families (EPPO, 2014;Gilligan & Epstein, 2014). In 1926, it was first reported in apple, rose and hawthorn plants in the North American region (Heinrich, 1926). From 1914 to 1960 it was considered the major pest in Prunus avium L. (Hoerner & List, 1952;Oatman & Ehlers, 1962). Finally, in 2014, EPPO mentioned G. packardi as the main host in cherry (P. avium) and as secondary hosts in plum (Prunus domestica L.), apple (Malus domestica Borkh), pear (Pyrus communis L.) and blueberry (Vaccinium macrocarpon Aiton). Moreover, it has been occasionally found in quince (Cydonia oblonga Miller) and peach (Prunus persica (L.) Stokes). Larvae make galleries in the interior of the fruits, with aesthetic damage that reduces the final fruit quality causing economic losses (Gilligan & Epstein, 2014;SAGARPA, 2014). The objective of the present research was to identify the tortricid G. packardi in fruits of C. mexicana in Mexico as well as its prevalence, to aid the implementation of prevention and control strategies in order to avoid its dispersion.

Fruit sampling
The monitoring was carried out weekly on mature hawthorn fruits and the visible damage caused by larvae was identified. Monitoring of adult specimens was performed during October and November 2014, 2015 and 2016 as well as during May, June and July 2015.  reported in the GenBank for this species, this is the first study of the amplification and sequencing of G. packardi. The nucleotide sequence of G. packardi was registered in GenBank under the accession number KX357711.
Tortricidae family includes pests in a wide variety of agricultural, horticultural, and forest crops. Even though it has a cosmopolitan distribution, the family is best represented in temperate, tropical and subtropical regions (Meijerman & Ulenberg, 2000;Valera-Fuentes et al., 2009). In general, members of the subfamily Tortricinae tend to be polyphagous. Dicotyledonous are the most common hosts, yet there are groups of Tortricinae species and genera specialized in gymnosperms. This study found that hawthorn trees are preferred by G. packardi, instead of cherry, the primary host in Veracruz, Mexico (Brown et al., 2003;Gilligan & Epstein, 2014).
An important aspect of tortricid dispersion and survival is that larvae employ a wide range of feeding G. packardi in creole fruits of apple and plum.
Therefore, monitoring the spread and damage caused by this potential pest in Mexico would help to prioritize epidemiological studies in order to prevent possible outbreaks in the state of Veracruz and even in the entire country.

ACKNOWLEDGEMENTS
To the Entomology Department of the National Reference Center of SENASICA for the traps, photographic material and the financial support given by the project UV-CA-354 PRODEP 2015 thematic network. Thanks to Dr. John Brown for his time in editing the written material.