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Incorporation of rhodamine B into male tobacco budworm moths Heliothis virescens to use as a marker for mating studies.

Blanco CA, Perera O, Ray JD, Taliercio E, Williams L - J. Insect Sci. (2006)

Bottom Line: The intake of this dye did not affect the life span, the production of eggs or the capacity of moths to copulate when compared with moths fed only a sucrose solution or water.The dye accumulation in internal abdominal organs was a better indicator of the presence of the pigment than external contamination of the moth.The use of the method described in this report can be a tool for the rapid incorporation of a low cost dye in the tobacco budworm for biological, behavioral and genetic studies.

View Article: PubMed Central - PubMed

Affiliation: USDA-ARS, Southern Insect Management Research Unit, Stoneville, MS 38776, USA. cblanco@ars.usda.gov

ABSTRACT
Rhodamine B, a dye commonly used in a variety of biological studies was incorporated into the bodies of male tobacco budworm moths, Heliothis virescens (Lepidoptera: Noctuidae), by allowing them to feed freely on 0.1% rhodamine dissolved in a 10% sucrose solution. After exposing males for one to three days to this pigment, rhodamine was clearly detectable in >82% of spermatophores extracted from untreated females. The intake of this dye did not affect the life span, the production of eggs or the capacity of moths to copulate when compared with moths fed only a sucrose solution or water. Rhodamine B was easily identifiable externally but was more apparent internally in males after only one day of exposure to the pigment. Even at this short feeding duration, rhodamine was detectable in >50% of males 5 days after feeding stopped. Longer exposure to the dye significantly increased the percentage stained. Detection of rhodamine was slightly enhanced by the use of ultraviolet light. The dye accumulation in internal abdominal organs was a better indicator of the presence of the pigment than external contamination of the moth. The use of the method described in this report can be a tool for the rapid incorporation of a low cost dye in the tobacco budworm for biological, behavioral and genetic studies.

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Related in: MedlinePlus

Percent rhodamine B detection in Heliothis virescens moths that were fed this solution for 1 to 4 days. Four different detection methods were employed described in upper graph legend box. Same letters by treatment day by sex are not significantly different at P < 0.05. (N = 48).
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i1536-2442-6-5-1-f04: Percent rhodamine B detection in Heliothis virescens moths that were fed this solution for 1 to 4 days. Four different detection methods were employed described in upper graph legend box. Same letters by treatment day by sex are not significantly different at P < 0.05. (N = 48).

Mentions: The percentages of rhodamine-stained spermatophores (≥82%, Figure 3) and males that showed rhodamine accumulated in their abdomen (≥95%, Figure 4) aid us in understanding the difference between the number of moths drinking the marker and its detection in sexual bundles. During fluorescent-light inspection of spermatophores from male moths that fed on rhodamine, spermatophores could be clearly distinguished (positive resolution) from those non-stained (negative resolution) (Illustration 1 (#I1)). The positive rhodamine resolution (≥82%) obtained from spermatophores dissected from the female bursa copulatrix indicates that this is a rapid and efficient way of marking H. virescens males. There were no significant differences in the percent detection of marked spermatophores between males fed for one, two or three days and mass mated for only one day. However, significantly lower detection (82%) of spermatophores occurred from males fed two days and mass mated two days (P> 0.0001, df = 9, t = 28.2) than males fed for the same amount of time and mass mated only one day (Figure 5), a fact that we attributed to experimental error. We did not detect any fluorescence in spermatophores of males fed on sucrose solution. These results indicate that incorporation of this marker into spermatophores is achieved rapidly (after only one day) with a success range of 82–100%.


Incorporation of rhodamine B into male tobacco budworm moths Heliothis virescens to use as a marker for mating studies.

Blanco CA, Perera O, Ray JD, Taliercio E, Williams L - J. Insect Sci. (2006)

Percent rhodamine B detection in Heliothis virescens moths that were fed this solution for 1 to 4 days. Four different detection methods were employed described in upper graph legend box. Same letters by treatment day by sex are not significantly different at P < 0.05. (N = 48).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2990291&req=5

i1536-2442-6-5-1-f04: Percent rhodamine B detection in Heliothis virescens moths that were fed this solution for 1 to 4 days. Four different detection methods were employed described in upper graph legend box. Same letters by treatment day by sex are not significantly different at P < 0.05. (N = 48).
Mentions: The percentages of rhodamine-stained spermatophores (≥82%, Figure 3) and males that showed rhodamine accumulated in their abdomen (≥95%, Figure 4) aid us in understanding the difference between the number of moths drinking the marker and its detection in sexual bundles. During fluorescent-light inspection of spermatophores from male moths that fed on rhodamine, spermatophores could be clearly distinguished (positive resolution) from those non-stained (negative resolution) (Illustration 1 (#I1)). The positive rhodamine resolution (≥82%) obtained from spermatophores dissected from the female bursa copulatrix indicates that this is a rapid and efficient way of marking H. virescens males. There were no significant differences in the percent detection of marked spermatophores between males fed for one, two or three days and mass mated for only one day. However, significantly lower detection (82%) of spermatophores occurred from males fed two days and mass mated two days (P> 0.0001, df = 9, t = 28.2) than males fed for the same amount of time and mass mated only one day (Figure 5), a fact that we attributed to experimental error. We did not detect any fluorescence in spermatophores of males fed on sucrose solution. These results indicate that incorporation of this marker into spermatophores is achieved rapidly (after only one day) with a success range of 82–100%.

Bottom Line: The intake of this dye did not affect the life span, the production of eggs or the capacity of moths to copulate when compared with moths fed only a sucrose solution or water.The dye accumulation in internal abdominal organs was a better indicator of the presence of the pigment than external contamination of the moth.The use of the method described in this report can be a tool for the rapid incorporation of a low cost dye in the tobacco budworm for biological, behavioral and genetic studies.

View Article: PubMed Central - PubMed

Affiliation: USDA-ARS, Southern Insect Management Research Unit, Stoneville, MS 38776, USA. cblanco@ars.usda.gov

ABSTRACT
Rhodamine B, a dye commonly used in a variety of biological studies was incorporated into the bodies of male tobacco budworm moths, Heliothis virescens (Lepidoptera: Noctuidae), by allowing them to feed freely on 0.1% rhodamine dissolved in a 10% sucrose solution. After exposing males for one to three days to this pigment, rhodamine was clearly detectable in >82% of spermatophores extracted from untreated females. The intake of this dye did not affect the life span, the production of eggs or the capacity of moths to copulate when compared with moths fed only a sucrose solution or water. Rhodamine B was easily identifiable externally but was more apparent internally in males after only one day of exposure to the pigment. Even at this short feeding duration, rhodamine was detectable in >50% of males 5 days after feeding stopped. Longer exposure to the dye significantly increased the percentage stained. Detection of rhodamine was slightly enhanced by the use of ultraviolet light. The dye accumulation in internal abdominal organs was a better indicator of the presence of the pigment than external contamination of the moth. The use of the method described in this report can be a tool for the rapid incorporation of a low cost dye in the tobacco budworm for biological, behavioral and genetic studies.

Show MeSH
Related in: MedlinePlus