VARIATION IN AGGRESSIVENESS AND AFLP AMONG Alternaria solani ISOLATES FROM INDONESIA

Alternaria solani is a necrotroph fungus that causes three-phased diseases in tomato. Management of the pathogen by using resistant cultivars requires knowledge on the aggressiveness and genetic diversity of the fungus. The aims of this study were to isolate A. solani from major tomato and potato producing areas in Indonesia and to study their aggressiveness and genetic variability. Twenty two A. solani isolates were recovered from early blighted tomato and potato in Central and West Java.  A. alternata was also isolated from tomato leaves in West Java and North Sumatra, indicating that early blight in Indonesia may be caused by more than one Alternaria species. Resistance tests of four tomato genotypes to selected A. solani isolates revealed that local isolates were more aggressive in inciting early blight and stem lesion than an imported isolate from USA. This implies that introduced breeding materials must be tested to local isolates to obtain effective resistance genes. Cluster analysis based on amplified fragment length polymorphism (AFLP) obtained from EcoRI+AG and MseI+C primer amplification separated 28 local and Taiwan isolates from the US isolate, which was coincided with aggressiveness separation between the local isolates and the US isolate. Three clusters of AFLP genotypes which did not associate with geographic origin were observed among tropical isolates. The low genetic diversity among the Indonesian isolates suggests clonal population structure with wide distribution. Successful local tomato breeding requires the availability of local A. solani collection with well-characterized aggressiveness level and molecular diversity to obtain effective resistance genes

Variation In Aggressiveness And Aflp Among Alternaria Solani Isolates From Indonesia

Alternaria solani is a necrotroph fungus that causes three-phased diseases in tomato. Management of the pathogen by using resistant cultivars requires knowledge on the aggressiveness and genetic diversity of the fungus. The aims of this study were to isolate A. solani from major tomato and potato producing areas in Indonesia and to study their aggressiveness and genetic variability. Twenty two A. solani isolates were recovered from early blighted tomato and potato in Central and West Java. A. alternata was also isolated from tomato leaves in West Java and North Sumatra, indicating that early blight in Indonesia may be caused by more than one Alternaria species. Resistance tests of four tomato genotypes to selected A. solani isolates revealed that local isolates were more aggressive in inciting early blight and stem lesion than an imported isolate from USA. This implies that introduced breeding materials must be tested to local isolates to obtain effective resistance genes. Cluster analysis based on amplified fragment length polymorphism (AFLP) obtained from EcoRI+AG and MseI+C primer amplification separated 28 local and Taiwan isolates from the US isolate, which was coincided with aggressiveness separation between the local isolates and the US isolate. Three clusters of AFLP genotypes which did not associate with geographic origin were observed among tropical isolates. The low genetic diversity among the Indonesian isolates suggests clonal population structure with wide distribution. Successful local tomato breeding requires the availability of local A. solani collection with well-characterized aggressiveness level and molecular diversity to obtain effective resistance genes

DNA Damage in Plant Herbarium Tissue

Dried plant herbarium specimens are potentially a valuable source of DNA. Efforts to obtain genetic information from this source are often hindered by an inability to obtain amplifiable DNA as herbarium DNA is typically highly degraded. DNA post-mortem damage may not only reduce the number of amplifiable template molecules, but may also lead to the generation of erroneous sequence information. A qualitative and quantitative assessment of DNA post-mortem damage is essential to determine the accuracy of molecular data from herbarium specimens. In this study we present an assessment of DNA damage as miscoding lesions in herbarium specimens using 454-sequencing of amplicons derived from plastid, mitochondrial, and nuclear DNA. In addition, we assess DNA degradation as a result of strand breaks and other types of polymerase non-bypassable damage by quantitative real-time PCR. Comparing four pairs of fresh and herbarium specimens of the same individuals we quantitatively assess post-mortem DNA damage, directly after specimen preparation, as well as after long-term herbarium storage. After specimen preparation we estimate the proportion of gene copy numbers of plastid, mitochondrial, and nuclear DNA to be 2.4–3.8% of fresh control DNA and 1.0–1.3% after long-term herbarium storage, indicating that nearly all DNA damage occurs on specimen preparation. In addition, there is no evidence of preferential degradation of organelle versus nuclear genomes. Increased levels of C→T/G→A transitions were observed in old herbarium plastid DNA, representing 21.8% of observed miscoding lesions. We interpret this type of post-mortem DNA damage-derived modification to have arisen from the hydrolytic deamination of cytosine during long-term herbarium storage. Our results suggest that reliable sequence data can be obtained from herbarium specimens