BIOLOGY AND LIFE-CYCLE OF LEAFMINER Napomyza (Phytomyza) gymnostoma Loew., A NEW PEST OF Allium PLANTS IN ROMANIA

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South Western Journal of Vol.2, No.1, 2011 Horticulture, Biology and Environment P-Issn: 2067-9874, E-Issn: 2068-7958 pp.57-64 BIOLOGY AND LIFE-CYCLE OF LEAFMINER Napomyza (Phytomyza) gymnostoma Loew., A NEW PEST OF Allium PLANTS IN ROMANIA Mihaela COMAN 1, *, Ioan ROSCA 2 1. Central Phytosanitary Laboratory, Voluntari, no. 11, Romania 2. University of Agronomic Sciences and Veterinary Medicine, Marasti no. 59, Bucharest, Romania * Corresponding author: M. Coman, E-mail: mihaela.coman@lccf.ro Abstract. In spring 2007, an allium leafminer was recorded for the first time in non-commercial onion crops in the areas of Arges and Ilfov in Romania. The pest was identify as Napomyza (Phytomyza) gymnostoma Loew based on morphological characters of the adults. It has been reported as an Allium pest from Central, Eastern and more recently in Western Europe. Elsewhere in Europe, Napomyza gymnostoma Loew has two generations per year. This paper presents the first description of the biology of Napomyza gymnostoma in Romania and the infestation rate on onions, leeks and garlic. The research was conducted in the Arges and Ilfov counties, during the period of 2008. It was found that Napomyza gymnostoma develops two generations annually the spring and autumn generations. Key words: Napomyza gymnostoma, appearing period, infestation, biology, life-cycle. INTRODUCTION Napomyza gymnostoma is the Allium leafminer pest of leek (Allium porrum), onion (Allium cepa), chives (Allium schoenoprasum L.), garlic (Allium sativum) and of ornamental Allium plants (Agallou et al. 2004). Napomyza gymnostoma was added to the European and Mediterranean Plant Protection Organisation (EPPO) Alert List, in April 2005, following reports in the scientific literature that the organism was spreading within the EPPO region and becoming a significant pest of Allium crops (EPPO Reporting Service 2005). Adults of Napomyza gymnostoma could be identified based on the following characters: small, grayish, mat flies of 3 mm long, with a head largely yellow (Zlobin 1994). The abdomen is also yellow on the ventral South west J Hortic Biol Environ Craiova, Romania http://anucraiova.3x.ro/swjhbe/index.html 2011

58 Coman, M., Rosca, I. side, halteres white, wing length varied from 2.9 in males to 4.0 mm in females, legs are dark with yellowish knees. The larvae are headless, white yellowish colored and rich 6-7 mm when fully grown. Puparia is dark brown, around 3,5 mm in length, have a pair of posterior spiracle each with 18-20 bulbs. The eggs are ovoid and translucent, whitish (Bouchery & Martinez 2004, Sionek 1999). The larvae of the first spring generation develop in garlic and onion stems and bulbs. First generation pupae diapause during the summer before adults emerge and lay eggs in leeks or other Allium species in the autumn, for the second generation. Pupae of the second generation overwinter in host plants, before adults emerge the following spring to lay eggs on hosts such as onions and garlic (Kahrer 1999, Collins & Lole 2005, Mesic et al. 2009). MATERIAL AND METHODS The study were conducted from 2008- in non insecticide treated field in two different regions: Arges and Ilfov. Laboratory experiments were made in the entomological laboratory of Central Phytosanitary Laboratory. The necessary observations to know the biology of the pest in Romania were made in both areas on the following plants of the genus Allium: onion, garlic and leek. Plants with symptoms were transported in the laboratory, placed in the laboratory condition in order to obtain adults. The obtained adults were observed in water or glycerin to check the external characters, mainly the coloration, but for an accurate identification the male genitalia was examined. In this way, males were placed in 10% KOH solution and leave for 24 h for tissue maceration. After that the specimens were washed in distilled water and the abdomens were dissected under a Leica MZ 12 5 stereomicroscope. For microscopic examination the genitalia were mounted in Hoyer solution and observed at Zeiss Axio Imager. A1. microscope. The research of the intensity of the attack of Napomyza gymnostoma were made yearly, in the garden area on onion, garlic and leek plants. In every fenologycal period was picked up ten plants for each experimental variant of Allium species. RESULTS AND DISCUSSION The research was conducted in the Arges region and in Ilfov during the period of 2008-. It was found that Napomyza gymnostoma develops two generations annually, in the spring and autumn generations. Adults emerge in the spring from pupae that over wintered within host plant or soil. The flight of adult starts in the second decade of March, and is continuous till the first decade of May. The second generation flight starts at the beginning

Biology and life-cycle of Napomyza gymnostoma Loew. 59 of September and last till the end of October (Table 1). A couple days after copulation, the females from the second generation lay eggs mostly on leek plants. Larvae hatched from those eggs feed inside leek plants till mid- November. After finishing their developments, the larvae transform into pupae. Pupae over winter till the next spring when emerge new imagos. Females make large numbers of feeding punctures using their ovipositors to feed on leaf exudates. These punctures are the first sign that the flies are active. Most of the feeding punctures are arranged in lines, parallel to the leaf (Figure 2). The male (Figure 1) does not produce damage because they are unable to puncture leaves but have been observed feeding at punctures produced by females. Table 1. Emerge and development adults Napomyza gymnostoma during 2009-. Region Year No Generation Period flight Duration flight (days) from to Argeş 2009 1 02.04 05.05 26 11.04 09.05 28 2009 2 15.09 20.10 43 20.09 30.10 40 Ilfov 2009 1 18.03 04.05 48 01.04 11.05 41 2009 2 03.09 15.10 43 20.09 29.10 40 The average duration of adults development (days) 38.6 Females lay eggs a few days after copulation. The eggs are ovoid and translucent, whitish (Figure 3). Larvae from first generation are present from the second decade of April till the end of May. Second generation larvae are present from late September to end of November, (Table 2). Larvae emerge and mine downwards the leaf or bulb feeding on the succulent layers of the bulbs. The larval stage produce the most important damage especially in untreated cultures. The main problem is the presence of numerous larvae and puparia - up to 100 in a single leek plant (Billen 1999). During the research, the highest infestation was recorded in 2009, when was found 22 larvae and pupae of Napomyza gymnostoma per one leek plant in the Arges area (Table 3). Napomyza gymnostoma is capable of severely infesting almost 100% of a crop. In Serbia around 20 pupae were found per leek stem. All plants were completely destroyed. Even at lower pest densities, the presence of mines on young plants may reduce the quality and marketability of produce

60 Coman, M., Rosca, I. (Spasic & Mihajlovic 1997). After finishing their development, larvae transform into the stage of pupae (Figure 6). This stage lasts till September for the first generation. Figure 1. The adult of Napomyza gymnostoma (male) Figure 2. Feeding punctures Allium sativum Figure 3. Egg of Napomyza gymnostoma.

Biology and life-cycle of Napomyza gymnostoma Loew. 61 Tabel 2. Emerge and development larval stage - Napomyza gymnostoma during 2009-. Region Year No. generation Period larval Duration of stage from to (days) Argeş 2009 1 22.04 28.05 36 30.04 31.05 32 2009 2 25.09 12.11 49 1 23.10 23.11 30 Ilfov 2009 10.04 25.05 46 20.04 18.06 59 2009 2 20.09 30.10 51 20.10 20.11 30 The average duration of larvae stage (days) 41.6 Host plant Allium cepa Allium sativum Allium porrum Table 3. Dynamics of larval stage - Argeş during 2009-. Period Average larvae/plant No. maximum larvae/plant Average larvae nonviable/plant 22.04.2009 1 2 0 28.05.2009 3,5 7 2 30.04. 1,2 3 0 31.05. 3,7 8 2 22.04.2009 0,5 1 0 24.05.2009 2 4 1 02.05. 0,5 1 0 25.05. 2 5 1 25.09.2009 1 2 0 12.11.2009 13,6 22 2 27.10. 1 3 0 Figure 4. Larvae of Napomyza gymnostoma.

62 Coman, M., Rosca, I. Figure 5. High larval infestation Allium porrum Figure 6. Pupae of Napomyza gymnostoma Table 4. Emerge and during the all stages - Napomyza gymnostoma Argeş area 2009-. Development No. During the stage During the stage stage generation 2009 (average/days) Pupae - 30.03-31.03 Adult 1 02.04-05.05 11.04-09.05 31 Egg 12.04-15.05 20.04-14.05 28,5 Larvae 22.04-28.05 30.04-31.05 33,5 Pupae 24.05-28.09 29.05-01.10 140,5 During the first generation 02.04-28.09 11.04-01.10 176 days (days) 179 days 173 days Adult 15.09-20.10 20.09-30.10 41,5 Egg 2 12.09-20.10 01.10-30.10 34 Larvae 25.09-12.11 23.10-23.11 40,5 Pupae 20.10-31.03. 03.11-163 During the second generation ( days ) 15.09-31.03. 197 ( days ) 20.09 197

Biology and life-cycle of Napomyza gymnostoma Loew. 63 Table 5 Emerge and during the all stages - Napomyza gymnostoma Ilfov area 2009-. Development No. During the stage During the stage stage generation 2009 (average/days) Pupae - 31.03-31.03 Adult 1 18.03-04.05 01.04-11.05 44,5 Egg 29.03-05.05 11.04-12.05 34,5 Larvae 10.04-25.05 20.04-18.06 52,5 Pupae 05.05-26.09 29.05-30.09 134 During the first generation 18.03-26.09 01.04-30.09 188 (days) 193 days 183 days Adult 03.09-15.10 20.09-29.10 41 Egg 2 08.09-16.10 27.09-20.10 31,5 Larvae 20.09-30.10 20.10-20.11 35,5 Pupae 20.10-31.03. 01.11 163 During the second generation ( days ) 03.09-31.03. 209 (days) 20.09-209 The stages development period for Napomyza gymnostoma in the two areas was different. Due to lower temperatures in the Arges region debut of flight was made later, and the period of the flight was shorter than Ilfov. Duration flight for the second generation was identical in both areas. The egg laying period for th e first generation in Arges is less than Ilfov. From the second generation the laying period was less in Ilfov than Arges. The larval stage period of both generations in Ilfov had a longer duration than Arges. The pupal stage has the highest duration. Reffering to life-cycle of leafminer based on obtained date we can state that in Romania this pest has two generations per year. Due to climatic conditions the periods of stages development of insect in the two areas had some differences. CONCLUSIONS The result of this study confirm the existence of pest in Romania. Due to the favourable climatic condition and host plants grown, we consider Napomyza gymnostoma as a new potential major pest of Allium crops. In Romania the pest has two generation per year, one in spring and the other is autumn. The highest infestation was recorded at untreated leek,

64 Coman, M., Rosca, I. with 22 larvae and pupae per plant. Plants can be completely destroyed or reduced in market value. The distribution of the pest at the national level it is not well known because of its new occurrence and also due to possible confusion with other Allium pest, especially Delia antiqua. It can be concluded that more attention should be paid to this potentially damaging pest of leek, onion, garlic and other Allium crops. ACKNOWLEDGEMENTS. Author is thankful to Mr. Michel Martinez from INRA (French National Institute for Agricultural Research) for confirmation of the pest. REFERENCES Aggalou, E., Cannon, R.D., Matthews, L. (2004): Allium leaf miner-phytomyza gymnostoma. Plant Pest Notice, no. 35, Central Science Laboratory, York (GB), 4 pp. Billen, W. (1999): A new pest of Allium species: Napomyza gymnostoma Loew (Diptera: Agromyzidae). Mitteilungen der entomologischen Gesellschaft Basel 49(1): 37pp. Bouchery, Y., Martinez, M. (2004): A new enemy of Allium in France : the leek mining fly Phytomyza gymnostoma. Phytoma 574: 5-7. Collins, D.W., Lole, M. (2005): Phytomyza gymnostoma Loew (Diptera: Agromyzidae), a leaf mining pest of leek and onion new to Britain. Entomologist's Monthly Magazine 141 (1694/96): 131-138. EPPO Reporting Service (2005): Phytomyza (Napomyza) gymnostoma is a new pest of Allium in Europe: addition to the EPPO Alert List. EPPO Reporting Service 2005/064. Kahrer, A. (1999): Biology and control of the leek mining fly, Napomyza gymnostoma. Bulletin OILB/SROP (22/ 5): 205-211. Mesic, A., Dupor, M., Igrc, B.J. (2009): Napomyza gymnostoma Loew (Agromyzidae: Diptera) biology in central Croatia. Entomologica Croatica 13(2):45-53. Sionek, R. (1999): Effect of leek planting dates on damage to plants by spring generation of the leek leaf miner (Napomyza gymnostoma Loew). Progress in Plant Protection 39(2): 500-502. Spasic, R., Mihajlovic, L.J. (1997): Napomyza gymnostoma (Loew) a pest on bulbed vegetables in Serbia and its parasitoids. ANPP Fourth International Conference on Pests in Agriculture, Montpellier, 6-8 January, 1977. Zlobin, V.V. (1994): Review of mining flies of the genus Napomyza Westwood (Diptera: Agromyzidae) V Phytomyza species currently placed in the Genus Napomyza Westwood. Dipterological Research 5: 289-311.