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Identification of Introgressed Rice Lines of BINAdhan-7×fl-378 Under Salt Stress Through Ssr Markers

M. A. Haque
Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

S. N. Begum
Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture, Mymensingh-2202, Bangladesh

M. M. Islam
Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture, Mymensingh-2202, Bangladesh

M. A. Mahmud
Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

Thirty two rice lines of BC₁F₁ population (Binadhan-7/FL 378) were used to identify introgressed rice lines for salt tolerance using SSR markers. Seeds of BC₁F₁ population of Binadhan-7/FL-378 were collected from backcross population of F₁ (Binadhan-7/FL 378). Salt tolerant genotype, FL-378 was crossed with high yielding variety, Binadhan-7. Randomly selected 32 BC₁F₁ progenies along with their two parents (Binadhan-7, FL-378) were genotyped with microsatellite or SSR markers for identification of introgressed rice lines. Parental polymorphism survey was assayed by 8 SSR markers and three polymorphic SSR markers viz., RM296, RM585 and OSR30 were selected to evaluate BC₁F₁ rice lines for salt tolerance. Primer RM585 indicated 16 lines as introgressed and 16 lines as susceptible in comparison with salt tolerant parent FL-378 and salt susceptible parent Binadhan-7. Primer RM296 identified 13 introgressed, and 19 susceptible lines. Nine introgressed and 23 susceptible lines were identified when BC₁F₁ lines were evaluated with marker OSR30. Line BC₁F₁-30, BC₁F₁-40, BC₁F₁-2, BC₁F₁-16 and BC₁F₁-34 were identified as introgressed in comparison with parent FL-378 (salt tolerant) when RM585 and RM296 markers were used but line BC₁F₁-1 and BC₁F₁-3 were identified as introgressed incase of 3 primers (RM585, RM296 and OSR30). Again the lines BC₁F₁-5, BC₁F₁-24, BC₁F₁-26, BC₁F₁-27, BC₁F_1-29, BC₁F₁-31, BC₁F₁-37 and BC₁F₁-38 were found susceptible as compared with parent Binadhan-7 when marker RM585, RM296 and OSR30 were used. The polymorphism information content (PIC) values ranged from 0.3290 to 0.3671 with an average of 0.3544. The overall gene diversity of SSR loci for the 32 BC₁F₁ rice lines along with two parents was 0.4614, ranging from 0.4152 to 0.4844. Positive correlations were found between the genetic diversity and the maximum number of repeats. Among the lines, all the loci were polymorphic and clearly distinct and the cluster analysis (NJ tree and UPGMA) showed nearly similar pattern of variation which could be used for improvement of salt tolerant rice lines of rice through SSR markers.

Keywords: Salt stress, SSR Markers, Binadhan-7×FL-378

Location: Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture (BINA), Mymensingh

Start Date: 00-07-2010

End Date: 00-09-2011

Program Area: Variety and Species

Commodity: Rice

Objectives:

1. To transfer salt tolerant gene(s) into high yielding rice variety (Binadhan-7)

2. To identify salt tolerant rice line of BC₁F₁ progenies of Binadhan-7× FL- 378 using SSR markers.

Methodology:

The experiment was carried out in Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture (BINA), Mymensingh from July, 2010 to September, 2011. On the basis of phenotypic and genotypic evaluation for salt tolerance, one rice genotype (FL-378) was selected as parent for transferring salt tolerant genes (Islam, 2004) from tolerant to high yielding rice variety (Binadhan-7). Several crosses were made for introgressing salt tolerant genes into popular rice variety. In this study, a backcross was made between BC₁F₁ lines (Binadhan-7/FL-378) with high yielding Binadhan-7. Based on desired agronomic characters 58 BC₁F₁plants were grown . But only 32 BC₁F₁ rice lines were utilized for the identification of salt tolerant lines and the genotyping was done using SSR markers. The seeds of BC₁F₁ line were collected and kept in the convection oven for 24 h at 30°C for germination. Among these seeds, 67 seeds were germinated and transplanted in the plastic bucket in which 58 seedlings were survived. Finally 32 seedlings were selected randomly to collect the leaf to make foreground selection genotypically at the molecular level by using SSR markers and 220 BC₂F₁seeds were developed through successful backcrossing. Young vigorously growing fresh leaf samples from the seedlings of Binadhan-7, FL-378 and BC₁F₁ population were collected to extract genomic DNA. The samples were stored in -80ºC freezer with marking. DNA were extracted from leaf samples following CTAB method and were stored at - 20⁰C temperature. Eight primers of random sequence were selected for amplification of the DNA sequences. Primers were evaluated based on intensity of bands, consistency within individual, presence of smearing and potentiality for population discrimination. In this experiment RM510, OSR30, RM21, RM127, RM140, RM9, RM296 and RM585 primers were used for parental survey (data not shown). Among them three polymorphic SSR markers viz., RM296, RM585 and OSR30 were selected to evaluate BC₁F₁(32) rice lines for salt tolerance.

Source of Information: J. Bangladesh Agril. Univ. 10(1): 49–54, 2012, ISSN 1810-3030

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Based on above result, it may be concluded that the tolerant germplasm grouped in same cluster due to lower genetic distance in comparison with FL-378. While, the susceptible germplasm grouped in same cluster due to higher genetic distance in comparison with FL-378.

Knowledge Management: Journal