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Induction of Somatic Embryogenesis in the Cultures of Zygotic Embryos of Lychee

M. N. AMIN
Department of Botany, University of Rajshahi, Rajshahi-6205, Bangladesh.

M.A. RAZZAQUE
1nstitute of Biological Sciences, University of Rajshahi, Rajshahi-6205, Bangladesh.

Abstract/ Executive Summary:

Zygotic embryos at different developmental stages were collected after 20-80 days of pollination and cultured on agar-gelled MS medium with half strength major salts and hlueherry mediumwith480mg/l KN0₃,400mg/l NH₄N0₃),41.3 mg/1 FeS0₄and55.7 mg/1 NaEDTA. Embryos 4 4 cultured on modified blueberry medium responded better than those cultured on modified MS medium and an addition of activated charcoal was beneficial in both the instances. The embryos excised at early stages of development (3-8 mm) showed direct somatic embryogenesis in almost 100% cases primarily from cotyledons and later from elongated radicles under the influence of 5.0 mg/1 BA and 1.0 g/1 activated charcoal. The primary embryos thus formed showed secondary and tertiary embryogenesis at the globular and heart stages. The repetitive embryogenic cultures had been maintained for more than a year on revised blueberry medium with BA and activated charcoal without loss of embryogenic competence. So far, 40% of in vitro formed embryos matured but none developed into a viable plantlet.

Keywords: Somatic embryogenesis, In vitro culture, Litchi chinensis.

Location: 1nstitute of Biological Sciences, University of Rajshahi

Start Date:

End Date:

Program Area: Variety and Species

Commodity: Litchi

Objectives:

To induce somatic embryogenesis in the cultures of zygotic embryos of lychee.

Methodology:

The developing fruits of an early maturing local cultivar were collected after 20-80 days of pollination. The fruits were first treated with I% ( v/v) Savlon (an antiseptic plus detergent)solution for about 10 min, washed thoroughly under running tap water, rinsed briefly with 80% ethanol and finally surface-sterilized with 0.1% HgCI₂ for 5 min. After three rinses with sterile distilled water, the fruits were dissected under a laminar flow cabinet using flame sterilized fine forceps and surgical knives. The embryos were taken out by removing the fruit pericarp and making a careful incision to the testa. The excised embryos were first collected in a sterile Petri dish lined with moistened sterile filter paper to protect the embryos from desiccation. Finally they were cultured on different nutrient media. For inducing somatic embryogenesis MS medium (Murashige and Skoog 1962) with half strength major salts (MMS) and blueberry medium (Zimmerman and Brome 1980) with 480 mg/1 KNO₃, 400 mg/1 NH₄N0₃_,41 .3 mg/1 FeS0₄and 55.7 mg/1 NaEDTA (MBM) were 4 3 4 used. Both the revised media were supplemented with 30 mg/1 sucrose, 0.5-5.0 mg/1 BA, I g/1 activated charcoal (AC) and gelled with 6 gil agar (BDH). For maturation and germination of the somatic embryos liquid MBM with filter paper bridge, 15% coconut water, I g/1 AC, 20-40 g/1 sucrose and 0-2.0 mg/1 GA₃was used. The media pH was adjusted to 5.7 ± 0. 1 and the media were sterilized at 121 °C under 1.1 kg/cm²pressure for 20 min. The cultures were grown at 25^oC with a light intensity of about 2000 lux.

Source of Information: Bangladesh J. Bot. 24(1): 25-29, 1995 (June): ISSN 0253-5416

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The embryos excised at early stages of development (3-8 mm) showed direct somatic embryogenesis in almost 100% cases primarily from cotyledons and later from elongated radicles under the influence of 5.0 mg/1 BA and 1.0 g/1 activated charcoal. The primary embryos thus formed showed secondary and tertiary embryogenesis at the globular and heart stages. The repetitive embryogenic cultures had been maintained for more than a year on revised blueberry medium with BA and activated charcoal without loss of embryogenic competence. So far, 40% of in vitro formed embryos matured but none developed into a viable plantlet.

Knowledge Management: Journal