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In Vitro Micropropagation Through Cotyledonary Node Culture of Castor Bean (ricinus Communis L.)

Iftekhar Alam
Division of Applied Life Science (BK21), Gyeongsang National University, Jinju 660-701, Republic of Korea

Shamima Akhtar Sharmin
Division of Applied Life Science (BK21), Gyeongsang National University, Jinju 660-701, Republic of Korea

Sanjoy Chandra Mondal
Department of Botany, Rajshahi University, Rajshahi-6205, Bangladesh

Md. Jahangir Alam
Department of Botany, Rajshahi University, Rajshahi-6205, Bangladesh

Muhammad Khalekuzzaman
Department of Genetic Engineering and Biotechnology, Rajshahi University, Rajshahi-6205, Bangladesh

M. Anisuzzaman
Department of Botany, Rajshahi University, Rajshahi-6205, Bangladesh

Mohammad Firoz Alam
Department of Botany, Rajshahi University, Rajshahi-6205, Bangladesh

Abstract/ Executive Summary:

An efficient plant regeneration protocol was described for castor (Ricinus communis L.) using whole cotyledonary nodes as explant. Seeds were surface sterilized with 0.1% (w/v) mercuric chloride and germinated in growth regulatorfree MS medium. Cotyledonary nodes were excised from 5-7 days old seedlings and were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of BAP, Kin singly or in combination with NAA. Use of BAP at 3.0 mg.l-1 induced the highest frequency (85%) of shoot induction as well as maximum number of shoots per explant (12.56). Proliferated shoot clusters were elongated in 1.0 mg.l-1 BAP in combination with 0.25 mg.l-1 GA3 . For root induction, in vitro shoots were transferred to rooting media containing NAA or IBA. The highest rooting frequency (87.5%) as well as highest number of roots (10.5) was observed in MS medium supplemented with 1.0 mg.l^-1 NAA. Regenerated plantlets were acclimatized successfully in the growth room for further development.

Keywords: In vitro, Micropropagation, Medicinal plant, Ricinus communis

Location: Division of Applied Life Science (BK21), Gyeongsang National University, Jinju 660-701, Republic of Korea

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Bean

Objectives:

To establish a high frequency plant regeneration system from whole cotyledonary explant.

Methodology:

Plant materials and culture conditions - Seeds collected from the Botanic Garden, Rajshahi University were partially decoated and surfacesterilized in 0.1% (w/v) mercuric chloride for 4 min followed by 4-5 times rinses in sterile deionized water. Five to seven seeds were placed on petri dish containing MS medium devoid of PGR and incubated in the dark condition for germination. After 5-7 days, primary leaves and the epicotyls were detached from the seedling using sterile surgical blades. Then the seedling radicle was excised, leaving approximately 3–5 cm long hypocotyls intact. Explants consisting of cotyledon and axillary meristem regions with hypocotyls (3–5 cm long) were inoculated vertically on different shoot induction medium to induce shoots. Cultures were maintained by periodic subculturing on fresh medium once every 3 weeks. After 3-6 weeks of culture the multiple shoot clusters were elongated in a medium containing 1.0 mg.l-1 BAP plus 0.25 mg.l-1 GA3. When the regenerated shoots were attained a height of 2-3 cm at approximately 3-4 weeks, adventitious shoots were excised and transferred to rooting medium for root induction. In all cases, 3% sucrose (w/v) was used as a carbon source. After adjusting the pH to 5.7±0.01 prior to gelling with 0.8% agar (w/v) (BHD, England), the media were sterilized by autoclaving at 1210 C for 20 min (1.06 kg cm-2). Cultures were maintained in a growth chamber at 25±1°C under a 16/8-h (light/dark) photoperiod with a light intensity of 58-60 μmol m^–2 s^–1 (supplied by cool-white fluorescent lamps). Acclimatization and Transfer to Soil - Plantlets with a well-developed root system were washed carefully to remove agar and then transferred to the pots containing sterile vermiculite. After watering, plantlets were maintained in a growth chamber at 27±1°C under 16 h illuminations (145-150 μmol m^–2 s^–1) with fluorescent lamps. After 3 weeks of acclimatization, plantlets were transferred to larger pots containing vermiculite for further growth. Data recording and statistical analysis - To test the efficiency of shoot multiplication media, frequency (percentage) of shoot induction, number of shoot per culture and mean of length were calculated. Frequency of root induction and number of root per shoots were recorded for determining rooting efficiency. Mean values and standard error were calculated for the total number of shoots or roots in a given treatment. Each treatment consists of at least 12 replications and the entire experiment was repeated twice.

Source of Information: AJCS 4(2):81-84(2010); ISSN:1835-2707

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Maximum rooting (87.5%) was observed on the medium supplemented with 1.0 mg.l^-1 NAA followed by that observed on the medium supplemented with 1.0 mg.l^-1 IBA (80%). Root induction was strengthened within 3 weeks of culture. Least callus formation occurred in all rooting medium.

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