CULTIVAR-SPECIFIC RESPONSES OF STEVIA REBAUDIANA TO MULTI-WALLED CARBON NANOTUBES: EFFECTS ON GROWTH, STEVIOL GLYCOSIDE PRODUCTION, AND GENE EXPRESSION
Abstract
This study investigated the effects of multi-walled carbon nanotubes (MWCNTs) on growth, steviol glycoside production, and gene expression in three Stevia rebaudiana cultivars: High Sugar, ShouA3-2, and Levan. The research aimed at understanding how MWCNTs impact these important aspects of stevia cultivation, potentially informing future agricultural practices and steviol glycoside production methods. MWCNTs nonsignificantly affect shoots number and shoots diameter means in the three cultivars, and showed significant differences for shoots length means across all cultivars and number of roots in Levan under T3 and number of leaves in all cultivars under T2 when applied to in vitro cultures at 1, 1.5, and 2 mg/L MWCNTs concentrations. HPLC analysis revealed cultivar-specific responses in steviol glycoside content. The High Sugar cultivar showed increased stevioside (2625.60 ppm) and rebaudioside A (1971.34 ppm) content with Aqua regia treatment. The ShouA3-2 cultivar exhibited maximum stevioside content (1681.86 ppm) at 1 mg/L MWCNTs, while the Levan cultivar produced the highest stevioside (2788.194 ppm) at 1.5 mg/L MWCNTs. Gene expression analysis of UGT74G1 and UGT85C2 demonstrated cultivar-specific modulation in response to MWCNTs. The two cultivars, High Sugar and ShouA3-2, showed higher expression than Levan. The study reveals that the effects of MWCNTs on Stevia rebaudiana are cultivar-specific, particularly regarding steviol glycoside production and gene expression. The relationship between gene expression and steviol glycoside content under MWCNT treatment is complex and not always directly correlated, highlighting the intricate nature of secondary metabolite biosynthesis in Stevia rebaudiana. The relative stability of both UGT74G1 and UGT85C2 expression across stevia cultivars suggests that steviol glycoside biosynthesis in these cultivars may be regulated by mechanisms that are not readily influenced by MWCNTs at the tested concentrations.
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