In vitro propagation and genetic fidelity evaluation in LA Lilium

Abstract

The present experiment was conducted at the Division of Floriculture and Landscaping, IARI, New Delhi, Indiaduring 2015–17 to develop a protocol for callus induction, PLB formation and plant regeneration from LA LiliumBrindisi using in vitro leaf segments, and to assess the genetic stability using SSR marker. Micropropagation of LALilium Brindisi led to compact calluses of dark brown to black colour. The in vitro regenerated leaves were inoculated at different concentration of 6-BAP and 2,4-D. Along with calluses, protocorm like bodies were also induced from the surface of cultured leaf segments, which further developed into shoots. MS medium fortified with 6-BAP (0.25 mg/l) and 2, 4-D (5 mg/l) recorded maximum callus formation. The mean number of shoot per callus clump ranged from 1.12 to 3.88, maximum number of shoots were recorded with 6-BAP (4 mg/l) and NAA (0.25 mg/l). Rooting ranged from 72–100% in IBA medium. Twenty regenerates were randomly selected for testing the fidelity. Out of 18 screened markers, only 10 produced clear and reproducible bands. A total of 244 bands were generated from 10 SSR primers in which seven primers were found polymorphic. Dendrogram generated by data analysis using Darwin 6 software package clearly indicated that the in vitro raised plants through leaf explant via callus phase were divided into three main clusters. The result of cluster analysis was supported by principal coordinate analysis (1/2 axis) where all the genotypes were distributed over different quadrangles. The total somaclonal variation was estimated to be 1.9% which indicated that even the plantlets raised through callus phase exhibited low frequency of somaclonal variation in case of LA hybrids of Lilium.