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European Journal of Applied Sciences – Vol. 12, No. 3
Publication Date: June 25, 2024
DOI:10.14738/aivp.123.17089
Hegazi, E., Khafagi, W. E., Zeid, A. A., Attia, M. A., Hasaneen, M., Shall, A. A., Aly, N., Taleb, H. K. A., El-Rahman, S. A., Showiel, S.
F., Al Eryan, M. A., Farag, M. M. A. & Mahmoud, A. K. (2024). Preliminary Evaluation of Diapause Intensity in Phytomyza
orobanchia Kalt. (Diptera, Agromyzidae) in Faba Bean Vicia faba Fields in Alexandria, Egypt. European Journal of Applied
Sciences, Vol - 12(3). 207-214.
Services for Science and Education – United Kingdom
Preliminary Evaluation of Diapause Intensity in Phytomyza
orobanchia Kalt. (Diptera, Agromyzidae) in Faba Bean Vicia faba
Fields in Alexandria, Egypt
Esmat. Hegazi
Faculty of Agriculture, Alexandria University, Alexandria, Egypt
W. E. Khafagi
Plant Protection Research Institute, Agricultural Research Center
(ARC), Sabahia, Alexandria, Egypt
A. Abou Zeid
Food Legumes Research Department, Field Crops Research Institute
(FCRI), Agricultural Research Center (ARC), Alexandria, Egypt
Manal A. Attia
Bioassay Research Department, Central Agricultural Pesticide
Laboratory (CAPL), Sabahia, Agricultural Research Center (ARC),
Alexandria, Egypt
Mervat Hasaneen
Plant Protection Research Institute, Agricultural Research Center
(ARC), Sabahia, Alexandria, Egypt
Amany Abu Shall
Faculty of Agriculture, Alexandria University, Alexandria, Egypt
Nagat Aly
Mammalian Toxcicity Department, Central Agricultural Pesticide
Laboratory (CAPL), Agricultural
H. K. Abou Taleb
Plant Protection Research Institute, Agricultural Research Center
(ARC), Sabahia, Alexandria, Egypt
Safaa. Abd El-Rahman
Bioassay Research Department, Central Agricultural Pesticide
Laboratory (CAPL), Sabahia, Agricultural Research Center (ARC),
Alexandria, Egypt
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European Journal of Applied Sciences (EJAS) Vol. 12, Issue 3, June-2024
Sania F. Showiel
Plant Protection Research Institute, Agricultural Research Center
(ARC), Sabahia, Alexandria, Egypt
M. Awad Al Eryan
Faculty of Agriculture, Alexandria University, Alexandria, Egypt
Marwa M. A. Farag
Economic Entomology and Pesticide Department, Cairo University,
Cairo, Egypt
Anter K. Mahmoud
Mammalian Toxcicity Department, Central Agricultural Pesticide
Laboratory (CAPL), Agricultural
ABSTRACT
This study was carried out to evaluate the diapause intensity in Phytomyza
orobanchia Kalt. (Diptera, Agromyzidae) in faba bean fields in Alexandria, Egypt.
Several visits were carried out in faba bean seasons of 2021 and 2022 during
January, February, March and April (cool to warm temperature) to collect spicks
from faba bean fields infested by Orobanche crenata Forsk (Orobanchaceae). The
spicks were dissected under binocular microscope and inspected for
Ph.orobanchia puparia. Total of 6663 and 2810 Ph.orobanchia puparia were
collected throughout the 1st and 2nd seasons, respectively. Ph.orobanchia puparia
were allowed hatching to evaluate diapause incidence in the fly .Monthly mean fly
emergence was recorded . This fly is a short-day species with a pupal summer -
winter diapause. For each faba bean season, the onset of pupal diapause was in
February increases in March and reached maximum in April. Large proportion of
diapausing puparia of 1year old, hatched in January, decreased gradually,
reaching its minimum in April. The live pupria that did not emerge after April,
were assumed to be prepupae-pupae in diapause for more longer period (2-3
years). Also, large proportion of remaining diapausing puparia were found
parasitized by Tetrastichus phytomyzae (Hymenoptera, Eulophidae) wasps. The
emerged wasps were observed to parasitized the old unparasitized diapaused fly
puparia and destroying the rearing fly culture during the two seasons. Females
outnumber the males among non-diapausing flies. However, males outnumber the
females for diapausing puparia hatched in March. While the sex ratio of emerged
flies of diapaused puparia hatched in January, February and April was in favor of
females.
INTRODUCTION
In Egypt, parasitic weed species, Orobanche crenata Forsk (Orobanchaceae) is noxious root
parasites which seriously constrain the production of many crops, including Leguminosae and
Solanaceae families in Egypt (Parker & Riches, 1993). Various control methods have been tested for
broomrape control, but none has proved entirely satisfactory (Saghir et al., 1980; Linke & Saxena,
1991). The production of a large number of seeds (30,000-200,000 per plant) (Schroeder,
1994), which remain viable in the soil for more than ten years, and the intimate physiological
interaction with their host plants limits the application of conventional weed control
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209
Hegazi, E., Khafagi, W. E., Zeid, A. A., Attia, M. A., Hasaneen, M., Shall, A. A., Aly, N., Taleb, H. K. A., El-Rahman, S. A., Showiel, S. F., Al Eryan, M. A.,
Farag, M. M. A. & Mahmoud, A. K. (2024). Preliminary Evaluation of Diapause Intensity in Phytomyza orobanchia Kalt. (Diptera, Agromyzidae) in
Faba Bean Vicia faba Fields in Alexandria, Egypt. European Journal of Applied Sciences, Vol - 12(3). 207-214.
URL: http://dx.doi.org/10.14738/aivp.123.17089
measures for Orobanche control. Almost 50 insect species are reported to feed on Orobanche
spp. Only one, Phytomyza orobanchia Kalt. (Diptera: Agromyzidae) is specific to Orobanche
(linke et al 1992). One generation takes about one month to develop, depending on the
climate conditions (Lekic 1970).
Several records show that biological control of Orobanche has been successfully achieved in the
field. The fly Ph. orobanchia, has been used successfully for Orobanche control in the former
USSR and Eastern Europe (Horvath & Wittmann, 1988), Turkey (Giray and Nemli, 1983), Syria
(Linke et al., 1990; Linke, 1992) and in India (Manjunath & Nagurkatti, 1977). This fly occurs
widely in Egypt, but it was too often killed by routine use of insecticides (Rezene Fessehaie &
Parker, 1992). Broomrape fly, Ph. orobanchia is oligophagous feeding only on Orobanche
species. It feeds mainly on the immature seeds destroying between 11% and 100% of seeds
(Sauerbom 1991). Three Ph. orobanchia larvae/capsule are able to destrory all immature
seeds (Girling et al., 1979). Despite the fact that the fly Ph. orobanchia, have been studied
extensively, gaps in the knowledge of their biology still remain. One issue is how the leaf
miner’ diapause, this distinct physiological event represents a primary part of the life-cycle in
some insect species. It is known that in obligate diapauses, all the individuals in generational
ways enter diapauses, while in facultative diapauses, only some members of a generation
enter diapauses (Denlinger, 2002). More knowledge of the Phytomyza activity will provide a
prediction for adult emergence and subsequent egg laying and potential damage by larvae.
This may help the biological control programs to justify the required numbers of Phytomyza
and adequate time for releases to obtain the maximum destruction of Orobanche seeds.
Diapause is a state of arrested growth or reproduction. It is an endogenously regulated
dormant state that provides a means for insects to survive seasons of adverse environmental
conditions and allows populations to synchronize periods of active development and
reproduction with seasons of optimal resource availability (Koštál, 2006; Lees, 1955; Tauber
et al., 1986). The aim of the present study is to evaluate the diapause incidence in Phytomyza
orobanchia in faba bean Vicia faba fields in Alexandria, Egypt.
MATERIALS AND METHODS
The larvae of Ph.orobanchia mine Orobanche shoots and feed on immature seed capsuples.
Pupation takes place in the capsules but most frequently in the stem base near the ground.
From January 1st to April end of the 2 seasons of 2021 and 2022 several samples of O.crenata
shoots were successively collected from 4 faba bean farms in Nubaria area in Alexandria
governorate. The range of field temperature during the sampling date was 12-15, 13-15,14-17
and 18-19°C, in January, February, Mach and April, respectively. By dissecting the 5390 and
3567infested capsule throughout the 1st and 2nd seasons a total of 6663 and 2810
Ph.orobanchia puparia were collected ,respectively. The Puparia were kept in glass vials
provided with wet pieces of cotton and covered with muslin kept in position. To identify the
diapausing pupraria, each sample was held for more than three weeks, to allow sufficient time
for the non-diapausing flies to complete their life cycles and emerge. The emerged adults were
counted and removed. When adult emergence was discontinued, the puparia that did not
emerge after three weeks, were assumed to be puparia containing prepupae-pupae in
diapause. The numbers of flies entering diapause and relevant notes were recorded. Each
sample of diapausing wasps were counted, dated and allowed to develop under controlled
conditions. The diapausing flies were monitored every day for eclosion. Emergence times in
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European Journal of Applied Sciences (EJAS) Vol. 12, Issue 3, June-2024
puparia samples collected in different dates (Januurary-April) were used to calculate diapause
intensity (DI). At emergence, they can be sexed by the slightly protruding ovipositor of the
females. The puparia were maintained at 25 ± 1°C in incubators (type Hann, Munden,
Germany), under photoperiod 12:12 (light: dark), RH 75% and light intensity 8000 Lx. Light
was measured with an available Weston light meter (Weston Electrical Instrument Company,
Newark, NJ), located in a controlled room. The puparia were examined every day for eclosion.
Monthly mean emergence Ph. orobanchia puparia was recorded and sexing.
RESULTS
Table 1 shows monthly diapausing and emergening of Ph. Orobancia puparia collected in
2021 and 2022 seasons. During the 1st season of 6663 puparia ,86.97% entered diapause. The
percentage of DI reached up 0.0, 0.7,1.4 and 97.9%, in January, February, March and April
(cool to warm), respectively. Of the total collected puaria some of these diapausing puparian
(16.87%) spent one-year diapausing and emergerd in 2022.The proportion of emerging flies
was 50.9, 37.0, 8.4 and 3.4% emerged in January, February, March and April, respectively. It
seems that the number of generations /seasons is 2-3 in faba bean fields. In 2023, two adult
females of Ph. orobanchia emerged in January and many of Tetrastichus phytomyzae Kost
wasps emerged and several dead and alive pupae P. orobanchia were found.
Table (1): Monthly diapausing and non-diapausing of Phytomyza Orobancia Kalt.
puparia collected in 2021 and 2022 seasons.
Season Collected
puparia
(NO.)
Diapaused Emerged
NO Month (%) Year NO Month (%)
Jan Feb March April Jan Feb March April
2021 6663 5795
(86.97%)*
0.0 0.74 1.4 97.9 2022 978
(16.8%)****
50.9 37 8.4 3.4
2022 2810 1390
(49.5%)*
0.0 11.1 12.1 76.76 2023 531
(38.2%)**
49.5 27.7 19.6 3.2
*Figures between Brackets refer to percentages of diapaused puparia among total number of collected puparia
** Figures between Brackets refer to percentages of emerged flies among total number of diapaused puparia
For puparia (n=2810) collected in 2022faba bean season 49.5%, entered diapause. Of this
total diapausing puparia (n=1390), the proportion of puparia entering diapause reached up
0.0,11.1,12.1 and 76.76% in January, February, March and April, respectively. In 2023,531
(38.2%) puparia emerged. Of these puparia 49.5,27.7,19.6 and 3.2 emerged in
January,February, March and April, respectively. The non-emerging puparia were mostly
parasitized by T. phytomyzae wasp, some were alive “diapausing” and some were dead. The
presence of alive non-emerging of 2 years old puparia in both seasons indicating that some
puparia entering in diapause for a length of more than 2 years, according to the climatic
records of faba bean fields during in 2021-2023the average range of temperature was 12-
15,13-15,14-17 and 18-19°C, respectively.
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Hegazi, E., Khafagi, W. E., Zeid, A. A., Attia, M. A., Hasaneen, M., Shall, A. A., Aly, N., Taleb, H. K. A., El-Rahman, S. A., Showiel, S. F., Al Eryan, M. A.,
Farag, M. M. A. & Mahmoud, A. K. (2024). Preliminary Evaluation of Diapause Intensity in Phytomyza orobanchia Kalt. (Diptera, Agromyzidae) in
Faba Bean Vicia faba Fields in Alexandria, Egypt. European Journal of Applied Sciences, Vol - 12(3). 207-214.
URL: http://dx.doi.org/10.14738/aivp.123.17089
Figure 1: Monthly diapausing and emerging of Phytomyza Orobanchia Kalt. Puparia
collected in 2021 &2022 seasons.
0
20
40
60
80
100
Jan. Feb. Mar. Apr.
Diapausing/ Emerging
%
Season 2021
Pupae diapausing in 2021
Adult emerging in 2022
0
20
40
60
80
100
Jan. Feb. Mar. Apr.
Diapausing/ Emerging
%
Season 2022
Pupae diapausing in 2022
Adult emerging in 2023
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Figure (2): Seasonal effects on proportions of diapausing and non-diapausing
Phytomyza Orobanchia flies Collected in (2021 & 2022).
As shown in Figs.1&2, the proportion of diapausing Ph. Orobanchia puparia gradually rose in
February increase in March and reached maximum in April. Adults emerge from the
hibernating puparia between early January and the end of April. Large proportion of
diapausing puparia hatched in January, decreased gradually, reaching its minimum in April.
The puparia that did not emerge after April, were assumed to be puparia containing
prepupae-pupae in diapause for more longer period (2-3 years). Also, large proportion of
1year diapausing puparia were found parasitized by T. phytomyzae wasp. The emerged wasps
were observed to parasitized the remaining old unparasitized diapaused fly puparia and end
the rearing fly culture during the two seasons. The sex ratio (♂: ♀) of emerged non- diapausing and diapausing Ph. orobanchia flies is shown in Table2. No Ph. Orobanchia puparia
were collected in January due to the absence of host plant. Females outnumber the males
among non-diapausing flies during faba bean season. However, males outnumber the females
for diapausing pupae hatched in March. While the sex ratio of emerged flies of diapaused
puparia hatched in January, February and April was in favor of females.
Table 2: Sex ratio (♂: ♀) of emerged non- diapausing and diapausing Phtromyza Orobanchia
flies.
Ph. Orobanchia pupae Sex ratio (♂: ♀) / month
Collected season Emergence season Status January February March April
2021 2021 Non-diapaused _ 0.8: 1
(155)
0.7: 1
(420)
0.5: 1
(215)
2021 2022 Diapaused 0.9: 1
(498)
0.8: 1
(361)
1.4: 1
(82)
0.8: 1
(37)
2022 2023 Diapaused 0.8: 1
(263)
0.8: 1
(147)
1.2: 1
(104)
0.5: 1
(17)
Figures between brackets refer to number of hatched puparia
0
2000
4000
6000
8000
Season-2021 Season-2022
No.
Collected puperia Diapausing flies Emerging flies after 1 year
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Hegazi, E., Khafagi, W. E., Zeid, A. A., Attia, M. A., Hasaneen, M., Shall, A. A., Aly, N., Taleb, H. K. A., El-Rahman, S. A., Showiel, S. F., Al Eryan, M. A.,
Farag, M. M. A. & Mahmoud, A. K. (2024). Preliminary Evaluation of Diapause Intensity in Phytomyza orobanchia Kalt. (Diptera, Agromyzidae) in
Faba Bean Vicia faba Fields in Alexandria, Egypt. European Journal of Applied Sciences, Vol - 12(3). 207-214.
URL: http://dx.doi.org/10.14738/aivp.123.17089
DISCUSSION
Cerny et al 2023 repopted that, rearing fly larvae of Ph.orobanchia under a 12-hours
photoperiod favours the quantity of diapause fly puparia. Therefore,the collected puparia
were maintained at 25 ± 1°C in incubators under photoperiod 12:12 (light: dark), RH 75%
and light intensity 8000 Lx located in a controlled room to evaluate diapause incidence in Ph.
orobanchia in faba bean Vicia faba fields in Alexandria, Egypt. It is known that diapause is the
period of suspended development or growth in Orobanchia puparia during stress conditions,
i.e during the warmer month (April vs. February ).The proportion of suspended puparia
development started in both seasons in February increase in March and reached up Maximum
in April. The first fly puparia sample was collected in mid-February. In both faba bean seasons
the largest proportions of diapausing flies emerge in January, i.e, the adult insects appear in
the fields with the first emergence of Orobance plant weed. The fly is long live adult and may
stay alive in the field up 2 months (unpuplished data) waiting for the capsules of the parasitic
plant in February. So the fly have intimate ecological interaction with its host plant. So this
facultative (February & March) and obligate (April) diapause plays a role in Ph.orobanchia life
cycles by synchronizes life cycles with the period of mate and food availability.
The present study suggested that the fly has prolonged diapause of different diapause
intensity. The polymorphism in the intensity of prolonged diapause split population of adult’s
flies of a single population to emerge in different years. The available data suggest that the
diapause is controlled by temperature, and lasts for 2-3 years. However, Klyueva and
Pamuckchi 1980. suggested that diapause in Orobanche is controlled by daylength; i.e.,
puparia developed under short day conditions may stay 4 years in diapause, while those
developed under long day conditions usually hatch in the next season. Unfortunately, the
effectiveness of fly population is drastically affected by parasitism of T. phytomyzae wasp.
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