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Seed health testing for bacterial infection

Seed health testing for bacterial infection

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Published by Babu Ram Panthi
Seed Health Testing for Bacterial Infection
Seed Health Testing for Bacterial Infection

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Published by: Babu Ram Panthi on Jul 11, 2011
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12/26/2012

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Compiled by:
Babu Ram PanthiBScAgIAAS, Rampur CampusRampur, Chitwan, Nepalcreation_babu@yahoo.com
 Table Of Contents
S.No
.
Contents
 
Page No.1. Introduction 12. Objectives 93. Materials and Method 104. Literature review 105. Summary and Conclusion 12
 
Plant Pathology
2010
C
ompiled By: Babu Ram Panthi Page 1
 
6
. References 13
 
1.
 
INTRODUCTION 
S
eed health
Seed heath refers mainly to the presence or absence of disease causing organism e.g. Fungi, bacteria, viruses, insects pest and even the physiological condition, for trace element deficiency.Seed health is one of the most important component of seed quality. Disease or pest infested seed play a major role in the spread of diseases causing pathogens and pest from one place to another.Fungi, bacteria, viruses, nematodes, eggs and /or larva of insect and even deficiency of microelements may cause diseases which may be transmitted to successive crops, affecting its yield potentiality. Health seed should, therefore, be free from such diseases, insect pest, other  pathogenic organisms and deficiency symptoms. Te importance of detection and control of seed borne diseases have been practiced for many years. Burning of wheat seed for controlling smutor tossing of sorghum and sugarcane seeds on straw flame were practiced by farmers in the 18
th
 century. Such indigenous method were gradually replaced by scientific methods andtechnologies for the use of fungicides and pesticides as prophylactic treatment were developed to prevent the occurrence of seed borne diseases. Production of healthy seeds, which don¶t carrydisease causing factors, help to maintain seed health. Seed health testing information is importantfor the following reasons:a)
 
Seed borne microbes may give rise to disease development in the field and reducecommercial value of the crop. b)
 
Imported seed lots may introduce new diseases and insect pests.c)
 
It may cause poor germination or field stand there by decimating yield.
S
eed Borne Diseases
The term µSeed Borne¶ merely indicates that the pathogen and seed are associated in oneof the following ways.
Accompanying
:- The pathogen may independently accompany the seed, without being attachedto it, for e.g. Ergot sclerotia smut balls etc.
External
:- The pathogen may be carried passively on the surface of the seeds, such as, fungusspores, nematode larvae or a virus.
Internal
: - It is carried embedded in plant parts essential for the production of the seedlings, for e.g. loose smut of wheat wherein the pathogen is imbedded in the embryo.
S
eed Health Testing
: Seed health testing is the science of determining the presence or absenceof disease causing agents, such as fungi, bacteria and viruses and animal pests such as eelwormsand insects in the seed samples. The extent of presence of disease infected seeds or the
 
Plant Pathology
2010
C
ompiled By: Babu Ram Panthi Page 2
 
infestation by insect pests determines the seed heath status of a seed sample and by inferenceseed lots.
Diagnostic
S
trategy
It is based for most diseases upon symptoms, isolation and practical identification schemesusing relatively few but important key tests. When testing for seed borne bacterial pathogen fromseeds, the following steps are usually involved:1)
 
Extraction of bacteria from seeds.2)
 
Isolation and purification.3)
 
Identification of the bacterial isolates.
Extraction of bacteria from seeds
Bacteria can be extracted from seeds by following methods:1.
 
Direct plating of seeds on various media followed by incubation at optimum conditions,especially temperatures ranging between 25 and 30°C, and darkness.2.
 
Soaking either whole seeds or ground-up seeds in sterile water or saline water for different durations followed by plating the water containing bacterial cells onto differentmedia including semi selective or selective ones.3.
 
growing seedlings from seeds on various substrates and then isolating bacteria from plantshowing symptoms followed by the streaking onto a medium, preferably general oneslike Nutrient agar (NA), Growth factor (GF), KB etc
Isolation and purification of bacteria
Bacteria thus extracted are isolated on media of special composition and purified in order to be able to identify them. The procedure is described below.1.
 
Surface sterilize infected plants by dipping into 95% ethanol before treating with NaOCl(0.5-1%) solutions for 2-5 minutes and rinse 2-3 times with sterile water. Leaf materialsor roots can also be washed in running tap water for 1-2 hours.2.
 
Cut small area of the affected tissues on a clean glass slide and mount in a drop of water.Put a cover slip and examine under compound microscope for streaming of bacterialcells. Only if bacterial ooze is observed proceed with further steps of isolation.3.
 
Remove the cover slip and put the infected tissue in a few more drops of water, teaseapart he tissue and leave it for 10-15 minutes to obtain more bacterial cells in the water.4.
 
Using a bacteriological loop streak the water containing bacteria a number of times on asuitable medium in a cross wise manner. Try to get single colony of bacterium.5.
 
Incubate the streaked plates in an inverted position at 25-30°C.6.
 
Start observing the plates after 1-2 days for pseudomonas and up to 4 days for xanthomonads. The colonies of pseudomonads on nutrient agar are white, or whitish

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