Method to screen compounds for antifungal activity and pharmaceutical compositions and methods to treat fungal diseases by inhibiting spore germination

A method of testing compounds for activity to inhibit germination of spores, the method comprising:

• (a) providing bacterial, fungal, or plant spores transformed to contain and express a detectable marker, wherein the marker when expressed, is operationally linked to a gene selected from the group consisting of SEQ. ID. NOS:
• 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 30, 31, 33, and 35, in a medium and under environmental conditions in which the spores will germinate, and measuring a first signal output generated by the marker prior to the spores initiating germination, wherein the detectable marker, when expressed, is a fluorophore or a chemiluminescent marker;
• (b) contacting the spores of step (a) with a compound whose activity to inhibit germination of spores is to be measured;
• (c) incubating the spores of step (b) under environmental conditions and for a time wherein spores not treated with the compound will germinate;
• and
• (d) determining extent of germination of the spores by measuring a second signal output generated by the marker, wherein a difference between the first signal output and the second signal output is proportional to the extent of germination of the spores.

A method of testing compounds for activity to inhibit germination of spores, the method comprising:

• (a) providing bacterial, fungal, or plant spores transformed to contain and express a detectable marker, wherein the marker when expressed, is operationally linked to a gene encoding a spore-specific protein selected from the group consisting of XP_567740.1 (SEQ. ID. NO:
• 2), XP_566791.1 (SEQ. ID. NO:
• 4), XP_570303.1 (SEQ. ID. NO:
• 6), XP_571089.1 (SEQ. ID. NO:
• 8), XP_571997.1 (SEQ. ID. NO:
• 10), XP_569295.1 (SEQ. ID. NO:
• 12), XP_569173.1 (SEQ. ID. NO:
• 14), XP_569068.1 (SEQ. ID. NO:
• 16), XP_569336.1 (SEQ. ID. NO:
• 18), XP_567136.1 (SEQ. ID. NO:
• 20), XP_568990.1 (SEQ. ID. NO:
• 22), XP_570610.1 (SEQ. ID. NO:
• 24), XP_571921.1 (SEQ. ID. NO:
• 26), XP_572925.1 (SEQ. ID. NO:
• 28), XP_570796.1 (SEQ. ID. NO:
• 30), XP_571548.1 (SEQ. ID. NO:
• 32), XP_570447.1 (SEQ. ID. NO:
• 34), and XP_571343.1 (SEQ. ID. NO:
• 36), in a medium and under environmental conditions in which the spores will germinate, and measuring a first signal output generated by the marker prior to the spores initiating germination;
• (b) contacting the spores of step (a) with a compound whose activity to inhibit germination of spores is to be measured;
• (c) incubating the spores of step (b) under environmental conditions and for a time wherein spores not treated with the compound will germinate;
• and
• (d) determining extent of germination of the spores by measuring a second signal output generated by the marker, wherein a difference between the first signal output and the second signal output is proportional to the extent of germination of the spores.

A method of testing compounds for activity to inhibit germination of spores, the method comprising:

• (a) providing bacterial, fungal, or plant spores transformed to contain and express a detectable marker, wherein the marker when expressed, is operationally linked to a gene selected from the group consisting of SEQ. ID. NOS:
• 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 30, 31, 33, and 35, in a medium and under environmental conditions in which the spores will germinate, and measuring a first signal output generated by the marker prior to the spores initiating germination;
• (b) contacting the spores of step (a) with a compound whose activity to inhibit germination of spores is to be measured;
• (c) incubating the spores of step (b) under environmental conditions and for a time wherein spores not treated with the compound will germinate;
• and
• (d) determining extent of germination of the spores by measuring a second signal output generated by the marker, wherein a difference between the first signal output and the second signal output is proportional to the extent of germination of the spores.

A method of testing compounds for activity to inhibit germination of spores, the method comprising:

• (a) providing bacterial, fungal, or plant spores transformed to contain and express a detectable marker, wherein the marker is operationally linked to a spore-specific or yeast-specific protein, in a medium and under environmental conditions in which the spores will germinate, and measuring a first signal output generated by the marker prior to the spores initiating germination;
• (b) contacting the spores of step (a) with a compound whose activity to inhibit germination of spores is to be measured;
• (c) incubating the spores of step (b) under environmental conditions and for a time wherein spores not treated with the compound will germinate;
• (d) determining extent of germination of the spores by measuring a second signal output generated by the marker, wherein a difference between the first signal output and the second signal output is proportional to the extent of germination of the spores, and
• (e) plotting area and aspect ratio of the spores and any germinated cells after the incubation of step (c).

A method of testing compounds for activity to inhibit germination of spores, the method comprising:

• (a) providing bacterial, fungal, or plant spores transformed to contain and express a detectable marker, wherein the marker when expressed, is operationally linked to a gene encoding a spore-specific protein selected from the group consisting of XP_567740.1 (SEQ. ID. NO:
• 2), XP_566791.1 (SEQ. ID. NO:
• 4), XP_570303.1 (SEQ. ID. NO:
• 6), XP_571089.1 (SEQ. ID. NO:
• 8), XP_571997.1 (SEQ. ID. NO:
• 10), XP_569295.1 (SEQ. ID. NO:
• 12), XP_569173.1 (SEQ. ID. NO:
• 14), XP_569068.1 (SEQ. ID. NO:
• 16), XP_569336.1 (SEQ. ID. NO:
• 18), XP_567136.1 (SEQ. ID. NO:
• 20), XP_568990.1 (SEQ. ID. NO:
• 22), XP_570610.1 (SEQ. ID. NO:
• 24), XP_571921.1 (SEQ. ID. NO:
• 26), XP_572925.1 (SEQ. ID. NO:
• 28), XP_570796.1 (SEQ. ID. NO:
• 30), XP_571548.1 (SEQ. ID. NO:
• 32), XP_570447.1 (SEQ. ID. NO:
• 34), and XP_571343.1 (SEQ. ID. NO:
• 36)), in a medium and under environmental conditions in which the spores will germinate, and measuring a first signal output generated by the marker prior to the spores initiating germination, wherein the detectable marker, when expressed, is a fluorophore or a chemiluminescent marker;
• (b) contacting the spores of step (a) with a compound whose activity to inhibit germination of spores is to be measured;
• (c) incubating the spores of step (b) under environmental conditions and for a time wherein spores not treated with the compound will germinate;
• and
• (d) determining extent of germination of the spores by measuring a second signal output generated by the marker, wherein a difference between the first signal output and the second signal output is proportional to the extent of germination of the spores.