Yaowachai Wipa, Luecha Prathan, Taratima Worasitikulya
Department of Biology, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand.
Department of Pharmacognosy and Toxicology, Faculty of Pharmaceutical Science, Khon Kaen University, Khon Kaen 40002, Thailand.
Biol Methods Protoc. 2023 Sep 19;8(1):bpad019. doi: 10.1093/biomethods/bpad019. eCollection 2023.
(L.) Kurz is used in Thai traditional medicine for the treatment of skin diseases, ringworm, and eczema. This research studied the effects of cytokinin and auxins on callus induction and evaluated antioxidant activity of . Nodes, young, and mature leaf explants were cultured on MS medium containing 0, 1, 2, 3, and 4 mg/l kinetin (6-furfurylaminopurine) and 0, 1 mg/l 1-naphthaleneacetic acid (NAA), indole-3-acetic acid (IAA), and 2,4-dichlorophenoxyacetic acid (2,4-D) for 6 weeks to induce callus. Calli derived from nodes, young and mature leaves, and other plant parts were ultrasonically extracted with methanol to determine total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity by ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhtdrazyl (DPPH), and 2,2'-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) assays. Nodal explants on MS medium containing 1 mg/l kinetin combined with 1 mg/l 2,4-D were most efficient in callus production with the average fresh and dry weight per explant of 2.29 ± 0.14 and 0.18 ± 0.01 g, respectively. Addition of kinetin combined with NAA or 2,4-D had a positive effect on callus induction from young and mature leaf explants. The leaf extract showed the highest TPC, TFC, FRAP, and IC of DPPH and ABTS assays (ca 113 mg GAE/g extract, 45 mg QE/g extract, 121 mg TE/g extract, 53 µg/ml and 14 µg/ml, respectively), followed by callus derived from nodes. Overall, phenolic content was higher than flavonoid content. A strong positive correlation was found between FRAP assay, TPC (=0.973), and TFC (=0.798), indicating that phenolic and flavonoid compounds are responsible for antioxidant activity of .
库尔兹（L. Kurz）在泰国传统医学中用于治疗皮肤病、癣和湿疹。本研究考察了细胞分裂素和生长素对愈伤组织诱导的影响，并评估了……的抗氧化活性。将节段、幼嫩和成熟叶片外植体培养在含有0、1、2、3和4毫克/升激动素（6-糠氨基嘌呤）以及0、1毫克/升1-萘乙酸（NAA）、吲哚-3-乙酸（IAA）和2,4-二氯苯氧乙酸（2,4-D）的MS培养基上6周以诱导愈伤组织。将来自节段、幼嫩和成熟叶片以及其他植物部位的愈伤组织用甲醇进行超声提取，以通过铁还原抗氧化能力（FRAP）、2,2-二苯基-1-苦基肼（DPPH）和2,2'-联氮-双-（3-乙基苯并噻唑啉-6-磺酸）（ABTS）法测定总酚含量（TPC）、总黄酮含量（TFC）和抗氧化活性。在含有1毫克/升激动素并结合1毫克/升2,4-D的MS培养基上的节段外植体在愈伤组织产生方面效率最高，每个外植体平均鲜重和干重分别为2.29±0.14克和0.18±0.01克。添加激动素并结合NAA或2,4-D对幼嫩和成熟叶片外植体的愈伤组织诱导有积极影响。叶片提取物在DPPH和ABTS法测定中显示出最高的TPC、TFC、FRAP和IC（分别约为113毫克没食子酸当量/克提取物、45毫克槲皮素当量/克提取物、121毫克 Trolox当量/克提取物、53微克/毫升和14微克/毫升），其次是来自节段的愈伤组织。总体而言，酚含量高于黄酮含量。在FRAP测定、TPC（=0.973）和TFC（=0.798）之间发现了很强的正相关，表明酚类和黄酮类化合物是……抗氧化活性的原因。
