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Mass Propagation of Bambusa Bambos (l.) Voss Through in Vitro Culture

Raihan I. Raju*
Department of Botany, Jahangirnagar University, Savar, Dhaka-1342, Bangladesh

Shyamal K. Roy
Department of Botany, Jahangirnagar University, Savar, Dhaka-1342, Bangladesh

Abstract/ Executive Summary:

Protocol for mass propagation of Bambusa bamboos (L.) Voss was developed through in vitro culture. Nodal segments containing pre-existing axillary bud, after surface sterilization, were inoculated on liquid Murashige and Skoog’s (MS) basal medium containing different concentrations and combinations of cytokinins (BAP, TDZ and Kn). The highest direct shoot induction (90%) was obtained in the MS liquid medium supplemented with 2.0 mg/l BAP and 1.0 mg/l TDZ with maximum average number of shoots (3.14 ± 0.06) per explant. Highest shoot multiplication (16.58 ± 0.24 shoots per culture) with highest average shoot length (9.21 ± 0.13 cm) was obtained when in vitro raised shoots were cultured in gelrite gelled MS medium in conjunction with 2.0 mg/l BAP and 1.0 mg/l TDZ. Incorporation of 10% coconut water with 4% sucrose in the above mentioned medium resulted satisfactory shoot growth and development with an average 26.7 ± 0.60 shoots per culture. For root induction, in vitro raised shoots were divided into clumps of 4-5 shoots in each clump and transferred onto both liquid and gelled half-strength MS medium containing different concentrations and combinations of auxins (IBA and NAA). Maximum rooting (86.67%) was achieved in half-strength of MS medium fortified with 2.5 mg/l IBA and 2.5 mg/l NAA with an average 8.72 ± 0.42 root per shoot. The rooted plantlets were then transferred to polybags containing garden soil, sand and compost mixture with 1:1:1 ratio. After a month the hardened plantlets were then transferred to the larger pots containing garden soil and compost with 1:1 ratio for sufficient growth and finally transplanted to the field. In this process, the highest 100% survivability was recorded from well-established rooted plantlets. The regenerated plants showed well developed root and shoot systems in field condition.

Keywords: Bambusa bambos, Mass propagation, In vitro culture.

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Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Bamboo

Objectives:

In this study, an efficient and reproducible micropropagation protocol has been established mass production and generating uniform clones for the bamboo species, Bambusa bambos (L.) Voss.

Methodology:

Nodal segments containing unsprouted axillary buds of Bambusa bambos (L.) Voss were collected from the lateral branches of young culm thicket grown in the campus of Jahangirnagar University, Savar, Dhaka. The nodes containing pre-existing axillary buds were used to initiate and establish the in vitro culture. Leaf sheath of nodal segments were removed with sharp blades and nodes containing lateral spines were removed by scraping carefully with scalpel. These nodal explants were thoroughly washed under running tap water to remove the dust particles. They were then washed with few drops of liquid detergent for 15 minutes followed by 3-4 times washing with distilled water. Further surface sterilization of the explants was done under laminar airflow by giving fungicide treatment (Bavistin) for 5-7 minutes followed by 3 times washing with autoclaved distilled water. Finally these explants were surface sterilized with (0.05%, 0.1% and 0.2%) HgCl₂ solution for 3-10 minutes to ensure contamination free culture and then thoroughly washed with autoclaved distilled water. Then both ends of the sterilized nodal segments were trimmed and cultured singly in liquid MS medium (Murashige and Skoog, 1962) supplemented with various concentrations and combinations of Cytokinins (BAP, TDZ and Kn). The pH of the medium was adjusted to 5.8 with the help of pH meter by adding 0.1N NaOH or 0.1N HCl accordingly prior to autoclaving at 15 lbs pressure and 121°C for 15 mins. Axillary buds of nodal segments were sprouted within 18-25 days. The sprouted buds were then sub-cultured in another bottle containing same gelled media. Within 4-weeks induced shoot elongated and developed into a number of multiple shoots. The elongated cluster (4/5 shoots) of shoots were used as explants for further shoot multiplication and root induction. At the end of each stage, shoot number per explant and shoot length (cm) were calculated. The effect of different concentrations of source (1-5%) and coconut water (5-20%) on shoot multiplication were also studied. An aseptic condition was maintained throughout the whole operation. Cultures were incubated at 24±2°C temperature under illumination with florescent light for 16-hours light and 8-hours dark per day. In vitro induced shoots (4/5 shoots) in multiplication medium were then used for root induction. Half-strength of both solid and liquid MS medium with various concentrations and combinations of IBA and NAA were used. After 30-45 days of culture, the number of roots per shoot cluster and rooting percentage was recorded. The plantlets with well developed root system were acclimatized. For acclimatization the plantlets with sufficient root inside the test tubes were kept in the room temperature for 5-7 days to bring them in contact of normal temperature. The roots of the plantlets were gently washed under running tap water to remove any chemical attached to the root area. Immediately after that they were transferred to small polybags containing the soil mixture garden soil, sand and compost with 1:1:1 ratio. The plants along with pots were covered with transparent polythene bags to prevent sudden desiccation. The inner side of the polythene bags was sprayed with water at every 8 hours to maintain high humidity around the plantlets. The polythene bags were gradually perforated to expose the plantlets to the outer normal environment and subsequently removed after 14 days.

Source of Information: Jahangirnagar University J. Biol. Sci. 5(2): 15-26, 2016 (December)

Article Link (Online Journal):

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Achievement/Findings:

The present study provides an efficient and cost effective protocol for in vitro propagation of Bambusa bambos (L.) Voss from the nodal segments of the field grown mature culm, with high multiplication efficiency, proper rooting and easy establishment in the field condition with normal growth performance of the in vitro propagated plants. This protocol will be helpful for large scale production of edible bamboo as well as for gene pool conservation.

Knowledge Management: Journal