I= PRODUCTION OF PHYTOTOXING FROM ASCOCHYTA RUBIEI, AND MOLECULAR STUDIES OF DEFENCE GENES IN BLIGHT INFECTED CHICKPEA
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Title of Thesis
PRODUCTION OF PHYTOTOXING FROM ASCOCHYTA RUBIEI, AND MOLECULAR STUDIES OF DEFENCE GENES IN BLIGHT INFECTED CHICKPEA

Author(s)
Ahmad Ali Shahid
Institute/University/Department Details
National Centre Of Excellence In Molecular Biology / University Of The Punjab
Session
2004
Subject
Molecular Biology
Number of Pages
121
Keywords (Extracted from title, table of contents and abstract of thesis)
chickpea, fungal blight, antifungal proteins, toxin, phytotoxic compounds, a.rabiei, antibodies, immunological studies

Abstract
The study presented in this thesis is aimed to investigate the production of phytotoxic compounds from Ascochyta rabiei and molecular studies of defence genes in blight infected chickpea. Blight, caused by A.rabiei is one of the most serious disease of chickpea in the world including Pakistan. When a.rabiei was grown on Czapek Dox broth by surface culture method supplemented with an aqueous extract of chickpea seeds, it produced at least three phytotxic compounds, identified as solanapyrone A,B and C. The addition of divalent metal cations such as Zn2+, Ca2+, Mn2+ and Cu2+ to Czapek Dox broth stimulated the production of solanapyrones A,B and C. These phytotoxic compounds were identical to the phytotoxins isolated previously from culture filtrates of the fungus altenaria solani. The compounds were separated by HPLC iwht a column of ODS silica as stationary phase and aqueous mixture of methanol (MeOH), acetonitrile (ACN) and terahydrofuran (THF) as mobile phase. Eluent form the column was monitored by a diode array detector operating in the UV mode.

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1086.12 KB
S. No. Chapter Title of the Chapters Page Size (KB)
1 0 Contents
96.6 KB
2 1 Introduction 01
65.79 KB
3 2 Literature Review 07
155.75 KB
  2.1 Chickpea And Fungal Blight 07
  2.2 Geographical Distribution 08
  2.3 Losses 09
  2.4 Symptoms 09
  2.5 Phytotoxins 10
  2.6 Plant Antifungal Proteins 18
4 3 Materials And Methods 23
145.38 KB
  3.1 Fungal Isolates 23
  3.2 Chickpea Seeds 24
  3.3 Fungal Growth Medium Of Toxin Production 24
  3.4 Extraction Of Sloanapyrones 24
  3.5 Minicolumn 25
  3.6 Toxin Analysis By HPLC 25
  3.7 Purification Of Solanapyrone B 25
  3.8 Cell Bioassay 26
  3.9 Estimation Of Toxins In Blight Infected Chickpea Plants 27
  3.10 Determination Of Antifungal Like Genes /Proteins In Chickpea 28
  3.11 Protein Isolation From Plants 28
  3.12 Western( Immuno ) Blotting 28
  3.13 Plant Genomic DNA Isolating 29
  3.14 RNA Isolation 30
  3.15 Isolation Of DNA Fragment From Agarose 30
  3.16 Southern Blotting 31
  3.17 Probe Labeling, Hybridization And Detection 32
  3.18 Probe Labeling 32
  3.19 Hybridization 32
  3.20 Detection 33
  3.21 Probe Removal From Membrane 33
  3.22 Northern Blotting 33
  3.23 Transfer Of RNA Membrane 33
  3.24 Probe Labeling, Hybridization And Detection 34
  3.25 Electroproation 34
  3.26 Miniplasmid Preparation 35
  3.27 Introduction And Expression Studies 35
  3.28 SDS-PAGE 36
  3.29 Isolation Bacteria Having Antifungal Activities 37
  3.30 Sample Collection 37
  3.31 Isolation Technique 37
  3.32 Antifungal Activity Of Protein On Ascochyta Rabiei 38