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57,223 randomly tagged Magnaporthe grisea lines with altered phenotypes,
including changes in pathogenicity towards rice, were created by the
University of Kentucky, University of Arizona and North Carolina State University.
Samples of the generated mutants are available for public
access at Fungal Genetics Stock Center
(Kansas City,KS).
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Growth Rate Assay:
Radial growth of monoconidial cultures was scored on complete medium containing
Hygromycin B (200mg/L) in 24-well plates. Growth was scored after 7 days of
growth at 28°C. The rate of growth is compared to the average rate of the
transformed lines instead of the parental strain (70-15), which is unable to
grow in the presence of Hygromycin.
: [ Total assays: 49248 (57223) ]
| Assay Type |
Description |
Percentage |
| NORMAL | mycelium fills most of the well (most strains fit this category). | 84.47 % | | NO GROWTH | | 2.80 % | | SLOW | mycelium fills 1/2-3/4 of well. | 5.64 % | | VERY SLOW | mycelium fills 1/4 of well. | 3.58 % | | FAST | mycelium fully fills the well. | 3.50 % |
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Conidiation Assay:
Defects in sporulation are scored on oatmeal agar in 24-well plates after 7 days
of growth at 25°C under constant fluorescent illumination.
: [ Total assays: 48264 (57223) ]
| Assay Type |
Description |
Percentage |
| NORMAL | Wild-type 70-15 growth with comparable pigmentation (in the case of pigmentation mutants, the intensity of the pigmentation as compared with mycelial growth) | 82.16 % | |
| LOW | Less pigmentation than 70-15. May be normal mycelial growth with little pigmentation, although not white as in albino mutants. | 4.87 % | |
| NO SPORULATION | no conidia are apparent. | 11.36 % | |
| HIGH | Generally greater mycelial growth and greater intensity of pigmentation as compared to wild-type 70-15 | 1.60 % |
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Pigmentation Assay:
Defects in the melanin biosynthetic pathway produce pigment mutants called albino,
buf or rosy. Pigment color was initially scored by the pigmentation of cultures
grown on oatmeal agar 24-well plates after 7 days of growth at 25°C under lights:
: [ Total assays: 48264 (57223) ]
| Assay Type |
Description |
Percentage |
| GRAY | Pigmentation like wild-type strain 70-15 | 99.89 % | | LIGHT GRAY | | 0.02 % | | BUFF | buff/ tan pigmentation | 0.06 % | | ALBINO | white pigmentation | 0.02 % | | LITTLE PIGMENT | | 0.02 % |
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Auxotrophy Assay:
Strains were screened for ability to grow on minimal medium and complete medium in
parallel. Assays were done in 24-well plates and scored after 10 days of growth at
28°C. Strains that failed to grow or exhibitied greatly reduced growth on miminal
medium compared to strain 70-15, while able to grow on complete medium, indicates
a putative nutritional defect. Additional tests to identify the specific
nutritional defect were performed. Tests first involved supplementing media with
groups of related compounds to place mutants into a category (vitamin, nuclei
acids or amino acids). Next mutants were supplemented with individual compounds
within the identified category to determine the specific defect.
: [ Total assays: 54408 (57223) ]
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| NO | NICOTINICACID | ISOLEUCINE, METHIONINE | LEUCINE | TESTING | INOSITOL | ARGININE, ORNITHINE | ORNITHINE | CYSTEINE, METHIONINE, THIOSULPHATE | ISOLEUCINE |
| 99.96 % | 0.01 % | 0.00 % | 0.00 % | 0.02 % | 0.00 % | 0.00 % | 0.00 % | 0.00 % | 0.00 % |
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Pathogenicity Assay:
: [ Total assays: 27705 (57223) ]
| Assay Type |
Description |
Percentage |
| 0 |
non-pathogenic, no lesions on leaves | 6.57 % | | 1 |
reduced pathogenicity compared to 70-15. Fewer lesions or smaller lesions
and limited or no effect on plant stature. | 20.19 % | | 2 |
lesion size and numbers equivalent to wild-type strain 70-15. | 73.23 % | | 3 |
hypervirulent, devastation of plants - greater than wild-type 70-15 | 0.01 % |
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[ Top ]
[1] Nicole M Donofrio, Ravi Rajagopalon, Douglas E Brown, Stephen E Diener,
Donald E Windham, Shelly Nolin, Anna Floyd, Thomas K Mitchell, Natalia Galadima,
Sara Tucker, Marc J Orbach, Gayatri Patel, Mark L Farman, Vishal Pampanwar, Cari
Soderlund, Yong-Hwan Lee, Ralph A Dean (2005). PACLIMS: A component LIM system
for high-throughput functional genomic analysis. BMC Bioinformatics 6:94
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