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Production of Breeder’s Propagule of Transgenic Late Blight Resistant Potato Clone Through Tissue Culture (2016--2017)

M. M. Khatun
Biotechnology Division, BARI, Joydebpur, Gazipur

S. C. Halder
Biotechnology Division, BARI, Joydebpur, Gazipur

D. Khanam
Biotechnology Division, BARI, Joydebpur, Gazipur

Abstract/ Executive Summary:

Transgenic D 951 (137), SP 951 and non-transgenic Katahdin were used in this study. Sprouts of these varieties were used as explant. Sprouts were cultured on PROP medium and multiplied in vitro. Initially, 30 tubers of D 951 (137) were used. Plantlets from the individual tuber of D 951 (137) were multiplied and tested for virus-free material. For that ELISA tests were performed and 10 materials were found virus-free. Only those materials were maintained and multiplied. From in vitro culture, a total of 2109 plantlets were produced of which 234 plantlets were non-transgenic Katahdin, 185 plantlets were SP 951 and 1690 plantlets were D 951 (137). Some plantlets were transferred to the greenhouse for mini tuber production. About 78- 86% of plantlets were survived in greenhouse

Keywords: ELISA test, Plantlets, Greenhouse, Potato, clone

Location:

Start Date: 00-00-2016

End Date: 00-00-2017

Program Area: Variety and Species

Commodity: Potato

Objectives:

The study has been undertaken to produce a large number of virus-free planting materials/mini tubes of that potato clone.

Methodology:

Sprouts of transgenic D 951 (137), SP 951 and non-transgenic Katahdin were used as explant sources. For inoculation of explants firstly potato sprouts were surface-sterilized under running tap water for 30 min. with jet and trix to remove the surface contamination outside the clean bench. After that, the sprouts were taken inside the laminar airflow cabinet in a autoclaved jar and dipped in 99% ethanol plus 2 drops of Tween-20 for 20 sec. and washed with autoclaved distilled water 2 to 3 times. Finally, sprouts were sterilized with clorox solution (60 ml clorox: 40 ml distilled water) for 1 min. and then washed for 3 to 4 times with autoclaved distilled water. After that the sterilized sprouts were excised with the help of the sterilized scalpel and forceps. Then the prepared explants were placed in text tubes/baby jars containing previously prepared media. The inoculated cultures were placed in a growth room at 25 ± 1⁰C under approx. 2000 lux by cool white fluorescent tubes for 16/8 light/dark cycle.

Source of Information: Annual Research Report 2016--2017, Biotechnology Division, BARI, Joydebpur, Gazipur

Article Link (Online Journal):

Funding Source:

1.

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Initiation of explant occurred within four weeks of culture. Shoots were produced straightway. Then single nodes were cut and put in the same medium for multiplication of these materials. Initially, 30 tubers of D 951 (137) were used. Plantlets from the individual tuber of D 951 (137) were multiplied and tested for virus-free material. For that ELISA test were performed and 10 materials 137(1), 137(2), 137(5), 137(7), 137(10), 137(12), 137(13), 137(19), 137(32), & 137(33) were found virus-free. Only those materials were maintained and multiplied in Laboratory. After three to four subcultures plantlets were multiplied in vitro and a good number of plantlets were produced. From in vitro culture, a total of 2109 plantlets were produced of which 234 plantlets were non-transgenic Katahdin, 185 plantlets were SP 951 and 1690 plantlets were D 951 (137). Some in vitro grown platelets were taken out from the growth room and acclimatized to greenhouse conditions for mini tuber production. About 78- 86% of plantlets were survived in the greenhouse. The plantlets were successfully established in potting soil and growing well. Among the varieties, the highest stem length was recorded from Katahdin (17.60 cm) and the lowest (10.80 cm) from 137(7). Maximum stem (5.40) was obtained from (137(7) and minimum from Katahdin (2.80). A higher number of nodes (10.60) was obtained from (137(12) and lower from SP 951(7.00) while the maximum leaf number (12.20) was recorded from (137(12) and lowest from SP 951(9.60).

Knowledge Management: Report/Proceedings