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Effect of Explants on Callus Formation of Potato

MS Huda
Farm Division, BARI Gazipur

MM Hossain
Dept. of Horticulture, Seed Science and Tech. Unit BSMRAU, Gazipur

MZ Haq
Seed Science and Tech. Unit BSMRAU, Gazipur

SS Zamal
Minor Irrigation BADC, Dhaka

MR Karim
Agric. and Food Security Programme, BRAC

Abstract/ Executive Summary:

Keywords: 2, 4-D, Dark condition, Alternative light and dark condition, Potato.

Location: Tissue Culture Laboratory of Tuber Crops Research Centre, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh

Start Date: 00-00-2011

End Date: 00-00-2012

Program Area: Variety and Species

Commodity: Potato

Objectives:

The study was undertaken to find out suitable explant for callus formation of potato.

Methodology:

The experiment was conducted at the Tissue Culture Laboratory of Tuber Crops Research Centre, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh during 2011-12. The different explants i.e. i. shoot ii. leaf iii. node and iv. internode were used in MS media supplemented with 2.0 ml L^-1 2,4-D (2,4-dichlorophenoxy acetic acid) and incubated at completely dark condition and alternative light and dark condition (1.83 m fluorescent tubes and was illuminated 16 h daily with a light intensity of 1500 lux). Preparation of expants: In vitro plantlets of 25-30 days old at solid medium were considered for the explants collection. For stem explants, only the first 5-6 internodes from the top of the plantlet excluding shoot apex of the each plantlet were excised. Internode and node segments were excised to avoid the axillary buds and divided into 0.5- 1.0 cm long segments. For leaf explants, thick and healthy leaves from the upper nodes of the plants were used. The leaf tips and basal portions, including the petiole were discarded and the entire leaf was cut into 5x5 mm pieces. The leaf explants were placed upside down on the medium. For shoot explants, only 8-10 mm apex shoot were used where side leaves were discarded. All the explants were placed in test tube and fuel paper were used and kept in a growth room at 25±2 °C. Only the first five internodes from the top of the plantlets were excised. Using alcohol, the laminar air flow chamber was thoroughly cleaned to maintain the aseptic condition. Under aseptic conditions, young and tender plantlets were taken out on a sterile glass plate using sterile forceps. The plantlets were not surface sterilized as those were already maintained under in vitro aseptic conditions. The roots of these plantlets were excised using a sterile scalpel and the leaves were removed. Preparation of media, inoculation of explants and incubation: The composition of the media was MS media supplemented with concentration of 2,4-D 2.0 mg L^-1 Fine agar powder of Loba, India brand was used. After pH adjustment 8 g L-1 agar was added to the solution to solidify the media. An amount of 10 ml media were dispensed into each test tube. The test tubes were autoclaved at l2l0 C for 20 minutes at 1.2 kg/m² and thereafter stored at 250 C. After 2-3 days, the media was checked for any type of contamination and the explants were inoculated after performing several sequential steps. The explants were placed in each plate for each treatment horizontally in contact with MS medium supplemented with 2,4-D. The treatment was replicated for three times under Completely Randomized Design (CRD) design. Every ten test tubes were used for one replication. The plates were sealed, labeled carefully and were finally kept in the culture room under constant temperature (25⁰C) and light (16 hrs. light, 8hrs. dark), and the growth of the inoculated explants was monitored regularly. Only plantlets with well developed roots were used for multiplication in net house. Data on callus initiation was recorded after two weeks and buds per explants after five weeks. The data were analyzed statistically and means were separated by Least Significance Difference method.

Source of Information: Eco-friendly Agril. J. 6(08): 146- 149, 2013 (August); ISSN 1999-7957

Article Link (Online Journal):

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The completely dark condition is suitable then alternative light and dark condition for callus formation within shortest time (10 to 13 days) but the alternative light and dark condition is suitable for largest or very massive degree of callus were produced. At the completely dark condition the explants of internode of potato (Asterix) is suitable but in case of alternative light and dark condition the explants of nodes is appropriate. and compact type of callus were produced from the node and inter node explants by using MS Media supplemented with 2 mg L^-12,4-D (2,4-dichlorophenoxy acetic acid).

Knowledge Management: Journal