BackgroundBlind and excessive application of fertilizers was found during the cultivation of Panax notoginseng in fields, as well as increase in root rot disease incidence.MethodsBoth “3414” application and orthogonal test designs were performed at Shilin county, Yunnan province, China, for NPK (nitrogen, phosphorus, and potassium) and mineral fertilizers, respectively. The data were used to construct the one-, two-, and three-factor quadratic regression models. The effect of fertilizer deficiency on root yield loss was also analyzed to confirm the result predicted by these models. A pot culture experiment was performed to observe the incidence rate of root rot disease and to obtain the best range in which the highest yield of root and saponins could be realized.ResultsThe best application strategy for NPK fertilizer was 0 kg/667 m2, 17.01 kg/667 m2, and 56.87 kg/667 m2, respectively, which can produce the highest root yield of 1,861.90 g (dried root of 100 plants). For mineral fertilizers, calcium and magnesium fertilizers had a significant and positive effect on root yield and the content of four active saponins, respectively. The severity of root rot disease increased with the increase in soil moisture. The best range of soil moisture varied from 0.56 FC (field capacity of water) to 0.59 FC, when the highest yield of root and saponins could be realized as well as the lower incidence rate of root disease.ConclusionThese results indicate that the amount of nitrogen fertilizer used in these fields is excessive and that of potassium fertilizer is deficient. Higher soil moisture is an important factor that increases the severity of the root rot disease.

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The SmERF6, which recognizes the GCC-box of SmCPS1 and SmKSL1 promoter in nucleus, regulates the tanshinone biosynthesis in Salvia miltiorrhiza hairy roots. Tanshinone, an important medicinal ingredient in Salvia miltiorrhiza, is best known for its use in medicine. However, the transcription factor regulation of tanshinone biosynthesis is unclear. Here, we isolated and identified a transcription factor in the ERF family of S. miltiorrhiza, SmERF6, which was screened from an S. miltiorrhiza cDNA library by the promoters of two key tanshinone synthesis genes (SmKSL1 and SmCPS1); this factor regulated tanshinone biosynthesis. The gene was highly expressed in the root and responded to ethylene treatment. SmERF6 modulated tanshinone biosynthesis by directly binding to an ethylene-responsive element (GCC-box) of the SmKSL1 and SmCPS1 promoters and activating their transcription. Overexpression of SmERF6 in the hairy roots increased their tanshinone accumulation, and SmERF6 silencing by RNAi led to a lower tanshinone content. Furthermore, tanshinone accumulation maintained homeostasis with the total phenolic acid and flavonoid contents in S. miltiorrhiza. These findings elucidated how SmERF6 directly co-regulates the transcription of SmCPS1 and SmKSL1 and modulates tanshinone synthesis to accelerate the metabolic flux of tanshinone accumulation in S. miltiorrhiza.

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