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University of Agricultural Sciences, Dharwad

The University of Agricultural Sciences, Dharwad was established on October 1, 1986. The University has 5 Colleges, 27 Research Stations, 6 Agriculture Extension Education Centers, 6 Krishi Vigyan Kendras and ATIC. The University has its jurisdiction over 7 districts namely Bagalkot, Belgaum, Bijapur, Dharwad, Gadag, Haveri, and Uttar Kannada in northern Karnataka. Greater diversity exists in soil types, climate, topography cropping and farming situations. The jurisdiction includes dry-farming to heavy rainfall and irrigated area. Important crops of the region include sorghum, cotton, rice, pulses, chilli, sugarcane, groundnut, sunflower, wheat, safflower etc. The region is also known for many horticultural crops. Considerable progress has been registered in the field of education, research and extension from this University.

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20101
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Subject: Plant Biotechnology × Author: Amithkumar.Verma ×
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    ThesisItemOpen Access
    Development and Characterization of IAC and DS Starter Lines For Gene Tagging in Sorghum
    (UAS, Dharwad, 2010) Amithkumar.Verma; Ramesh.S.Bhat
    Transposon-mediated mutagenesisin Sorghum bicolour L. (Moench) was attempted with the development and characterization of Ds starter lines carrying pUR224NA and pUbiDs were generated for insertional inactivation and activation tagging, respectively, and iAc lines with pKU352NA. In vitro regenerated plants that rooted in liquid medium (MS +0.5 ppm NAA +0.5 ppm IBA, 6% sucrose) recorded significantly high survivability and establishment. In total, 6, 7 and 33 PCR confirmed plants were established for pUR224NA, pUbiDs and pKU352NA, respectively. Six Ds lines previously developed with pNU435 were also included in this study. T1 progenies of pNU435 plants showed 3:1 segregation ratio with Basta application, indicating single copy T-DNA/Ds integration. PCR confirmed T1 progenies of pKU352NA, pUR224NA and pUbiDs also showed 3:1 segregation. Southern hybridization for EcoRI digested DNA of pNU435-T0(1)-T1(27) showed a single band of expected 2567 bp with CaMV 35S probe. A 202 bp right border flanking sequence showed homology to a hypothetical mRNA (XM_002459886) of sorghum. Nine regenerants were obtained by co-infecting the calli derived from pNU435 [pNU435-T0(1)] with iAc construct (pKU435NA) for transient expression of transposase (TET). Out of nine, four had intact Ds; while five plants had transposed Ds. Of the four plants with intact Ds, one plant carried iAc, indicating its mutagenic nature. Of the five plants with intact Ds, four plants contained iAc indicating that are not stable mutants. One plant IDs- T0(8) showed Ds transposition, but did not show the presence of iAc, so it was regarded as a putative stable Ds insertion line. Crosses were made for pNU435 and pUR224NA launch pad lines with iAc lines. Seed set was seen in both the crosses involving pNU435 and in one cross involving pUR224NA.The starter lines generated in this study can be employed to generate mutagenic population, while Ds tagged mutant can be studied for mutant phenotype.