INTRODUCTION

The traditional method of identifying plant pathogens is through visual examination. This is often possible only after major damage has already been done to the crop, so treatments will be of limited or no use. To save plants from irreparable damage by pathogens, farmers have to be able to identify an infection even before it becomes visible. This can be reached by using very sensitive methods for early diagnosis of disease

Diagnostic

Non Diagnostic

These symptoms may be diagnostic or non-diagnostic

By follow these symptoms can be identified that causes disease

1- Become acquainted with the environmental Conditions surrounding the plant 2- Review of agricultural operations 3- examination Of the plant sample 1- Soil analysis and measurement of pH 2- hypothesis Koch 3- Biochemical methods 4- microscopic examination 5- serological methods 6- the use of methods rely on DNA

Local symptoms

Systemic symptoms

As mentioned before , the laboratory methods are: 1- Soil analysis 2- Koch's postulates 3- Biochemical methods 4- Microscopic examination 5- Immunological methods 6- The use of methods rely on DNA

Nucleic acid techniques included :

Direct visualization of Entire genomes

Restriction digestion And electrophoresis Of DNA

Hybridization of DNA and RNA

The polymerase Chain reaction (PCR)

The polymerase chain reaction (PCR)

PCR technique based on the three-step thermal: 1. Denaturation 2. Annealing 3. Extension

The primary materials, or reagents, used in PCR

The primary materials, or reagents, used in PCR are: 1- Template DNA, the DNA sequence that you want to amplify 2- DNA nucleotides, the building blocks for the new DNA 3- Primers, single-stranded DNAs between 20 and 30 nucleotideslong (oligonucleotides) that are complementary to a short region on either side of the template DNA 4- DNA polymerase, a heat stable enzyme that drives, orcatalyzes, the synthesis of new DNA 5- buffer (containing Mg)

Thermocycler or PCR machine

Mg++

Mg++

Mg++

Exponential increase of the number of copies during PCR

To diagnose plant diseases by PCR can you follow these steps

1-DNA extraction 2-PCR amplification 3-Gel Electrophoresis and analysis

DNA extraction

Gel Electrophoresis and analysis

Summary
The use of the most sensitive and modern methods helps a lot in early identification of disease These modern technologies include PCR beside serological methods

And through which , access to the real cause of the disease in a short time is possible before significant losses in yield are encountered