Efficiency of RAPD,ISSR and ITS markers in detecting genetic variability among Salacia species sampled from the Western Ghats of Karnataka

Diversity and phylogenetic relationship between four closely related Salacia species, i.e., Salacia chinensis, Salacia macrosperma, Salacia fruticosa and Salacia oblonga, collected from the Western Ghats of Karnataka, India, was assessed. Ten each of RAPD and ISSR primers generated a total of 76 and 68 loci, generating polymorphisms of 92.21 and 89.71%, respectively. Maximum likelihood analysis of the ITS sequences revealed three clades. Dendrogram analyses of RAPD and ISSR revealed two and four clusters, respectively. Overall polymorphism revealed by RAPD was 41.45?±?10%, ISSR was 33.58?±?6.52%, and ITS was 25.50?±?17.25%. Molecular variance revealed significant variance within and among the Salacia species. Tajima’s D neutrality test and Fu’s Fs were negative for all four species, implying presences of rare alleles and population expansion. Comparative study of RAPD, ISSR and ITS for Salacia species has given an insight into the efficiency of each technique in detecting diversity within and among the population sampled in the Western Ghats of Karnataka.

     

Morphological and molecular characterization of Neopestalotiopsis vitis associated with leaf blight disease of Manilkara zapota—a new record from India

Sapota is an important horticultural crop grown in India, and Karnataka is a major producer of sapota. A characteristic leaf blight disease was observed in Southern Karnataka during field surveys conducted in 2019 with an incidence of 13–22% in approximately 45 ha of sapota field. The leaf blight-associated pathogen was isolated on the potato dextrose agar medium. A total of 12 isolates obtained from each location were identified culturally and morphologically. Based on the morphological and cultural features, the pathogen was identified as Pestalotiopsis or Neopestalotiopsis, which was further confirmed by molecular identification using a representative isolate (MZ03). The ITS rDNA and β-tubulin genes were amplified and sequenced using ITS1/ITS4 and T1/T22 primer pairs respectively. nBLAST search analysis and concatenated (ITS-rDNA and TUB2 loci) phylogenetic analysis confirmed the pathogen identity as Neopestalotiopsis vitis. Pathogenicity tests conducted on detached leaves by inoculation with a conidial suspension of N. vitis produced typical blight symptoms after 4–5 days and progressed to cover the entire leaf lamina after 10–12 days. The pathogen’s identity was confirmed after re-isolation by cultural and morphological features. Although Pestalotiopsis clavispora and Pestalotiopsis versicolor causing diseases on sapota seedlings and trees have been reported, no reports are available for the occurrence of N. vitis to sapota from India. This is the first report of N. vitis associated with leaf blight disease of sapota from India.