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Marker-assisted Introgression of Saltol Locus Into Genetic Background of BRRI Dhan-49

A.B.M. Zahidul Hoque
Department of Biotechnology, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur-1706, Bangladesh

Md. Ashraful Haque*
Department of Biotechnology, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur-1706, Bangladesh

Md. Ruhul Amin Sarker
Plant Breeding Division, Bangladesh Rice Research Institute (BRRI), Gazipur, Bangladesh

Mohammad Akhlasur Rahman
Plant Breeding Division, Bangladesh Rice Research Institute (BRRI), Gazipur, Bangladesh

Abstract/ Executive Summary:

The major purpose of this research was to introgress Saltol QTL into the genetic background of BRRI dhan49 through marker-assisted backcrossing. FL478 was used as a donor parent of Saltol QTL. Marker assisted backcrossing strategies were applied to develop BRRI dhan49-Saltol lines. A primer polymorphism survey was carried out between the two parental genotypes viz. BRRI dhan49 and FL478. A total of 363 SSR and InDel primers were surveyed and a total of 96 markers (27%) were found polymorphic. A cross was made between BRRI dhan49 and FL478 to produce F1 seeds and F1 was confirmed by RM493. Foreground selection was carried out using RM493 in all generations. Fifty six SSR markers were used for background selection. The Graphical Genotype (GGT) software was used to estimate the percentage recovery rate of the recurrent parent genome and to find out the genome ratio of the parents in the selected progenies of the backcross populations. From BC1F1 generation five best plants were selected based on background recovery of recurrent parent and backcrossed with recurrent parent. From BC2F1 generation, six plants were selected based on the highest background recovery of 80.7% to 89.5%. BC2F2 seeds were produced by selfing of selected BC2F1 individuals. The Saltol introgressed lines performed better in salt stress condition in hydroponic screening. The developed BRRI dhan49-Saltol lines would be useful for developing salt tolerant rice varieties or for using in other breeding programs.

Keywords: MABC, PCR, Gel electrophoresis, Saltol, QTL.

Location:

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Rice

Objectives:

This study deals with converting the early T. Aman rice variety BRRI dhan49 into a salinity tolerant variety by incorporating Saltol using marker assisted backcrossing (MABC) approach. MABC works were done in BC1F1 and BC2F1 generations.

Methodology:

The experiment was conducted at the experimental fields of Bangladesh Rice Research Institute (BRRI), Gazipur and Marker Assisted Selection Laboratory of Plant Breeding Division, BRRI, Gazipur. Recurrent parent and Donor parent: BRRI dhan49 is an early maturing Transplanted Aman variety which was used as recurrent parent. DNA extraction, PCR and gel electrophoresis Leaf samples were collected from young leaves from the plants at 20-25 days after transplanting. DNA was extracted following modified Miniscale method (Zheng et al., 1995). PCR was performed in 10 μl reactions containing around 25 ng of DNA template (3 μl DNA with 10X dilution factor), 1 μl 10X TB buffer (containing 200 mM Tris–HCl pH 8.3, 500 mM KCl), 1.35 μl 25 mM MgCl₂, 0.2 μl of 10 mM dNTP, 0.5 μl each of 10 μM forward and reverse primers and 0.1 μl of Taq DNA polymerase (5 U/μl) using thermal cycler (Chen et al., 1997; Neeraja et al., 2007). Allele scoring The band having same level of BRRI dhan49 was scored as ‘A’ which indicated the homozygous allele of the recipient parent for the particular SSR marker. Again, the band having same level of FL478 was scored as ‘B’ which indicated the homozygous allele of the donor parent for the particular SSR marker. However, heterozygous alleles were scored as ‘H’ having both the bands of two parents. Importantly, heterozygous alleles always had the extra bands in most cases with bigger size. Unidentified or missing alleles were scored as ‘U’. Data analysis: The marker data was analyzed using computer software Graphical Genotyper (GGT 2.0) (VanBerloo, 2007). This software was freely available from: http://www.dpw.wau.nl/PV/. Analysis of variance and mean comparisons based on Least Significant Difference (LSD) for phenotyping at screenhouse used in this study were performed using SAS (Statistical Analysis System) (ver. 08) package. Parental polymorphism survey: A primer survey was carried out between the two parental genotypes viz. BRRI dhan49, the recurrent or recipient parent and FL478, the donor parent of the MABC breeding program. A total of 363 microsatellite and InDel markers were used to screen the parents for identifying polymorphic markers. Marker assisted backcrossing scheme: The marker-assisted backcrossing scheme used several cycles of crossing back to the recurrent parent. 56 SSR markers were used for background selection and 1SSR marker RM493 was used for foreground selection.

Source of Information: International Journal of Biosciences | IJB | ISSN: 2220-6655 (Print), 2222-5234 (Online) Vol. 6, No. 12, p. 71-80, 2015

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

From BC2F1 generation, six plants were selected based on the highest background recovery of 80.7% to 89.5%. BC2F2 seeds were produced by selfing of selected BC2F1 individuals. The Saltol introgressed lines performed better in salt stress condition in hydroponic screening. The developed BRRI dhan49-Saltol lines would be useful for developing salt tolerant rice varieties or for using in other breeding programs.

Knowledge Management: Journal