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Talapati Aruna Chenna Vydyanad
MSc Agri Plant Pathology
Virulence Analysis of Citrus canker caused by
Xanthomonas axonopodis pv. citri in Madhya Pradesh
Region
Introduction
Citrus is a member of the Rutaceae family and the Aurantiodeae subfamily.
Citrus is thought to have originated in the region of South East Asia surrounded by
North East India, Myanmar (Burma), and China's Yunnan Province.
India ranks sixth in the production of citrus fruit in the world.
Madhya Pradesh is the country's fourth largest producer of citrus fruits, accounting
for 10.8 percent of total country share, with a production of 0.80 MT of citrus on
an area of 0.05 M. ha.
.
Objectives:
1. Survey for incidence and severity if citrus canker in different citrus growing area
and collection of disease samples.
2. Isolation, purification and identification of plant pathogen from collected samples
of different citrus growing areas.
3. Evaluation of bactericide agents under In-vitro condition.
4. Evaluation of botanical agents under In-vitro condition.
Symptomatology
The symptoms usually appear on leaves, stems, twigs and Fruits.
Leaves – Ultimately evolve Tan and then brown, with water-soaked margins and a
yellow halo. Lesions will also emerge and old lesions eventually fall out
Stems and Twigs – Corky lesions that are brown in color. These are too
water-soaked. Lesions on the stems and twigs are often dry and scabby as
they age
Fruit – Dark brown lesions appears and similar to those found in stems and
twigs. Fruits also having yellow halo lesions.
Symptoms on Leaves, Stems and Fruit
RESULT
Sr.No Tehsil Village Age of
Orchard
Variety Disease %
Incidence Severity
Gwalior district
1 Bhitarwar Adampur 2 Local 33 30
2 Bagwai 4 Local 37 28
3 Bajna 3 Local 22 28
Tehsil average 30.67 28.67
2 Bhatigaon 3 Local 32 30
Tehsil average 32 30
3 Morar Aaroli 4 Local 29 18
Adupura 4 Local 20 28
Arora 2 Local 32 26
Tehsil average 27 24
Shivpuri district
1 Pohri Bhilodi 4 Local 38 36
Umari 2 Local 26 27
Gohara 3 Local 31 30
Tehsil average 31.67 31
2 Kolaras Dangora 3 Local 22 30
Atara 1 Local 29 27
Anantpur 2 Local 18 16
Tehsil average 23 24.3
Canker incidence and severity of acid lime orchards in Gwalior, Shivpuri,
Bhind, and Guna districts during 2022.
Bhind district
1 Lahar Ajnar 4 Local 16 14
Madori 5 Local 12 19
Sijroli 3 Local 10 12
Tehsil average 12.7 15
2. Mehgaon Bahera 2 Local 27 40
Bahua 3 Local 40 37
Baharpura 1 Local 15 10
Tehsil average 27.3 29
3 Ater Arjunpura 3 Local 36 39
Rohinda 2 Local 34 32
Modhana 3 Local 30 28
Tehsil average 24 16.66
Guna district
1 Raghogarh Amber 2 Local 22 17
Badarpur 3 Local 26 21
Balakheda 4 Local 24 12
Tehsil average 24 16.7
2 Aron Arjunpura 5 Local 6 4
Amoda 4 Local 18 16
Ankkheda 6 Local 24 30
Tehsil average 16 16.67
3. Bamori Ajronda 3 Local 38 36
Akoda 2 Local 26 22
Am kheda 4 Local 32 34
Tehsil average 32 30.7
Tehsil wise incidence and severity of citrus canker on acid lime in the
orchard of Gwalior, Shivpuri, Bhind and Guna districts.
Sr.
No
Districts Tehsil No.of Villages
surveyed
Disease %
Incidence Severity
1 Gwalior Bhitarwar 2 29 35
Bhatigaon 2 23.5 28.5
Morar 2 34 30.5
Total 6
Mean 28.8 31.3
2 Shivpuri Pohri 2 22 24
Umari 2 30.5 28
Gohara 1 26 12
Total 5
Mean 26.1 21.3
3 Bhind Lahar 2 00 00
Mehgaon 2 35 36.5
Ater 2 24 19
Total 6
Mean 19.66 18.5
4 Guna Raghogarh 2 12 16.5
Aron 2 14 9
Bamori 2 12.5 11
Total 6
Mean 12.83 12.16
Sr.No Isolates Isolates from Host Infected Plant parts
1 Xac 1 Aron village C. aurantifolia Leaf
2 Xac 2 Arroli village C. aurantifolia Leaf
3 Xac 3 Bahera village C. aurantifolia Leaf
4 Xac 4 Morar village C. aurantifolia Leaf
5 Xac 5 Bhitarwar village C. aurantifolia Leaf
Isolation of the casual organism
Table3. Different isolates use in the study of morphological, cultural,
biochemical, pathogenicity and In-vitro efficacy of Plant extracts and
antibiotics/antibacterial chemicals.
Virulence analysis of different leaves isolate
Test isolates Incubation period(Day) Symptoms on leaves
Xac1 12 Lesion surrounded by yellow halo
Xac2 12 Crater like lesion
Xac3 14 Lesion surrounded by yellow halo
Xac4 16 Crater like lesion
Xac5 14 Lesion surrounded by yellow halo
Table.4 Type of symptoms and Incubation period developed in pathogenicity test.
Table.4 Cultural characteristic of X. campestris pv.citri isolates
Sr.
No.
Isolates Colony
colour
Colony
shape
Margin Surface Elevation Cell
shape
After 72
hrs colony
diameter(
mm)
1 Xac1 Yellow Circular Entire Mucoid Convex Rod 4.5
2. Xac2 Light
yellow
Circular Entire Mucoid Convex Rod 4.2
3. Xac3 Light
yellow
Circular Entire Mucoid Convex Rod 5.1
4. Xac4 yellow Circular Entire Mucoid Convex Rod 6.4
5. Xac5 yellow Circular Entire Mucoid Convex Rod 5
Biochemical characteristics of Xanthomonas axonopodis pv.citri.
Test
isolates
KOH
test
H2S
test
Catal
ase
test
Starch
hydrolysi
s
Gram
staining
Xac1 +ve +ve +ve +ve -ve
Xac2 +ve +ve +ve +ve -ve
Xac3 +ve +ve +ve +ve -ve
Xac4 +ve +ve +ve +ve -ve
Xac5 +ve +ve +ve +ve -ve
Antibiotics/Anti
bacterial chemicals and
Botanicals
Concentration Inhibition zone (mm)
Treatments Xac1 Xac2 Xa3 Xac4 Xac5 Mean
Streptomycin sulphate 0.0025%25ppm 25.1 24.3 14.4 21.4 17.5 20.5
Tetracycline
hydrochloride
0.02% 23.6 20.3 18.8 19.6 17.9 20.04
Bordeaux mixture 0.40% 19.7 16.9 19 21.9 18.2 19.1
Streptocycline 0.02% 16.4 9.1 4.2 0 11.1 8.2
Copper oxychloride 0.30% 16.4 16.7 14.2 14.3 15.5 15.4
Tulsi extract 10% 18.5 17 19.2 15.2 17.8 17.5
Neem extract 5% 17.5 18.6 18.2 8.9 15.7 15.8
Jasmine extract 12% 10.6 11.7 13.5 12.8 7 11.1
Control Water(H2O) 0 0 0 0 0 0
Total 16.4 14.9 13.5 12.7 13.4
CD at 5% 4.17
S.Em 1.45
Invitro evaluation of Antibiotic/ Anti bacterial chemicals and Botanicals against
Xanthomonas axonopodis pv. citri isolates.
CONCLUSION
Findings of survey showed that maximum average percent disease incidence &
disease severity were found in the Bamori Tehsil (32 & 32.7%) and lowest average
percent disease incidence & disease severity was founded in Lahar Tehsil (12.7 &
15 %, respectively).
A bacterium is gram-negative, round ended, rod-shaped, with only one polar
flagellum. isolates, Xac1, Xac4, and Xac5 were yellow, while the other two, Xac2
and 3, were yellow to light yellow in color.
Streptomycin sulphate was effective against the pathogen with maximum
inhibition zone of 20.5mm. Lower efficacy was exhibited by streptocycline 8.2
mm
THANK YOU

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Virulence Analysis of Citrus canker caused by Xanthomonas axonopodis pv. citri in Madhya Pradesh Region

  • 1. Talapati Aruna Chenna Vydyanad MSc Agri Plant Pathology Virulence Analysis of Citrus canker caused by Xanthomonas axonopodis pv. citri in Madhya Pradesh Region
  • 2. Introduction Citrus is a member of the Rutaceae family and the Aurantiodeae subfamily. Citrus is thought to have originated in the region of South East Asia surrounded by North East India, Myanmar (Burma), and China's Yunnan Province. India ranks sixth in the production of citrus fruit in the world. Madhya Pradesh is the country's fourth largest producer of citrus fruits, accounting for 10.8 percent of total country share, with a production of 0.80 MT of citrus on an area of 0.05 M. ha. .
  • 3. Objectives: 1. Survey for incidence and severity if citrus canker in different citrus growing area and collection of disease samples. 2. Isolation, purification and identification of plant pathogen from collected samples of different citrus growing areas. 3. Evaluation of bactericide agents under In-vitro condition. 4. Evaluation of botanical agents under In-vitro condition.
  • 4. Symptomatology The symptoms usually appear on leaves, stems, twigs and Fruits. Leaves – Ultimately evolve Tan and then brown, with water-soaked margins and a yellow halo. Lesions will also emerge and old lesions eventually fall out Stems and Twigs – Corky lesions that are brown in color. These are too water-soaked. Lesions on the stems and twigs are often dry and scabby as they age Fruit – Dark brown lesions appears and similar to those found in stems and twigs. Fruits also having yellow halo lesions.
  • 5. Symptoms on Leaves, Stems and Fruit
  • 6. RESULT Sr.No Tehsil Village Age of Orchard Variety Disease % Incidence Severity Gwalior district 1 Bhitarwar Adampur 2 Local 33 30 2 Bagwai 4 Local 37 28 3 Bajna 3 Local 22 28 Tehsil average 30.67 28.67 2 Bhatigaon 3 Local 32 30 Tehsil average 32 30 3 Morar Aaroli 4 Local 29 18 Adupura 4 Local 20 28 Arora 2 Local 32 26 Tehsil average 27 24 Shivpuri district 1 Pohri Bhilodi 4 Local 38 36 Umari 2 Local 26 27 Gohara 3 Local 31 30 Tehsil average 31.67 31 2 Kolaras Dangora 3 Local 22 30 Atara 1 Local 29 27 Anantpur 2 Local 18 16 Tehsil average 23 24.3 Canker incidence and severity of acid lime orchards in Gwalior, Shivpuri, Bhind, and Guna districts during 2022.
  • 7. Bhind district 1 Lahar Ajnar 4 Local 16 14 Madori 5 Local 12 19 Sijroli 3 Local 10 12 Tehsil average 12.7 15 2. Mehgaon Bahera 2 Local 27 40 Bahua 3 Local 40 37 Baharpura 1 Local 15 10 Tehsil average 27.3 29 3 Ater Arjunpura 3 Local 36 39 Rohinda 2 Local 34 32 Modhana 3 Local 30 28 Tehsil average 24 16.66 Guna district 1 Raghogarh Amber 2 Local 22 17 Badarpur 3 Local 26 21 Balakheda 4 Local 24 12 Tehsil average 24 16.7 2 Aron Arjunpura 5 Local 6 4 Amoda 4 Local 18 16 Ankkheda 6 Local 24 30 Tehsil average 16 16.67 3. Bamori Ajronda 3 Local 38 36 Akoda 2 Local 26 22 Am kheda 4 Local 32 34 Tehsil average 32 30.7
  • 8. Tehsil wise incidence and severity of citrus canker on acid lime in the orchard of Gwalior, Shivpuri, Bhind and Guna districts. Sr. No Districts Tehsil No.of Villages surveyed Disease % Incidence Severity 1 Gwalior Bhitarwar 2 29 35 Bhatigaon 2 23.5 28.5 Morar 2 34 30.5 Total 6 Mean 28.8 31.3 2 Shivpuri Pohri 2 22 24 Umari 2 30.5 28 Gohara 1 26 12 Total 5 Mean 26.1 21.3 3 Bhind Lahar 2 00 00 Mehgaon 2 35 36.5 Ater 2 24 19 Total 6 Mean 19.66 18.5 4 Guna Raghogarh 2 12 16.5 Aron 2 14 9 Bamori 2 12.5 11 Total 6 Mean 12.83 12.16
  • 9. Sr.No Isolates Isolates from Host Infected Plant parts 1 Xac 1 Aron village C. aurantifolia Leaf 2 Xac 2 Arroli village C. aurantifolia Leaf 3 Xac 3 Bahera village C. aurantifolia Leaf 4 Xac 4 Morar village C. aurantifolia Leaf 5 Xac 5 Bhitarwar village C. aurantifolia Leaf Isolation of the casual organism Table3. Different isolates use in the study of morphological, cultural, biochemical, pathogenicity and In-vitro efficacy of Plant extracts and antibiotics/antibacterial chemicals.
  • 10. Virulence analysis of different leaves isolate Test isolates Incubation period(Day) Symptoms on leaves Xac1 12 Lesion surrounded by yellow halo Xac2 12 Crater like lesion Xac3 14 Lesion surrounded by yellow halo Xac4 16 Crater like lesion Xac5 14 Lesion surrounded by yellow halo Table.4 Type of symptoms and Incubation period developed in pathogenicity test.
  • 11. Table.4 Cultural characteristic of X. campestris pv.citri isolates Sr. No. Isolates Colony colour Colony shape Margin Surface Elevation Cell shape After 72 hrs colony diameter( mm) 1 Xac1 Yellow Circular Entire Mucoid Convex Rod 4.5 2. Xac2 Light yellow Circular Entire Mucoid Convex Rod 4.2 3. Xac3 Light yellow Circular Entire Mucoid Convex Rod 5.1 4. Xac4 yellow Circular Entire Mucoid Convex Rod 6.4 5. Xac5 yellow Circular Entire Mucoid Convex Rod 5
  • 12. Biochemical characteristics of Xanthomonas axonopodis pv.citri. Test isolates KOH test H2S test Catal ase test Starch hydrolysi s Gram staining Xac1 +ve +ve +ve +ve -ve Xac2 +ve +ve +ve +ve -ve Xac3 +ve +ve +ve +ve -ve Xac4 +ve +ve +ve +ve -ve Xac5 +ve +ve +ve +ve -ve
  • 13. Antibiotics/Anti bacterial chemicals and Botanicals Concentration Inhibition zone (mm) Treatments Xac1 Xac2 Xa3 Xac4 Xac5 Mean Streptomycin sulphate 0.0025%25ppm 25.1 24.3 14.4 21.4 17.5 20.5 Tetracycline hydrochloride 0.02% 23.6 20.3 18.8 19.6 17.9 20.04 Bordeaux mixture 0.40% 19.7 16.9 19 21.9 18.2 19.1 Streptocycline 0.02% 16.4 9.1 4.2 0 11.1 8.2 Copper oxychloride 0.30% 16.4 16.7 14.2 14.3 15.5 15.4 Tulsi extract 10% 18.5 17 19.2 15.2 17.8 17.5 Neem extract 5% 17.5 18.6 18.2 8.9 15.7 15.8 Jasmine extract 12% 10.6 11.7 13.5 12.8 7 11.1 Control Water(H2O) 0 0 0 0 0 0 Total 16.4 14.9 13.5 12.7 13.4 CD at 5% 4.17 S.Em 1.45 Invitro evaluation of Antibiotic/ Anti bacterial chemicals and Botanicals against Xanthomonas axonopodis pv. citri isolates.
  • 14. CONCLUSION Findings of survey showed that maximum average percent disease incidence & disease severity were found in the Bamori Tehsil (32 & 32.7%) and lowest average percent disease incidence & disease severity was founded in Lahar Tehsil (12.7 & 15 %, respectively). A bacterium is gram-negative, round ended, rod-shaped, with only one polar flagellum. isolates, Xac1, Xac4, and Xac5 were yellow, while the other two, Xac2 and 3, were yellow to light yellow in color. Streptomycin sulphate was effective against the pathogen with maximum inhibition zone of 20.5mm. Lower efficacy was exhibited by streptocycline 8.2 mm