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Salinity Screening and Molecular Analysis to Identify Salt Tolerant Rice (oryza Sativa L.) Genotypes

A. Sultana
MS, GPB,
BAU, Mymensingh

M.M. Hasan Sohel
Scientist
BINA, Mymensingh

M.M. Islam
Scientist
BINA, Mymensingh

S.N. Begum
Scientist
BINA, Mymensingh

A. K. Patwary
Professor
GPB, BAU, Mymensingh

Abstract/ Executive Summary:

Twenty seven rice genotypes were used to evaluate salinity tolerance at the seedling stage and to identify salt tolerant lines using SSR markers. Salinity screening was done at the seedling stage using hydroponic system following IRRI standard protocol. Based on 1-9 scale scoring, four tolerant, eight moderately tolerant and fifteen susceptible rice genotypes were identified. Three selected polymorphic markers viz., RM 10772, RM 296 and RM 10701 were used to evaluate salt tolerant rice lines. The marker RM 10772 identified eight lines as salt tolerant, sixteen lines as susceptible and three lines as moderately tolerant. Marker RM 296 identified seven genotypes as tolerant, seventeen lines as susceptible and three lines as moderately tolerant in comparison with Binadhan-7 and Binadhan-8. Whereas RM 10701 identified seven lines as tolerant, seventeen linens as susceptible and three lines as salt tolerant. SET 37 L-29, SET 37 L-41 and PBRC-37 were identified as salt tolerant in comparison with Binadhan-8 (salt tolerant) when RM 10772, RM 296 and RM 10701 markers were used. The tested markers RM10772, RM296 and RM10701 could be used to identify salt tolerant lines and marker-assisted selection (MAS) breeding for developing salinity tolerant rice varieties.

Keywords: Salinity screening, Hydroponic, SSR, Salinity tolerant.

Location: Bangladesh Institute of Nuclear Agriculture, Mymensingh

Start Date: 00-00-0000

End Date: 00-00-0000

Program Area: Risk Management in Agriculture

Commodity: Rice, Soil salinity

Objectives:

To observe phenotypic performance of rice lines under saline and non-saline condition at seedling stage and to use of microsatellite markers for identification of salt tolerant rice genotypes at seedling stage.

Methodology:

The experiment was carried out at the glasshouse and biotechnology laboratory of Bangladesh Institute of Nuclear Agriculture (BINA), Mymensingh. Twenty seven rice genotypes with diversified genetic background were used in this study, of which twenty five were advanced lines and two were cultivars IRRI standard protocol in hydroponic system was used at the glasshouse to evaluate salt tolerance rice lines. Nutrient solution was used in hydroponic system for screening salinity tolerance at the seedling stage. Each macronutrient 450- ml and 450 ml of micronutrient solutions were mixed with distilled water (dH₂O) to prepare 360 L of nutrient solution,. The pH of the solution was measured by pH meter. The nutrient solution was changed after every eight days. The modified standard evaluation score (SES) of IRRI was used to assess the visual symptoms of salt toxicity. This scoring discriminates the tolerant, moderately tolerant and susceptible rice lines. Initial scoring was started at 15-day after salinization and final scoring was done at 21-day after salinization. Highly tolerant, tolerant, moderately tolerant, susceptible and highly susceptible lines were scored by 1, 3, 5, 7 and 9, respectively. CTAB method was used for DNA extraction for 25-day-old seedling (IRRI, 2006). Eight primers were used for this study. Among these primers, three primers were showed polymorphic and clear bands (Fig. 2, Fig. 3 and Fig. 4). Each PCR reaction carried out with 15.0µl reactions containing 1.5 µl 10 X buffer, 0.75 µl dNTPs, 1µl primer forward, 1µl primer reverse, 0.5 µl Taq polymerase, 8.25 µl ddH₂O and 2.0 µl of each template DNA samples. PCR profile was maintained as initial denaturation at 94^oC for 5 min, followed by 34 cycles of denaturation at 94^oC for 1 min, annealing at 55^oC for 1 min and polymerization at 72^oC for 2 min; and final extension by 7 min at 72^oC. Then electrophoresis in 1.5% agarose gel was done after polymorphism in the PCR products and stained in ethidium bromide for staining and documentation. Banding patterns were visualized with UV gel documentation system. The banding patterns of genotypes were scored compared with check variety.

Source of Information: Bangladesh J. Seed Sci. & Tech. 16 (1 &2): 125-132 (2012), ISSN 1029 - 8800

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Among the twenty seven rice genotypes salt tolerant Binadhan-8, Set 37 L-29, Set 37 L-41, PBRC-37 were identified as salt tolerant and Binadhan-7, Set 10 SL-10, Set 39 L-28, Set 39 L-1, Set 10 SL-15, Set 39 L-3, Set 39 L-25, Set 10 SL-2, Set 37 L-23, Set 39 L-22, Set 37 L-36, PBRC-5 were identified as salt susceptible based on SES scoring and phenotypic performance. Binadhan-8, Set 37 L-41, Set 37 L-29, PBRC-37 showed no symptoms of salt injury. On the other hand Binadhan-7, Set 10 SL-10, Set 39 L-28, Set 39 L-1, Set 10 SL-15, Set 39 L-3, Set 39 L-25, Set 10 SL-2, Set 37 L-23, Set 39 L-22, Set 37 L-36, PBRC-5 were severely injured by salt injury. According to modified SES system four tolerant, eight moderately tolerant and fifteen susceptible genotypes were identified. The lowest % reduction in TDM was recorded in Set 37 L- 29 (13.1), Set 37 L-41 (16.8), Binadhan-8 (18.6), and PBRC-37 (19.2). Considering percent reduction in performance based on their plant height, root length and total dry matter production four genotypes (Set 37 L-29, Set 37 L-41, Binadhan-8 , PBRC-37) were recorded tolerant and twelve genotypes (Binadhan-7, Set 10 SL-10, Set 10 SL-2, Set 37 L-36, Set 39 L-25, Set 10 SL-15, Set 37 L-23, Set 39 L-29 , Set 39 L-28, Set 39 L-22, Set 39 L-13, Set 39 L-10, Set 39 L-3) were recorded susceptible. In molecular study, marker RM 10772 identified eight lines as salt tolerant, sixteen lines as susceptible and three lines as moderately tolerant. Seven tolerant, seventeen susceptible and three lines were showed moderately tolerant when tested with the marker RM 296. RM 10701 identified seven tolerant, seventeen susceptible and three lines as moderately tolerant. Binadhan-8, Set 37 L-29, PBRC-37 Set 37 L-41 showed salt tolerant when genotyped with marker RM10772 and RM 296, RM10701 and Binadhan-7, Set 10 SL(10), Set 39 L-28, Set 39 L-22, Set 39 L-3, Set 39 L-1, Set 10 SL(15), Set 39 L-25, Set 10 SL(2), Set 37 L-23, Set 37 L-36, PBRC-5 showed salt susceptibility when genotyped with primer RM10772, RM296 and RM10701. The markers RM10772, RM296 and RM10701 could be used to identify salt tolerant lines in rice and can also be used in marker-assisted selection (MAS) for developing salinity tolerant rice varieties.

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