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Breeding Studies on Stable Production of Essential Oil Compounds in Atractylodes lancea De Candolle

津坂, 宜宏 筑波大学 DOI:10.15068/00160465

2020.07.22

概要

Atractylodes lancea is a medicinal plant distributed across East Asia. Its rhizomes are used in traditional Japanese medicines (Kampo) and Chinese medicines to treat a lot of diseases and disorders because they contain many pharmacologically active compounds. A. lancea is one of the most important botanical raw materials for Kampo medicine due to its overwhelming use. In recent years, the demand for mass production of A. lancea with consistent quality has increased as the use of Kampo has increased.

The major active compounds in A. lancea are essential oil compounds, namely, β-eudesmol, hinesol, atractylon, and atractylodin, and the contents of these compounds are important quality criteria in A. lancea production. Moreover, important quality criteria for pharmacognosy are not only limited to the contents of essential oil compounds but also include the content ratio between β- eudesmol and hinesol. The quality of A. lancea rhizomes is closely related to their natural habitat. Furthermore, content and content ratio of essential oil compounds in A. lancea reportedly vary based on geographical factors. However, it is not clear whether the geographical variations are based on genetic or environmental factors. Therefore, this study aims to (1) evaluate the influence of genetic and environmental factors on the content of essential oil compounds in A. lancea and (2) evaluate heritability of β-eudesmol/hinesol content ratio in A. lancea.

A. lancea is a perennial plant, and it is propagated by division of rhizomes. A clonal line was propagated from a rhizome of a single plant, and 25 clonal lines were used in this study. The clonal lines were cultivated in different years (2016, 2017) and different locations (Hokkaido, Ibaraki), and heritability, G×E interactions, and influence of environmental factors on the content and content ratio of essential oil compounds in A. lancea were evaluated. To estimate the broad-sense heritability, calculations were performed using a model for vegetatively propagated crops like herbage.

In this population, broad-sense heritability of β-eudesmol, hinesol, atractylon, and atractylodin contents were 0.84, 0.77, 0.86, and 0.87, respectively. The effects of interannual variability on the contents of the compounds were lower than those of genotype. In addition, the cultivated environmental factors were assessed by different locations, and the correlations between Hokkaido and Ibaraki grown plants based on β-eudesmol, hinesol, atractylon, and atractylodin contents were 0.94, 0.94, 1.00, and 0.83, respectively. Furthermore, the broad-sense heritability of the β- eudesmol/hinesol content ratio was revealed to be high at 0.92. The effects of cultivation year and location were smaller than that of genotype, and few G × E interactions were observed. The results suggest that the content and content ratio of essential oil compounds in A. lancea are largely influenced by genetic factors, indicating that selective breeding and clonal propagation could be an effective strategy for obtaining populations with high content of essential oil compounds. Furthermore, the contents of β-eudesmol, hinesol, atractylon, and atractylodin in A. lancea exhibited few correlations with rhizome yields. A. lancea cultivars with not only high content of essential oil compounds but also high rhizome yield could be developed through selective breeding.

Breeding elite cultivars is only useful if they can be distributed for cultivation, and current propagation techniques are limited in how efficiently they can increase A. lancea seedlings. To increase A. lancea seedlings efficiently, in vitro propagation has been explored, but previous studies have only investigated in vitro propagation on solid media, not in liquid media. In general, liquid culture is an effective method for mass propagation of plant seedlings. In this study, the effect of liquid culture for A. lancea shoot propagation was investigated with the goal of developing mass production methods for A. lancea seedlings.

The shoot propagation rate of A. lancea was higher in liquid culture than on solid media, and the rate of shoot propagation was 3.9 shoots per initial shoot in liquid media with an optimal BAP concentration after 8 weeks. A caveat of this method is that it is necessary to select a suitable strain for liquid culture. Under optimal culture condition, it is possible to produce thousands of plantlets from a single shoot in 1 year depending on the selected line. In addition, the rooting of shoots from in vitro culture on a rooting medium was consistent. However, liquid culture tended to induce the vitrification of leaves, warranting further investigation and optimization to reduced leaf vitrification. In this study, I demonstrated that the content and content ratio of essential oil compounds in A. lancea were largely influenced by genetic factors, indicating that selective breeding and clonal propagation are effective methods for reducing the variability of A. lancea quality. Furthermore, I showed that in vitro shoot propagation by liquid culture is an effective method for mass clonal propagation of A. lancea. I believe that the knowledge acquired in the present study will provide basic information for the development of elite cultivars and seedling production, and contribute to a stable production and supply of high quality A. lancea.

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