Inter Simple Sequence Repeat Analysis to Confirm Genetic Stability of Micropropagated Plantlets in Three Grape (Vitis spp) Rootstock Genotypes

Abstract

Three grape rootstock genotypes — Dogridge (Vitis champini), SO4 (V. beriandieri × V. rupestris) and H-144 (V. vinifera × V. labrusca), and their 30 in vitro regenerated plantlets were subjected to Inter Simple Sequence Repeat (ISSR) analysis in order to ascertain the genetic stability of micropropagated plantlets. Out of 35 primers screened initially with three mother plants, 10 were finally selected based on sufficient polymorphism and appearance of clear and scorable banding patterns. Each primer generated a unique set of amplification products ranging in size from 100 to 1800 bp. These ten ISSR primers produced 81 distinct and scorable band classes with an average of 8.1 bands per primer. Based on similarity matrix and cluster analysis the rootstock genotypes and their tissue culture derivatives formed three distinct genetic groups indicating their genetic relationships. Furthermore, no variation was detected among in vitro regenerated grape plantlets and their field-grown mother plants corroborating the high level of clonal fidelity of the in vitro regenerated plantlets and supporting the multiplication protocol utilizing nodal segments as in vitro culture initiation material.