Production of CMV-free <I>Gloriosa rothschildiana</I> O’ Brien Using Tissue Culture Techniques

Abstract

Gloriosa rothschildiana O’ Brien tubers from Nong Khiaw Royal Project Development Center were collected and planted under greenhouse condition. After planting for 90 days, stripe mosaic and distortion of leaves, mosaic pale on flower petals were clearly observed. Infections were proved by RT-PCR of Cucumber mosaic virus (CMV) Five criteria severity levels 0-4 (healthy normal-severest) were used to evaluate the diseases in 100 leaf-samples, no healthy normal detected, severity level 3 was the major percentage found at 33.75% followed by level 4, 2, 1 and 0 at 25, 20, 13.75 and 7.5 % respectively. Additionally, more severe of the symptom was less flower number per plant. Virus type checking by ELISA technique was proved for CMV infection. Then RT-PCR showed CMV-free treatments studies 100% succeed, excision of meristem at size 0.3 mm and 40 mg/l ribavirin treatment of in vitro shoot removal on tuber at size 0.5 cm with the survival rates at 75 and 100%, respectively. Culture medium, MS supplemented with BAP 3.0 mg/L and NAA 0.5 mg/L was proper for production of gloriosa plantlets at rate 2.27 shoots and 2.20 mini-tuber per explant after culture for 12 weeks.