Institutional Repository
Thesis Issued 2026-09-06 EN

In Vitro Regeneration, Genetic Analysis and Field Evaluation of Tropical Cultivars of Sugarbeet (Beta vulgaris L.)

Author: Md. Anisur Rahman

Abstract

The experiments were carried out at Biotechnology Laboratory and farm of Bangladesh Sugarcrop Research Institute (BSRI), Ishurdi, Pabna, Bangladesh during 2012-2013, 2013- 2014 and 2014-2015 cropping seasons for in vitro plant regeneration of tropical sugarbeet and evaluate the genetic fidelity as well as field performance of regenerated plants over their parents. Tropical cultivars of sugarbeet CS 0327, CS 0328, HI 0044, HI 0473, Shubhra and Cauvery were used for plantlet regeneration and field evaluation. On the basis of different shoot characters cultivar HI 0044 was most responsive for shoot regeneration which produced highest number of shoot (96.67%), shoot per explant (2.50) and shoot length (4.31 cm) cultured in medium containing BAP 1.0 mgl-1 + NAA 0.5 mgl-1. Different explants viz. shoot tip, hypocotyl, cotyledon, leaf and leaf petiole were also incubated on the same medium. Explant shoot tip produced the highest shoot (93.33%) and shoot length (4.40 cm) while leaf petiole explant produced the highest number of shoot per explant (3.67). Among five concentrations of BAP and two explants the highest shoot induction% was found in medium containing 1.0 mgl-1 of BAP for both types of explants. The highest number of rooted plant was found in half strength MS medium supplemented with 2.0 mg1-1 NAA from genotype HI 0044 and lowest from genotype CS 0328. MS media containing 0.1 mgl-1 NAA + 1.0 mgl-1 TDZ produced the highest per cent callus (93.67) as well as highest amount of callus (1.43 g) under the explant ofof hypocotyl. This treatment combination also produced the highest number of shoot per callus and longest shoot. The highest per cent explants induced callus (94.33%) in MS medium fortified with 1.0 mg1-1 2, 4-D + 1.0 mgl-1 BAP by the leaf petiole explant. Out of eighteen primers, four primers OPG03, UBC283, UBC431 and UBC467 generated 37 distinct bands among them 36 bands were polymorphic and percentage of polymorphic loci was 97.92%. The average gene diversity across all varieties for all loci studies was 0.2399. High level of Nei's gene diversity value was found 0.4986 and the lowest one was found 0.0000. The value of the highest genetic distance (0.903) was found among sugarbeet genotypes CS 0328 vs Shubhra, CS 0328 vs CC-Shubhra-6 and CS 0328 vs CC-Shubhra-7. The lowest value (0.000) was observed in genotypes H10044, HIO473, Shubhra and Cauvery vs their somaclones. Dendogram based on Nei's (1972) genetic distance using the segregated nineteen genotypes into two main clusters. Eighteen of them were formed cluster and the remaining one genotype CC-Shubhra-7 was in cluster 2. The developed somaclones and their parents on field evaluation exhibited variation regarding different yield attributes. Cultivar CS 0328 produced the highest beet yield (tha-¹) but HI 0044 consisted higher sugar content. All regenerated somaclones produced lower yield and higher sugar compared to their parents and matured earlier compared to their parents. From the above findings, it could be concluded that genotype HI 0044 would be high responsive for in vitro regeneration and high sugar production. The variation assessed among genotypes by RAPD markers could be used in future breeding program for tropical sugarbeet improvement while in vitro regeneration protocol developed has the potential use in improvement of the species through genetic transformation.