EP0240332A2
Expired / Lapsed
EP
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Virus resistant plants having antisense RNA
Assignee: Pioneer Hi-Bred International Inc
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Filing: 1987-04-01
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Granted: —
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Expires: 2007-04-01
Abstract
Abstract The present invention discloses the making of plant cells which contain antisense RNA (aRNA) complementary to the mRNA of a target plant virus. Construction of aRNA genes and transformation of aRNA genes into plant cells is also taught. Such cells are relatively resistant to infection by the target virus when compared with cells not containing aRNA. Furthermore, methods and DNA molecules useful for producing plant cells containing aRNA are also disclosed.
Inventors
- 👤 Sue L. Loesch-Fries
- 👤 Donald J. Merlo
- 👤 Nancy P. Jarvis
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-04 21:59
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/11; C12N15/113; C12N15/1131; C12N15/63; C12N15/79; C12N15/82; C12N15/8241; C12N15/8261; C12N15/8271; C12N15/8279; C12N15/8283
Claims Analysis
Total Claims: 54
Claim 1
Independent Claim
52 words
A DNA molecule comprising a promoter, a viral cDNA encoding an aRNA, and a transcript terminator, wherein the promoter, the cDNA, and the terminator have such position and orientation with respect to each other that the aRNA may be transcribed in a plant cell under control of the promoter and the terminator.
Claim 2
Dependent Claim
52 words
A DNA molecule comprising a promoter, a viral cDNA encoding an aRNA, and a transcript terminator, wherein the promoter, the cDNA, and the terminator have such position and orientation with respect to each other that the aRNA may be transcribed in a plant cell under control of the promoter and the terminator.
Claim 3
Dependent Claim
22 words
A DNA according to claim 1, wherein the cDNA is a cDNA to a plant virus having single-stranded, plus-stranded, tripartite RNA genome.
Claim 4
Dependent Claim
14 words
A DNA according to claim 2, wherein the cDNA is a cDNA to RNA4.
Claim 5
Dependent Claim
21 words
A DNA according to claim 2, wherein the cDNA is a cDNA to a virus of the alfalfa mosaic virus group.
Claim 6
Dependent Claim
11 words
A DNA according to claim 4, wherein the virus is AMV.
Claim 7
Dependent Claim
13 words
A DNA according to claim 5, wherein the virus is AMV strain 425.
Claim 8
Dependent Claim
14 words
A DNA according to claim 5, wherein the cDNA is an AMV RNA1 cDNA.
Claim 9
Dependent Claim
14 words
A DNA according to claim 5, wherein the cDNA is an AMV RNA2 cDNA.
Claim 10
Dependent Claim
14 words
A DNA according to claim 5, wherein the cDNA is an AMV RNA3 cDNA.
Claim 11
Dependent Claim
15 words
1O. A DNA according to claim 5, wherein the cDNA is an AMV RNA4 cDNA.
Claim 12
Dependent Claim
18 words
A DNA according to any preceding claim, wherein the aRNA is complementary to a viral translation initiation site.
Claim 13
Dependent Claim
17 words
A DNA according to any preceding claim, wherein the aRNA is complementary to a viral 5ʹ-untranslated sequence.
Claim 14
Dependent Claim
17 words
A DNA according to any preceding claim, wherein the aRNA is complementary to a viral 3ʹ-conserved sequence.
Claim 15
Dependent Claim
15 words
A DNA according to any preceding claim, wherein the cDNA is essentially a full-length cDNA.
Claim 16
Dependent Claim
15 words
A DNA according to any preceding claim, wherein the promoter is a CaMV 19S promoter.
Claim 17
Dependent Claim
15 words
A DNA according to any preceding claim, wherein the transcript terminator is a polyadenylation site.
Claim 18
Dependent Claim
15 words
A DNA according to claim 16, wherein the polyadenylation site is a CaMV polyadenylation site.
Claim 19
Dependent Claim
15 words
A DNA according to any preceding claim, further comprising a plant-expressible selectable or screenable marker.
Claim 20
Dependent Claim
13 words
A DNA according to claim 18, wherein the selectable marker encodes neomycin phosphotransferase.
Claim 21
Dependent Claim
13 words
2O. A DNA according to any preceding claim, further comprising an ocs gene.
Claim 22
Dependent Claim
16 words
A DNA according to any preceding claim, wherein the DNA further comprises the T-DNA of pH4OO.
Claim 23
Dependent Claim
11 words
A DNA according to claim 21, wherein the DNA is pH4OOA4I.
Claim 24
Dependent Claim
12 words
A DNA according to any preceding claim, further comprising T-DNA border repeat.
Claim 25
Dependent Claim
16 words
A DNA according to any preceding claim, wherein the DNA is contained by a bacterial cell.
Claim 26
Dependent Claim
13 words
A bacterial cell containing the DNA of any of claims 1 to 23.
Claim 27
Dependent Claim
19 words
A DNA according to any of claims 1 to 23, wherein the promoter/cDNA/terminator combination is flanked by plant DNA.
Claim 28
Dependent Claim
21 words
A DNA according to claim 26, wherein the DNA is contained by a plant cell, plant tissue, plant seed or plant.
Claim 29
Dependent Claim
15 words
A plant cell, plant tissue, plant seed, or plant containing the DNA of claim 26.
Claim 30
Dependent Claim
26 words
A plant cell, plant tissue, plant seed, or plant that is uninfected by a virus and that contains an aRNA to an mRNA of the virus.
Claim 31
Dependent Claim
26 words
A plant cell, plant tissue, plant seed, or plant that is uninfected by a virus and that contains an aRNA to an mRNA of the virus.
Claim 32
Dependent Claim
21 words
3O. A cell, tissue, seed, or plant according to claim 29, wherein the virus has a single-stranded, plus-stranded, tripartite RNA genome.
Claim 33
Dependent Claim
20 words
A cell, tissue, seed, or plant according to claim 3O, wherein the virus is of the alfalfa mosaic virus group.
Claim 34
Dependent Claim
15 words
A cell, tissue, seed, or plant according to claim 31, wherein the virus is AMV.
Claim 35
Dependent Claim
17 words
A cell, tissue, seed, or plant according to claim 32, wherein the virus is AMV strain 425.
Claim 36
Independent Claim
56 words
A method for producing a virus-resistant plant comprising the steps of making a viral cDNA segment that encodes an aRNA and ligating the cDNA segment to a promoter DNA segment, the promoter and the cDNA having such position and orientation with respect to each other that the aRNA may be transcribed under control of the promoter.
Claim 37
Independent Claim
56 words
A method for producing a virus-resistant plant comprising the steps of making a viral cDNA segment that encodes an aRNA and ligating the cDNA segment to a promoter DNA segment, the promoter and the cDNA having such position and orientation with respect to each other that the aRNA may be transcribed under control of the promoter.
Claim 38
Dependent Claim
62 words
A method according to claim 34, further comprising the step of ligating the cDNA segment to a transcript terminator DNA segment, the promoter, the cDNA, and the terminator having such position and orientation with respect to each other that the aRNA may be transcribed in a plant cell under control of the promoter and the terminator, whereby an aRNA gene is formed.
Claim 39
Independent Claim
31 words
A method for producing a virus-resistant plant comprising the steps of ligating (i) a viral cDNA segment that encodes an aRNA, (ii) a promoter DNA segment, and (iii) a transcript terminator,
Claim 40
Independent Claim
31 words
A method for producing a virus-resistant plant comprising the steps of ligating (i) a viral cDNA segment that encodes an aRNA, (ii) a promoter DNA segment, and (iii) a transcript terminator,
Claim 41
Dependent Claim
13 words
to form a DNA molecule according to any of claims 1 to 23.
Claim 42
Dependent Claim
81 words
A method according to claim 35 or claim 36, further comprising the step of transforming a first plant cell to contain a DNA molecule comprising a promoter, a viral cDNA encoding an aRNA, and a trans cript terminator, wherein the promoter, the cDNA, and the terminator have such position and orientation with respect to each other that the aRNA may be transcribed under control of the promoter and the terminator in a second plant cell descended from the first plant cell.
Claim 43
Dependent Claim
18 words
A method according to claim 37, wherein the DNA molecule further comprises a plant-expressible selectable or screenable marker.
Claim 44
Dependent Claim
13 words
A method according to claim 38, wherein the selectable marker encodes neomycin phosphotransferase.
Claim 45
Dependent Claim
32 words
4O. A method according to claim 38, comprising the step of culturing the first or second plant cells in the presence of a selective agent to which the selectable marker confers resistance.
Claim 46
Independent Claim
16 words
A method according to claim 4O, wherein the selective agent is kanamycin or an analog thereof.
Claim 47
Dependent Claim
54 words
A method according to claim 37, further comprising the step of introducing the DNA into an Agrobacterium cell, wherein the step of transforming is accomplished by infecting the first plant cell with an Agrobacterium cell containing the DNA, and wherein the aRNA gene-bearing DNA of the transforming step further comprises a T-DNA border repeat.
Claim 48
Dependent Claim
13 words
A method according to claim 42, wherein the DNA further comprises ocs .
Claim 49
Dependent Claim
27 words
A method according to claim 43, further comprising the step of culturing the first plant cell or the second plant cell in the presence of aminoethyl cysteine.
Claim 50
Dependent Claim
12 words
A method according to claim 43, wherein the DNA further comprises pH4OO.
Claim 51
Dependent Claim
11 words
A method according to claim 45, wherein the DNA is pH4OOA4I.
Claim 52
Dependent Claim
16 words
A plant cell, plant tissue, plant seed, or plant produced by the method of claim 35.
Claim 53
Dependent Claim
21 words
A plant cell, plant tissue, plant seed, or plant descended from the first cell produced by the method of claim 37.
Claim 54
Dependent Claim
20 words
A plant cell, plant tissue, plant seed, or plant containing the second cell produced by the method of claim 37.
References Cited (Backward Citations)
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