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

Regeneration of Plantlets Through Somatic Embryogenesis from Nucellus Tissue of Citrus Macroptera Mont. Var. Anammensis ('sat Kara')

M. N. Miah
Institute of Life Sciences, National University, Gazipur, Bangladesh

Sahina Islam
IFRD, BCSIR, Dr. Kudrat-E-Khuda Road, Dhanmondi, Dhaka-1205, Bangladesh.

Syed Hadiuzzaman
Department of Botany, University of Dhaka, Dhaka-1000, Bangladesh.

Abstract/ Executive Summary:

Somatic embryogenesis and plantlet regeneration were achieved in callus cultures of nucellus derived from undeveloped ovule of immature fruits of Citrus macroptera. Four types of media were used but only modified MS medium supplemented with malt responded well. Calli produced in malt supplemented MS medium were embryogenic in nature. After transfer of this embryogenic callus in hormone free MS medium, somatic embryos were developed. Independent plantlets were developed from these somatic embryos by further subculture in the same medium. After five weeks of culture the plantlets were transferred to pot soil mixed with cow-dung derived biogas slurry and survived well.

Keywords: Nucellus, Somatic embryogenesis, Plantlet regeneration, Citrus macroptera.

Location: Citrus Research Station, BARI, Jaintapur, Sylhet.

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Satkara

Objectives:

To develop the in vitro suitable protocols for micropropagation of Sat Kara plants from nucellus through somatic embryogenesis.

Methodology:

Four to five weeks old immature fruits were collected from Citrus Research Station, BARI, Jaintapur, Sylhet. Fruits were washed thoroughly under running tap water to reduce dust and surface contaminants. Then they were dipped in 90 % alcohol for one minute followed by 4 % sodium hypochlorite solution for 10 min and finally rinsed four times with sterile distilled water under laminar airflow cabinet. The fruits were then placed on an autoclaved ceramic tile and cut open by sharp sterilized knife and then undeveloped ovules/immature seeds were separated. For somatic embryogenic callus induction the seeds were cut by scalpel and nucellus halves were separated and cultured on a semi-solid modified MS medium supplemented with 500 mg/l malt extract called it somatic embryogenic medium1 (SEM1) after Tisseret and Murashige (1977). At the same time nucellus halves were cultured into MS + 0.1 mg/l 2,4-D + 0.05 mg/l Kn (SEM2), MS + 0.5 mg/l 2,4-D + 0.1 mg/l Kn (SEM3) and MS + 1 mg/l 2,4-D + 0.5 mg/l Kn (SEM4) media. The nucellus halves were taken in conical flasks and containing five samples with three replicates for each medium.The responded calli were further subcultured on same media and simultaneously into hormone free MS medium. The pH of all media was adjusted to 5.7 before addition of agar and sterilized by autoclaving for 20 minutes at 1.05 kg/cm² (15 psi) pressure at 121_C. An amount of 10 gm/l agar (BDH) was used for SEM1 and 0.8 gm/l for other three media. The flasks containing explants were incubated on culture racks. The cultures were maintained at 25_C ± 2 under the cool white fluorescent lights for 16 hr photoperiod.

Source of Information: Plant Tissue Cult. 12(2) : 167-172, 2002 (December)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The results obtained from this experiment are presented in among all the media tested, calli developed in SEM1 were only embryogenic in nature. These calli were loose light green and having good growth. Calli obtained in other media were compact and initially green but with the lapse of time they turned brown. Embryogenic calli when transferred to hormone free MS medium somatic embryos were developed. Complete plantlets were developed from these somatic embryos by further subculture in the same hormone free MS medium. Regeneration of plants via somatic embryogenesis has been preferred as a method for multiplication of viable germplasm in many woody plants. The somatic embryogenic callus and plantlets were obtained from nucellus of many citrus species on malt supplemented SEM1 medium. This medium was suitable to develop a protocol for somatic embryogenesis from nucellus of citrus species. Successful embryogenic calli were developed only from SEM1 medium. In case of auxin-cytokinin supplemented media initially calli were induced but these were not embryogenic. Single primary root growth was observed in each plantlet and the mean height of plantlets was 13 mm. However, the observed GA₃ was effective for embryoid to plantlet regeneration. The plantlets thus obtained through somatic embryogenesis were transferred into soil mixed with biogas slurry 1 : 1 ratio and the rate of survival was 100%.

Knowledge Management: Journal