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Identification of Qtls for Salinity Tolerance at Both Seedling and Reproductive Stage(2009-10)

Dr.Mrs Hosneara Hossain
SSO
Biotechnology Division, BRRI, Gazipur

Md.Monirul Islam
SSO
Biotechnology Division, BRRI, Gazipur

Ripon Kumar Roy
SO
Biotechnology Division, BRRI, Gazipur

Dr. Md. Shamsher Ali
CSO
Biotechnology Division,Brri-Gazipur

Dr,Md.Enamul Hoque
CSO and Head (In Charge )
Biotechnology Division, BRRI, Gazipur

Abstract/ Executive Summary:

Salinity is one of the important constraints to rice production in coastal region of Bangladesh, particularly during dry season. Around one million hector area is directly 10 affected by salinity. The seedling and reproductive stage are most vulnerable at salt stress in rice. The present study was conducted to identify the QTLs for salt tolerance at seedling and reproductive stage. A total number of 203 F₁ seeds and 2108 F₂ seeds have been collected and 69 simple sequence repeat (SSR) markers were screened for polymorphism between the parents. Among them 11 primers showed polymorphism Markers that detected polymorphism between the two parents will be used for molecular analysis of the population.

Keywords: QTLS, Seeds, Salinity, SSR markers

Location: Biotechnology Division, BRRI-Gazipur

Start Date: 01-07-2009

End Date: 01-12-2011

Program Area: Variety and Species

Commodity: Rice

Objectives:

To develop high yielding variety with salt stress tolerance for the coastal saline region of Bangladesh
To study the genetic basis of tolerance to salinity at seedling/ reproductive stage and to identify QTLs responsible for salt stress tolerance.
To introgress identified QTL regions into advance breeding lines/variety to develop salt tolerant genotypes

Methodology:

Two hundred fifty F₂ individuals derived from the cross IR4630/BRRI dhan29 are grown for the study. Phenotyping: Seedling stage- Two pre-germinated seeds will be sown per hole in the styrofoam seedling float. The seedling float will be first suspended on distilled water in 10 L plastic trays for 3 days, then filled with culture solution (Yoshida et al. 1976) and will be left until the plants grow will be 14 days old. At 14 days, the solution will be salinized with NaCl at a concentration of 12 dS/m and silicon will be added in the form of sodium metasilicate 9 hydrate (4.5 mg Li-1) to avoid lodging. A randomized complete block design will be used with two replications. Nutrient solution will renewed every 7 day and will be maintained the pH @ 5.0. Reproductive stage- Seeds will be placed in a convection oven at 50ºC for 5 days to break the seed dormancy. Pre-germination of the seeds will be initiated by placing them in a Petri dish containing distilled water, which will be jousted enough to soak the seeds, and will besoaked for 24 hours. The distilled water in the Petri dish will be replaced with fresh distilled water and the seeds will be placed on tissue paper. Once the seed radicle and root emerged, then pre germinated seeds will be sown in perforated plastic pots filled with fertilized soil (50N, 25P, and 25K mg kg-1) and will be kept in concrete tanks. Level of water will be maintained at 3 cm below the soil surface. Salinization will be started (EC 10dS/m) immediate after flag leaf appearance. Spikelets and flag leaf (for Na/K analysis) will be collected 10 days after salinization. Biomass will be harvested on individual plant basis and will be kept in oven at 70⁰C for a week and weight will be recorded gram. Parental polymorphism and QTL analysis: A set of 500 microsatellite markers spanning all the 12 rice chromosomes will be screened between IR4630 and BRRI dhan29 parents. Microsatellite or Simple sequence repeat (SSR) primers were used for selection (Temnykh et al. 2001; McCouch et al. 2002; IRGSP 2005). A total of 100 polymorphic microsatellite markers will be separated which will be evenly spaced through out the genome. To identify the QTLs for salinity tolerance, QTL analyses will be performed by using Window based WinQTL Cartographer version 2.5 software (Wang et al., 2005). Each marker will be analyzed to determine the regions associated with salt tolerance. Data to be collected- Phenotyping: Plant height, tiller number, panicle length, spikelet fertility grain yield and biomass (individual plant basis), flag leaf samples (reproductive stage), third leaf (seedling stage) for Na+ and K+ analysis, overall phenotypic performance (SES score). Genotyping: polymorphic primers will be identified. After that, detection of QTLs and nature of inheritance of salt stress tolerance will be done.

Source of Information: Proceedings of the BRRI Annual Internal Review for 2009-10

Article Link (Online Journal):

Funding Source:

1. Government

Budget:

Total Budget (Tk.) :

Achievement/Findings:

A total number of 203 F₁ seeds and 2108 F₂ seeds have been collected and 69 simple sequence repeat (SSR) markers were screened for polymorphism between the parents. Among them 11 primers showed polymorphism Markers that detected polymorphism between the two parents will be used for molecular analysis of the population.

Knowledge Management: Report/Proceedings