DOI: 10.5061/dryad.bzkh189gn
title: Asymmetric reproductive interference of male Drosophila suzukii caused by both male and female Drosophila melanogaster

description: Reproductive interference, leading to negative effects on fitness, usually
 occurs within two closely related species which sharing overlapping
 ecological niches. Our preliminary experiments showed D. suzukii
 population decreased when reared with D. melanogaster. However, the
 explanation of this phenomenon is not much clear. Based on the overlapping
 ecological niches and courtship behaviours of D. suzukii and D.
 melanogaster, we deduced that reproductive interference was the dominant
 reason. We found that the proportion of heterospecific orientation time is
 significantly greater in male D. suzukii (7.5 ± 4.183 %) than in male D.
 melanogaster (4.439 ± 4.936 %, U=194.5, Z=2.223, P=0.026). The successful
 mating rate of D. suzukii was significantly lower when the species was
 reared with D. melanogaster (F1,31=4.303, p=0.046). The egg production of
 D. suzukii was significantly lower in the combined rearing condition
 (F1,343=98.623, p&lt;0.001), for D. melanogaster, the egg production
 in combined condition was significantly greater than in single rearing
 conditions (F1,354=4.202, p=0.041). The reproductive interference exerted
 by D. melanogaster increased the heterospecific courtship behaviours of
 male D. suzukii, causing the successful mating rate to decline and
 ultimately reducing the number of eggs.
description: <em>Male conspecific and
 heterospecific</em><em> cour</em><em>ting
 behaviours</em> To ensure that the flies used
 for courting observations were unmated, we placed each pupa into 2-ml
 Eppendorf tubes for emergence. All tubes were maintained in incubators
 with a light:dark photoperiod of 14:10 h, temperature of 25±1°C, and
 humidity of 60±5%. All flies were kept in Eppendorf tubes for 48 h from
 eclosion until observation started. By comparing the male courtship
 behaviours of <em>D. suzukii</em> and <em>D.
 melanogaster </em>(Revadi et al. 2015; Spieth 1974), we selected
 seven specific behaviours—orientation, tapping, wing scissoring and
 spreading, circling, successful mating, heterospecific mating attempts and
 premating duration—for observation (<strong>Table 2</strong>).
 Proportion of misdirected courting time to total courting time in the two
 files were also recorded. Based on the rearing conditions, <em>D.
 suzukii</em> and <em>D. melanogaster</em> were divided
 into three groups (<strong>Figure 1</strong>): (1)
 <em>D. suzukii</em> single rearing
 (<strong>DSS</strong>): two pairs of <em>D.
 suzukii</em>; (2) <em>D. melanogaster</em> single
 rearing (<strong>DMS</strong>): two pairs of <em>D.
 melanogaster</em>; (3) <em>D. suzukii</em> and
 <em>D. melanogaster</em> combined rearing
 (<strong>DC</strong>):one pair of <em>D.
 suzukii</em> and one pair of <em>D. melanogaster</em>.
 <em>Drosophila </em>were transferred from Eppendorf tubes into
 perforated Petri dishes (355 mm) containing food, using a
 custom-made <em>Drosophila</em> aspirator. We recorded a
 40-min video of courting behaviour with a microscope (VHX-5000, Osaka
 Japan, Keyence Corporation) for 20 replicates. During the data collection
 process, courtship time or frequency was calculated starting from the
 occurrence of the first courtship behaviour and ending with either mating
 occurrence or the specified time limit. 
 <strong><em>Successful mating
 rate</em></strong> For obtaining virgin
 <em>D. suzukii</em> and <em>D. melanogaster</em>,
 the method is the same as described in the previous section. Based on the
 rearing conditions, <em>D. suzukii</em> and <em>D.
 melanogaster</em> were divided into four groups
 (<strong>Figure 1</strong>): (1) <em>D.
 suzukii</em> single rearing (<strong>DSS</strong>): two
 pairs of <em>D. suzukii</em>; (2) <em>D.
 melanogaster</em> single rearing (<strong>DMS</strong>):
 two pairs of <em>D. melanogaster</em>; (3) <em>D.
 suzukii</em> and <em>D. melanogaster</em> combined
 rearing (<strong>DC</strong>):one pair of <em>D.
 suzukii</em> and one pair of <em>D. melanogaster</em>;
 (4) <em>D. suzukii</em> and <em>D.
 melanogaster</em> combined rearing in dark
 (<strong>DCD</strong>): one pair of <em>D.
 suzukii</em> and one pair of <em>D. melanogaster</em>
 were combined rearing under red lights (wavelength:620-630 nm).
 <em>Drosophila </em>were transferred from Eppendorf tubes into
 perforated Petri dishes (355 mm) containing food, using a
 custom-made <em>Drosophila</em> aspirator. We then recorded
 40-min videos (VHX-5000, Osaka Japan, Keyence Corporation) of the two
 flies under the two rearing conditions with a microscope, and 20
 replicates were performed. Frequency of heterospecific mating attempts,
 rate of successful mating and rate of successful mating in dark of each
 pair involved were recorded.
 <strong><em>Number of
 eggs</em></strong> For obtaining virgin
 <em>D. suzukii</em> and <em>D. melanogaster</em>,
 the method is the same as described in the second section of Methods.
 Based on the rearing conditions, <em>D. suzukii</em> and
 <em>D. melanogaster</em> were divided into three groups
 (<strong>Figure 1</strong>): (1) <em>D.
 suzukii</em> single rearing (<strong>DSS</strong>): two
 pairs of <em>D. suzukii</em>; (2) <em>D.
 melanogaster</em> single rearing (<strong>DMS</strong>):
 two pairs of <em>D. melanogaster</em>; (3) <em>D.
 suzukii</em> and <em>D. melanogaster</em> combined
 rearing (<strong>DC</strong>):one pair of <em>D.
 suzukii</em> and one pair of <em>D. melanogaster</em>.
 <em>Drosophila </em>were transferred from Eppendorf tubes into
 <em>Drosophila</em> tube containing food, using a custom-made
 <em>Drosophila</em> aspirator. The next day, the adult
 flies<em> </em>in each tube were<sub>
 </sub>transferred to another clear tube with fresh solid food and
 maintained for ten days. Because <em>D. melanogaster
 </em>larvae have a deterring effect on <em>D.
 suzukii</em> oviposition (Tungadi et al., 2022), the number of eggs
 was counted daily starting from the egg stage. Twenty replicates of each
 fly species and rearing type were established.
description: # Asymmetric reproductive interference of male Drosophila suzukii caused
 by both male and female Drosophila melanogaster
 [https://doi.org/10.5061/dryad.bzkh189gn](https://doi.org/10.5061/dryad.bzkh189gn) ## Description of the data and file structure 80 vedios and a supplementary materia file about reproductive interference between *Drosophila suzukii* and *Drosophila melanogaster* are included. ### Files and variables File: DSS-1.avi Description: D. suzukii single rearing group 1 File: DSS-2.avi Description: D. suzukii single rearing group 2 File: DSS-3.avi Description: D. suzukii single rearing group 3 File: DSS-4.avi Description: D. suzukii single rearing group 4 File: DSS-5.avi Description: D. suzukii single rearing group 5 File: DSS-6.avi Description: D. suzukii single rearing group 6 File: DSS-7.avi Description: D. suzukii single rearing group 7 File: DSS-8.avi Description: D. suzukii single rearing group 8 File: DSS-9.avi Description: D. suzukii single rearing group 9 File: DSS-10.avi Description: D. suzukii single rearing group 10 File: DSS-11.avi Description: D. suzukii single rearing group 11 File: DSS-12.avi Description: D. suzukii single rearing group 12 File: DSS-13.avi Description: D. suzukii single rearing group 13 File: DSS-14.avi Description: D. suzukii single rearing group 14 File: DSS-15.avi Description: D. suzukii single rearing group 15 File: DSS-16.avi Description: D. suzukii single rearing group 16 File: DSS-17.avi Description: D. suzukii single rearing group 17 File: DSS-18.avi Description: D. suzukii single rearing group 18 File: DSS-19.avi Description: D. suzukii single rearing group 19 File: DSS-20.avi Description: D. suzukii single rearing group 20 File: DC-1.avi Description: D. suzukii and D. melanogaster combined rearing group 1 File: DC-2.avi Description: D. suzukii and D. melanogaster combined rearing group 2 File: DC-3.avi Description: D. suzukii and D. melanogaster combined rearing group 3 File: DC-4.avi Description: D. suzukii and D. melanogaster combined rearing group 4 File: DC-5.avi Description: D. suzukii and D. melanogaster combined rearing group 5 File: DC-6.avi Description: D. suzukii and D. melanogaster combined rearing group 6 File: DC-7.avi Description: D. suzukii and D. melanogaster combined rearing group 7 File: DC-8.avi Description: D. suzukii and D. melanogaster combined rearing group 8 File: DC-9.avi Description: D. suzukii and D. melanogaster combined rearing group 9 File: DC-10.avi Description: D. suzukii and D. melanogaster combined rearing group 10 File: DC-11.avi Description: D. suzukii and D. melanogaster combined rearing group 11 File: DC-12.avi Description: D. suzukii and D. melanogaster combined rearing group 12 File: DC-13.avi Description: D. suzukii and D. melanogaster combined rearing group 13 File: DC-14.avi Description: D. suzukii and D. melanogaster combined rearing group 14 File: DC-15.avi Description: D. suzukii and D. melanogaster combined rearing group 15 File: DC-16.avi Description: D. suzukii and D. melanogaster combined rearing group 16 File: DC-17.avi Description: D. suzukii and D. melanogaster combined rearing group 17 File: DC-18.avi Description: D. suzukii and D. melanogaster combined rearing group 18 File: DC-19.avi Description: D. suzukii and D. melanogaster combined rearing group 19 File: DC-20.avi Description: D. suzukii and D. melanogaster combined rearing group 20 File: DCD-1.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 1 File: DCD-2.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 2 File: DCD-3.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 3 File: DCD-4.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 4 File: DCD-5.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 5 File: DCD-6.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 6 File: DCD-7.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 7 File: DCD-8.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 8 File: DCD-9.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 9 File: DCD-10.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 10 File: DCD-11.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 11 File: DCD-12.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 12 File: DCD-13.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 13 File: DCD-14.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 14 File: DCD-15.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 15 File: DCD-16.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 16 File: DCD-17.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 17 File: DCD-19.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 19 File: DCD-18.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 18 File: DCD-20.avi Description: D. suzukii and D. melanogaster combined rearing in dark group 20 File: DMS-1.avi Description: D. melanogaster single rearing group 1 File: DMS-2.avi Description: D. melanogaster single rearing group 2 File: DMS-3.avi Description: D. melanogaster single rearing group 3 File: DMS-4.avi Description: D. melanogaster single rearing group 4 File: DMS-5.avi Description: D. melanogaster single rearing group 5 File: DMS-6.avi Description: D. melanogaster single rearing group 6 File: DMS-7.avi Description: D. melanogaster single rearing group 7 File: DMS-8.avi Description: D. melanogaster single rearing group 8 File: DMS-9.avi Description: D. melanogaster single rearing group 9 File: DMS-10.avi Description: D. melanogaster single rearing group 10 File: DMS-11.avi Description: D. melanogaster single rearing group 11 File: DMS-12.avi Description: D. melanogaster single rearing group 12 File: DMS-13.avi Description: D. melanogaster single rearing group 13 File: DMS-14.avi Description: D. melanogaster single rearing group 14 File: DMS-15.avi Description: D. melanogaster single rearing group 15 File: DMS-16.avi Description: D. melanogaster single rearing group 16 File: DMS-17.avi Description: D. melanogaster single rearing group 17 File: DMS-18.avi Description: D. melanogaster single rearing group 18 File: DMS-19.avi Description: D. melanogaster single rearing group 19 File: DMS-20.avi Description: D. melanogaster single rearing group 20 File: Supplementary_material.xlsx Description: The supplementary material file contains data on the conspecific and heterospecific courtship and egg-laying numbers for D. suzukii and D. melanogaster. D.suzukii combined rearing 1 and 2: The sheet contains courtship or pre-mating behaviors observed in heterospecific interactions (interactions between individuals of D. suzukii of different strains) across 8 different experimental groups. Each row represents one group, and the columns record counts or durations of specific behaviors. D.mel combined rearing1 and 2: The sheet contains courtship or pre-mating behaviors observed in heterospecific interactions (interactions between individuals of D. melanogaster of different strains) across 8 different experimental groups. Each row represents one group, and the columns record counts or durations of specific behaviors. D.suzukii single rearing: This sheet contains courtship behaviors observed during conspecific interactions—that is, interactions between individuals of the same D.suzukii species—across 5 experimental groups. Each row represents a different group or experimental condition, and the columns record how often specific courtship behaviors occurred, as well as the total premating duration. D.melanogaster single rearing: This sheet contains courtship behaviors observed during conspecific interactions—that is, interactions between individuals of the same D.melanogaster species—across 5 experimental groups. Each row represents a different group or experimental condition, and the columns record how often specific courtship behaviors occurred, as well as the total premating duration. heterospecific behaviours (%): This sheet represents a comparison of courtship behaviors between two species—Drosophila suzukii and Drosophila melanogaster—under combined rearing conditions (i.e., when individuals of both species are raised or tested in the same environment). The values seem to represent percentages or frequencies of specific behaviors during these interspecies interactions, possibly averaged across replicates, with one row likely representing a different experimental group or replicate. Egg production: This sheet appears to track courtship or mating behavior counts between different Drosophila species or strains, where each column likely corresponds to a specific interaction type or species pairing. Variables: D.suzukii combined rearing 1: D.suzukii heterospecific behavior in combined rearing conditions. D.suzukii combined rearing 2: D.suzukii conspecific behavior in combined rearing conditions. D.mel combined rearing 1: D. melanogaster heterospecific behavior in combined rearing conditions. D.mel combined rearing 2: D. melanogaster conspecific behavior in combined rearing conditions. D.suzukii single rearing: D.suzukii conspecific behavior in single rearing conditions. D.melanogaster single rearing: D. melanogaster conspecific behavior in single rearing conditions. heterospecific behaviours (%): The proportion of heterospecific courtship to total courtship behaviour of male Drosophila melanogaster and male Drosophila suzukii in combined rearing condition. Egg production: Egg production of D. melanogaster and D. suzukii under single and combined rearing conditions ## Code/software IQIYI can be used to view the video and installation package is uploaded. Use Microsoft Office software to open the Supplementary material. ## Access information Other publicly accessible locations of the data: * None Data was derived from the following sources: * Author's own experimental videos.