| Salviae Miltiorrhizae Radix et Rhizoma is the dried root and rhizome of Salvia miltiorrhiza Bge. Salvia is a model plant for medicinal plant research, has a wide range of adaptability to soil, can form of geoherbs in different soil environment. It shows that the interaction mechanism of Salvia and soil environment in the interpretation of formation of the quality of the medicinal plant has important significance, and root exudates is the medium of communication between plant and soil environment, it has an important role in regulating plant nutrient absorption, preventing and controlling soil borne diseases, easing environmental poison has an important value. On the basis of previous research that found the quality of Salvia was affected by both genetic and environment, the tissue culture seedlings of Salvia from 3 different geographical provenances served as materials, collectting the root exudates under hydroponics culture, pot experiment and field cultivation, UV spectrophotometer, amino acid analyzer, GC-MS and HPLC technique were applied to analyze the changes in the composition of root exudates under different cultivation patterns of Salvia, including polysaccharide, amino acid, protein, volatile compounds and organic acids, comparing the similarities and differences of root exudates under different provenances of Salvia and soil interaction cultivation. To explore the law of the interaction between root exudates and cultivation environment, in order to explore the ecological and physiological significance of root exudates, Lay the foundation for establishing the regulation and control technology of the rhizosphere nutrition The main results are as follows:?1? The rapid propagation systemof tissue culture seedlings of 3 different geographical provenance of Salvia was established, and its genetic stability was proved by field experiments. The leaves of shoot tips as explants, MS and 1/2MS as basic medium, the effect of different hormones on callus induction, adventitious buds differentiation and adventitious roots induction were studied. The best medium of callus induction was MS+2.0 mg·L-1 16-BA+1.0 mg·L"1 NAA; best buds differentiation medium was MS+1.0 mg·L-1 16-BA+0.1 mg·L-1 NAA; best adventitious roots induction medium 1/2MS+0.2 mg·L-1 NAA+0.5 mg·L-1 IBA. Tissue culture seedling of Salvia from Linqu Shandong, Nanyang, Henanand Zhongjiang, Sichuan grow robustly after potted and field transplanting, maintaining the origin characteristics of Salvia.?2? The root exudates of Salvia contained polysaccharides, amino acids, proteins or peptides, volatile components and organic acids. The results have shown that the rhizosphere polysaccharide mainly come from soil microorganism; free amino acids in root exudates including aspartic acid, threonine, serine, glycine, cysteine, valine, leucine, methionine, phenylalanine, histidine, lysine, leucine, isoleucine, tyrosine etc; the kinds and contents of acid hydrolyzed amino acids were increased than the free amino acids, adding glutamic acid, alanine, pro line etc, Which was the same as the amino acids composition of acid hydrolyzed protein in rhizosphere soil. The cysteine, phenylalanine, histidine, methionine were common amino acids under different cultivation patterns from normal secretion of Salvia, nothing to do with the environment, and the remaining amino acids were different due to different provenances and cultural environment of Salvia. The volatile components in root exudation of Salvia included hydrocarbons, alcohols, esters, phenols, aldehydes, acids, amines and hydrocarbons, alcohols, esters are the most abundant, high content. Organic acids included oxalic acid, tartaric acid, formic acid, malic acid, malonic acid, lactic acid, acetic acid, citric acid, fumaric acid.?3? Roots secrete behavior of Salvia is mainly affected by soil environment. Salvia of the same provenances were cultivated in different soils, Pentosans and hexosans in rhizosphere soil of Salvia showed increased successively from Henan soil, Sichuan soil, Shandong soil potting and field cultivation, consistent with the variation of blank soil; which showed the polysaccharide of rhizosphere soil was primarily affected by soil microorganisms in the environment. The total amount of free amino acids in rhizosphere soil varied in the same trend with rhizosphere polysaccharide, except for leucine, phenylalanine, lysine, various amino acid content decreased successively from Sichuan soil, Shandong soil, Henan soil potting, the remaining glutamic acid, threonine, serine, glycine, isoleucine, tyrosine were different due to different potting soil, indicating that the the 6 amino acids were mainly affected by soil environment. The types and contents of amino acids from acid hydrolyzed protein in rhizosphere soil were significantly higher than free amino acids, but only the content changed of each amino acid, the percentage was more stable, overall consistent with the variation of blank soil, which suggested that soil environment mainly affected content of rhizosphere protein, rather than the composition and structure. The results of organic acids in rhizosphere soil showed, except for oxalic acid, tartaric acid, the organic content in rhizosphere soil of Salvia from Henan and Sichuan decreased successively from Shandong soil, Henan soil, Sichuan soil potting, but in Salvia of Shandong changed contrastly, and the content of formic acid, malic acid, malonic acid, lactic acid in rhizosphere soil of three provenances Salvia changed greatly due to different potting soil. The hydroponic experiments showed that, under water cultivate, the kinds and content of acid hydrolyzed amino acid and organic acids were higher than 1 /2MS cultivate, and except for histidine and cysteine, methionine, phenylalanine of Shandong Salvia, the percentage of the remaining amino acids decreased, which indicated that the kinds and content of amino acids, peptides and proteins from root exudates of Salvia were related to the nutritional status of environment.?4? Roots secrete behavior of Salvia also related to the genetic characteristics of Salvia. Salvia of different provenances were cultivated in the same soil, hexosans, pentosan and total polysaccharides were overall highest in the rhizosphere soil of Salvia from Henan, followed by Sichuan. Similarly, under hydroponic conditions, the content of free amino acid and the percentage of acid hydrolyzed amino acid were different in water culture solution of different provenances Salvia. The difference was greater of acid hydrolyzed amino acid in 1/2MS culture solution of different provenances Salvia, the rhizosphere of Shandong Salvia only detected 7 kinds of acid hydrolyzed amino acids, and Salvia from Henan and Sichuan detected 14 kinds and the content were similar, both were higher than Shandong Salvia, it showed the genetic characteristics of Henan and Sichuan Salvia were more similar. However, soil culture environment, glutamic acid, threonine, serine, glycine, isoleucine, tyrosine in free amino acids of rhizosphere soil were different among different provenances of Salvia. The percentages of aspartic acid, glutamic acid, leucine, isoleucine, glycine, histidine, lysine, arginine in amino acids from acid hydrolyzed protein have a bigger difference among different provenances of Salvia. It suggested that these amino acids and its related proteins showed provenance differences. The composition and percentage of volatile components in root exudates is not the same among different provenances of Salvia. And the content of oxalic acid in water culture solution of different provenances Salvia were significantly different, the water culture solution of Shandong Salvia contained unique formic acid, Henan and Sichuan Salvia contained unique apple acid; and in 1/2MS culture solution, the kinds of amino acids were similar among different provenances of Salvia, the content of amino acids decreased successively from Henan Salvia, Sichuan Salvia, Shandong Salvia, but malonic acid changed contrastly. In the soil culture, the secretion of malonic acid, formic acid and malic acid was greatly influenced by the provenance of Salvia. Obviously, the secretion of amino acids, peptides and proteins, volatile components and organic acids also showed genetic characteristics, nutritional stress promoted the secretion of amino acids, proteins and organic acids.In summary, the behavior of root secretion of Salvia is dual regulation ofboth genetic and environmental, nutritional stress promoted the secretion of amino acids, proteins and organic acids. It showed root exudates of Salvia ware related to soil nutritional regulation, and the regulatory mechanisms were different among different genotypes Salvia. Which Further indicated that the change of root exudates caused by the environment maybe physiological and ecological behavior mechanism in the rhizosphere of environment modification in traditional Chinese medicine quality. This study laid a foundation for exploring the adaptive mechanisms ofroot exudates of Salvia on the different soil and the regulation of rhizo sphere behavior on the quality of traditional Chinese medicine. |
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