| Gastrodia elata Bl.is a perennial completely heterotrophic herb of Orchidaceae,and its dried tuber is a traditional and precious medicinal material.Gastrodia elata Bl.has no roots and leaves,Armillaria mellea(Vahl.ex fr.)quel provides nutrition for its growth,and some studies have found that there are microorganisms in the rhizosphere soil of Gastrodia elata Bl.that can promote the growth of Armillaria mellea and Gastrodia elata Bl..Moreover,the output of Gastrodia elata Bl.will also be affected by the micro-ecological effects of Armillaria mellea and soil chemicals.To solve these problems,this paper studied the microbial diversity in rhizosphere soil of sexual reproduction Gastrodia elata Bl.and asexual reproduction Gastrodia elata Bl.,the correlation between soil physical and chemical properties and yield and quality,and the chemical composition of soil-Armillaria mellea.The main results are as follows:1.The rhizosphere soil of Gastrodia elata Bl.is rich in microbial diversity,and the growth of Gastrodia elata Bl.does not affect the microbial community structure and diversity of hemp hole soil,but the environment and soil physical and chemical properties have a great influence on it.The rhizosphere soil of Gastrodia elata Bl.,which has been sexual reproduction for 18 months,was analyzed by 16 s RNA and ITS amplicon sequencing.The results showed that 37 phylum,20 classes,29 orders,27 families,and 22 genera of bacterial communities,and 15 phylum,11 classes,21 orders,26 families,25 genera,and 16 species of fungal communities.The 16 s RNA and ITS amplicon sequencing analysis of the treatment group(wood materials+Armillaria mellea+Gastrodia elata Bl.seeds)and the control group(wood materials+Armillaria mellea)showed that there was no significant difference in the number of species with abundance higher than 0.5%,and Mycena was not found in the genus with species abundance higher than 0.5%,but Armillaria was found,and Gastrodia elata and Mycena affected the soil microorganisms in hemp pits.T-test and LEf Se analysis showed that 26,13,and 9 different microorganisms in the rhizosphere soil of gastrodia elata from difference habitats,altitudes,and growth periods,respectively.The correlation analysis between these difference microorganisms and Gastrodia elata Bl.yield showed that 14 of them were positively correlated with Gastrodia elata Bl.yield and 9 were negatively correlated with Gastrodia elata Bl.yield,indicating that these florae positively correlated with Gastrodia elata Bl.yield may be the key microorganisms to promote Gastrodia elata Bl.growth and improve its yield,on the contrary,they may be important to inhibit Gastrodia elata Bl.growth and reduce its yield.In addition,the physical and chemical indexes of the rhizosphere soil of Gastrodia elata Bl.,soil microorganisms,and the yield of Gastrodia elata Bl.are highly correlated,and 19 different microorganisms are affected by soil p H,organic matter,available phosphorus,available potassium,and alkali-hydrolyzable nitrogen.It is speculated that the microbial diversity of the rhizosphere soil of Gastrodia elata Bl.can be improved by controlling soil physical and chemical properties during the planting process of Gastrodia elata Bl.,to promote the growth and high yield of Gastrodia elata Bl..2.39 kinds of microbial gather were identified from the rhizosphere soil around three high-yield Gastrodia elata Bl.,while 30 kinds of microbial gather in low-yield Gastrodia elata Bl.,and the dominant bacteria were all different.Besides,through T-test and LEf Se analysis,it was found that there were Nitrosotalea,Russula-mariae,Archeaeorhizomyces,Ilyonectria,Coprinellus＿disserminatus,and Fusarium in the rhizosphere soil of high-yield and low-yield Gastrodia elata Bl..The correlation analysis between the microorganisms gathered in the rhizosphere soil and the yield of Gastrodia elata Bl.showed no significant correlation between the bacterial community and the yield of Gastrodia elata Bl..Only in the fungal community,were five genera to be positively correlated with the yield of Gastrodia elata Bl.,and two genera were negatively correlated with the yield of Gastrodia elata Bl..Among them,Archaeorhizomyces also to be positively correlated with the yield in the rhizosphere soil of sexually propagated Gastrodia elata Bl.,that is,this genus of microorganisms can promote the high yield of Gastrodia elata Bl.,which should be further studied.3.Controlling the content of chemical components such as apocynin in the soil can increase the microbial diversity in the rhizosphere soil of Gastrodia elata Bl.,thus promoting the high yield of Gastrodia elata Bl..A total of 31 chemical components were detected from the rhizosphere soil of Gastrodia elata Bl.by GC-MS,among which apocynin was only detected in the soil of the treatment group(wood materials+Armillaria mellea+Gastrodia elata Bl.seeds),but not in the hemp pit of the blank control group(wood materials+ Armillaria mellea).Crotonic acid,Butanoic acid,3-hydroxy-,and apocynin are common chemical components in different habitats,different altitudes,and different periods.Among them,Bai Yang Village in Huichuan District contains compounds such as acetic acid,benzoic acid,acetic acid butyl ester,acetic acid hydroxy-methyl ester,propanoic acid 2-hydroxy-methyl ester(.+/-.)-,butyrolactone,propanoic acid 2-oxo-methyl ester,hexanoic acid methyl ester,and benzene acetic acid,methyl ester,which are significantly different from the other two producing areas.At the same time,it was found that the content of butyrolactone decreased with the elevation,while the content of crotonic acid increased with the elevation,and the content of benzoic acid and vanillin decreased gradually with the growth time of Gastrodia elata Bl..It was speculated that Gastrodia elata Bl.might secrete chemical components such as apocynin from the soil to inhibit allelochemicals such as benzoic acid and vanillin.The correlation analysis between the chemical composition of the rhizosphere soil of Gastrodia elata Bl.and the microorganisms with an abundance greater than 0.5% showed that the microorganisms significantly related to the yield of Gastrodia elata Bl.(except IMCC26256)were all affected by the chemical composition of the soil.4.A total of 59 compounds were isolated and identified from Armillaria mellea fermentation broth and its mycelium(liquid culture)and Armillaria mellea production strain(Armillaria mellea co-cultured with branches),among which 3 new compounds were isolated from the production strain.The results of activity screening showed that two steroidal components inhibited the inflammatory reaction of BV2 cells induced by LPS,12 triterpenoids showed cytotoxicity to HEL cancer cells,1 triterpenoid showed cytotoxicity to H1975 cancer cells,and 1 triterpenoid showed cytotoxicity to HEL,H1975,A549,H1299,and MDA-MB231 cancer cells.Linoleic acid,ethyl linoleate,p-hydroxy benzyl methyl ether,uracil,proline,5α,8α-epidioxyergosta-6,22-dien-3β-ol and other components isolated this time were also isolated in the study of chemical constituents of Gastrodia elata Bl.,which may be the material basis of the symbiotic ecological relationship between Gastrodia elata Bl.and Armillaria mellea.In summary,the growth of Gastrodia elata Bl.has no significant effect on the microbial diversity in the hemp point soil,but the different microorganisms in the rhizosphere soil of Gastrodia elata Bl.have a great influence on the yield of Gastrodia elata Bl.,and the differences in soil physical and chemical properties and chemical composition are important factors for the formation of different microorganisms in the rhizosphere soil.Therefore,Gastrodia elata Bl.-rhizosphere microorganisms-soil physical and chemical properties-chemical components constitute a micro-ecosystem that affects Gastrodia elata Bl.yield.The research results of this paper reveal the interaction of Gastrodia elata Bl.-soil-microorganism from different levels,which lays a theoretical foundation for constructing the ecological planting model of Gastrodia elata Bl.. |
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