Mutants Data Description

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).

  Statistics     References  

Mutant Assay Statistics:
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.

Assay Scoring: [ Total assays: 49248 (57223) ]

Assay Type Description Percentage
NORMALmycelium fills most of the well (most strains fit this category). 84.47 %
NO GROWTH 2.80 %
SLOWmycelium fills 1/2-3/4 of well. 5.64 %
VERY SLOWmycelium fills 1/4 of well. 3.58 %
FASTmycelium fully fills the well. 3.50 %

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.

Assay Scoring: [ Total assays: 48264 (57223) ]

Assay Type Description Percentage
NORMALWild-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 %
LOWLess pigmentation than 70-15. May be normal mycelial growth with little pigmentation, although not white as in albino mutants. 4.87 %
NO SPORULATIONno conidia are apparent. 11.36 %
HIGHGenerally greater mycelial growth and greater intensity of pigmentation as compared to wild-type 70-15 1.60 %

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:

Assay Scoring: [ Total assays: 48264 (57223) ]

Assay Type Description Percentage
GRAYPigmentation like wild-type strain 70-15 99.89 %
BUFFbuff/ tan pigmentation 0.06 %
ALBINOwhite pigmentation 0.02 %

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.

Assay Percentage: [ Total assays: 54408 (57223) ]

99.96 %0.01 %0.00 %0.00 %0.02 %0.00 %0.00 %0.00 %0.00 %0.00 %

Pathogenicity Assay:

Assay Scoring: [ Total assays: 27705 (57223) ]

Assay Type Description Percentage
0 non-pathogenic, no lesions on leaves6.57 %
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 70-150.01 %

   References[ 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