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Plant Regeneration from the Cotyledonds of Tossa Jute (corchorus Olitorius L.)

Asma Khatun
Genetics and Seed Division, Bangladesh Jute Research Institute, Manik Mia Avenue, Dhaka 1207, Bangladesh.

Chandan Kumar Saha
Genetics and Seed Division, Bangladesh Jute Research Institute, Manik Mia Avenue, Dhaka 1207, Bangladesh.

Zabun Naher
Genetics and Seed Division, Bangladesh Jute Research Institute, Manik Mia Avenue, Dhaka 1207, Bangladesh.

Shirin Mahbub
Genetics and Seed Division, Bangladesh Jute Research Institute, Manik Mia Avenue, Dhaka 1207, Bangladesh.

Abu Bakar Siddique
Genetics and Seed Division, Bangladesh Jute Research Institute, Manik Mia Avenue, Dhaka 1207, Bangladesh.

Shahana Bilkis
Genetics and Seed Division, Bangladesh Jute Research Institute, Manik Mia Avenue, Dhaka 1207, Bangladesh.

Abstract/ Executive Summary:

Plant regeneration from the cotyledons of C. olitorius has recently been optimized at Bangladesh Jute Research Institute (BJRI). One of the major constraints of getting plant regeneration from the explants of C. olitorius was the production of healthy seedlings in vitro. An efficient seed germination system and healthy seedling production have been established at BJRI. Seeds of different varieties (O4, O9897, OM1 and O72) of C. olitorius germinated on both agar supported hormone free MS medium and cotton supported hormone free liquid MS medium. The percentage of seeds germinated on cotton supported medium was found to be much higher than seeds germinated on agar supported medium. This system ensured maximum of an average of 98% seed germination. Whereas, in case of agar-supported medium an average of 46% seeds could be germinated with very slow growth. The seedlings grown on cotton supported medium were found to be much more healthily than seedlings grown on agar supported medium. Plant regeneration was obtained from the cotyledonary petioles of the varieties (O4, O9897, OM1 and O72) of C. olitorius on MS agar solidified medium supplemented by IAA 0.5 and BAP 3.0 mg l^-1. The efficiency of plant regeneration from the cotton supported seedlings was found to be as good as the agar supported seedlings. The plant lets produced roots on hormone free MS medium readily. After transfer to soil the plants grew into maturity and produced fruits. No morphological changes were noticed.

Keywords: Tossa jute,Bio technology, Shoot regeneration, cotyledons, Genotypes, Hormones

Location: Genetics and Seed Division, Bangladesh Jute Research Institute

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Jute

Objectives:

To develop a protocol for gene insertion in tossa jute through Agrobacterium vectors, the cotyledons

Methodology:

Seeds of C. olitorius L. (vars. O4, O9897, OM1 and O72) were surface sterilized by immersion in 0.1% (w/v) Mercuric Chloride for 20 min, followed by 6 washes with sterile deionized water. Seeds were placed on the surface of 50 ml aliquots of hormone free agar-solidified (0.8%, w/v) MS basal medium contained in 100 ml capacity of conical flasks. In another set of experiment, surgical cotton (1 g approx. in each flask) was used instead of agar in association with MS basal liquid medium to obtain optimum seedling production. Surgical cotton was placed at the bottom of 100 ml flasks. Each flask contained 20 ml of hormone free MS liquid medium. Cultures were placed in a growth room with 28°C under 1.0 Wm-2 of daylight fluorescent tubes with 12 h photo period. The cotyledons (with attached petioles) of 7 day old in vitro grown seedlings of C. olitorius (vars. O4, O9897, OM1 and O72) were excised and cultured on MS agar (0.8%, w/v) solidified medium with 3% (w/v) sucrose and pH 5.8 with the supplement of IAA 0.5 mg l^-1 and with BAP (3.0 mg l^-1). Explants were cultured in 250 ml conical flasks and maintained at 28°C under a 1.0 Wm-2 of daylight fluorescent tubes with 12 h day length. The experiments were repeated 3 times with 3 replications. The results were scored 6 weeks after culture. Excised single shoots (1.5-2.5 cm in length) were transferred to hormone free MS medium for root production and maintained under the same culture conditions as above. For transfer, sterilized dairy soil (Savar Dairy) and sand mixture was used followed the methods. All data were analyzed by using the analysis of variance (ANOVA) method of Minitab Statistical Package, Version-13. Test of differences between means were made at 5% probability level when a significant F value was obtained for varietal effect. A two sample T-test was also performed on the data to compare agar-supported and cotton-supported media. Different varietal means were compared by calculating a Least Significant Difference (LSD).

Source of Information: Biotechnology, 2(3): 206-213, 2003, ISSN 1682-2978

Article Link (Online Journal): DOI: 10.3923/biotech.2003.206.213, URL: http://scialert.net/abstract/?doi=biotech.2003.206.213

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The experiments reported in this paper demonstrate a dramatic result for seed germination from the varieties of C. olitorius on clinical cotton supported liquid MS medium which is very high compared to agar solidified medium. This system also ensured very healthy seedling production. This result could be a valuable addition for tissue culture system as cotton supported seed germination system is comparatively cheaper than agar supported system. A repeatable plant regeneration protocol has been established from the cotyledons (with attached petioles) of C. olitorius in BJRI Laboratory. This experiment is still being continued to increase the average number of shoot regeneration cotyledon-1 and also to improve the percentage of cotyledons producing shoots. This plant regeneration system can be used for genetic transformation in C. olitorius.

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