Micro Propagation of Several Potted Anthurium Accessions UsingSpathe Explants

The introduction of new varieties and production system of anthuriums has faced some problems due to the low multiplication rates in conventional vegetative and genotypic alteration problem in reproductive propagation. Since then, in vitro propagation technique became important to be investigated. The research was carried out from September 2006 to August 2007 at the Indonesian Ornamental Crops Research Institute. A complete factorial experiment was designed to accomplish two chronological in vitro activities. The first step dealt with different incubation sites, i.e. dark and light conditions for callus induction of three potted anthurium accessions, namely clone no. SM. 001, cv. Alphine and cv. Bonito. The best callus obtained from the incubation treatment were then, transferred into defined media with different formulations to get free-callus plantlets in the second steps. The results showed that more progressive callus development from spathe explant was detected on the media of modified Nitsch and Nitsch + 1 mg/l 2,D + 1 mg/l Kin + 1 mg/l BA stored at dark after 60 days incubation. After 45 days transferring the callus in the same media on light conditions, torpedo-shaped callus was detected. Complete planlets were obtained after 75 days reculturing the buds in Nitsch and Nitsch medium containing 2,4 D with various BA concentrations. Clone SM. 001 and cv. Bonito showed higher number of visible shoot and root developments compared to cv. Alphine. In general, better shoot and root developments and higher planlets survived after acclimatization were observed on the BA concentrations of 2 and 3 mg/l.


INTRODUCTION
To accelerate the incoming varieties to be released, the proper propagation of selected clones should be established to preserve sufficient amount of healthy seedlings. In other crops, in vitro propagation has shown to be an excellent tool for rapid upbuilding for marketable quantities of plants with selected characters. These method guaranteed the identical reproduction of the parents tested and selected, and prevented genotypic alteration which would occur after the generative multiplication (Chen et al, 2002).
The successful in vitro propagation of cut flower anthurium using young leaf explant to reduce contamination and oxidative browning of explants after culture inspiring the use of whirled and young spathe was also possible as source of explant. The steps of culture followed the methods of Budiarto and Handayati (2007). The first step was the establishment of aseptic explant culture to encourage cell division for callus formation. Modification of nutrient and vitamins, supplemental hormones and dark condition were usually applied to facilitate this meristematic activity (Chand et al, 1999). The next step was then, induction of the multiplied cells to undergo further differentiation and Telp/Fax: (0263) 512607/514138 Email: bud1arto@yahoo.com Budiarto proliferation to form shoot and root until the complete planlets were obtained. In this period, the existence of light was usually needed to serve the organogenesis process (Martin et al, 2001). In this paper, the study of in vitro propagation in three accessions of anthurium using spathe explant was described. The presence of light for callus induction and hormone modification for further callus proliferation were investigated.

MATERIALS AND METHODS
The research was conducted in the tissue culture  In contrast with these situation, the callus from the explants incubated in dark condition continued to grow and the appearance of torpedo callus (initial bud) was detected in 60 days (Figure 1). Forty five days after transferring the callus in the same media and incubating them in light condition, the buds showed more advance growth with initial leaves appearance ( Figure 2). Rosario & Lapitan (1981)  to Martin et al, (2002), the growth characteristic among accessions was genotypicly indigenous. Such cultivars could show faster or slower shoot and root formation even in the same media during in vitro culture. The complete planlets of the three accessions of anthurium were presented in Figure 3.
The presence of BA also affected on the number of newly developed shoots and roots of planlets. Table   3 showed that number of shoots increased in line with the higher BA concentration in the media. The highest shoot formation was observed in Nitsch and Nitsch media supplemented with 3 mg/l BA, though the value was not significantly different with those in 2 mg/l. The number of roots, however, was found highest in 2 mg/l BA with slight decrease compared to those in the media of 3 mg/l BA. While, the least shoot and root formations were observed in the media containing 1 mg/l BA.
Further organogenesis process, reflected from higher shoot and root formation on the planlets given with 2 and 3 mg/l BA, indicated that the process of shoot and root development was accelerated with the increase of BA concentration. The negligible differences between the 2 and 3 mg/l BA also inferred the light-BA interaction in the certain range was influenced by the endogen hormonal balance and the maximum response of the cells to the BA concentration (Villalobos et al, 1984). These findings implied that BA concentration ranging from 2 to 3 mg/l was specific for these three anthurium accessions in Nitsch and Nitsch medium for optimal shoot and root development. The optimal growth of shoot and root then, might contributed to better adaptation and higher survival rate of planlets during acclimatization (Table 3).