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Identification of Salt Tolerant Rice Lines in F3 Population of Pnr-519 X Ir-29 Using Ssr Markers

H. Sohrawardy
Department of Biotechnology Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

M. M. Islam
Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture, Mymensingh-2200, Bangladesh

S. N. Begum
Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture, Mymensingh-2200, Bangladesh

M. H. Kabir
Department of Biotechnology Bangladesh Agricultural University, Mymensingh-2202, Bangladesh

Abstract/ Executive Summary:

Twenty six F₃rice lines of PNR-519 x IR-29 cross population were used to evaluate salinity tolerance at the seedling stage and to identify salt tolerant lines via SSR markers. Salinity screening was done at the seedling stage using hydroponic system at the glasshouse following IRRI standard protocol. Among the lines, tested lines 05-28, 05-33 and 05-47 were found to be tolerant, other 4 lines were moderately tolerant, 11 lines were susceptible and rest of the lines were highly susceptible. Twenty six F₃lines along with parents (PNR-519 and IR-29) were genotyped with SSR markers for identification of salt tolerant rice lines. Parental polymorphism survey was done with eight SSR markers. Out of 8 markers, four polymorphic SSR markers viz., RM21, RM51, RM127 and RM510 were selected to evaluate F₃lines for salt tolerance. Out of 26 F₃lines, twelve, six, five and seven were found as salt tolerant (compare to banding pattern of their tolerant parent) against the primers RM21, RM51, RM127 and RM510, respectively. Thus the tested markers could be efficiently used for tagging salt tolerant genes in marker-assisted breeding programme.

Keywords: Rice, Salinity, SSR markers.

Location: Mymensingh

Start Date: 00-12-2006

End Date: 00-11-2007

Program Area: Variety and Species

Commodity: Rice

Objectives:

1.To screen salt tolerant F₃ rice lines of PNR-519 x IR-29 through phenotypic selection at the seedling stage

2.To identify salt tolerant rice lines in F₃population of PNR-519 x IR-29 using SSR markers.

Methodology:

The study was conducted from December 2006 to November 2007 in the experimental field and Biotechnology Laboratory of Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture. Based on previous screening of several genotypes, some rice genotypes were selected as parents for transferring salt tolerant genes from tolerant to high yielding rice genotypes. A cross was made between the parents PNR-519 and IR-29. Based on desired agronomic characters 37 F₂ plants were selected out of 1350 F₂ plants. In next season, seeds of 37 selected plants were grown in plant progeny rows and 26 F₃lines were selected based on their better agronomic performances. These 26 F₃ lines were tested for salt tolerance both phenotypically (using IRRI standard protocol) and genotypically (using SSR markers). Phenotyping of F₃ rice lines for salinity tolerance at the seedling stage. The salinity screening was done using IRRI standard protocol (Gregorio et al., 1997) and nutrient solution was prepared according to Yoshida et al. (1976). Hydroponic system was used in this study. Pregerminated seeds were sown in this system with tap water. After 3 days of seedling, tap water was replaced with nutrient solution. Salinity level was maintained EC 12 dSm^-1 at this level and P^H was monitored every day and maintained at 5.5. Nutrient solution was replaced in every 8 days. The modified Evaluation System (SES) of IRRI was followed to assess the visual symptoms of salt toxicity Initial and final scoring was done at 15 and 21 day after salinization. Genotyping the F₃ rice lines for salinity tolerance. Modified CTAB mini prep was used for DNA extraction for 25-day-old seedling (IRRI, 1997). Eight primers were used for this study. Among these primers, four primers showed polymorphic and clear bands . Each PCR reaction was carried out with 15.0 µl reactions containing 1.5 µl 10 X buffer, 0.75 µl dNTPs, 1.0 µl primer forward, 1.0 µl 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 for 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. Banding patterns were visualized with ultraviolet gel documentation system. The banding patterns of segregating F₃population were scored compared with tolerant and susceptible parents. The segregating population having similar banding pattern to PNR-519 were considered as salt tolerant, similar to the susceptible parent IR-29 were considered as salt susceptible and having both banding pattern (PNR-519 and IR-29) were considered as heterozygous.

Source of Information: Bangladesh J. Seed Sci. & Tech. 12(2): 225–230 (2008) ISSN 1029-8800

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

These four markers RM21, RM51, RM127 and RM510 could be efficiently used to identify salt tolerant lines in rice and can also be used in marker-assisted selection (MAS) for breeding, quantitative trait loci (QTL) mapping and gene pyramiding in rice salinity breeding. The selected salt tolerant rice lines can be further tested in saline areas for developing high yielding salt tolerant verities.

Knowledge Management: Journal