Insect viruses and their uses in protecting plants

Details Insect viruses and their uses in protecting plants Insect viruses and their uses in protecting plants

1995-06-07

2001-01-23

Wortman, Donna C. (Department: 1642)

Drug, bio-affecting and body treating compositions

Whole live micro-organism, cell, or virus containing

Genetically modified micro-organism, cell, or virus

C424S093600, C435S235100

Reexamination Certificate

active

06177075

ABSTRACT:

FIELD OF THE INVENTION
The present invention relates to insect viruses useful in control of insect attack on plants. It particularly relates to biological insecticides, especially those comprised of insect viruses. In particular applications, the invention also provides recombinant viruses and transgenic plants.
BACKGROUND OF THE INVENTION
There is increasing awareness of the desirability of insect pest control by biological agents. Considerable effort in recent years has been devoted to the identification and exploitation of DNA viruses with large genomes, especially the baculoviruses. These viruses generally require extensive genetic manipulation to become effective insecticides, and the first such modified viruses are only now being evaluated.
In contrast, very little effort has been devoted to the study and use of small viruses with RNA genomes.
Four main groups of small RNA viruses have been isolated from insects. These include members of the Picornaviridae, the Nodaviridae, the Tetraviridae and the unclassified viruses. Descriptions of these groups can be found in the Atlas of Invertebrate Viruses (eds J. R. Adams and J. R. Bonami) (CRC Press, Boca Raton, 1991) and Viruses of Invertebrates (ed. E. Kurstak) (Marcel Dekker, New York, 1991). These disclosures relating to these viruses concern their pathology and biology, not their use in biological control.
Further information regarding small RNA viruses of insects an be found in P. D. Scotti et al (1981). “The biology and ecology of strains of an insect small RNA virus complex”
Advances in Virus Research
26, 117-143. This review describes the insect picornaviruses cricket paralysis virus and Drosophila C virus (diameters estimated at 27-30 nm with one RNA component of 7.5-8.5 kb). N. F. Moore & T. W. Tinsley (1982). The small RNA viruses of insects. Brief review
Archives of Virology
72, 229-245. This review included viruses of the following families:
Nodaviridae (diameter 29-30 nm, 2 RNA components totalling 4.5 kb)
Picornaviridae (diameter 27-30 nm, one RNA component of 7.5-8.5 kb)
Nudaurelia &bgr; family (now called Tetraviridae) (diameter around 35 nm, either one RNA of 5.5 kb or two totalling 8 kb)
N. F. Moore, B. Reavy & L. A. King (1985) General characteristics, gene organisation and expression of small RNA viruses of insects.
Journal of general Virology
66, 647-659. This reference defines small RNA viruses of insects as being those less than 40 nm in diameter. The review covers Picornaviridae, Nodaviridae and the Nudaurelia &bgr; family (now called Tetraviridae).
D. Hendry, V. Hodgson, R Clark and J Newman (1985) Small RNA viruses co-infecting the pine emperor moth (
Nudaurelia cytherea capensis
).
Journal of general Virology
66, 627-632 described viruses with mean diameters of 40 nm and 38 nm and one or two RNA components up to 5.5 kb in length.
Most recently, the term insect small RNA viruses has been used by one of the present inventors to cover three main recognised toxic groups: the Picornaviridae, the Tetraviridae and the Nodaviridae (P. Scotti & P. Christian (1994) The promises and potential problems of using small RNA insect viruses for insect control.
Sains Malaysiana
23, 9-18).
These references illustrate a long standing usage of the term in this field of the term “small RNA virus” for viruses with certain characteristics as listed above. Another important characteristic of these virus groups is that they are not occluded, in contrast to many large viruses like the cytoplasmic polyhydrosis (RNA) viruses or the DNA baculoviruses, granulosis viruses and entomopox viruses. The term would also be applied to viruses not members of the three families listed above, as long as they satisfied the definition of being up to 40 nm in size. There are reports of such unclassified viruses (eg in Hendry et al. 1985). Moreover, the taxonomic status of some members of the Tetraviridae still requires clarification and it might even be possible for this family to be split, with
Ha
SV and other members with two RNA components in their genome being separated from those with only one component, like the type member Nudaurelia &bgr; virus, which has not yet been sequenced. The above definition of “small RNA virus” would still cover all members of such virus families.
SUMMARY OF THE INVENTION
In a first aspect of the present invention there is provided an isolated small RNA virus wherein the virus is up to 40 nm in size, is not occluded and infects insect species including Heliothis species.
In one particular embodiment, the present invention provides an isolated preparation of
Heliothis armigera
stunt virus referred to as “
Ha
SV” herein.
In a further aspect of the present invention there is provided an isolated nucleic acid molecule comprising a nucleic acid sequence hybridizable with RNA 1 (SEQ ID No: 39) or RNA 2 (SEQ ID No: 47) described herein under low stringency conditions.
In still a further aspect the invention provides a vector comprising a nucleic acid molecule, the sequence of which is hybridizable with RNA 1 (SEQ ID No: 39) or RNA 2 (SEQ ID No: 47) as described herein. These vectors include expression and transfer vectors for use in animals including insect, plant and bacterial cells.
In a further aspect the invention provides an isolated protein or polypeptide preparation of the proteins or polypeptides derivable from the isolated virus of the present invention. The invention also extends to antibodies specific for the protein and polypeptide preparations.
In a yet further aspect the invention provides a recombinant insect virus vector incorporating all or a part of the isolated virus of the present invention.
In a still further aspect of the present invention there is provided a method of controlling insect attack in a plant comprising genetically manipulating said plant so that it is capable of producing
Ha
SV or mutants, derivatives or variants thereof or an insecticidally effective portion of
Ha
SV, mutants, variants or derivatives thereof such that insects feeding on the plants are deleteriously effected. The present invention also provides a transgenic plant so manipulated.
In another aspect of the present invention there is provided a preparation of
Ha
SV or a mutant variant or derivative thereof, or an insecticidally effective portion of
Ha
SV, mutant, variant or derivative thereof, suitable for application to plants, wherein the preparation is capable of imparting an insect protective effect.


REFERENCES:
patent: 5135917 (1992-08-01), Burch
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patent: 5508186 (1996-04-01), Young et al.
McCutchen, B.F., et al., “Development of a Recombinant Baculovirus Expressing an Insect-Selective Neurotoxin: Potential for Pest Control.”BioTechnology, 9:848-852 (1991).
Dasmahapatra, B., et al., “Infectious RNA Derived by Transcription from Cloned cDNA Copies of the Genomic RNA of an Insect Virus.”Proc. Natl. Acad. Sci. USA, 83:63-66 (1986).
Vaeck, M., et al., “Transgenic Plants Protected from Insect Attack.”Nature, 328:33-37 (1987).
Fischhoff, D.A., “Insect Tolerant Transgenic Tomato Plants.”Biotechnology, 5:807-813 (1987).
Rubinstein, R., et al., “The Nucleic Acids of Viruses InfectingHeliothis armigera.” Virology, 69:323-326 (1976).
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Garzon, Simon and Guy Charpentier, “Nodaviridae.”Atlas of Invertebrates Virus, (Eds. J.R. Adam and J.R. Bonami), CRC Press, pp. 351-593 (1991).
Moore, Norman F., “TheNudaurelia&bgr; Family of Insect Viruses.”The Viruses of Invertebrates(Ed. E. Kurtak), pp. 277-299.
Hendry, Donald A., “Nodaviridae of Invertebrates.”Viruses of Invertebrates, (Ed. E. Kurtak), pp. 227-275.
Desmier de Chenon and Sipayung, “Use of Microbial Pesticides for Field Crops, the Case of Viruses to Control Oil Palm and Coconut Leaf-Eating Caterpillars,”Proceedings of the Biotechnology for Tropical Plant Protection Conference, Abstract, Kuala Lumpur, Aug

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