Abstract
Anoectochilus roxburghii is a traditional Chinese herb with high medicinal value, with main bioactive constituents which are flavonoids. It commonly associates with mycorrhizal fungi for its growth and development. Moreover, mycorrhizal fungi can induce changes in the internal metabolism of host plants. However, its role in the flavonoid accumulation in A. roxburghii at different growth stages is not well studied. In this study, combined metabolome and transcriptome analyses were performed to investigate the metabolic and transcriptional profiling in mycorrhizal A. roxburghii (M) and non-mycorrhizal A. roxburghii (NM) growth for six months. An association analysis revealed that flavonoid biosynthetic pathway presented significant differences between the M and NM. Additionally, the structural genes related to flavonoid synthesis and different flavonoid metabolites in both groups over a period of six months were validated using quantitative real-time polymerase chain reaction (qRT-PCR) and high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS). The results showed that Ceratobasidium sp. AR2 could increase the accumulation of five flavonol-glycosides (i.e., narcissin, rutin, isorhamnetin-3-O-beta-d-glucoside, quercetin-7-O-glucoside, and kaempferol-3-O-glucoside), two flavonols (i.e., quercetin and isorhamnetin), and two flavones (i.e., nobiletin and tangeretin) to some degrees. The qRT-PCR showed that the flavonoid biosynthetic genes (PAL, 4CL, CHS, GT, and RT) were significantly differentially expressed between the M and NM. Overall, our findings indicate that AR2 induces flavonoid metabolism in A. roxburghii during different growth stages, especially in the third month. This shows great potential of Ceratobasidium sp. AR2 for the quality improvement of A. roxburghii.
Highlights
Anoectochilus roxburghii (Wall.) Lindl., called “Jin Xianlian” and “Jin Xianlan” is a perennial herb of the genus Anoectochilus of the Orchidaceae family
AR2 would soon grow over the whole substrate for about two weeks. Both morphological observation and toluidine blue staining, which showed no hyphae in the control group (Figure 1A) and that the intracellular pelotons existed in the treatment (Figure 1B), indicated that a symbiotic relationship had been established between A. roxburghii and mycorrhizal fungus Ceratobasidium sp
For arbuscular mycorrhizal (AM) fungi, they can penetrate and colonize the root of the host to form intracellular haustoria-like structures known as arbuscules, which are the principal sites of metabolic exchange between the two organisms [39,40]
Summary
Anoectochilus roxburghii (Wall.) Lindl., called “Jin Xianlian” and “Jin Xianlan” is a perennial herb of the genus Anoectochilus of the Orchidaceae family. It is widespread in southern China and considered a famous drug in the provinces of Jiangxi, Taiwan, Guangdong, Guizhou, Zhejiang, and Fujian which is its main markets [1]. Due to the fact of its high medical values, A. roxburghii is widely used in medicine and health care and in beauty and drinking products with an increasing market demand. The vigorous market demand results in excessive harvesting and exploitation of the plant, leading to resource depletion. Tissue culture has become the main source of commercial A. roxburghii
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