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Seedling Salinity Tolerance Report

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Seedling Salinity Tolerance Evaluation
A total of 138 introgression lines were screened for salinity tolerance at seedling stage in greenhouse following the standard protocol of IRRI with some modifications (Gregorio et al. 1997) (Supplementary Figure S1). The screening experiment was conducted in a randomized complete block design with three replications. All lines were germinated in the laboratory and were transferred to nutrient solution which contained 1g/L of Jack’s Professional fertilizer 20-20-20 (J.R. Peters Inc.) and 200mg/L ferrous sulfate. When the seedlings were at two leaf stage, they were subjected to salt stress of EC 6dSm-1 for two days followed by EC 12 dSm-1. Three uniform plants were selected and the phenotypic …show more content…

The final concentrations of Na+ and K+ ions were computed using the standard curve developed using different dilutions.
Statistical Analyses
The analysis of variance (ANOVA) for each trait was computed by GLIMMIX procedure. The introgression line was entered as a fixed effect and replication as a random effect. Pearson correlation coefficients were computed to determine the relationship among different morphological and physiological traits. Statistical Analysis System
(SAS) software version 9.4 for Windows (SAS 2012) was used for the data analysis. The histograms were constructed in Microsoft Excel 2010 to analyze the distribution of ILs for each phenotypic trait.
Genotyping of Introgression Lines Using SSR Markers
Leaf tissue was collected from all the 138 ILs along with parents grown in control condition with no salt stress. The genomic DNA was extracted from the ground tissue using CTAB method (Chen and Ronald 1999). The concentration of DNA in each sample was estimated by a ND-1000 spectrophotometer (Thermo Fisher Scientific, Wilmington, USA). The DNA concentration of all samples was adjusted to a final concentration of 25ng/µl for PCR amplification. Genotyping of the ILs was performed using SSR markers. The PCR reactions were performed with 15 μl final volume containing 50 ng (2 µl) of genomic DNA, 1.5 µl of 10X buffer containing 25mM MgCl2, 1.5 µl of 2.5 mM dNTP mix, 1 µl each of forward and reverse primers (5 mM), and 1U of Taq polymerase. The

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