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

Xanthomonas axonopodis pv.citri is a bacterium that attacks citrus trees and flourishes in locations with high temperatures, rains and strong winds. In places with these features, X. axonopodis pv.citri produces citrus canker, resulting in significant economic losses for citrus companies. It spreads by the unintentional movement of diseased citrus fruits and seedlings to uninfected areas.

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 H 2 S test Catalase test Starch hydrolysis 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 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(H 2 O) 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

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