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In Vitro Regeneration of Corchorus Capsularis

M. Hoque
Department of Agronomy and Seed Science, Sylhet Agricultural University, Sylhet

K.M. Nasiruddin
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202

M.S. Haque
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202

M.K. Islam
OFRD, BARI, Rangpur, Bangladesh

G.C. Biswas
Department of Biotechnology, Bangladesh Agricultural University, Mymensingh-2202

Abstract/ Executive Summary:

The experiment was conducted during May to December 2008 in the Biotechnology Laboratory of the department of Biotechnology, Bangladesh Agricultural University, Mymensingh to observe the callus induction, regeneration potentiality and to establish a suitable in vitro plantlet regeneration protocol of Corchorus capsularis. MS medium supplemented with different concentrations of phytohormone and their combinations were used to observe the callus induction, shoot regeneration and root formation ability of the cotyledon with attached petiole derived explant of three genotypes viz. CVL-1, CVE-3 and BJC-7370. The highest callus induction (96.43%) was observed in CVE-3 in the MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA. Genotype CVE-3 in MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA produced the highest percentage of shoot regenerants (87.50%) followed by CVL-1 (75.00%) in the media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA. The root formation from regenerants was the best on half-strength of MS media supplemented with 0.6 mg/L IBA in genotype CVE-3 (75.00%). The in vitro regenerated plantlets from the genotypes CVE-3 and CVL-1 were established in the field successfully. Therefore, genotypes CVE-3 of C. capsularis in MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA for callus induction and shoot regeneration and ½ MS + 0.6 mg/L IBA for root formation would be used for further research work.

Keywords: Regeneration, Phytohormone, Corchorus capsularis

Location: Biotechnology Laboratory of the department of Biotechnology, Bangladesh Agricultural University, Mymensingh

Start Date: 00-05-2008

End Date: 00-12-2008

Program Area: Variety and Species

Commodity: Jute

Objectives:

1. to optimize the hormonal concentration for in vitro regeneration performance of different genotypes of C. capsularis.

Methodology:

The experiment was conducted during May to December, 2008 in the Biotechnology laboratory of Department of Biotechnology, Bangladesh Agricultural University, Mymensingh to observe the regeneration potentiality of three genotypes of C. capsularis viz. CVL-1, CVE-3 and BJC-7370. Culture Media: Half strength MS medium supplemented with clinical cotton or agar was used for seed germination. For callus induction and shoot regeneration, MS media supplemented with a single concentration of IAA (0.5 mg/L) and four concentration of BAP (1.5, 2.5, 3.5 and 4.5 mg/L) were used in four combinations. Half strength MS media supplemented with four concentration of IBA (0.2, 0.4, 0.6 and 0.8 mg/L) were used for root initiation. Soil containing 25% garden soil, 50% sand and 25% cow dung was used for transplanting of plantlets from culture vessel to pot. Culture Techniques i) Axenic culture: Twenty five sterilized C. capsularis seeds were placed into sterilized seed germination medium in each vial. The culture was then incubated in dark till the germination of seeds and then transferred to 16 hours light for normal seedling growth. Seven days old seedlings were used as source of explants. ii) Explant culture: Cotyledons with attached petiole were cut off from the seedlings and incubated to culture vial. Seven explants from each genotype were inoculated in each culture vial. The culture vials containing the explants were placed under fluorescent light in a room with controlled temperature (22 ± 2°C) using 16 hours photoperiod. iii) Subculture of the callus for shoot regeneration: Six calli of 20-25 mm in diameter from each genotype were used in this step in MS media containing different concentration and combination of IAA and BAP. The sub-cultured vials were incubated at 22 ± 2°C with 16 hours photoperiod. iv) Subculture of the regenerated shoot for root initiation: Shoot regenerated five calli of each genotype were cultured in vials with freshly prepared root induction medium to form root. The vials were again incubated at 22 ± 2°C with 16 hours photoperiod. v) Preparation of pot and transplantation: The plantlets of 5-7 cm in length having enough shoot and root system were taken out from the vials and then transplanted to pots containing of garden soil, sand and cow dung. To resist sudden stress, the pots were kept in a growth room for 10-15 days under controlled environment covered with moist polythene. After two to three days the polythene bags were partially removed and completely removed when the complete plantlets were seems to be self-sustainable. Recording Data i) Callus initiation: Data on days required for callusing and per cent callus induction were recorded after five to seven days of incubation of explants. The mean value of the data was considered as the days required for callusing. The percentage of callus induction was calculated by the following formula. Per cent callus induction = {(Number of explants induced callus) ÷ (Number of explants incubated)} × 100. ii) Shoot regeneration: The number of shoot proliferated over a number of days were recorded. The mean value of data provided the days required for shoot initiation. The percentage of shoot regeneration was calculated by the following formula. Percent shoot regeneration = {(Number of calli with plantlets) ÷ (Number of inoculated calli)} × 100. iii) Root formation: Days required for initiation of root from the day of implantation was recorded. The number of roots proliferated over a number of days were recorded. The mean value of the data provided the days required for root initiation. The percentage of established plants was calculated by on the following formula. Per cent plant establishment = {(Number of established plantlets) ÷ (Total number of plantlets)} × 100.

Source of Information: J. Agrofor. Environ. 7 (1): 41-44, 2013; ISSN 1995-6983

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The highest callus induction (96.43%) was observed in CVE-3 in the MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA. Genotype CVE-3 in MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA produced the highest percentage of shoot regenerants (87.50%) followed by CVL-1 (75.00%) in the media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA. The root formation from regenerants was the best on half-strength of MS media supplemented with 0.6 mg/L IBA in genotype CVE-3 (75.00%). The in vitro regenerated plantlets from the genotypes CVE-3 and CVL-1 were established in the field successfully. Therefore, genotypes CVE-3 of C. capsularis in MS media supplemented with 2.5 mg/L BAP + 0.5 mg/L IAA for callus induction and shoot regeneration and ½ MS + 0.6 mg/L IBA for root formation would be used for further research work.

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