US2002115849A1PendingUtilityA1
Plant promoter activated by fungal infection
Est. expiryMay 5, 2015(expired)· nominal 20-yr term from priority
C12N 15/8239C07K 14/415C12N 15/8282
44
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Claims
Abstract
The present invention relates generally to genetic sequences which are useful in the diagnosis and treatment of fungal infections in plants which involve a susceptible interaction between a fungal pathogen and the host plant. In particular, the present invention provides genetic sequences which confer, activate, or enhance expression of a gene in a plant, in response to infection of said plant by a plant fungal pathogen in a susceptible interaction the invention further provides genetic sequences such as structural genes, the expression of which is induced in response to a susceptible interaction between a plant and a fungal pathogen. The present invention further provides methods for the detection of infection by a fungal pathogen in a susceptible interaction and for the production of transgenic plants with improved resistance to said fungal pathogen. Transgenic plant material which expresses an artificial resistance gene against a fungal pathogen is also within the scope of the invention claimed.
Claims
exact text as granted — not AI-modified1 .(amended) An isolated nucleic acid molecule comprising 5′ upstream or promoter sequences which, when operably connected to a structural gene, confer, activate, enhance or otherwise increase the expression of said structural gene in response to a susceptible interaction between a host plant and a fungal pathogen.
2 . An isolated nucleic acid molecule according to claim 1 , originating from a crop plant selected from the list comprising flax, maize, rice. barley, rye and oats. amongst others.
3 . An isolated nucleic acid molecule according to claim 1 , wherein said nucleic acid molecule is genetically-linked in vivo as hereinbefore defined, to an SR structural gene which is at least 40% identical to the nucleotide sequence set forth in SEQ ID NO:1 or a complementary strand thereof.
4 . An isolated nucleic acid molecule according to claim 1 , wherein said nucleic acid molecule is genetically-linked in vivo as hereinbefore defined, to an SR structural gene which is at least 60-65% identical to the nucleotide sequence set forth in SEQ ID NO:1 or a complementary strand thereof.
5 . An isolated nucleic acid molecule according to claim 1 , wherein said nucleic acid molecule is genetically-linked in vivo as hereinbefore defined, to an SR structural gene which is at least 70-75% identical to the nucleotide sequence set forth in SEQ ID NO:1 or a complementary strand thereof.
6 . An isolated nucleic acid molecule according to claim 1 , wherein said nucleic acid molecule is genetically-linked in vivo as hereinbefore defined, to an SR structural gene which is at least 80-90% identical to the nucleotide sequence set forth in SEQ ID NO:1 or a complementary strand thereof.
7 . An isolated nucleic acid molecule according to claim 1 , wherein said nucleic acid molecule is genetically-linked in vivo as hereinbefore defined, to an SR structural gene which hybridises under at least low stringency conditions to the nucleotide sequence set forth in SEQ ID NO:1 or to a complementary strand thereof.
8 . An isolated nucleic acid molecule according to claim 7 , wherein said SR structural gene further comprises a sequence of nucleotides which is at least 40% identical to SEQ ID NO:1 or a complementary strand thereof.
9 . An isolated nucleic acid molecule according to claim 7 , wherein said SR structural gene further comprises a sequence of nucleotides which is at least 60-65% identical to SEQ ID NO:1 or a complementary strand thereof.
10 . An isolated nucleic acid molecule according to claim 7 , wherein said SR structural gene further comprises a sequence of nucleotides which is at least 70-75% identical to SEQ ID NO:1 or a complementary strand thereof.
11 . An isolated nucleic acid molecule according to claim 7 , wherein said SR structural gene further comprises a sequence of nucleotides which is at least 80-90% identical to SEQ ID NO:1 or a complementary strand thereof.
12 . An isolated nucleic acid molecule according to claim 1 , wherein said nucleic acid molecule comprises a sequence of nucleotides of a least 2 kilobases in length including at the 5′ end the sequence of nucleotides set forth in SEQ ID NO:3 or a homologue, analogue or derivative thereof and at the 3′ end the sequence of nucleotides set forth in SEQ ID NO: 4 or a homologue, analogue or derivative thereof.
13 . An isolated nucleic acid molecule according to claim 12 , further capable of hybridising under at least low stringency conditions to the Flax Fis1 gene promoter contained in the microorganism deposited under AGAL Accession No. N96/027087.
14 . An isolated nucleic acid molecule according to claim 13 , wherein said nucleic acid molecule is of plant origin.
15 . An isolated nucleic acid molecule according to claim 14 , wherein the plant is a crop plant selected from the list comprising flax, maize barley, rice, rye and oats, amongst others.
16 .(amended) An isolated nucleic acid molecule which comprises an SR structural gene induced in response to a susceptible interaction between a host plant and a fungal pathogen.
17 . An isolated nucleic acid molecule according to claim 16 , wherein said nucleic acid molecule originates from a crop plant selected from the list comprising flax, maize, rice. barley, rye and oats, amongst others.
18 . An isolated nucleic acid molecule according to claim 16 , wherein said SR structural gene is at least 40% identical to the sequence set forth in SEQ ID NO:1 or a homologue, analogue or derivative thereof.
19 . An isolated nucleic acid molecule according to claim 16 , wherein said SR structural gene further comprises a sequence of nucleotides which is at least 60-65% identical to SEQ ID NO:1 or a complementary strand thereof.
20 . An isolated nucleic acid molecule according to claim 16 , wherein said SR structural gene further comprises a sequence of nucleotides which is at least 70-75% identical to SEQ ID NO:1 or a complementary strand thereof.
21 . An isolated nucleic acid molecule according to claim 16 , wherein said SR structural gene further comprises a sequence of nucleotides which is at least 80-90% identical to SEQ ID NO:1 or a complementary strand thereof.
22 . An isolated nucleic acid molecule according to claim 16 , wherein said nucleic acid molecule is capable of hybridising under at least low stringency conditions to the nucleotide sequence set forth in SEQ ID NO:1 or to a complemental strand thereof.
23 .(amended) An isolated nucleic acid molecule comprising a sequence of nucleotides which encodes or is complementary to a sequence which encodes an SR gene product induced in response to a susceptible interaction between a host plant and a fungal pathogen.
24 . An isolated nucleic acid molecule according to claim 23 , wherein said SR gene product comprises an amino acid sequence which is at least 40% identical to the polypeptide set forth in SEQ ID NO:2.
25 . An isolated nucleic acid molecule according to claim 24 , originating from a crop plant selected from the list comprising flax maize, rice, barley, rye and oats, amongst others.
26 . An isolated nucleic acid molecule according to claim 25 , wherein if said nucleic acid molecule originates from maize it encodes the Mis1 polypeptide.
27 . An isolated nucleic acid molecule according to claim 25 , wherein if said nucleic acid molecule originates from flax it encodes the Fis1 polypeptide.
28 . An isolated nucleic acid molecule capable of hybridising under at least low stringency conditions to the flax Fis1 promoter sequence contained in the microorganism deposited under AGAL Accession No.N96/027087.
29 . An isolated nucleic acid primer molecule of at least 10 contiguous nucleotides in length derived from a flax Fis1 nucleotide sequence set forth in any one of SEQ ID NOS: 1, 3, 4.5, 6 OR 7.
30 . A genetic construct which comprises a nucleic acid molecule according to any one of claims 1 to 15.
31 . A genetic construct according to claim 30 , wherein said nucleic acid molecule is operably liked to a structural gene.
32 . A genetic construct according to claim 31 , wherein said structural gene is a reporter gene.
33 . A genetic construct according to claim 32 , wherein said reporter gene is selected from the list comprising GUS, chloramphenicol acetyltransferase and firefly luciferase genes
34 . A genetic construct deposited under AGAL accession no. N96/027087.
35 . A genetic construct according to claim 31 wherein said structural gene comprises a nucleotide sequence which is at least 40% identical to SEQ ID NO:1 or a homologue, analogue or derivative thereof
36 . A genetic construct according to claim 31 , wherein said structural gene is a cytotoxin gene.
37 . A genetic construct. according to claim 36 , wherein said cytotoxin gene is a barnase or other ribonuclease gene.
38 . A genetic construct according to claim 30 , wherein said nucleic acid molecule is operably linked to a ribozyme, antisense or co-supression molecule which is capable of inhibiting the expression of a plant gene required for viability of a plant cell.
39 . A transgenic plant transformed with a genetic construct according to claim 30 .
40 . A transgenic plant transformed with a genetic construct according to claim 31 .
41 . A transgenic plant transformed with a genetic construct according to claim 32 .
42 . A transgenic plant transformed with a genetic construct according to claim 33
43 . A transgenic plant transformed with a genetic construct according to claim 34 .
44 . A transgenic plant transformed with a genetic construct according to claim 35
45 . A transgenic plant transformed with a genetic construct according to claim 36 .
46 . A transgenic plant transformed with a genetic construct according to claim 37 .
47 . A transgenic plant transformed with a genetic construct according to claim 38 .
48 . A genetic construct comprising a nucleic acid molecule according to any one of claims 16 to 27 .
49 . A recombinant SR gene product comprising a sequence of amino acids which is at least 40% identical to SEQ ID NO:2.
50 . A recombinant SR gene product according to claim 49 further comprising at least one of the amino acid sequence motifs selected from the list comprising GPL, GSG, GQG and EEP as hereinbefore defined in Table 2.
51 . An isolated antibody molecule capable of binding to the recombinant SR gene product of claim 49 or 50 .
52 . A method of producing a plant with improved resistance to the spread of a fungal pathogen in a susceptible interaction, said method comprising the steps of transforming a plant cell with a genetic construct according to any one of claims 36 to 38 and regenerating a whole plant therefrom.
53 . A method according to claim 5 . wherein said plant and respective fungal pathogen is selected from the list of pathogen:host plant combinations comprising:
PATHOGEN
HOST PLANT
Puccinia graminis
wheat, barley and rye
Puccinia striformis
wheat and rye
Puccinia recondita
rye and wheat
Puccinia hordei
barley
Puccinia coronata
oat
Puccinia sorghi
maize
Puccinia polysora
maize
Puccinia purpurea
sorghum
Puccinia sacchari
sugar cane
Puccinia kuehnii
sugar cane
Puccinia arachidis
peanut
Puccinia stachmanii
cotton
Uromyces striatus medicaginis
alfalfa
Uromyces phaseoli
phaseolus beans
Hemileia vastatrix
coffee rust
Melampsora lini
flax
Gymnosporangium juniperi-virginianae
cedar and apple
Cronartium ribicola
white pine
Cronartium fusiforme
loblolly and slash pine
54 . A method according to claim 53 , wherein said plant is selected from the list comprising flax, maize, wheat, barley, oats, rye and rice.
55 . (amended) A method of detecting infection of a plant by a fungal pathogen in a susceptible interaction, said method comprising detecting increased expression of an SR structural gene in said plant by contacting a nucleic acid molecule derived from said plant with an isolated SR structural gene or a fragment of at least 10 contiguous nucleotides in length derived therefrom for a time and under conditions sufficient to allow a double-stranded nucleic acid molecule to form.
56 .(amended) The method according to claim 55 , wherein the isolated SR structural gene comprises the nucleotide sequence set forth in SEQ ID NO:1 or a fragment of at least 10 contiguous nucleotides in length derived therefrom.
57 .(amended) A method according to claim 56 . wherein the isolated SR structural gene is labelled with a reporter molecule.
58 . A method according to claim 57 , further comprising the step of detecting the amount of bound reporter molecule.
59 . A method according to claim 55 when used for the detection of an infection in a plant by a fungal pathogen, wherein said plant and respective fungal pathogen is selected from the list of pathogen:host plant combinations comprising:
PATHOGEN
HOST PLANT
Puccinia graminis
wheat, barley and rye
Puccinia striformis
wheat and rye
Puccinia recondita
rye and wheat
Puccinia hordei
barley
Puccinia coronata
oat
Puccinia sorghi
maize
Puccinia polysora
maize
Puccinia purpurea
sorghum
Puccinia sacchari
sugar cane
Puccinia kuehnii
sugar cane
Puccinia arachidis
peanut
Puccinia stachmanii
cotton
Uromyces striatus medicaginis
alfalfa
Uromyces phaseoli
phaseolus beans
Hemileia vastatrix
coffee rust
Melampsora lini
flax
Gymnosporangium juniperi-virginianae
cedar and apple
Cronartium ribicola
white pine
Cronartium fusiforme
loblolly and slash pine
60 . A method according to claim 59 , wherein said plant is selected from the list comprising flax, maize, wheat, barley, oats, rye and rice.
61 . A progeny plant derived from the transgenic plant according to any one of claims 27 to 33 .Join the waitlist — get patent alerts
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