Platycodon grandiflorum, generally known as the bellflower or balloon flower, is the only species in the genus Platycodon of the family Campanulaceae. Platycodon plants have been traditionally used as a medicinal crop in East Asia for their antiphlogistic, antitussive, and expectorant properties. Despite these practical uses, marker-assisted selection and molecular breeding in platycodons have lagged due to the lack of genetic information on this genus. In this study, we performed RNA-seq analysis of three platycodon accessions to develop molecular markers and explore genetic diversity. First, genic simple sequence repeats (SSRs) were retrieved and compared; dinucleotide motifs were the most abundant repeats (39%-40%) followed by trinucleotide (25%-31%), tetranucleotide (1.5%-1.9%), and pentanucleotide (0.3%-1.0%) repeats. The result of in silico SSR analysis, three SSR markers were detected and showed possibility to distinguish three platycodon accessions. After several filtering procedures, 180 single nucleotide polymorphisms (SNPs) were used to design 40 cleaved amplified polymorphic sequence (CAPS) markers. Twelve of these PCR-based markers were validated as highly polymorphic and utilized to investigate genetic diversity in 21 platycodon accessions collected from various regions of South Korea. Collectively, the 12 markers yielded 35 alleles, with an average of 3 alleles per locus. Polymorphism information content (PIC) values ranged from 0.087 to 0.693, averaging 0.373 per locus. Since platycodon genetics have not been actively studied, the sequence information and the DNA markers generated from our research have the potential to contribute to further genetic improvements, genomic studies, and gene discovery in this genus.
The eukaryotic translation factors, including eIF4E which plays a major role in host translation initiation by recruiting messenger RNAs to the ribosomal complex, have repeatedly been identified as host factors required for viral infection. The cDNA sequence of eIF4E and eIF(iso)4E from 16 different genotypes, including 8 Nicotiana species and 8 Nicotiana tabacum cultivars, were determined in this study. The consensus Nicotiana eIF4E cDNA is 669 nucleotides long and codes for 222 amino acids. A high level of similarity was detected among eIF4E proteins of Nicotiana species sharing over 97% nucleotide identity and 95% amino acid identity in most cases. The eIF4E from tobacco (Nicotiana tabacum) shares a protein identities of 75.7% with pepper-eIF4E, 74.3% with tomato-eIF4E, 73.4% with potato-eIF4E, 73.1% with melon-eIF4E, 72.6% with cucumber-eIF4E, 73.1% with lettuce-eIF4E, and 71.5% with pea-eIF4E. In the case of eIF(iso)4E, the cDNA is 603 nucleotides long and codes for 200 amino acids. In this study, we investigated variations which exist in eIF4E or eIF(iso)4E, and attempted to correlate the variations which exist in these translation initiation factors with the resistance phenotype against several potyviruses in Nicotiana plants. We detected amino acid substitutions at the 18 positions for eIF4E and substitutions at the 11 positions for eIF(iso)4E. We attempted to correlate the resistance against Potato virus Y (PVY) or Pepper mottle virus (PepMoV) with the natural variations detected within the Nicotiana genotypes. However, no variation responsible for the gains of potyviral resistance was detected in this study. The result suggests that the potyviral resistance observed in Nicotiana species against PVY and PepMoV was conferred by an unknown resistance factor other than eIF4E or eIF(iso)4E, unlike general potyviral resistance observed in other plant species of the Solanaceae family.Additional key words: host factors, Solanaceae, tabacco, wild species Hort.
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