Department of Agronomy and Plant Breeding, Agricultural and Natural Resources College, University of Tehran, Karaj, Iran.
Department of Agronomy and Plant Breeding, Agricultural and Natural Resources College, University of Tehran, Karaj, Iran; Department of Agrobiotechnology, Institute of Agriculture, RUDN University, 117198, Moscow, Russia.
Plant Physiol Biochem. 2024 Mar;208:108440. doi: 10.1016/j.plaphy.2024.108440. Epub 2024 Feb 22.
Due to low amounts of withanolides produced in some plants and high demand for various applications, their biotechnological production is widely researched. The effects of two explant types (i.e., leaf and stem from the in vitro seedlings of three genotypes of Withania somnifera) and four Rhizobium strains (i.e., LBA 9402, A4, ATCC 15834, and C58C1) to improve hairy root formation efficiency was studied. Furthermore, the combined effects of β-cyclodextrin (β-CD) and methyl jasmonate (MeJA) on withaferin A production after 48 h exposure time was examined. Four hairy roots having the maximum percentage of induced roots and mean number of induced roots to analyze their growth kinetics and identified G3/ATCC/LEAF culture having the maximum specific growth rate (μ = 0.036 day) and growth index (GI = 9.18), and the shortest doubling time (T = 18.82 day) were selected. After 48 h exposure of G3/ATCC/LEAF culture to different elicitation conditions, maximum amounts of withaferin A were produced in samples co-treated with 0.5 mM β-CD + 100 μM MeJA (9.57 mg/g DW) and 5.0 mM β-CD + 100 μM MeJA (17.45 mg/g DW). These outcomes represented a 6.8-fold and 12.5-fold increase, respectively, compared to the control. Similarly, combined β-CD/MeJA elicitation increased gene expression levels of HMGR, SQS, SMT-1, and SDS/CYP710A involved in withanolides biosynthetic pathway, of which just SMT-1 had significant correlation with withaferin A production. These results demonstrated the superiority of G1-leaf explant and ATCC 15834 for hairy root induction, and revealed synergistic effect of MeJA and β-CD on withaferin A production.
由于某些植物中所含有的醉茄内酯数量较低，而各种应用的需求量又很高，因此人们广泛研究其生物技术生产。本研究分别以 3 种基因型（G1、G2 和 G3）的睡茄离体幼苗的叶片和茎作为外植体，同时选用 4 种根瘤菌菌株（LBA 9402、A4、ATCC 15834 和 C58C1），研究了它们对毛状根形成效率的影响。此外，还研究了β-环糊精（β-CD）和茉莉酸甲酯（MeJA）在 48 小时暴露时间后对诱导生成醉茄内酯 A 的协同作用。通过比较诱导生根的外植体和平均诱导生根数量，选择了 4 株具有最大诱导生根百分比和平均诱导生根数量的毛状根，用于分析其生长动力学。结果表明，G3/ATCC/LEAF 培养物的最大比生长速率（μ=0.036 天）和生长指数（GI=9.18）最高，倍增时间（T=18.82 天）最短。在将 G3/ATCC/LEAF 培养物暴露于不同的诱导条件 48 小时后，在同时用 0.5 mM β-CD+100 μM MeJA（9.57 mg/g DW）和 5.0 mM β-CD+100 μM MeJA（17.45 mg/g DW）处理的样品中产生了最大量的醉茄内酯 A。与对照相比，这两种处理方式分别提高了 6.8 倍和 12.5 倍。同样，β-CD/MeJA 联合诱导提高了涉及醉茄内酯生物合成途径的 HMGR、SQS、SMT-1 和 SDS/CYP710A 基因的表达水平，其中只有 SMT-1 与醉茄内酯 A 的产生具有显著相关性。这些结果表明，G1 叶片外植体和 ATCC 15834 更适合诱导毛状根，并且 MeJA 和 β-CD 对醉茄内酯 A 的产生具有协同作用。
