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Effect of Explant Type on Direct Organo Genesis of Tomato (lycopersicon Esculentummill.)

N Sultana*
Associate Prof., Dept. of Genetics and Plant Breeding PSTU, Patuakhali

Irin Sultana
Lecturer, Dept. of Agriculture, Ghiore Govt. College, Manikganj

Abstract/ Executive Summary:

The direct in vitro regeneration of six types of explants (segments from hypocotyl, cotyledons, apical meristem, leaf disc, internode and petiole) was evaluated in tomato (Lycopersicon esculentum Mill.) variety BARI tomato-15. Fifteen days old explants were cultured on MS medium supplemented with 1.0 mg / L kinetin and 2.0 mg / L BAP for shoot regeneration and with 1.0 mg/L IAA for root induction. In terms of shoot regeneration percentage, hypocotyl explant performed the best (83.33%) in association with large number of shoots (9.0) and the maximum shoot length (3.8 cm). Explant apical meristem exhibited the second the highest performance in terms of all these characters. The lowest performance was shown by the explant internode. Explant cotyledon was close to apical meristem for shoot regeneration percentage (75%) and average number of shoot (7.9). Explant leaf disc generated medium result for all the characters. In terms of root induction, explant hypocotyl showed the highest percentage (100%) followed by cotyledon, apical meristem, internode, leaf disc, petiole but for average root number, cotyledon showed the highest (17.00) whereas hypocotyl explant ranks the second (15.13). For mean root length, leaf disc showed the highest length (7.9 cm) and the lowest length (4.8 cm) by the apical meristem. In a nut cover, explant hypocotyl and apical meristem performed better than the other types of explant.

Keywords: In vitro, Tomato, Explant, Regeneration, Root induction

Location: Tissue culture laboratory of Lal Teer Seed Ltd. during May to August 2018

Start Date: 00-05-2018

End Date: 00-08-2018

Program Area: Variety and Species

Commodity: Tomato

Objectives:

The present study was undertaken to achieve the following objectives-1. To determine the most appropriate explant type for plantlet regeneration 2. To establish a reproducible protocol for in vitro regeneration tomato.

Methodology:

The experiment was conducted at the tissue culture laboratory of Lal Teer Seed Ltd. during May to August 2018. The seeds of tomato variety (namely BARI tomato 15)were kindly supplied by Bangladesh Agricultural Research Institute (BARI). Six types of explants (viz. segments from hypocotyl, cotyledons, apical meristem, leaf disc, internode and petiole) were used. Explants were collected from 15 days old seedlings. Sodium hypochlorite(5%)and 70% ethanol were used as disinfectant. Basal nutrient salt solution (stock solution) like macronutrient, micronutrient, iron source and vitamins were used to produce MS medium. A single concentration of Plant Growth Regulator combination (1.0mg/L kinetin, 2.0 mg/L BAP and 1mg/L IAA) was used so that the differences in regeneration frequency due to explant type can be assessed and result may not be affected by variation in growth regulators. 0.8% (8g/L) agar was used as solidifying agent in the media. 3% (30g/L) sugar was used as Carbon source. MS medium (Murashige and Skoog, 1962) was prepared using the legislative formula and the cultures were maintained in a 16h photoperiod and 24±2 temperature. For shoot regeneration medium, MS medium was supplemented with cytokinin 1.0 mg/L kinetin and 2.0 mg/L BAP (Gubis et al 2004, Chaudhry et al 2010). For root induction media, MS medium was supplemented with 1mg/L IAA (Gubis et al 2004). The pH of the media was kept in 5.8. The medium was prepared by using the stock solution which concentration was 10 times than the operating media. All the growth regulators were added to media after autoclaving and under laminar air flow cabinet. The entire experiment was carried out in a Completely Randomized Design (CRD). The tomato seeds were sterilized with 70% ethanol for 2 minutes and then put on half MS media for germination. When the seedlings were 15 days old, the explants were collected and put on to shoot regeneration medium and kept under 16 hrs light and 24±2 temperature. Sub culturing was done at 3 weeks interval to avoid nutrient deficiency and space competition. When the shoot reached at 4-5cm length with 4-5 leaves, they were put on root induction medium. It took 10-15 days for shoot regeneration for all of the explants. When the shoots were 6 -8 week of age, they were transferred to root induction medium and it took 7 to 14 days to root induction. Data were collected on days to shoot initiation, percent shoot initiation, shoot length, number of shoot per explant, days to root initiation, percent root initiation, root length, number of root per shoot.The average number of shoots per explant were calculated by counting each shoot, making summation and dividing by total no. of explants.The data were analyzed statistically using MSTATC software. The analysis of variance for the characters under study was performed and means were compared by DMRT (Duncan’s Multiple Range Test).

Source of Information: Eco-friendly Agril. J. 11(10):110-113, 2018 (October)

Article Link (Online Journal): www.efaj-international.com

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

The explant type is an important phenomenon in case of tissue culture experiment and the present investigation has been able to find out appropriate explant type for successful in vitro culture of tomato plant which will lead to successful transformation programs, micro-propagation projects and different in vitro stress tolerance experiments of tomato plants. To conclude, explant hypocotyl and apical meristem were the better performing than the other types of explant.

Knowledge Management: Journal