Characterisation of resistance in African cassava genotypes against Indian Cassava mosaic begomoviruses
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1 4 th Triennial ISTRC Symposium 2006, Thiruvananthapuram, India Plant Plant Viruses Viruses Characterisation of resistance in African cassava genotypes against Indian Cassava mosaic begomoviruses Alfred Dixon Ivan Ingelbrecht Beate Stein Marianne Koerbler Anja Butgereitt Oluwole Ariyo Hassan Were Konstanze Dietrich Stephan Winter Plant Virus Department German Collection of Microorganisms & Cell Cultures Outline Current virus situation of cassava Terms & definitions of virus resistance Techniques used in resistance screening Correlating virus with host response Quantifying resistance Cassava responses to begomovirus infections
2 Cassava mosaic disease can lead to failure of root formation and total loss of harvest Cassava mosaic disease is caused by whitefly transmitted Geminiviruses AC 4 AV2 Rep ssdna-a ca 2800nt CP ssdna B ca 2800nt TrAP MP NSP REn ssdna viruses with bipartite genomes replicate through a dsdna intermediates in nuclei of infected cells unique cassava viruses Virus situation in Africa to date Cassava mosaic disease in India Mali Niger Chad Sudan Guinea Ivory Coast ACMV Nigeria Cameroon Ethiopia Uganda Kenya Democratic Republic of Congo Tanzania Somalia Tanzania ACMV -UG (UgV) EACMCV -UG (UgV) EACMZV EACMMV EACMCV SACMV Angola Malawi Zambia Zimbabwe South Africa Mozambique Madagascar Ndunguru et al., 2005 Indian cassava mosaic virus Sri Lankan cassava mosaic virus 2
3 Genome variation in Cassava mosaic begomoviruses (DNA A) Consideration for deployment of begomovirus resistant cassava AV 2 Coat Protein Ren Rep TraP AC 4 There are currently 6 distinct cassava virus species causing CMD in Africa & 2 distinct begomovirus species cause CMD in India There is a strong evolutionary force towards the development of new virus strains/variants with a focal point and centre of diversity in East Africa (Tanzania) ACMV The distribution of the viruses is at local focal points EACMZV/EACMMV -UG EACMCV EACMMV EACMZV SACMV ICMV The virus EACMCV from Cameroon is present throughout West Africa and now also found in East Africa. The epidemiological significance is unclear. The epidemiological significance of the recombinant virus -UG, Uganda variant, is evident! Is this the most severe and dangerous virus problem? In India, the Sri Lankan cassava mosaic virus is the dominant virus type There is only little genome variation Few ICMV incidences in mixed infections highlight as the key target to be considered in resistance development! Definition of virus resistance Screening for resistance against CMD a host plant is resistant if it can suppress multiplication of a virus and consequently suppresses the development of disease symptoms is very difficult under field conditions? Virus types present in a given geography? Whitefly vector populations in time at location Resistance evaluation by scoring for incidence & severity of symptoms and disease progress is ambiguous and can be misleading Resistance ranges from very high immune to moderate or low but is always associated with lower levels of virus replication Tolerance is a unique feature with negligible or mild disease symptoms but with normal levels of virus replication Deployment of natural virus resistance in Cassava at this stage is the only promising approach to counteract the Cassava Mosaic Disease 3
4 Approaches to generate defined infections in cassava with begomoviruses Inoculation of cassava genotypes with cloned Cassava mosaic viruses Microprojectile bombardment Rep ORI BamHI HindIII ORI EcoRI ApaI BV genomic DNA from virus infected plants cloned infectious virus genomic DNA TraP ReN CP -UG2 DNA A BC -UG2 DNA B Grafting BamHI ApaI ORI Ca2A fragment EcoRI HindIII ORI Ca08B fragment Establishing mixed defined Cassava mosaic virus infections Introducing pseudo-recombinant viruses through inoculation of virus genomic component DNA A and DNA B mixes Ca23A/ -UG Ca055B / -UG Ca55A/ B Ca23 A/ B 4
5 Evaluation of virus resistance in cassava CMD resistant cassava varieties recover from infection after an initial phase of symptom development and virus replication IV III Transplant Evaluate virus infected plant response cuttingi Inoculate scion with defined virus by approach grafting II Monitor disease development Response of TME3, TME 4 to single & mixed infections with TMS 96/0304 TME 4 TME ACMV / ICMV / A virus resistance status in cassva is expressed as recovery from symptoms & virus? Leaf -Nr. 9 Leaf Nr. Leaf Nr relative virus conc relative virus conc. 3 0,0 0, relative virus conc. only in symptomatic leaves negative in non symptomatic plant parts distribution /symptoms homogenous 5
6 Response of TME4 to TME4 responding to virus inoculations East African Cassava mosaic Zanzibar virus East African Cassava mosaic Uganda virus Virus resistance in cassava is a function of virus type High resistance Non recovery Recovery resistance Response of 96/0529A to East African Cassava mosaic virus Uganda virus Characterisation of resistance against CMD in cassava Resistant cassava genotypes - after initial infection phase - recover from symptoms and newly formed leaves remain symptomless with less, little or no virus in recovered plant parts; Virus resistance in cassava is a characteristics of the cassava genotype In symptomless leaves (plants), virus can still be present and symptomatic phases with usually mild symptoms can re-emerge! Susceptible cassava lines also can show recovery, however, this is a transient feature followed by (severe) symptomatic phases; In susceptible cassava lines mixed virus infections always lead to serious disease with no recovery; In resistant cassava, mixed infections result in recovery for some virus combinations, with lead to breakdown of resistance with other combinations. 6
7 Sources of resistance against CMD begomoviruses from Africa and India Resistance in cassava against cassava mosaic viruses Accession TME TME I3052 TME TMS/0304 TME 3 96/060 TME 4 96/089A Resistance single No ACMV// EACMCV/ICMV/ ALL ALL Resistance multiple No No ACMV// ACMV// ICMV/ Type S Rec.R HR HR Susceptibility HS virus + Rec.HR EACMZV Rec.HR EACMCV/ A good number of improved IITA varieties respond with recovery from infections with ACMV or ICMV as a non symptomatic phase of infection Most of the varieties were susceptible to species TME 4, 96/089A, 96/0529A are excellent sources of resistance with differential resistance characters Resistance is directed against all cassava viruses and their strain(s)incl. ICMV &/or infections in India 96/0529A ACMV// EACMCV/ ICMV/ ACMV// HR R Response of TME4 to mixed virus infections with CBSV /
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