Agricultural Research Management Information System

Home
Research Summary
Search
About Us
- ARMIS
- Brochure
Contact Us
Report
User Request
Data Input
Help
- Operation Manual
  - PDF
  - Video
- Program Area & Commodity
We have reached 37600 number of research entries at this moment.

- Logout

Research Detail

Home
Research
Detail

Microspore Culture and the Performance of Microspore Derived Doubled Haploid in Brassica Juncea (l.)

M. M. ALI
Oilseed Research Centre, BARI

M.A. KHALEQUE MIAN
Department of Genetics and Plant Breeding, BSMRAU, Bangladesh

J.B.M. CUSTERS
Plant Research International, The Netherlands.

M.M.H. KHURRAM
Oilseed Research Centre, BARI

Abstract/ Executive Summary:

The experiment was conducted at the Tissue Culture Laboratory as well as in the net house of Oilseed Research Centre, Bangladesh Agricultural Research Institute (BARI), Joydebpur, Gazipur on microspore culture and the performance of microspore derived doubled haploid (DH) in Brassica juncea (L.). The variety BARI Sarisha II of B. junccea was used for microspore culture and 8 DH genotypes viz., BJDH 01, BJDH 05, BJDH ii, BJDH 12, BJDH 14, BJDH 17, BJDH 18, and BJDH 20 along with their parent Rajat collected from the Plant Research International, the Netherlands were used for performance evaluation. Four-day cold treatment at 6°C along with bud size 3.0-3.1 mm of BARI Sarisha II showed better response in the embryonic development. Significant variation was observed for the different yield contributing characters among the DH genotypes along with their parent Rajat. All the DH genotypes were late both for days to flower and days to maturity compared to the parent. Most of the DH genotypes produced higher seed yield than the parent. The genotypes BJDII 14 (7 g) and BJDH 01(7 g) were bold seeded and almost double or more in 1000-seed weight compared to the parent. Total oil content of the DH genotypes (40.64-42.28) were significantly different from the parent (3 g). Oil content of BJDH 01 was almost 3% higher than the parent could be selected to incorporate in the future breeding program.

Keywords: Microspore culture, Doubled haploid, Brassica juncea.

Location: Tissue Culture Laboratory and the net house of Oilseed Research Centre, Bangladesh Agricultural Research Institute

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Mustard

Objectives:

The present research work was undertaken to develop a suitable protocol for microspore embryogenesis in B. juncea and to comparative study between amphidiploid Rajat and its microspore derived DH generation for different agronomic traits.

Methodology:

The experiment was conducted in the Tissue Culture Laboratory and the net house of Oilseed Research Centre, Bangladesh Agricultural Research Institute. The genotypes BARI Sarisha 11 of B. juncea was used for microspore embryogenesis, while the DH genotypes collected from Business unit, Plant Research International, The Netherlands under the project International Co-operation for the Developing Countries (FNCO-DC) were used for performance evaluation. a) Microspore culture: For microspore culture, inflorescences of BARI Sarisha 11 having 1-3 buds flowering were collected from the field and exposed to 1-6 days at 6°C in refrigerator taking in a beaker with water. For microspore isolation 6-8 inflorescences were taken from the beaker. Desired buds (mostly 2.8- 3.2 mm) were picked with a fine tweezers and classified in 3 lengths group by using a measuring gauge. Ten buds per length group were selected. The laminar flow hood was switched on 30 minutes in advance and disinfected with 70% Ethyl alcohol (EtOH). Hands were also sterilized with EtOH. The buds were transferred in to tea-sieves for sterilization 10 mm in 2% NaOC1 making sure there were no air bubbles left. The buds were rinsed in sterile tap water for 1, 4 and 10 minutes. Adding 2 ml of sterile NLN 13 medium in 50 ml beaker, the buds were homogenized with injection piston by a turning pressure movement. Rinsing the piston on the 50 ml beaker, the suspension was poured over the sterile filter funnel with 10 ml centrifuge tubes. The volume of all the tubes adjusted upto 10 ml. The suspensions with microspores were centrifuged 3 mm at 800 rpm in 4°C adjusted. Supernatant was poured off and resuspended the pellet with loml medium. Centrifugation and resuspension were repeated twice. After the last washing,, microspores were resuspended in I ml NLN 13. Taking a sample of 15 μl microspores suspensions were counted by Fuchs Rosenthal counting chamber. The number (X) of microspores in 5 sub cells multiplied by 16000 were the density of microspore per ml. NLN 13 medium was added in the tube until the right density for cultivation (4x104 microspore/mi). The petridishes(TC quality) of 3 cm in diameter (for I ml culture) were filled out and tapped with Paraffin. The cultures were placed 35°C for 2 days and then transferred to 25°C continuously in dark. The embryo development was observed by an inverted microscope regularly. After 14 days globular, embryo development was found. Finally, after 21 days, the embryos were counted. b) Performance evaluation of DH of B. juncea: The seeds of eight DH genotypes viz., BJDH 01, BJDH 05, BJDH 11, BJDH 12, BJDH 14, BJDH 17, BJDH 18, BJDH 20 and along with the parent Rajat were sown in the pots (8? × 12?) on November 2004. The pots were arranged in a randomized block design with 3 replications. Ten seeds were sown in each pot. The seedlings emerged three to four days after sowing. After 15 days only, 4 plants were allowed to grow well in each pot. The data days to flower, days to maturity, plant height, primary branches per plant, secondary branches per plant, siliquae per plant, siliqua length, seeds per siiiqua, 1000-seed weight, yield per plant were recorded from selected four plants. Total oil content and fatty acid composition were estimated using by Soxhelt’s extraction apparatus and Gas Liquid Chromatograph (GLC), respectively. Analysis of variance and mean separation was done by Duncan’s Multiple Range test (DMRT) following the LStat program.

Source of Information: Bangladesh J. Agril. Res. 33(3) : 571-578, December 2008 ISSN 0258-7122

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

There was a significant variation observed for the character oil content among the DII genotypes along with their parent Rajat . Total oil content of the DH genotype BJDH 01(42.28%) was the maximum followed by the genotype BJDH 14 (41.62%) and were statistically different from the remaining genotypes along with the parent, which contained 39.49%. Oil content of the DH genotypes BJDH 12, BJDH 17 and BJDH 18 were similar. A range of variation was observed for fatty acid composition in oil of DH genotypes along with their parent. The variation for palmitic (C16 2.33 - 3.69%) acid content for all the genotypes along with parent was low. On the contrary, the ranges of other fatty acids C18:1 (5.83-10.36%), C18:2 (15.3222.28%) C18:3 (13.3120.16%), C201 (4.52-11.93%), and C22.1 (41 .89-53.57%) were found high. Though the erucic acid content was high in all the DH genotypes including parent, minimum of 41.89% was found in the parent Rajat, while the maximum in the DH genotype BJDH 18 of 53.57% followed by BJDH 05 of 53.07%. The DH genotype BJDH 01 could be exploited in future breeding programme for its higher oil content in the seed.

Knowledge Management: Journal