Vanilla Case Study

Vanilla is a vine, generally propagated by stem cutting from mature vine. But this method is slow, time consuming and labour intensive. With the high demand in the world market, conventional technique is not able to accommodate the demand and hence this technique is currently being replaced by tissue culture. Tissue culture technique are not depending on the season and require only a limited quantity plant tissues as source of initial explants. The acclimatization stage involves the acclimatization stage involves conversion of heterotropic to autotropic growth condition and gradual return to natural growth characteristics of the plant. This is because survival rate in nature to be low. Therefore the use of PGPR is need to overcome the difficulties

Explant (3- 3.5) were disinfected with detergent and washer under a running tap water for 45 min. For sterilization , there were 3 treatments. Treatment 1 used 0.15 HgCl2 and 60% Clorox. Treatment 2 used 0.15 HgCl2 , 60% and 40% Clorox. For treatment 3, the solution was 0.15 HgCl2 , 60%, 40% and 20% Clorox. A final cutting for each explant was 1.5 -2cm. The explants were transferred into NDM with 0.2% activated charcoal and 1mg/L BAP hormones. Data free contamination, survival and shoot initial rate percentage were recorded after 60 day of culture. The next experiment, studied the effect of BAP (0, 0.5, 1, 3, 6 and 9mg/L) on shoot regeneration. Node (1.5cm) from in vitro derived shoot were used as explant for this experiment and transferred into NDM with o.2% activated charcoal. Each flask contained one explant. Percentage of shoot regeneration, day to shoot formation, number of shoot per explant and shoot length was recorded after 60 days of culture. For multiplication experiment, the combination of BAP (0.5 and 1mg/L) with NAA (0, 0.5, 0.1 0.2, 0.3mg/L) were

For first experiment, treatment 3 was the best for explant sterilization and best explant was Explant 1. The second experiment showed that the 1mg/L of BAP was best concentration in shoot regeneration and increased the shoot length. For third experiment, explant in media combination 1mg/L BAP with 0.3mg/L NAAwas produced shoot in short time and followed by 1mg/L BAP with 0.1mg/L NAA and also found that explant in all treatments produced one shoot per explant. Meanwhile explant in media with 1mg/L BAP and 0.2mg/L NAA produced highest length of shoot.Result of decapitated in vitro-derived shoot after 60 days cultured, the highest number of shoot was explant in media with 1mg/L BAP and 0.2mg/L NAA and result of shoot length was showed that explant in media with 0.5mg/L BAP and 0.3mg/L NAA produced a highest length of shoot. The forth experiment were showed that all explants in NDM with different concentration of IBA and NAA were produced root. Media containing 2mg/L IBA was a highest number of roots and explant in media with 1mg/L IBA was produced a longest root.For acclimatization, the result was indicated that the all plant was survived after 80 days transfer to ex vitro condition among