Genetic evaluation of cold atmospheric plasma, Jasmonic acid and Spermine treatments on Catharanthus roseus (L.) seeds by TRAP marker

Catharanthus roseus (L.) from the Apocynaceae family produces more than 130 terpenoid indole alkaloids (TIAs), including vinblastine and vincristine which are used as anti-cancer drugs. These alkaloidal drugs are still in clinical use and this plant is the only source of them. In the present study, we used cold atmospheric plasma Jet to treat C. roseus seeds and investigated the impact of Cold Atmospheric Plasma (CAP) on genetic diversity using Target Region Amplification Polymorphism (TRAP) as a molecular marker. TRAP-PCR was performed by fixed primer designed for TDC or STR genes which have a key role on alkaloids biosynthesis pathway, and arbitrary primers that target the ORF sequences. The C. roseus seeds were divided into six groups: 1-untreated seeds as controls, 2-treated group by cold plasma with 50 seconds (the 50s), 3-treated seeds by Jasmonic acid, 4-treated seeds by a combination of Jasmonic acid and cold plasma, 5-treated seeds by spermine and 6-treated seeds by a combination of spermine and cold plasma. Our results showed genetic differentiation between 6 studied groups. Among these groups, Cold plasma+ Jasmonic acid-treated plants showed the highest genetic diversity values including a number of effective alleles, Nei’s genetic diversity, Shannon index and percentage of polymorphism (Ne= 1.332, He= 0.249, I= 0.176 and P%= 39%). NJ clustering showed three main groups. Each group consisted of plants from different groups. Although TRAP marker could show genetic variations, more molecular markers are necessary to bring more genetic differentiation among samples. However, hormone and cold atmospheric plasma treatments revealed genetic changes on C. roseusand may be potential sources for introducing genetic variation in this medicinal plant.

Keywords:

Catharanthus roseus , Genetic diversity , Cold Atmospheric Plasma Jet , TRAP , Jasmonic acid , Spermine