Institutional Repository
Thesis Issued 2026-09-03 EN

Short Duration, Stem Borer Tolerant Sugarcane Genotypes Development and Molecular Validation

Author: Md. Noor Alam

Abstract

Sugarcane is an important sugar producing crop in the world and also in Bangladesh. It is a long duration crop and prone to different abiotic and biotic stresses. These two constrains limit farmer's interest towards its cultivation. Therefore, it is necessary to develop short duration and biotic stress tolerance, like stem borer resistance sugarcane genotypes with high yielding potential of %brix content. In this study attempts have been made to develop desired genotypes following germplasm screening, creation of somaclonal variants using tissue culture, and radiation during morphogenesis in tissue culture. Forty sugarcane accessions including local and exotic were screened for selecting the materials for tissue culture experiment. Four genotypes (Isd 15, Amrita, HoCP 91-555 and Isd 40) were selected based on their morphophysiological attributes, biochemical attributes and diversity analysis. In the tissue culture experiment, successful regeneration and somaclonal variations were achieved by using different concentrations and combinations of phytohormones. BAP (1 mgL-1) and Kn (0.25 mgL-1) were maintained along with 2, 4-D (3 mgl-1) for maximum shoot regeneration. IBA and NAA (1 mgL1+1 mgL-1) with MS media produced noticeable number of roots in Isd 15 (30 plantlet¹) and Amrita (13 plantlet1). Regeneration efficacy was influenced by genotypes, in where Isd 15 produced the highest number of shoots (29.6), longest shoots (9.37 cm), maximum roots (22.20), and longest roots (9.21 cm). The highest rate of mortality (%) of plantlets was found in both Amrita (78.27%) and Isd 15 (75.75%) at 9 Gy irradiation and it was proportionate to the increase of radiation dosage. In addition, number of shoots plantlet¹, number of roots plantlet1, length of shoots and roots were significantly higher in control (0 Gy) than irradiated plantlets (at 3, 6, and 9 Gy). It has been marked that radiation had significant effect on in vitro growth and development of sugarcane plantlets. All together 140 tissue culture and irradiated variants were successfully produced after proper hardening; those were evaluated in a field trial following Augmented design, where four parental clones were deployed as check. The genotypes evaluated based on their morpho-physiological attributes, level of insect infestation (%), biochemical contents (Zinc, iron, and proline) and finally characterized by molecular markers. There was significant variation among the genotypes in terms of %brix content and insect infestation (%). The yield contributing parameters cane diameter, stalk length and internode length was significant and positively correlated with stalk weight. Cane diameter was the prime yield contributing factor to get higher cane yield followed by internode length and stalk length. Brix content (%) with >17 was used as the indicator of the maturity, the somaclonal variants НоСР 91-555R,03 attained brix content >17% at the age of 9/10 months (September/October), which was selected as short duration genotype. Iron and zinc had negatively correlation with stem borer infestation. Proline increased the %brix content in cane and positively correlated to the rate of stem borer infestation. Twenty nine EST-SSR markers targeted to insect tolerance and short duration in maturity were used, among them four markers were polymorphic and able to characterize the desired somaclones. EST-SSR31-1 maker confirmed the variants HoCР 91- 555Ro03 was a short duration genotype. EST-SSR markers EST-SSR41-1, EST-SSR41-2 and ESTSSR80 also confirmed the genotypes Isd 40R01, Isd 40R04, and Isd 40R 09 were tolerant against stem borer. The materials developed in this study could be use as either breeding materials for future breeding programme to bring both these trait together in a single genotyрe or could be cultivated in the farmers' field as per their demand.