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DNA construct to confer multiple traits on plants

a technology of dna and plants, applied in the field of dna constructs to confer multiple traits on plants, can solve the problems of complex structure, high cost and time consumption, and difficulty in incorporating multiple virus resistance in a given plant, and achieve the effect of distributing resistan

Inactive Publication Date: 2005-07-07
PANG SHENG ZHI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This approach allows for the creation of transgenic plants resistant to multiple viruses using shorter gene fragments and a silencer DNA, enhancing resistance and reducing the complexity and cost of genetic transformation, as demonstrated by examples in papaya and potato.

Problems solved by technology

It is well known that particular plant types are often susceptible to more than one virus.
Although PDR is an excellent approach to controlling the damaging effects of plant viruses, incorporating multiple virus resistance in a given plant can be challenging.
For example, identifying and producing full length viral genes to transform plants can be expensive and time consuming.
Further, such genes may be so large that they need to be incorporated in different expression systems which must be separately incorporated in plants.

Method used

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  • DNA construct to confer multiple traits on plants
  • DNA construct to confer multiple traits on plants
  • DNA construct to confer multiple traits on plants

Examples

Experimental program
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Effect test

example 1

Cloning and Transformation

[0065] The N gene (Pang, S.-Z., et al., “Resistance to Heterologous Isolates of Tomato Spotted Wilt Virus in Transgenic Plants Expressing its Nucleocapsid Protein Gene,”Phytopathology, 82: 1223-29 (1992), which is hereby incorporated by reference) of the lettuce isolate of tomato spotted wilt virus (TSWV-BL) was used as the template for construction of the N gene fragments of various lengths using the primers listed in Table 1.

TABLE 1Primers used in cloningNamePositions1SequenceFor the N gene fragments91-842776-27445′-AGCTAATCTAGAACCATGGATGACTCACTAAGGAAAGCATTGTTGC(SEQ. ID. NO. 1)93-892669-26505′-TACAGTTCTAGAACCATGGTCTGGAAAACCTTGACCAG(SEQ. ID. NO. 2)93-852576-25565′-TACAGTTCTAGAACCATGGTAAAGCGATTTTACTTTTGGTA(SEQ. ID. NO. 3)92-552373-23545′-AGATTCTCTAGACCATGGTGACTTGATGAGCAAAGTCTGTGAGGCTTGC(SEQ. ID. NO. 4)93-912266-22485′-TACAGTTCTAGAACCATGGAAAATACAAGGATCTCGGG(SEQ. ID. NO. 5)93-872153-21335′-TACAGTTCTAGAACCATGGTAGAAGGGGAAAGAGTATGCTG(SEQ. ID. NO. 6)90-471918-...

example 2

ELISA, Northern Blot Analyses of Transgenic Plants

[0069] Double antibody sandwich enzyme-linked immunosorbent assay (“DAS-ELISA”) was used to detect the nptII enzyme in transgenic plants using an nptII ELISA kit (5 Prime to 3 Prime, Inc). Northern blot was performed as described previously (Pang, S. Z., et al., “Different Mechanisms Protect Transgenic Tobacco Against Tomato Spotted Wilt and Impatiens Necrotic Spot Tospoviruses,”Bio / Technology, 11: 819-24 (1993), which is hereby incorporated by reference).

example 3

Inoculation of Transgenic Plants

[0070] Inoculations were done as described previously (Pang, S.-Z., et al., “Resistance to Heterologous Isolates of Tomato Spotted Wilt Virus in Transgenic Plants Expressing its Nucleocapsid Protein Gene,”Phytopathology, 82: 1223-29 (1992), which is hereby incorporated by reference). Systemic symptoms were recorded every other day for at least two months.

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Abstract

The present invention is directed to a DNA construct formed from a fusion gene which includes a trait DNA molecule and a silencer DNA molecule. The trait DNA molecule has a length that is insufficient to impart a desired trait to plants transformed with the trait DNA molecule. The silencer DNA molecule is operatively coupled to the trait DNA molecule with the trait and silencer DNA molecules collectively having sufficient length to impart the trait to plants transformed with the DNA construct. Expression systems, host cells, plants, and plant seeds containing the DNA construct are disclosed. The present invention is also directed to imparting multiple traits to a plant.

Description

[0001] This application is a divisional of U.S. patent application Ser. No. 09 / 025,635, filed Feb. 18, 1998, which is hereby incorporated by reference in its entirety and claims the benefit of U.S. Provisional Application Ser. No. 60 / 035,350, filed Feb. 19, 1997, and 60 / 062,870, filed Oct. 21, 1997, which are hereby incorporated by reference.FIELD OF THE INVENTION [0002] The present invention is directed to a DNA construct to confer multiple traits on plants. BACKGROUND OF THE INVENTION [0003] Control of plant virus diseases took a major step forward in the last decade when it was shown in 1986 that the tobacco mosaic virus (“TMV”) coat protein gene that was expressed in transgenic tobacco conferred resistance to TMV (Powell-Abel, P., et al., “Delay of Disease Development in Transgenic Plants that Express the Tobacco Mosaic Virus Coat Protein Gene,”Science, 232: 738-43 (1986)). The concept of pathogen-derived resistance (“PDR”), which states that pathogen genes that are expressed in...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N1/21C12N15/63C12N15/82
CPCC12N15/63C12N15/8283C12N15/8218C12N15/8216
Inventor PANG, SHENG-ZHIGONSALVES, DENNISJAN, FUH-JYH
Owner PANG SHENG ZHI
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