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In Vitro Cormel Producttion of Gladiolus (gladious Sp.) (2016-2017)

M. K. Jamil
Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

M. M. Khatun
Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

M. K. Hasan
Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

S. C. Halder
Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

D. Khanam
Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

Abstract/ Executive Summary:

To develop an efficient protocol for invitro cormel production of BARI gladiolus-4, corm sections with buds and nodal segments of inflorescence at heading stage were used as explant which were cultured in MS medium with BAP (1,2,3,4 and 5) mg L^-1 and NAA at (0.5 and 1.0) mgL^-1. When corm sections were used as explant, callus was initiated in the cultures after four to eight weeks. The maximum callus induction of 80% was observed on MS medium supplemented with BAP at 4 mg L^-1 and NAA at 1 mg L^-1. The efficient shoot regeneration from proliferated calli was observed in the presence of BAP from 1 to 2 mg L^-1 and also enhanced the earliest shoot explant induction (9.4 days). Shoot proliferation was maximum in MS medium supplemented with BAP 2 mg L^-1 and NAA 1 mg L^-1. On the other hand, shoot initiation started in the nodal culture within two weeks (13.8 days) of inoculation. The highest number of shoots per explant (3.6) was obtained in MS medium supplemented with 4 mg L^-1 BAP and 1 mg L^-1 NAA. The longest shoot (8.5 cm) and leaves per shoot (3.6) was also found in the same treatment. The earliest rooting (11.6 days) and longest roots (5.3 cm) was observed in T₂ treatment i.e. MS media supplemented with IBA 2 mgL^-1 and 5 % sucrose. After successfull regeneration of the shoots, the plantlets were transferred into the cormel production media which consists of five treatments. Among the treatments, C₂ (MS+ IBA 2 mgL^-1 + sucrose 5%) performed better than all other treatments in case of cormel production,. The treatment C₄ (MS+ IBA 1.0 mgL^-1 + BAP 1.0 mgL^-1) did not produce any cormel during the study period.

Keywords: Flower, Gladiolus, Agar, autoclave temperature, Culture of explant, Flower spike

Location: Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur, Bangladesh

Start Date: 00-00-2016

End Date: 00-00-2017

Program Area: Crop-Soil-Water Management

Commodity: Gladiolus

Objectives:

To establish a suitable protocol for in vitro cormel formation of BARI gladiolus-4 using different type of explant.

Methodology:

Corms of BARI gladiolus-4 and flower spike at heading stage were collected from the field of floriculture division of Horticulture Research Center, BARI, Gazipur. Terminal and axillary buds of corms and nodal segments of inflorescence has been used as explant and culture on Murashige and Skoog basal medium supplemented with different concentration of BAP and NAA. The nodal segments were washed thoroughly with distilled water 3-5 times and surface sterilized by dipping the plant material into 70% (v/v) ethanol for 10 minutes. Then it was transferred to a solution of 1% sodium hypochlorite containing 2-3 drops of Tween-20 for 10 minutes followed by 4-5 times washing with sterile distilled water in a laminar airflow cabinet. In case of corms, the outer scale of corms was removed with surgical blade. The descaled corms were soaked in running tap water for 30 minutes to remove any sticky material present on the corms following 4-5 washings with distilled water. The corms were then surface sterilized with Ridomil Gold MZ 68 WG for another 20 minutes to remove the fungal contamination. Then the explant were sterilized with Jet and Trix for 30 minutes followed by 3-5 times washing with distilled water. After that the explants were treated with Clorox solution(60 ml clorox: 40 ml distilled water) with 2 drops of Tween-20 for 10 minutes followed by 3-5 times washing with sterile distilled water and again dip in 70% (v/v) ethanol for 15 minutes followed by 5-6 rinses in sterile distilled water under a laminar airflow cabinet. MS containing standard salts and vitamins, 3% sucrose and 0.85% agar was used and pH of the each medium was adjusted separately to 5.7 prior to addition of Agar. Medium was sterilized in autoclave at temperature of 121⁰C and pressure 15 psi for 20 minutes. The medium was placed in growth room for one week to check any medial contamination before use for culture of explant. The explants were incubated in a culture room where the temperature was maintained at 25-27ºC under continuous photoperiod of 16h light and 8h dark.

Source of Information: Annual Research Report 2016-2017, Biotechnology Division, Bangladesh Agricultural Research Institute, Joydebpur, Gazipur-1701, Bangladesh

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

From the experiment it can be concluded that the maximum callus induction of 80% was observed when corm sections used as explant and cultured on MS medium supplemented with BAP at 4 mg L^-1 and NAA at 1 mg L^-1. The earliest shoot induction (9.4 days), the maximum shoot proliferation (3.8), the longest shoot (5.6 cm) and the maximum average leaf per shoot (2.4) was found in T₄ treatment i.e. MS medium supplemented with BAP 2 mg L^-1 and NAA 1 mg L^-1.

When nodal segment used as explant,the highest number of shoots per explant (3.6), the longest shoot (8.5 cm) and the maximum leaves per shoot (3.6) was obtained from T₈ treatment i. e. MS medium supplemented with BAP 4 mg L^-1 and NAA 1 mg L^-1.On the other hand, the earliest rooting (11.6 days), the longest roots per plant (5.3 cm) and the maximum roots per plant (14.2) was observed in T₂ treatment i.e. MS media supplemented with IBA 2 mgL^-1 and 5 % sucrose.

Among the cormel production treatments, C₂(MS+ IBA 2 mgL^-1 + sucrose 5%) performed better than all other treatments. The treatment C₄(MS+ IBA 1.0 mgL^-1 + BAP 1.0 mgL^-1) did not produce any cormel during the study period.

Knowledge Management: Report/Proceedings