acp project 1.3
DESCRIPTION
ACP Project 1.3. International quarantine facility for the exchange of sugarcane germplasm among ACP countries. Mid-Term Review 1 October 2012 MSIRI Réduit Mauritius. International quarantine facility for the exchange of sugarcane germplasm among ACP countries. - PowerPoint PPT PresentationTRANSCRIPT
![Page 1: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/1.jpg)
ACP Project 1.3
International quarantine facility for the exchange of sugarcane
germplasm among ACP countries
Mid-Term Review1 October 2012
MSIRIRéduit
Mauritius
![Page 2: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/2.jpg)
International quarantine facility for the exchange of sugarcane germplasm among ACP countries
Implementing Institution: Mauritius Sugarcane Industry Research InstituteCountries targeted: ACP sugar producing countriesDuration of project: 4 yearsCost: Euros 918 510
![Page 3: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/3.jpg)
INTRODUCTIONObjective:To enhance the productivity and efficiency of the sugarcane industry in ACP countries by promoting germplasm exchange, making their industries more responsive to the challenges in the global economy
Purpose of the International quarantine
To provide ACP countries with sugarcane plants free from detectable pathogens in order to safeguard their industry from potentially damaging diseases
![Page 4: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/4.jpg)
Sugarcane diseases Some 120 diseases recorded on sugarcane Most important ones are those known to be
transmitted by cuttings, referred as systemic diseases
Their most common mode of spread is through exchange of varieties between countries
28 systemic diseases are recorded The risks represented by them, if introduced,
can vary from minor to severe
![Page 5: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/5.jpg)
How to minimize risk of accidental introduction of sugarcane diseases?
• Safe movement of sugarcane germplasm• Quarantine of germplasm• Effective testing (indexing) of material - use of
molecular tools• Movement of tissue culture plants
![Page 6: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/6.jpg)
Sugarcane Quarantine in MauritiusMauritius has been regularly importing germplasm since 1914
A National closed quarantine is in existence since1946
The responsibility for the national quarantine has been vested on the MSIRI since 1953
Setting-up of an internationalquarantine station in Mauritius, which any member of the ACP countries can use to facilitate exchange of disease-free germplasm
![Page 7: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/7.jpg)
International Quarantine Benefits:
Minimizing the risks of introducing potentially devastating diseases into ACP countries
Avoiding duplication of resources by establishing an international facility in one state
Using up-to-date disease detection technologies and tissue culture to speed availability of germplasm
![Page 8: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/8.jpg)
Main Project Components
• Renovation of an existing quarantine to meet biosecurity level 3
• Setting up and equipping of a plant pathology Lab
• Renovation of a tissue culture facility and equipping
• Training of staff• Training of ACP members quarantine officials
![Page 9: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/9.jpg)
project 1.3
Activity 1.1- Existing glass houses renovation
International quarantine facility designed to meet biosecurity level 3
Near completion
GH for imported germplasm & tissue culture plants
![Page 10: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/10.jpg)
Activity 1.2 – Setting up of a new Plant Pathology Lab
1 2 3 4 5 6 7 8 9 10 11 12 13
352 bp
Near completion
![Page 11: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/11.jpg)
Activity 1.3 – Upgrading of an existing Tissue culture laboratory
Completed
Tissue culture Laboratory designed & equipped
![Page 12: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/12.jpg)
project 1.3
Activity 2.1
Training in molecular diagnostic of diseases
Major diseases not present in Mauritius: SCMV, SCSMV, Fiji disease, Downy mildewCIRAD provided training to staff in molecular detection of SCMV,SCSMV, and red leaf mottle virus
![Page 13: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/13.jpg)
Activity 2.2 Diagnostic tools development
![Page 14: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/14.jpg)
DEVELOPMENT OF DISEASE TESTING AND ELIMINATION PROCEDURE FOR POTENTIAL DISEASES
Based on: • Country of origin of germplasm• Requirements of importing country• Risk assessment analysis
![Page 15: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/15.jpg)
Sugarcane Systemic DiseasesUNKNOWN ETIOLOGY
FUNGI VIRUSES BACTERIA
Ramu stunt Downy mildew Fiji leaf gall Gumming
Chlorotic streak Dry top rot Leaf fleck Leaf scald
Fusarium sett Mild mosaic Mottles stripe
Pineapple disease Mosaic Ratoon stunt
Red rot Red leaf mottle Red stripe
Smut Streak PHYTOPLASMA
Sclerophthora Streak mosaic Grassy shoot
Wilt Striate mosaic Green grassy shoot
Yellow leaf White leaf
Leaf yellows
![Page 16: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/16.jpg)
Detection of Xanthomonas albineans
Primers and Probe designed from the
albicidin gene cluster sequence of
X albilineans
Primers/Probes checked for specificity
Conventional PCR & Real-time PCR tests
optimized Leaf scald disease
![Page 17: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/17.jpg)
Xa specific fragment amplified by PCR
No amplification from Xcv
Highly specific as compared to ITS based primers
Xcv
isol
ates
Xcv
isola
tes
Wat
er c
ontr
ol
XaXa
![Page 18: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/18.jpg)
Taqman ® Real-Time PCR for X albilineans detection
Inclusion of probe - added specificity guarding against non-specific annealing of PCR primer pairs
Amplification curve observed from all isolates of Xa tested
Flat line for Xcv isolates & water control
![Page 19: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/19.jpg)
Sugarcane Mosaic • Application of RT-PCR test for detection of
SCMV, SrMV, JgMV using Primer pair Oligo1n/2n
327 bp fragment amplified from poaceae potyviruses
![Page 20: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/20.jpg)
Detection of Sugarcane yellow leaf virus
Multiplex RT-PCR optimized
Real-Time RT-PCR optimized
The virus displays high genetic diversity
Development of real-time RT-PCR tests for screening genotypes in progress Leaf yellows disease
![Page 21: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/21.jpg)
Multiplex RT-PCR for CUB, BRA-PER & REU genotypes of SCYLV
BRA-
PER CU
B REU
M
ultip
lex
Wat
er R
T-St
ep
Wat
er P
CR S
tep
Primers: REU-F/CPR, CUB-F/R & BRA-PER F/R
589 bp
450 bp
360 bp
![Page 22: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/22.jpg)
Real-time RT-PCR specific for REU genotypes
REU genotypes amplified
No amplification from water controls, disease free plantlet, and plant infected with genotype BRA-PER
![Page 23: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/23.jpg)
Activity 2.3
Tissue culture for elimination of diseases
![Page 24: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/24.jpg)
Tissue culture/elimination of SCYLV- optimizedMonth 0
Month 1Callus formation
3 subculturesof callus
Infected
Months 2-4
Months 5-6 RegenerationMultiplication & Rooting
Months 7-8
Diagnosis
![Page 25: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/25.jpg)
Success rate of SCYLV elimination in vitro
Period Infected varieties
Successful regeneration
Virus elimination
% success for virus cleaning
1991-2001 18 16 16 1002001-2003 38 29 25 862003-2005 12 11 11 1002006-2008 13 10 10 1002008-2010 13 13 13 1002011-2012 8 6 6 100
![Page 26: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/26.jpg)
Activity 2011 2012 2013 2014
1.1. Setting up of a glasshouse for imported germplasm
1.2 Setting up of a diagnostic lab
1.3 Upgrading of a tissue culture laboratory for disease elimination & multiplication of plantlets1.4 International advice on setting up the quarantine
1.5 Certification of the facility
2.1 Training of staff
2.2 Development of disease testing
3.0 Reception of germplasm in quarantine
4.0 Training of Plant health officials of ACP countries
International quarantine facility for the exchange of sugarcane germplasm among ACP countries
![Page 27: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/27.jpg)
Expected Project Outputs• Quarantine facility available for ACP
countries to allow safe movement of germplasm
• Create awareness on diseases of quarantine importance
• Diagnostic protocols available to ACP countries
• Capacity building of plant health officials in ACP countries
![Page 28: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/28.jpg)
Collaborators
• Dr Salem Saumtally• Mr Sonalall Dhayan• Mr Guy Triton• Dr Asha Dookun-Saumtally• Mr Nawshad Joomun• Mr Miguel Antoine
![Page 29: ACP Project 1.3](https://reader035.vdocuments.mx/reader035/viewer/2022062521/56816689550346895dda41ad/html5/thumbnails/29.jpg)
Acknowledgements