Representative appressorium stage cDNA library of Magnaporthe grisea

Abstract

A mature appressorium cDNA library of rice blast fungus, Magnaporthe grisea, was constructed in a lambdaTriplEx2 vector by SMART cDNA library containing 2.37x10(6) independent clones about 100% of which harbor foreign cDNA inserts with average size of 660 bp. Of 9 randomly selected clones, 2 expressed sequence tags (ESTs) sequences did not have homologous EST sequences of M. grisea in GenBank. The appressorium cDNA library is suitable for gene expression analysis and function analysis of the late stages of appressorium formation and the early stages of penetration of M. grisea.

Fig. 1

Appressoria of M. grisea GUY11 on a projection membrane inoculated for 23.5–24.5 h at 25 °C

Fig. 2

Appressorium total RNA (5 μl) on a 1.2% denaturing formaldehyde/agarose gel

Fig. 3

Ten μl ds cDNA product synthesized from appressorium RNA electrophoresed on a 1.1% agrose/EtBr gel Lane M, 1-kb DNA size marker (Fermentas, Lithuania) (0.2 μg loaded)

Fig. 4

Sfi I-digested plasmids of 12 clones on a 1% agrose/EtBr gel Lane M, 1-kb DNA marker size (Fermentas, Lithuania). 1~12: clones 1~12