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

Effect of Different Genotype and Hormonal Combination on in Vitro Shoot Regeneration of Hybrid Mustard (brassica Rapa L.)

M.A. Halim
Department of Biotechnology, Sher-e-Bangla Agricultural University (SAU), Sher-e-BanglaNagar,Dhaka-1207,Bangladesh

M.S. Islam
Student, Department of Biotechnology, SAU, Sher-e-Bangla Nagar,Dhaka-1207,Bangladesh

F. Mahmud
Department of Genetics and Plant Breeding, SAU, Sher-e-Bangla Nagar, Dhaka-1207, Bangladesh

M.E. Hoque
Department of Biotechnology, SAU, Sher-e-Bangla Nagar,Dhaka-1207,Bangladesh

M.S.R. Bhuiyan
Department of Genetics and Plant Breeding, SAU, Sher-e-Bangla Nagar, Dhaka-1207, Bangladesh

Abstract/ Executive Summary:

Keywords: Anther, Hormone, Genotype, Generation

Location: Biotechnology Laboratory of the Department of Biotechnology, Sher-E-Bangla Agricultural University, Dhaka

Start Date: 00-08-2011

End Date: 00-03-2013

Program Area: Variety and Species

Commodity: Mustard

Objectives:

1. The present research work has, therefore, been planned and executed for screening the appropriate days to shoot initiation and length of shoot at different days after inoculation of callus on tissue culture media in Brassica rapa L. for their good regeneration potentiality through anther culture.

Methodology:

The experimental materials were F1 regeneration of SAU sharisha 1×Tori 7, BARI sharisha 6×Tori 7 & BARI sharisha 15×Tori 7.The experiment was conducted in the Biotechnology Laboratory of the Department of Biotechnology, Sher-E-Bangla Agricultural University, Dhaka during the period of August 2011 to March 2012. Seeds were germinated and grown under controlled environment in the net house. Unopened flower buds were collected from the donor plants having the different nucleated stages (observed using 1%accetocarmine under compound microscope).The 3.01-3.50 mm size of buds were selected and wrapped with aluminum foil and kept at 4?C chamber 4 days. Full green and healthy flower buds were used for culture. The following MS (Murashige and Skoog) tissue culture medium supplemented with following concentrations of 2,4-D and BAP were used as phytohormone: T₁=Normal MS, T₂=1.0 BAP, T₃=1.5 BAP, T₄=2.0 BAP, T₅=2.5 BAP, T₆=1.02,4-D, T₇=1.0 BAP + 1.0 2,4-D, T₈=1.5 BAP + 1.0 2,4-D, T₉=2.0 BAP + 1.0 2,4-D, T₁₀=2.5 BAP + 1.0 2,4-D, T₁₀=2.5 BAP + 1.0 2,4-D, T₁₁=0.5 2,4-D, T₁₂=1.0 BAP + 0.5 2,4-D, T₁₃=1.5 BAP + 0.5 2,4-D, T₁₄=2.0 BAP + 0.5 2,4-D and T₁₅=2.5 BAP + 0.5 2,4-D. Sterilization - Sterilization of flower bud was practiced by sterile water then 70% ethyl alcohol for 2 minute, again washed by sterile water and then 0.1% w/v HgCl₂ for 1 minute and finally washed by DDH₂O twice. Different phytohorme were sterilized by using filter (Sieve size 0.45μm). Growth medium and required glassware and small instrument were sterilized with autoclave (at 121?C, 15psi and for 20 minute). Measurement of flower Bud - By using digital slide calipers flower bud size was measured and expressed in millimeter (mm).It is well established that uninucleated stage is most responsive to regeneration. The uninucleated satge was identified by 1% accetocarmine test of microspore under compound microscope. Growth of vegetative nucleus in microsporogenesis should be retarded as it were pollen grain can not formed during anther culture. This is important for development of haploid. Duration of cold pretreatment (4?C) was counted for 4 days. Inoculation of anthers - Anthers were removed by using sterilized fine Tweezers (forceps) and inoculated on the sterile culture vials containing medium and supplemented with growth hormone. The inoculated vials were cultured at 32?C growth chamber for 2 weeks in complete dark condition and next 1-2 weeks in 5000 lux light intensity with 70% relative humidity and 14/10 hour light-dark condition. Sixty anthers of each flower bud group were inoculated. The culture vials showing signs of contamination were discarded. Day to day observations was carried out to record the responses on callus induction. Inoculation of callus - Callus were sliced by using sterilized fine sterilized surgical blade and inoculated on the sterile culture vials containing medium and supplemented with growth hormone. The inoculated vials were cultured at 25?C growth chamber for different days in 5000 lux light intensity with 70% relative humidity and 16/8 hour light-dark condition. Each piece callus was inoculated. The culture vials showing signs of contamination were discarded. Day to day observations was carried out to record the responses on shoot initiation and length of shoot.

Source of Information: BANGLADESH RESEARCH PUBLICATIONS JOURNAL; ISSN: 1998-2003, Volume: 8, Issue: 2, Page: 159-165, March - April, 2013

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The genotype F1 generation of BARI Sharisha 15 x Tori7 required minimum ( 7.933 days) and the genotype F1 of SAU Sharisha 1 x Tori7 required maximum ( 8.120 days) to initiate shoots from callus. The maximum (14.80 days) and the minimum (4.667 days) time was required to initiate shoot from callus with the hormonal treatment T₂ (1.0 mg/L BAP) and T₁₂ (1.0 mg/L BAP + 0.5 mg/L 2,4-D). The minimum (4.200 days ) time was required to initiate shoot in the genotype of F1 generation of BARI Sharisha 15 x Tori7 with the combination of T₁₂ (1.0 mg/L BAP + 0.5 mg/L 2,4- D) hormonal treatment. The maximum length of shoot (0.2707, 0.9160, 1.116 and 1.197 cm at 14, 21, 28 and 35 DAI respectively) was observed from the genotype F1 generation of BARI Sharisha 6 x Tori7. The longest shoot (1.220, 2.007, 2.257 and 2.357 cm at 14, 21, 28 and 35 DAI respectively) was observed from the hormonal treatment T₁₄(2.0 mg/L BAP + 0.5 mg/L 2,4-D). The highest length ( 1.34, 2.120, 2.370 and 2.470 cm at 14, 21, 28 and 35 DAI respectively) was observed from the genotype F1 generation of BARI Sharisha 6 x Tori7 with from the hormonal treatment combinationT₁₄ ( 2.0 mg/L BAP + 0.5 mg/L 2,4- D). So, the genotype F1 generation of BARI Sharisha 15 x Tori7 and the hormonal treatment combination T₁₂ (1.0 mg/L BAP + 0.5 mg/L 2,4-D) are best to initiate shoot and ; the genotype F1 generation of BARI Sharisha 6 x Tori7 and the hormonal treatment combinationT₁₄ ( 2.0 mg/L BAP + 0.5 mg/L 2,4- D) are best for length of shoot.

Knowledge Management: Journal