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Functional Analysis of Eleven Predicted Unique Genes of Magnaporthe Oryzae Isolate FJ81278

Author: WangXingXing
Tutor: WangZongHua；WangBaoHua
School: Fujian Agriculture and Forestry University
Course: Of Plant Pathology
Keywords: M. grisea Avirulence gene Specific genes Molecular markers
CLC: S435.111.41
Type: Master's thesis
Year: 2011
Downloads: 18
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Abstract


In order to clone avirulence genes in the rice blast fungus strains FJ81278 Avr-Pi1, Avr-Pi2 Avr-Pi4a, map-based cloning position in the BAC clones 805L15 BAC clone sequencing, the sequence may predict The gene encoding the sequenced strain 70-15 genomic sequences obtained by comparing the specific genes in the strains of some FJ81278. Of this study, on which the four specific gene function. Meanwhile, the analysis FJ81278 strain genome sequence, 7 prediction of the signal peptide-specific gene function. Constructed the 11 specific genes that MoUng01 MoUng02 MoUng03 MoUng04 MoUng05 MoUng06 MoUng07 MoUng08 excessive MoUng09 MoUng10, MoUng11 expression vector, and then transferred to M. grisea wild-type strain GUY11 protoplast access to these specific gene overexpression transformants. These transformants by a conidia suspension of the living body was spray-inoculated CO39 nearly isogenic lines rice varieties are to C101LAC the Pi-1 (t), C101A51 the Pi-2 (t), C104PKT Pi-3 (t), C101PKT Pi -4a and C105TTP-4L-23 the Pi-4b. Learned by the experimental results, the M. grisea overexpression pathogenic transformants with the wild-type strain GUY11, speculated avirulence gene Avr-Pi1, Avr-Pi2 the Avr-Pi4a does not exist in these specific genes or These specific genes contain three avirulence genes but not normal expression. These rice blast fungus mutant phenotype analysis shows that rising MoUng05 MoUng03 MoUng07 the overexpression (OE) transformants sporulation, and MoUng04, MoUng06 the contrary; the overexpression conversion of specific genes MoUng01, MoUng09 sub onion epidermal infection capacity declined virulence of rice has not changed. Multiple molecular markers avirulence gene Avr-Pi1 Avr-Pi2 and Avr-Pi4a,, located in the BAC clone 805L15. However, the results of this experiment show that the four specific genes are not derived from the BAC clone three avirulence gene. The laboratory pre using the same method, ruled out the possibility of other predicted genes in the BAC clone as avirulence gene. We analyze the reasons for this result may be the original sequence assembly of BAC clones 805L15 wrong. The last 805L15 clone splicing method using Southern hybridization analysis confirmed the BAC clone sequences spliced. The should improve 805L15 clone of stitching, re-sequence the gene prediction, analyze the function of specific genes in FJ81278 strains to last avirulence gene Avr-Pi1, Avr-Pi2 and Avr-Pi4a.

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CLC: > Agricultural Sciences > Plant Protection > Pest and Disease Control > Crop pests and diseases and their prevention > Cereal crop pests and diseases > Rice pests and diseases > Disease > Transgression ( pass ) an infectious agent harmful > Rice blast
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