EP0404890A4 - Defective wing medfly sex selection - Google Patents

Defective wing medfly sex selection

Info

Publication number
EP0404890A4
EP0404890A4 EP19890913040 EP89913040A EP0404890A4 EP 0404890 A4 EP0404890 A4 EP 0404890A4 EP 19890913040 EP19890913040 EP 19890913040 EP 89913040 A EP89913040 A EP 89913040A EP 0404890 A4 EP0404890 A4 EP 0404890A4
Authority
EP
European Patent Office
Prior art keywords
wing
phenotype
wild
medflies
medfly
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP19890913040
Other languages
English (en)
Other versions
EP0404890A1 (fr
Inventor
Stephen H. Saul
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
HAWAII MEDFLY BIOTECHNOLOGY PARTNERS
Original Assignee
HAWAII MEDFLY BIOTECHNOLOGY PARTNERS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by HAWAII MEDFLY BIOTECHNOLOGY PARTNERS filed Critical HAWAII MEDFLY BIOTECHNOLOGY PARTNERS
Publication of EP0404890A1 publication Critical patent/EP0404890A1/fr
Publication of EP0404890A4 publication Critical patent/EP0404890A4/en
Withdrawn legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
    • A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates

Definitions

  • the subject invention concerns production of sterilized male medflies free of female medflies using genetic lesions for separation.
  • the Mediterranean fruit fly (medfly), Ceratitis capitata (Wiedemann), is a major agriculture pest in temperate and sub-tropical regions worldwide. The medfly attacks over 150 varieties of cultivated fruit crops. It is found throughout the world in a variety of climes. Current control practice in most areas consist of the use of bait sprays, i.e., an attractant such as yeast hydrolysate mixed with an insecticide such as malathion. In addition, the United States Department of Agriculture has used the sterile insect release method (SIRM) in California and Texas and has participated in or consulted on SIRM in Central America and other places throughout the world.
  • SIRM sterile insect release method
  • the SIRM would have greater world wide use if not for several current and potential problems. Released females, though sterile, will sting or puncture fruit while attempting to oviposit, allowing the entrance of decay organisms and causing considerable, sometimes severe, damage. The large number of sterile females released in conjunction with sterile males, where there is no separation, can monopolize the matings with the released males needed for the sterile control of wild fertile females. There is evidence that sterile females mate with sterile males (intrastrain mating) more readily than with wild males (interstrain mating). Since released females will outnumber wild females up to a 50 to 1 ratio, they may be involved in the vast majority of matings by released males. This decrease in the likelihood of matings between sterile males and native fertile females greatly reduces the efficiency of SIRM.
  • a genetic sexing method would not only help avoid the problems described above, but could add several distinct benefits to a SIRM program. Sterilization technologies could be fined tuned to maximize sterile males without regard for effects on females, so that male competitiveness might be easier to improve.
  • a genetic marker could be provided in some programs that would identify inadvertent releases of fertile males as a result of accidental failure to sterilize and reduce opportunities for unexplained failures of some programs.
  • Ceratitis capitata (Medfly) and the resulting medflies are provided for controlling the medfly population in an environment subject to infestation.
  • the method involves providing breeding stocks of male and female medflies, where the males have the normal wing pheno ⁇ type and include an autosomal Y-translocation involving one arm of the autosome containing the ap_ and d_c genes with the Y-chromosome, where the wild-type gene allele of the v-wing mutated gene phenotype is involved in a translocation with the Y-chromosome.
  • the normal wing phenotype male containing the Y-chromosomal translocation is mated with homozygous v-wing females.
  • Lines may then be propagated and bred through a plurality of generations, where the progeny contain only normal wing phenotype males and v-wing phenotype females.
  • the progeny containing medflies of both sexes may then be sterilized in accordance with conventional ways, for example, gamma radiation employing a total dosage of least about 10, usually 15Krads.
  • the v-wing mutant may be obtained in a variety of ways. Particularly, routine screening of laboratory stocks can uncover the v-wing mutant.
  • fly stocks can be mutagenized by standard techniques, which include but are not limited by the following: Medflies may be irradiated with dosages in the range of about 2 to 8 Krad to induce mutations, ' followed by mating and screening the progeny for the v-wing phenotype.
  • Larvae may be subjected to a wide variety of mutagens, such as nitrosoguanidine, methyl methanesulfonate, formalin, or the like and the resulting medflies are screened for the v-wing phenotype.
  • embryos may be transformed with P-type transposable elements, which include a selective marker.
  • Introduction of the P-type element into the embryos can be achieved by injection, employing a potassium chloride phosphate-buffered solution having from about 50 to 500 ⁇ g/ml of P-type element DNA.
  • the embryos which contain the P-type element are detected by resistance to a selective medium.
  • the resistance may be to a cytotoxic agent, such as an antibiotic, methotrexate, heavy metal or other toxin.
  • P-type transposable elements are available for use with some diptera. By transforming a sufficient number of embryos, one may then screen the progeny for the v-wing mutation.
  • DNA from translocation stock and from homozygous v-wing stock is isolated by the method described by Poustka and Lehrach, Trends in Genetics (1986) 2:174-179.
  • the DNA is cloned into a large insert 40kb cosmid library.
  • the clones are probed with total Y-chromosome Medfly DNA isolated by a fluorescence activated cell sorter based on the fluorescence of the Y-chromosome.
  • the library is reprobed with total genomic Medfly DNA or DNA of the X-chromosome plus the autosome containing the wild-type wing gene complementary to the v-wing mutant, which can be obtained by fluorescence activated cell sorting.
  • Clones which hybridize to both probes contain the translocation junction points.
  • the v-wing mutant autosome chromosome is walked down from the junction towards the v-wing gene using techniques descsribed in Poustka and Lehrach, supra. Clones obtained in the walking procedure are used to transform wild-type flies. Clones which are specific for the v-wing mutant medfly are transformed into wild-type larvae in accordance with the procedure described in Rubin and Spaulding, Nucleic Acids Res. (1983) 11:6341-6351. See also, Thomas et al.. Nature (1986) 324:34-38; Song et al., Proc. Natl. Acad. Sci. (1987) 84: 6820-6824; and U.
  • the v-wing phenotype may be variable at one temperature and uniformly extreme at a different temperature.
  • the v-wing mutant employed in the subject invention is found to provide about 10 to 20% of medflies with stubby remnants when reared at room temperature, while 100% of the medflies have stubs and are totally unable to fly when reared at 30°C.
  • the larvae may be grown in any conventional larval food recipe.
  • the conventional nutrient media will usually include a base of wheat bran plus sucrose, yeast r a vitamin fortification, as well as other additives.
  • Adults may be raised on sucrose, yeast hydrolysate, casein hydrolysate, and as appropriate, essential amino acids, e.g., L-methionine.
  • the pupae are irradiated to sterility one day before adult eclosion. Sterility can be achieved by irradiation at a total dose of up to about 18Krads of gamma irradiation in a nitrogen atmosphere.
  • the resulting sterile fruit flies may then be introduced into the environment to be protected, in accordance with conventional methods.
  • the flies may be placed in an open container, where the females are incapable of flying over the walls of the container, while the males may freely leave the container.
  • the sterile males may then compete with wild-type males for the fertile females to substantially diminish the medfly population.
  • the v-wing stock was produced from a single female fly found in routine screening of laboratory stocks. After a true breeding stock was produced, crosses revealed that this apparently spontaneous mutant is an autosomal recessive located on the same chromosome as ap and dc genes.
  • Homozygous v-wing flies reared at room temperature show a wide range of phenotypic expressions from a few percent of flies with essentially wild-type wings to 10-20% with only stubby remnants. At 30°C 100% of the flies have only stubs and are totally unable to fly.
  • Larval food consisted of 250 g sucrose, 250 g torula yeast, 500 g of wheat bran and 10 g of Vanderzant vitamin fortification for insects (U.S. Biochemical Co.), 1 L distilled water containing 15 ml of cone. HC1 and 10 ml of a solution containing 750 mg of methylparaben and 1 g of ascorbic acid dissolved in 95% ethanol.
  • Translocations were produced by irradiating 24-48 hr old male flies in air with a dose of 4800 rads in the Hawaii Research Irradiator fcobalt-60 source). The mating scheme to isolate Y-autosome translocations then followed the plan used by Saul (1984). After irradiation the males were held for 1 day and then mass mated to virgin v-wing females. Each male offspring from this cross was pair mated to several female v-wing flies. Those crosses which gave progeny suggestive of a desired translocation, i.e., all wild type males and all v-wing females, were saved and propagated. One line (#28) from the first 102 male lines has proven to have just such a Y-autosome translocation exhibiting Y- linked inheritance of the wild-type V-wing allele. Additional lines are still being investigated.
  • a genetic sexing method for medflies' where females are produced having a physical deficiency which allows for the automatic separation of females from males and prohibits the females from affecting fruit or large scale mating with the sterile males.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Environmental Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Zoology (AREA)
  • Animal Husbandry (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
EP19890913040 1988-11-22 1989-10-20 Defective wing medfly sex selection Withdrawn EP0404890A4 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US27498988A 1988-11-22 1988-11-22
US274989 1988-11-22

Publications (2)

Publication Number Publication Date
EP0404890A1 EP0404890A1 (fr) 1991-01-02
EP0404890A4 true EP0404890A4 (en) 1992-01-08

Family

ID=23050441

Family Applications (1)

Application Number Title Priority Date Filing Date
EP19890913040 Withdrawn EP0404890A4 (en) 1988-11-22 1989-10-20 Defective wing medfly sex selection

Country Status (5)

Country Link
EP (1) EP0404890A4 (fr)
AU (1) AU4635289A (fr)
IL (1) IL92290A0 (fr)
PT (1) PT92371A (fr)
WO (1) WO1990005779A1 (fr)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
SG152938A1 (en) * 2007-11-21 2009-06-29 Erich Dollansky Automated insect breeding system
CN115088681B (zh) * 2022-08-02 2024-03-08 浙江省农业科学院 一种获得番茄潜叶蛾不育雄虫的方法及其在该虫防治中的应用

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
ANN. ENTOMOL. SOC. AM., vol. 78, no. 3, 1985, pages 265-269; Y. RÖSSLER: "Effect of genetic recombination in males of the Mediterranean fruit fly (Diptera: Tephritidae) on the ingegrity of "Genetic Sexing" strains produced for sterile-insect releases" *
See also references of WO9005779A1 *
Z. ANGEW. ENTOMOL., vol. 95, no. 2, 1983, pages 181-188; R.J. STEFFENS: "Methodology of translocation production and stability of translocations in the Mediterranean fruit fly, Ceratitis capitata Wied. (Dipt., Tephritidae)" *

Also Published As

Publication number Publication date
PT92371A (pt) 1990-05-31
AU4635289A (en) 1990-06-12
IL92290A0 (en) 1990-07-26
EP0404890A1 (fr) 1991-01-02
WO1990005779A1 (fr) 1990-05-31

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