<![CDATA[Abstract. Basri AHH, Chailani SR, Rahardjo BT, Widyasari WB. 2026. Pilot evaluation of in vitro culture methods for ScSMV reduction in sugarcane PS 864 confirmed by RT-PCR. Asian J Agric 10 (2): g100217. https://doi.org/10.13057/asianjagric/g100217. Sugarcane Streak Mosaic Virus (ScSMV) is transmitted mainly through infected vegetative planting material, yet the feasibility of in vitro culture for producing RT-PCR-negative PS 864 planting material has not been well established. This study aimed to compare three in vitro regeneration pathways (callus, shoot, and meristem culture) in PS 864, with RT-PCR indexing of ScSMV in the regenerated plantlets. Visually symptomless and infected PS 864 plants were grown for 4-5 months as explant sources. Callus culture achieved the highest regeneration rate (~95%), whereas shoot culture was severely limited by contamination (90%), and meristem culture produced no plantlets during the study period. Thirteen samples (six conventional controls and seven in vitro plantlets) were indexed by RT-PCR using SM1/SM2 primer pair. Of the six callus-derived plantlets, four were ScSMV-positive, and two were negative; the single shoot-derived plantlet was also negative. Because no internal amplification control was included, the negative results are provisional and cannot confirm the true absence of the virus. The dataset was small and unequal across pathways (callus, n = 6; shoot, n = 1; meristem, n = 0), precluding robust comparative inference. These findings indicate that callus culture is the most reproducible route for regeneration of PS 864 and that molecular indexing is essential, as ScSMV was detected even in visually symptomless plants. Confirmatory studies with larger, balanced samples and an internal control are needed before these pathways can support virus-free seed-cane programs.]]>
