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ACP Project 1.3

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.

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ACP Project 1.3

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  1. 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

  2. International quarantine facility for the exchange of sugarcane germplasm among ACP countries Implementing Institution: Mauritius Sugarcane Industry Research Institute Countries targeted: ACP sugar producing countries Duration of project: 4 years Cost: Euros 918 510

  3. Objective: 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 INTRODUCTION

  4. 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

  5. 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

  6. Mauritius 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 international quarantine station in Mauritius, which any member of the ACP countries can use to facilitate exchange of disease-free germplasm Sugarcane Quarantine in Mauritius

  7. International Quarantine Benefits:Minimizing the risks of introducing potentially devastating diseases into ACP countriesAvoiding 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

  8. 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

  9. project 1.3Activity 1.1- Existing glass houses renovation GH for imported germplasm & tissue culture plants

  10. Activity 1.2 – Setting up of a new Plant Pathology Lab Near completion

  11. Activity 1.3 – Upgrading of an existing Tissue culture laboratory Tissue culture Laboratory designed & equipped Completed

  12. project 1.3Activity 2.1

  13. Activity 2.2 Diagnostic tools development

  14. DEVELOPMENT OF DISEASE TESTING AND ELIMINATION PROCEDURE FOR POTENTIAL DISEASES Based on: • Country of origin of germplasm • Requirements of importing country • Risk assessment analysis

  15. Sugarcane Systemic Diseases

  16. 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 Detection of Xanthomonasalbineans Leaf scald disease

  17. Xa specific fragment amplified by PCR Xcv isolates Xcv isolates Water control Xa Xa No amplification from Xcv Highly specific as compared to ITS based primers

  18. Taqman ® Real-Time PCR for X albilineansdetection 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

  19. Sugarcane Mosaic • Application of RT-PCR test for detection of SCMV, SrMV, JgMV using Primer pair Oligo1n/2n 327 bp fragment amplified from poaceaepotyviruses

  20. 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

  21. Multiplex RT-PCR for CUB, BRA-PER & REU genotypes of SCYLV Primers: REU-F/CPR, CUB-F/R & BRA-PER F/R Water PCR Step Multiplex Water RT-Step REU CUB BRA-PER 589 bp 450 bp 360 bp

  22. 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

  23. Activity 2.3 Tissue culture for elimination of diseases

  24. Tissue culture/elimination of SCYLV- optimized Infected Month 0 Month 1 Callus formation Months 2-4 3 subcultures of callus Months 7-8 Months 5-6 Regeneration Multiplication & Rooting Diagnosis

  25. Success rate of SCYLV elimination in vitro

  26. International quarantine facility for the exchange of sugarcane germplasm among ACP countries

  27. 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

  28. Collaborators • Dr Salem Saumtally • Mr SonalallDhayan • Mr Guy Triton • Dr Asha Dookun-Saumtally • Mr NawshadJoomun • Mr Miguel Antoine

  29. Acknowledgements

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