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Morphological and Molecular Characterization of Some Nerica Mutant Lines Under Drought Condition

MN Hasan
Department of Biochemistry and Molecular Biology, Bangladesh Agricultural UniversityMymensingh-2202, Bangladesh

MJH Bhuiyan
Department of Biochemistry and Molecular Biology, Bangladesh Agricultural UniversityMymensingh-2202, Bangladesh

SN Begum
Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture, Mymensingh 2202 , Banghadesh

SA Raffi
Department of Genetics and Plant Breeding, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh

S. Razia
Plant Breeding Division, Bangladesh Institute of Nuclear Agriculture, Mymensingh 2202 , Banghadesh

Abstract/ Executive Summary:

To ensure the productivity of rice in drought condition, ‘NERICA’ varieties of African origin has been introduced in Bangladesh and subsequently mutant lines are also developed for improving the performance further. The present study was undertaken to screen out better drought tolerant mutant lines of NERICA having other desirable traits. Thirteen NARICA mutant lines, four susceptible (BINA dhan8, BINA dhan10, BINA dhan11 and BINA dhan12) and one tolerant (BRRI dhan56) varieties were included in the experiment and selection was based on morphological and molecular characteristics. The plants were grown in pot containing 9 kg of soil and when they were 60 days old, drought was imposed by adding 1L of distilled water at every four days interval till harvesting. The quantity of water applied to control pots was double. The results showed that important yield contributing morphological attributes like days of 50% flowering, plant height, total number of tiller hill^-1 , panicle length, 100 seed weight, fresh weight of plant and dry weight of plant were significantly positively regulated due to drought in N₁₀ /300/P -2(1) -3 -5, N₁ /250/P - 6-2 -7 and N₁₀ /300/P -2(1) - 6-13 mutant lines. The findings of genetic diversity analysis through SSR markers RM32, RM202, RM351 and RM510 for N₁₀ /300/P - 2(1) - 3- 5, N₁ /350/P - 2- 2-6 and N₁ /250/P - 6- 2-7 lines appeared to resemble to that of control (BRRI dhan56) from the UPGMA dendrogram . Based on morphological, molecular characteristics and UPGMA dendrogram study, mutant lines N₁₀ /300/P -2(1) -3 -5 and N₁ /250/P - 6-2 -7 lines appeared superior as genetic material for developing better drought tolerant rice varieties.

Keywords: NERICA, Mutant lines, Drought tolerance

Location: Field laboratory of Bangladesh Institute of Nuclear Agriculture

Start Date: 00-00-2013

End Date: 00-00-2014

Program Area: Variety and Species

Non-commodity: Rice

Objectives:

To select the superior mutant lines for drought tolerance based on the study of morphological traits, followed by the genetic diversity analysis through the simple sequence repeat (SSR) markers.

Methodology:

The experiment was carried out with 18 rice germplasms in which promising 13 mutant lines of NERICA. BRRI dhan56 was used as control genotype. The seeds of NERICA were treated with physical (250, 300 and 350 Gamma - rays, unit is roentgen, R) mutagens and were grown as M₁ generation. Populations were grown as advanced. Finally , 13 NERICA mutant lines were selected from M₄ to M₅ generations for this experiment. The pot experiment was conducted during 2013- 2014 in Boro season at the field laboratory of Bangladesh Institute of Nuclear Agriculture. For imposition of water stress ‘soil pH and moisture tester’ model DM -15 (made by Takemura Electric Works, Ltd. Tokyo, Japan) was used to determine the moisture level of soil. After 60 days of sowing when sufficient tiller was noticed, DM -15 was used and moisture level was recorded (scale range 1-8). For rice proper moisture scale is 3-5 (DM -15). After four days of water stress another reading was taken and it was in scale of DM -15 less than 2. Restricted amount of water was added to impose water stress. For stress plant 1000 ml of fresh water was added at 96 hours interval for having moisture scale reading ranged as 1.5 to 2 (DM-15). Again for control, 2000 ml of fresh water was added with same interval to maintain normal moisture level with DM - 15 as 3 to 5 reading. During rainy days the pots were kept under plastic paper shade to avoid rainwater. Data on days to 50 percent flowering, plant height, total number of tillers hill^-1 , panicle length and 100 - seed weight were recorded as follows from the trials conducted under two different hydrological conditions. DNA extraction was done by the modified Cety1 Trimethy1 Ammonium Bromide (CTAB) mini- prep method. DNA samples were evaluated both quantitatively and qualitatively using spectrophotometer and λ (lamda) DNA (concentration marker) respectively. Based on previous references five SSR markers namely RM510, RM351, RM215, RM202, and RM32 were taken in this research work to screen on a sub sample of one randomly chosen individual from 18 rice genotypes to evaluate their suitability for amplifying DNA sequences, which could be accurately scored. Out of 5 primers, 4 primers (RM510, RM351, RM202 and RM32) showed clear polymorphism which was used in selection of the rice lines for early maturing drought tolerance. The PCR cocktail was prepared and 25 bp ladder was used and then placed in the PCR tubes and run in the DNA thermal cycler according. After completion of electrophoresis the gel was soaked in ethidium bromide (10 ml) solution for 20 -25 minutes then placed on high performance ultraviolet light box (UV transluminator) for checking the DNA bands and photographed using a Gel Documentary System of Biometra Company. For SSR Data Analysis the size (in nucleotide base pairs) of the amplified band for each microsatellite marker was determined based on its migration relative to a molecular weight size marker (25 bp DNA Ladder) with the help of Alpha Ease FC 5.0 software. Results The summary statistics including the number of alleles per locus, major allele frequency, gene diversity and Po lymorphism Information Content (PIC) values were determined using POWER MARKER version 3.23, a genetic analysis software. The 18 lines were clustered based on the matrix of genetic similarities using the Unweighted Pair Group Method with arithmetic averages (UPGMA) the cluster analysis and dendrogram construction were performed with NTSYS- PC.

Source of Information: Progressive Agriculture 26:15-21, 2015, ISSN: 1017-8139

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Considering important morphological characters and from UPGMA dendrogram it is concluded that N₁₀ /300/P -2(1) -3 -5 and N₁ /250/P -6 -2-7 genotypes were the best among 13 NERICA mutant lines. These genotypes can be used for field trial for suggesting new drought tolerant early maturing genetic materials.

Knowledge Management: Journal