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Identification of Self-incompatible Lines and Their Hybrids of Radish Through Isozyme Analysis

N. A. Ivy
Associate Professor
Genetics and Plant Breeding, BSMRAU, Gazipur-1706.

M. S. Biswas
Lecturer
Department of Horticulture, BSMRAU, Gazipur-1706.

G. Rasul
Professor
Genetics and Plant Breeding, BSMRAU, Gazipur-1706.

T. Hossain
Professor
Crop Botany, BSMRAU, Gazipur-1706

M. A. K. Mian
Professor
Genetics and Plant Breeding, BSMRAU, Gazipur-1706.

Abstract/ Executive Summary:

The experiment was conducted to determine the hybridity of 48 radish genotypes among the crosses of 30 parental lines of radish using esterase and peroxidase isoenzymes. Peroxidase enzyme system was utilized for confirming the authenticity of 48 hybrids with their parents. Five bands at different Rf values varied from 0.00 to 0.569. All the hybrids showed the inheritance of few paternal as well as maternal bands. On the other hand, esterase enzyme system was utilized for confirming the authenticity of 48 hybrids with their parents. Thirty bands at different Rf values ranging from 0.04 to 0.93 in the parental lines and hybrids. All the hybrids showed the inheritance of paternal and maternal bands here also. The results of the experiment revealed that peroxidase showed four to five distinct bands in hybrids where as parental line produced three bands. The total number of bands in peroxidase analysis was only five and esterase analysis was thirteen. The inheritance pattern among the parental bands by all the hybrids indicated the confirmation of the hybrids obtained.

Keywords: Radish, Hybrid, Peroxidase, Esterase isoenzyme

Location: Department of Genetics and Plant Breeding, BSMRAU

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Radish

Objectives:

To know the banding pattern among the parents and hybrids and also the inheritance pattern of parental bands by all the hybrids indicated the authenticity of the hybrids of radish.

Methodology:

The plant materials for this study are comprised of 78 genotypes of radish where 30 are parental lines and 411 are hybrids. Young leaf material were taken from plant sample into a small eppendorf tube with a small amount of sea sand as a crushing agent. Exactly 0.70 µl extraction buffer solutions were added and crushed the sample using a glass rod. The sample was dissolved by vortex until become homogenization. Dissolved samples were centrifuged at 15000 rpm for 4°C for 3 minutes. Samples were kept in a refrigerator until it used. Gels of different concentrations were prepared using the stock solution. The stock solutions were stored in dark and cold condition. Freshly prepared ammonium per sulphate (APS) solution was used. The glass plates (one was notched and the other was plain) and the gaskets with the clips were assembled first to make plate sandwich. A comb was inserted into the sandwich. A level of marking was made on the glass plate, 4-5 mm lower than the teeth of the comb, by using marker pen. The comb was then removed. The solution of separation gel except APS and tetramethylene diamine (TEMED) were gently mixed in a 50 ml conical flask. After words APS and TEMED were added and mixed very carefully. Immediately after adding APS and TEMED, the separation gel solution was poured with its notched glass plate upside. After removing the clips and gaskets, the glass plate with gel was attached to the electrode assembly. About 430 ml of the electrode buffer was poured in the electrophoresis chamber slowly and gently to avoid trapping of air bubbles at the bottom of the gel. About 70 ml of electrode buffer was poured on to the upper portion of the electrode assembly and the level of the buffer would be 2-3 ml lower from the top of the glass plates to submerge the top of the upper gel. Ten to twenty micro liter of the homogenized sample was loaded into each well with a micro syringe. Electrophoresis was performed by connecting the electrode assembly with the power supply for 34 hours depending on the concentration of gels at 20 mA constant current per gel. After removing the gel sandwich from the electrode assembly, the gel was separated from the glass plate gently with a steel spatula keeping under slow running tap water. The gel was than put into the staining solution. The isozyme, which was stained to observe the variation in banding pattern are esterase and peroxidase. Solution A was prepared by mixing 40 ml methanol and 200 mg of 0-phynyl-diamine in 50 ml of 1 M Na-acetate buffer. Isozyme banding patterns were recorded on the basis of number and the relative front (Rf) values of the bands. The Rf value of each respective bands on schematic isozyme patterns was determined to allow precise comparisons among the various germ plasm. The Rf value is the mobility of each isozyme band that traveled from the origin divided by the distance traveled by the front tracking dye. An electrophoretic pattern comprised genotype with the same isozyme patterns.

Source of Information: J. Bangaldesh Soc. Agric. Sci. Technol., 6 (1&2): 147 - 150, 2009; ISSN 1811-6221.

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Peroxidase enzyme system was utilized for confirming the authenticity of 48 hybrids with their parents. Five bands at different Rf values varied from 0.00 to 0.569. All the hybrids showed the inheritance of few paternal as well as maternal bands. On the other hand, esterase enzyme system was utilized for confirming the authenticity of 48 hybrids with their parents. Thirty bands at different Rf values ranging from 0.04 to 0.93 in the parental lines and hybrids. All the hybrids showed the inheritance of paternal and maternal bands here also. The results of the experiment revealed that peroxidase showed four to five distinct bands in hybrids where as parental line produced three bands. The total number of bands in peroxidase analysis was only five and esterase analysis was thirteen. The inheritance pattern among the parental bands by all the hybrids indicated the confirmation of the hybrids obtained.

Knowledge Management: Journal