Dr Mohammad Khalequzzaman,
Chief Scientific Officer and Head,
GRS Division, Bangladesh Rice Research Institute, Gazipur- 1701, Bangladesh.
Dr Md. Shamsher Ali,
Former Director (Research),
Bangladesh Rice Research Institute, Gazipur-1701, Bangladesh.
Md. Abubakar Siddique,
Scientific Officer,
GRS Division, Bangladesh Rice Research Institute, Gazipur- 1701, Bangladesh.
Md. Monirul Islam,
Senior Scientific Officer,
IAPP, BRRI R/S Barisal, Bangladesh Rice Research Institute, Bangladesh.
Morphological and molecular characterization, Important plant genetic resources, Bangladesh.
Variety and Species
Morphological Characterization: A total of 266 rice germplasm accessions including 20 Geographical Indication (GI) rice viz. Biruin, Kalijira, Topa, Begunbichi,Chinigura, Balam, Dadkhani, Binni, Pashusail, Kataribhogh, Chamara, Digha, Bashful, Tulsimala, Horkoch, Birpala, Sadamota, Galon, Radhunipagal and Joina were grown in Aus (2012), T. Aman (2012) and Boro (2012-13) seasons for characterization using the rice germplasm descriptors and evaluation form. Seedlings were transplanted in single row of 5.4 m long with a spacing of 25 x 20 cm between rows and plants, respectively. Fertilizers were applied @ 60:60:40 kg NPK/ha in T. Aman & Aus, and @ 100:60:40 kg NPK/ha in Boro seasons, respectively. Appropriate control measures were taken for pests, diseases and weeds whenever needed. Table 1 presents the data recorded on 53 morpho-agronomic characters. Molecular characterization: Plant materials- Ninety six T. Aman, 48 Aus, 96 Boro landraces and 20 GI rice landraces were used as test materials for this study. Five gram seeds from each of the accession was first germinated and then sown in the earthen pots. Genotyping: DNA was extracted from young leaves of three weeks old plants following a simple and modified protocol of Zheng et al. 1995 to isolate total genomic DNA for polymerase chain reaction (PCR) analysis. PCR was performed in 12.5 µl reaction containing 5-25 ng of DNA template, 1.25 µl of MgCl2 free 10X PCR buffer (100 mM Tris-HCl pH 9.0 at 25°C, 500 mM KCl, 0.1% Triton® X-100 and H2O), 1.5 µl of 25 mM MgCl2, 0.25 µl of 10mM dNTP, 0.25 µl of 5 U/µl Taq polymerase enzyme, 0.625 µl each of 10 µM forward and reverse primers using a MJ Research single 96 well thermal cycler. After initial denaturation for five minutes at 94°C, each cycle comprised one min denaturation at 94°C, 1 min annealing at 55°C, and 2 min extension at 72°C with a final extension for 7 min at 72°C at the end of 35 cycles. The PCR products were mixed with bromophenol blue gel loading dye and were analyzed by electrophoresis on 8% polyacrylamide gel using mini vertical polyacrylamide gels for high throughput manual genotyping (CBS Scientific Co. Inc., CA, USA). The gels were stained in 0.5 mg/ml ethidium bromide and were documented using UVPRO (Uvipro Platinum, EU) gel documentation unit. Data analysis: Molecular weight for each amplified allele was measured in base pair (bp) using Alpha-Ease 5.0 software. The summary statistics including the number of alleles per locus, major allele frequency, gene diversity, polymorphism information content (PIC) values were determined using Power Marker version 3.25 (Liu and Muse, 2005). A similarity matrix was calculated with the Simqual subprogram using the Dice coefficient followed by cluster analysis with the SAHN subprogram using the UPGMA clustering method as implemented in NTSYS-pc.
M Khalequzzaman and M A Siddique. 2014. SPGR Sub-Project-Completion Report on Coordinated Sub Project on “Characterization of Important Plant Genetic Resources: BRRI component”. Genetic Resources and Seed Division, Bangladesh Rice Research Institute. Juydebpur, Gazipur-1701.
96,83,000.00 Tk.
Report/Proceedings