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Para Sexual Recombination Between Blast Fungi Pyricularia Oryzae and P. Grisea on Double Inoculated Lesion

M A Monsur
Senior Scientific Officer
Plant Pathology Division, Bangladesh Rice Research Institute, Gazipur-1701, Bangladesh

Abstract/ Executive Summary:

An investigation was carried out to know the recombination between Pyricularia oryzae and P. grisea. Two rice blast isolates, TP106 and TP022 were used in this study. Conidial suspension of SA13-1ME was sprayed on it's host. Seven days after first inoculation , an agar plug with conidia of TP106 or TP022 was put on blasts lesions of leaves. At least five monoconidial isolates were collected from each of the double inoculated lesions. Internal transcribed spacer (ITS) region was amplified from the monoconidial isolates. The amplicons was digested with Dra1, And separated on 1.5% agarose gel. The monoconidial isolates were also subjected to MAGGY-DNA fingerprint analysis. An investigation was carried out to know the recombination between Pyricularia oryzae and P. grisea. A total of 520 isolates collected from the double inoculated lesions were subjected to PCR-RFLP analysis of the ITS region to identify subcultures of the inoculated rice blast isolates. As a result, four isolates from the double inoculated lesions with SA13-1ME and TP 106 were identified as subcultures of TP106. The 319 isolates from the 48 double inoculated lesions with SA13-1ME and TP106, four isolates Ca32-1, Ca32-7, Ca42-4 and Ca44-4 had the PCR product that could be digested with DraI. SA13-1ME and TP106 differed in mycelial color, i.e., SA13-1ME produced colony with black while TP106 produced white colour. The four isolates had colonies with the same color as that of colony of TP106. On the other hands, the other 315 isolates had colonies with black. These results indicate that the four isolates were derived from TP106. In other words, TP106 succeeded in colonization in the three lesions, Ca32, Ca42 and Ca44. On the contrary, in the 201 isolates from the 28 double inoculated lesions with SA13-1ME and TP022, isolates were not derived from TP022. All of the 201 isolates had PCR product that could not be digested with DraI. A total of 17 isolates from three double inoculated lesions were subjected to MAGGY- DNA fingerprint analysis to access the recombination genotypes. However, recombinant DNA fingerprint patterns between TP106 and SA13-1ME were not detected among the 17 isolates. Although TP022 was not recovered from the double inoculated lesions, the fact that TP106 was recovered from the double inoculated lesion indicated that rice blast fungus could invade and colonized in blast lesions on leaves. Therefore, it can be expected that the recombinant event occurs in nature only when the rice blast pathogen opportunistically infects a blast lesions on it’s host in which the pathogen preexist.

Keywords: Rice, Blast, Pathogen, Parasexual recombination, Pyricularia oryzae, Pyricularia grisea.

Location: Molecular lab, Plant Pathology

Start Date: 01-07-2014

End Date: 30-06-2015

Program Area: Pest Management

Commodity: Rice

Objectives:

To know the recombination between Pyricularia oryzae and P. grisea.

Methodology:

Two rice blast isolates, TP106 and TP022 and one crabgrass isolate SA13-1ME were used in this study. Conidial suspension of SA13-1ME was sprayed on it's host accession, SACB-1 as the first inoculation. Seven days after first inoculation , an agar plug with conidia of TP106 or TP022 was put on blasts lesions on the crab grass leaves. At least five monoconidial isolates were collected from each of the double inoculated lesions. Internal transcribed spacer (ITS) region was amplified from the monoconidial isolates. The amplicons was digested with Dra1, And separated on 1.5% agarose gel. The monoconidial isolates were also subjected to MAGGY-DNA fingerprint analysis.

Source of Information: BRRI Annual Report 2014-15.

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

It can be expected that the recombinant event occurs in nature only when the rice blast pathogen opportunistically infects a blast lesions on it's host.

Knowledge Management: Report/Proceedings