Wang Dong, Wei Jiansheng, Yuan Xiaolong, Chen Zhonghua, Wang Lei, Geng Yunfen, Zhang Jinfeng, Wang Yi
College of Forestry, Southwest Forestry University, Kunming, 650224, China.
Laboratory of Forest Plant Cultivation and Utilization, The Key Laboratory of Rare and Endangered Forest Plants of State Forestry Administration, Yunnan Academy of Forestry and Grassland, Kunming, 650201, China.
Heliyon. 2024 Feb 28;10(5):e27223. doi: 10.1016/j.heliyon.2024.e27223. eCollection 2024 Mar 15.
Paclitaxel is a potent anti-cancer drug that is mainly produced through semi-synthesis, which still requires plant materials as precursors. The content of paclitaxel and 10-deacetyl baccatin III (10-DAB) in has been found to differ from that of other species, but there is little research on the mechanism underlying the variation in paclitaxel content in of different provenances. In this experiment, the contents of taxoids and precursors in twigs between a high paclitaxel-yielding individual (TG) and a low paclitaxel-yielding individual (TD) of were compared, and comparative analyses of transcriptomes as well as chloroplast genomes were performed. High-performance liquid chromatography (HPLC) detection showed that 10-DAB and baccatin III contents in TG were 18 and 47 times those in TD, respectively. Transcriptomic analysis results indicated that genes encoding key enzymes in the paclitaxel biosynthesis pathway, such as taxane 10-β-hydroxylase (T10βH), 10-deacetylbaccatin III 10-acetyltransferase (DBAT), and debenzoyl paclitaxel -benzoyl transferase (DBTNBT), exhibited higher expression levels in TG. Additionally, qRT-PCR showed that the relative expression level of T10βH and DBAT in TG were 29 and 13 times those in TD, respectively. In addition, six putative transcription factors were identified that may be involved in paclitaxel biosynthesis from transcriptome data. Comparative analysis of plastid genomes showed that the TD chloroplast contained a duplicate of , leading to a longer plastid genome length in TD relative to TG. Fifteen mutation hotspot regions were identified between the two plastid genomes that can serve as candidate DNA barcodes for identifying high-paclitaxel-yield individuals. This experiment provides insight into the difference in paclitaxel accumulation among different provenances of individuals.
紫杉醇是一种强效抗癌药物,主要通过半合成生产,而半合成仍需要植物材料作为前体。已发现红豆杉中紫杉醇和10-去乙酰巴卡亭III(10-DAB)的含量与其他红豆杉物种不同,但关于不同种源红豆杉中紫杉醇含量变化的潜在机制研究较少。在本实验中,比较了高产紫杉醇个体(TG)和低产紫杉醇个体(TD)的红豆杉嫩枝中紫杉烷类化合物和前体的含量,并进行了转录组和叶绿体基因组的比较分析。高效液相色谱(HPLC)检测表明,TG中10-DAB和巴卡亭III的含量分别是TD中的18倍和47倍。转录组分析结果表明,紫杉醇生物合成途径中编码关键酶的基因,如紫杉烷10-β-羟化酶(T10βH)、10-去乙酰巴卡亭III 10-乙酰转移酶(DBAT)和去苯甲酰紫杉醇-苯甲酰转移酶(DBTNBT),在TG中表现出更高的表达水平。此外,qRT-PCR表明,TG中T10βH和DBAT的相对表达水平分别是TD中的29倍和13倍。此外,从转录组数据中鉴定出六个可能参与紫杉醇生物合成的假定转录因子。叶绿体基因组的比较分析表明,TD叶绿体包含一个重复序列,导致TD的叶绿体基因组长度相对于TG更长。在两个叶绿体基因组之间鉴定出15个突变热点区域,可作为鉴定高产紫杉醇个体的候选DNA条形码。本实验为不同种源红豆杉个体间紫杉醇积累差异提供了见解。