Assessment of Bacterial Leaf Blight Incidence and Severity in Rice Growing Areas of Pakistan
Rafia Ahsan1, Saif Ullah1, Ijaz Yaseen1, Faisal Sohail Fateh1, Muhammad Fayyaz1, Shahzad Asad1, Atif Jamal1, Muhammad Sufyan2 and Muhammad Zakria1*
(A), (B) and (C) Rice fields infected with BLB in KPK, Sindh and Punjab province respectively. (D), (E); The leaves showedyellowish lesions/stripes at the margins and along midrib with wavy edges.
(A); Surface sterilization of BLB leaves. (B); Leaves cut into small pieces with sterilized scissors in a tube containing ddH2O and the bacteria were allowed to be ooze out. (C); Streaking of bacterial suspension with wooden stick on Wakimoto’s media.
(A) Isolation of Xoo on Wakimoto’s media; Yellow, smooth, shiny circular Xoo colonies after 3-5 days of incubation at 28-30ºC. (B); Single colonies sub cultured on fresh plates for colony PCR. (C), Agarose gel electrophoresis: Multiplex colony PCR of samples from KP (1-4) and Punjab (5-8) showing band size of 1500bp, 331bp and 162bp with 16SrDNA, Xo3756 (F, R) and Xoo3866 (F, R) primers respectively. 100bp+ Ladder between line 5 and 6, 9 line is PXO-99 (positive control), 10 is Negative control.
(A, B and C); Typical symptoms of BLB observed on Basmati leaves after 21 days of inoculation.