Biswas Pritom, Kumari Ankita, Modi Arpan, Priyam Amiya, Haque Rizwanul, Ola Mohammad Shamsul, Kumar Sanjeev, Kumar Nitish
Department of Biotechnology, Central University of South Bihar, Gaya, Bihar, India.
Department of Agricultural Biotechnology, Anand Agricultural University, Anand, Gujarat, India.
Front Plant Sci. 2025 Aug 21;16:1566037. doi: 10.3389/fpls.2025.1566037. eCollection 2025.
The plant (Asteraceae) is gaining popularity as a zero-calorie natural sugar substitute. This paper investigates the regeneration of from callus, emphasizing steviol glycoside (SGs) production and the evaluation of genetic similarity. The highest rate of callus induction (89.20%) and maximum biomass were obtained from leaf explants using Murashige and Skoog (MS) medium, optimized with the addition of Naphthalene acetic acid (NAA) and 2,4-Dichlorophenoxyacetic acid (2,4-D). MS medium containing NAA and 6-Benzylaminopurine (BAP) was most effective for shoot regeneration, yielding the highest shoot induction rate (87.77%) and robust plant growth. Rooting efficiency was significantly enhanced by using a quarter-strength MS medium with Indole-3-acetic acid (IAA), which produced the highest rooting percentage (88.40%) and longest roots (3.41 cm). The acclimatized plantlets demonstrated a survival rate of 77-78% and closely resembled the parent plants in morphology. It was indicated by HPLC analysis that SGs concentrations were significantly higher in the leaves of regenerated plants compared to callus, while leaves showed the highest content of both the SGs. The consistent amplification profiles observed in the genetic analysis, conducted using not only Random Amplified Polymorphic DNA (RAPD) but also Inter Simple Sequence Repeats (ISSR) markers, revealed no polymorphic bands, suggesting minimal somaclonal variation. This study highlights the effectiveness of callus culture for enhancing steviol glycoside production and maintaining genetic stability in .
这种植物（菊科）作为一种零热量的天然糖替代品越来越受欢迎。本文研究了甜叶菊愈伤组织的再生，重点关注甜菊糖苷（SGs）的产生以及遗传相似性评估。使用添加了萘乙酸（NAA）和2,4-二氯苯氧乙酸（2,4-D）优化的Murashige和Skoog（MS）培养基，从叶片外植体获得了最高的愈伤组织诱导率（89.20%）和最大生物量。含有NAA和6-苄基腺嘌呤（BAP）的MS培养基对芽再生最有效，产生了最高的芽诱导率（87.77%）和健壮的植株生长。使用含有吲哚-3-乙酸（IAA）的四分之一强度MS培养基显著提高了生根效率，该培养基产生了最高的生根率（88.40%）和最长的根（3.41厘米）。驯化后的植株成活率为77 - 78%，形态上与亲本植株非常相似。高效液相色谱分析表明，再生植株叶片中的SGs浓度明显高于愈伤组织，而甜叶菊叶片中SGs的含量最高。不仅使用随机扩增多态性DNA（RAPD）而且使用简单序列重复区间（ISSR）标记进行的遗传分析中观察到的一致扩增图谱显示没有多态性条带，表明体细胞无性系变异最小。本研究强调了愈伤组织培养在提高甜菊糖苷产量和保持甜叶菊遗传稳定性方面的有效性。
