Occurrence of Banana Blood Disease in Malaysia Datin Jatil Aliah Timin 1 , Irene Lah 2 and Saifful Bahri Abd Mutalib 3 Plant Biosecurity Division, Department of Agriculture, Kuala Lumpur, Malaysia 1 Plant Pathology Unit. E-mail: [email protected]; [email protected]; 2 Plant Biotechnology Unit. Email: [email protected]; [email protected]; 3 Plant Pathology Unit. E-mail: [email protected]; [email protected]; 8th ICPPT 8-10 April 2014 K. Lumpur Malaysia 1
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Occurrence of Banana Blood Disease
in Malaysia
Datin Jatil Aliah Timin 1, Irene Lah2 and Saifful Bahri Abd Mutalib3
Banana Vascular Wilt Incidence in Different States
Bacterial wilt
Fusarium/ Panama disease
Both
[Source: Laporan Status Serangan Penyakit Layu Bakteria (bakteria Ralstonia solanacearum ras 2) dan Layu Fusarium atau 'Panama Disease’ (kulat Fusarium oxysporum f.sp cubense) di Semenanjung Malaysia 2010 (Mengikut laporan diagnostik dan pemantauan1 / bancian status2 di negeri-negeri tahun 2008-2009) - Laporan Unit Patologi Tumbuhan, Bahagian Biosekuriti Tumbuhan, Kuala Lumpur]
0
500
1000
1500
2000
2500
Are
a af
fect
ed
(h
a)
State
Fig 3: Banana Vascular Wilt Incidence in Different States
Bacterial wilt
Fusarium/ Panama disease
Both
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• banana wilt disease in Johor , N. Sembilan & Malacca
- ProMED mail July 2009: Professor Emeritus Ivan Buddenhagen, University
of California, Davis suggested that BDB may be the possible cause of the
banana disease that appears to be emerging in south-western Malaysia.
However confirmation by molecular methods would be needed for a conclusive
diagnosis.
Banana Vascular Wilt in Malaysia
• In 2012-2013 plants with bacterial wilt symptoms were reported in Negeri Sembilan, Selangor, Penang, Pahang, Johor and Sabah. Infected plants were tested for R. solanacearum spesies complex and BDB using molecular methods.
8th ICPPT 8-10 April 2014 K. Lumpur Malaysia
In 2007- 2009, identification of banana bacterial wilt was done using ooze test and BIOLOG system –BIOLOG identification only to R. solanacearum level thus cannot determine the exact phylotype or race.
13
Laboratory Analysis : Identification of Ralstonia solanacearum species complex
A phylotype multiplex PCR protocol was used to verify the phylotypes of Ralstonia solanacearum species complex and BDB in infected banana samples.
All R. solanacearum strains should produce a band (282 bp) with the 759/760 primers, as well as another band for each of the Phylotypes
I = 144 bp II = 372 bp III = 91 bp IV = 213 bp
The Blood Disease Bacterium (BDB) diagnostic protocol uses the BDB primer set: 121F CGT ATT GGA TGC CGT AAT GGA and 121R AAG TTC ATT GGT GCC GAA TCA – amplifies a 317 bp amplicon from BDB strains.
8th ICPPT 8-10 April 2014 K. Lumpur Malaysia 14
Location
No of
infected
samples
analysed*
No of positive samples
Ralstonia solanacearum species complex Blood Disease
Bacterium (BDB) R. s
General Phylot I Phylot II Phylot III Phylot IV
Sabak Bernam,
Selangor 6 3 1 0 0 4 4
Gemas, N.S 6 6 0 0 0 6 6
Putrajaya, WP 4 4 0 0 0 4 4
Tuaran, Sabah 18 6 0 0 0 6 4
Sg. Burong,
Selangor 4 2 0 0 0 2 0
Batu Pahat, Johor 4 4 0 0 0 4 4
Jerantut, Pahang 3 3 0 0 0 3 3
Serdang, Selangor 3 3 0 0 0 3 3
Sekinchan
Selangor. 3 3 0 0 0 3 3
S. Prai, Penang. 5 5 0 0 0 5 5
Laboratory Analysis : RESULTS Identification of Ralstonia solanacearum species complex
* Ooze test positive 8th ICPPT 8-10 April 2014 K. Lumpur Malaysia 15
8th ICPPT 8-10 April 2014 K. Lumpur Malaysia 16
Bil Sample R. solanacearum species complex Notes
Ref Sample General Phylotype IV BDB
1. SP3048 Petiole Positive Positive Positive R. solanacearum sp. complex
Blood Disease of Banana
2. SP3050 Petiole Positive Positive Negative R. solanacearum sp. complex
unknown
3. SP3055
Pseudostem (inner
part)
Positive Positive Positive R. solanacearum sp. complex
Blood Disease of Banana
4. Pulp Positive Positive Positive
5. Fruit skin Positive Positive Positive
6. SP3056
Pseudostem (inner
part)
Positive Positive Negative R. solanacearum sp. complex.
7. Pulp Positive Positive Positive R. solanacearum sp. complex
Blood Disease of Banana
8. Fruit skin Positive Positive Negative R. solanacearum sp. complex.
9. SP3057
Pulp Positive Positive Negative R. solanacearum sp. complex
unknown 10. Fruit skin Positive Positive Negative
11. SP3058
Pulp Positive Positive Positive R. solanacearum sp. complex
Blood Disease of Banana
12. Fruit skin Positive Positive Negative R. solanacearum sp. complex.
Laboratory Analysis : RESULTS - SABAH Identification of Ralstonia solanacearum species complex
• BD in Sg Burung, Selangor 2012
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• BD in Sabak Bernam, Selangor 2012
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• BD in Sabak Bernam, Selangor 2012
8th ICPPT 8-10 April 2014 K. Lumpur Malaysia 19
• BD in Federal Territory Putrajaya 2012
Amplication of 317 bp amplicon from banana samples infected with BDB. Lane 1-mol wt marker, lane 2- negative control, 3-14 banana samples, 15 positive control
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
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• BDB in Batu Pahat, Johor 2013
8th ICPPT 8-10 April 2014 K. Lumpur Malaysia
Amplication of 317 bp amplicon from banana samples infected with BDB. Lane 1-mol wt marker, lane 2- negative control, 3-14 banana samples, 15 positive control
Amplication of 282 bp and 213 bp amplicon from banana samples infected with R. s and phylotype IV. Lane 1-mol wt marker, lane 2- negative control, 3-14 banana samples, 15 positive control
21
• BDB in Raub Pahang 2013
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• Banana Wilt R. solanacearum in Johor 2007
8th ICPPT 8-10 April 2014 K. Lumpur Malaysia 23
Date
Disease/ pathogen Detection Method
Location
2007-2010 Bacterial Wilt Ralstonia solanacearum Ooze test and BIOLOG
Johor, N. Sembilan, Malacca, Kedah, Kelantan, Perak
2011-2014 Bacterial Wilt Ralstonia solanacearum species complex , BDB
Ooze test, PCR
Selangor, Johor, Sabah, Pahang, Penang, W.P Putrajaya, N. Sembilan
Banana Bacterial Wilt in Malaysia ● ● ● Summary ● ● ●
8th ICPPT 8-10 April 2014 K. Lumpur Malaysia 24
Infected banana samples from Peninsular Malaysia and Sabah were found to be infected by:
(2009-present) R. solanacearum phylotype IV and BDB
– N. Sembilan , Johor, Selangor, WP Putrajaya,
Penang, Pahang, Sabah
Banana Bacterial Wilt in Malaysia ▪ ▪ ▪ CONCLUSION ▪ ▪ ▪
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R. solanacearum
R. s phylotype IV + BDB
Banana Bacterial Wilt in Malaysia ▪ ▪ ▪ CONCLUSION ▪ ▪ ▪
8th ICPPT 8-10 April 2014 K. Lumpur Malaysia
26
• Blood disease of banana which previously limited to Indonesia has spread to Peninsular Malaysia and Sabah.
• To the best of our knowledge, this is the first confirmation of Banana Blood Disease in Malaysia.
• Department of Agriculture has developed a Standard of Procedure which stated the action plans and various control measures to mitigate this disease from affecting the banana industry in Malaysia. DOA imposed new import regulation for importation of banana fruits into Malaysia.
Banana Bacterial Wilt in Malaysia ▪ ▪ ▪ CONCLUSION ▪ ▪ ▪
8th ICPPT 8-10 April 2014 K. Lumpur Malaysia 27
We would like to thank the following people for their support and contribution in making this Banana Vascular Wilt and BDB detection program possible.
• collegues and staffs of Plt Patho Unit & Plt Biotech Unit, Plant Biosecurity Division HQ and State Plant Biosecurity Division
• Ms Julia Lamdin & Ms Evenni Poili (DOA Sabah),Dr Lily Eng (DOA Sarawak), Dr Noor Hana Hussain (UiTM)