The Key Laboratory of Dai and Southern Medicine of Xishuangbanna Dai Autonomous prefecture, Yunnan Branch of Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences, Peking Union Medical College, Jinghong, China.
Guangxi Botanical Garden of Medicinal Plants, Nanning, China.
PLoS One. 2018 Jun 28;13(6):e0199259. doi: 10.1371/journal.pone.0199259. eCollection 2018.
Uncaria macrophylla Wall. is an important Chinese medicinal herb. Rhynchophylline (RIN) and isorhynchophylline (IRN) are its major active compounds. We investigated the influence of genetic differentiation and environmental factors on the RIN and IRN to find the main influencing factors of their contents and lay the foundation for the following cultivation and breeding. We used inter-simple sequence repeat (ISSR) markers to investigate the genetic diversity, and high-performance liquid chromatography (HPLC) to measure the contents of RIN and IRN in 200 samples of U. macrophylla obtained from nine natural populations, and then to analyze the correlation between genetic differentiation, environmental factors of sampling sites and the contents of RIN and IRN. We found that High intra-population (80.05%) and low inter-population (19.95%) genetic diversity existed in the samples of U. macrophylla. To some extent, genetic differentiation and the contents of RIN and IRN had correlation in individual populations (such as JH, MH, XM, and ML). The RIN and IRN contents were significant negatively correlated with the precipitation in May (RIRN = -0.771, p = 0.015) and June (RRIN = -0.814, p = 0.008; RIRN = -0.921, p = 0.000), indicating that precipitation was the main affecting factor of their contents. Interestingly, the analysis results showed that the RIN content had a significant positive correlation (r = 0.585, p = 0.000) with the IRN content (they are isomers); the proportion of RIN had a significant negative correlation with the sum of the two (r = -0.390, p<0.0001), while the proportion of IRN had a significant positive correlation (r = 0.390, p<0.0001). It meant that, with the total quantity of the two compounds increased, the proportion of RIN decreased and the proportion of IRN increased, illustrating that their conversion exist some regularity. Moreover, the content ratio of RIN and IRN was significant positively correlated with the January precipitation (r = 0.716, p = 0.030), implying that January may be the key period for the mutual transformation of RIN and IRN.
钩藤是一种重要的药用植物,其主要活性成分为钩藤碱(RIN)和异钩藤碱(IRN)。本研究旨在探讨遗传分化和环境因素对 RIN 和 IRN 的影响,以找到影响其含量的主要因素,为后续的栽培和选育奠定基础。我们采用简单重复序列间扩增(ISSR)标记分析了 9 个自然种群 200 份钩藤样本的遗传多样性,并用高效液相色谱法(HPLC)测定了 RIN 和 IRN 的含量,并分析了遗传分化、采样点环境因子与 RIN 和 IRN 含量的相关性。结果表明,钩藤种群内遗传多样性较高(80.05%),种群间遗传多样性较低(19.95%)。在个别种群(如 JH、MH、XM 和 ML)中,遗传分化与 RIN 和 IRN 含量存在一定相关性。RIN 和 IRN 含量与 5 月(RIRN = -0.771,p = 0.015)和 6 月(RRIN = -0.814,p = 0.008;RIRN = -0.921,p = 0.000)的降水呈显著负相关,表明降水是影响其含量的主要因素。有趣的是,分析结果还表明,RIN 含量与 IRN 含量(二者互为同分异构体)呈显著正相关(r = 0.585,p = 0.000);RIN 所占比例与二者总量呈显著负相关(r = -0.390,p<0.0001),IRN 所占比例与二者总量呈显著正相关(r = 0.390,p<0.0001)。这表明,随着两种化合物总量的增加,RIN 比例降低,IRN 比例升高,说明二者的转化存在一定规律。此外,RIN 和 IRN 的含量比与 1 月降水呈显著正相关(r = 0.716,p = 0.030),表明 1 月可能是 RIN 和 IRN 相互转化的关键时期。