In Vitro Callus Induction from Leaf Explants of Vanda sp Stimulated by 2,4-D

Iman Budisantoso,Kamsinah Kamsinah,Nurul Amalia

doi:10.15294/biosaintifika.v9i3.11018

Iman Budisantoso, Kamsinah Kamsinah + Show 1 more

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https://doi.org/10.15294/biosaintifika.v9i3.11018

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Abstract

The addition of growth regulator is one of the critical success factors in in vitro cultures. 2,4-D as a plant regulator in media can stimulate the cell division, enlargement of the explants and promotes the formation and growth of callus. The purpose of this study was to determine the time of callus formation and to determine the best concentration of 2.4-D in inducing the growth of callus from leaf explants of Vanda sp. This research was conducted by experiment with completely randomized design, which consists of six levels of treatment concentration of 2,4-D i.e. 0 ppm; 1 ppm; 1.5 ppm; 2 ppm; 2.5 ppm; and 3 ppm. The parameters observed were the percentage of callus formation and the form of callus from Vanda sp leaf explants. The results were statistically analyzed by using MINITAB program version 17. Analysis of variance (ANOVA) was performed and the difference between means score/value was separated by F test at p &lt; 0.05. The results showed that 2,4-D treatment give significant effect (F 5,12 = 3,20; p = 0,046 &lt; 0,05) on the callus growth time and its percentage. Application of 2 ppm 2.4-D was the best concentration for accelerating the callus growth time (14.3 days after planting) and increasing the percentage of callus formation (83.3%). Most of callus type were proliferative callus (36.11%) and senescence callus (11.11%). The results of this research are very important to grow the callus from Vanda leaves orchid explant because it is very diffucult to grow.

Highlights

Vanda as one of orchid species is cultivated because it has beautiful flowers and valuable, so the seedlings need to be produced in large quantities
The shortest time of callus formation was at the treatment of 2 ppm 2,4-D (14,3 ± 1,5) and the longest time was at a control group (20 ± 2,0)
The data shows that the highest percentage of callus formation is found in the treatment with 2 ppm of 2,4-D

Summary

Introduction

Vanda as one of orchid species is cultivated because it has beautiful flowers and valuable, so the seedlings need to be produced in large quantities. According to Dohling et al (2012), the multiplication of an orchid can be done in various ways, conventional and non conventional (in vitro culture). The multiplication of orchid can be done by using vegetative and generative organ. Generative propagation by using seed has low success rate, because the orchid seed have no endosperm (Jainol & Gansau, 2017). Conventional method through seed propagation is sometimes problematic as seed germination in natural environment is a slow process and it requires mycorrhizal or fungal contribution (Parthibhan et al, 2015). Germination of orchid seed must be in symbiosis with mycorrhiza (Nasiruddin et al, 2003), so to solve this problem, it can be done by using in vitro culture propagation (Hasanah et al, 2014)

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