Effect Of Herbicide In Quality Of Spargnii Rhizoma

Research on plant physiological changes and defense reactions under adversity environment has become compared mature,but the molecular mechanisms of all physiological and biochemical changes have not been studied thoroughlly.Herbicides affect physiological and biochemical processes of aquatic organisms,then further affect its' morphological characteristics,primary and secondary metabolism accumulation and concentration of hazardous substances.Domestic and overseas study focus on the determination of harmful and effective substances,it is seldom to study on mutual influence of pesticides in the aquatic environment and herbs quality.To study the effecfs of herbicide metsulfuron and oxyfluorfen on quality and molecular mechanisms of aquatic herbs Sparganium stoloniferum,the methods of plant physiology,plant metabolomics and proteomics environment were used to explore how the herbicides water affect the physiological-biochemical characteristic,secondary metabolite and related indicators,it laid the foundation for fully revealing the quality formation of Sparganium stoloniferum.The main results were as follows:1.To investigate the effect of oxyfluorfen adversity stress on the physiological and biochemical index of Sparganium stoloniferum Buch.-Ham,different concentration of oxyfluorfen was used for studying some related index.The results showed that the variationtrend in SOD activity of sparganium stoloniferum plant applied oxyfluorfen decreased firstly and then increased,CAT and POD activities showed to rise firstly and then to decline.MDA content increased firstly and then gradually declined,which were not significantly different with control.Proline,soluble protein and soluble sugar as the main osmolytes had the same trend,increased firstly and then gradually returned to the control level.Oxyfluorfen decreased chlorophyll content significantly,while stimulated the root activity of Sparganium stoloniferum.It was concluded that herbicide oxyfluorfen had different effects on plant physiology.Plants mitigated the harmful effect of oxyfluorfen stress through antioxidant enzyme activities and osmolytes,which showed up some tolerant abilities of Sparganium stoloniferum in oxyfluorfen conditions.2.The response of Sparganium stoloniferum plants to oxyfluorfen stress was investigated through simulating natural state growth,a variety of physiological and biochemical parameters were measured,We determined the related content of phenols and the activities of PAL,C4H,4CL in Sparganium stoloniferum under different concentration of oxyfluorfen after different days cultivating.The results showed that after the initial dose,SOD activity,CAT and POD activity increased firstly and then decreased,oxyfluorfen aggravated membrane lipid peroxidation and inhibited the synthesis of chlorophyll,to keep the normal physiological function of cells,plant accumulated osmolytes(proline,soluble protein,soluble sugar).In the late growing period,MDA content were lower than the control,chlorophyll content increased to maintain the normal growth of plants.It clarified the physiological and biochemical characteristics of Sparganium stoloniferum under oxyfluorfen stress.Oxyfluorfen restrained PAL activity at first and then promoted,C4H activity gradually increased while 4CL activity increased first and then decreased.The content of rutin was the highest,then followed by vanilla acid and r-coumaric acid.PAL and C4H had negative correlation with phenolic compounds and total phenols while had positive correlation with the three flavonoids.There was positive correlation between 4CL and formononetin,ferulic acid and total phenols while had negative correlation between 4CL and other phenols.3.To study the stress of oxyfluorfen on Sparganii Rhizoma,we investigated physiological and biochemical characteristics,including the antioxidant enzyme systems and osmolytes,the results showed that Sparganii Rhizoma grew normally under oxyfluorfen stress,SOD,CAT and POD activities were lower than the control.Oxyfluorfen could significantly reduce the content of proline,while increased soluble sugar and soluble protein content to maintain the stability of plant cells.Oxyfluorfen made the activities of three secondary metabolism enzymes(PAL,C4H and 4CL)stronger than control,but it did not find a significant correlation between enzymes activity and the seven phenolics.Oxyfluorfen inhibited the generation and accumulation of?-sitosterol,total amino acids,polysaccharides and total saponins,but promoted the content of nucleoside and nucleotide ingredients,with the concentration of oxyfluorfen increasing,the promoting effection became weaker.Low and high concentrations of oxyfluorfen promoted the generation and accumulation of mannitol while middle concentrations of oxyfluorfen inhibited mannitol content.4.Sparganii Rhizoma protein under different concentrations of oxyfluorfen was measured By LC-MS/MS technology,483 kinds of proteins,801 peptides and 41 kinds of different proteins was founded,which mainly related to metabolism,protein synthesis,photosynthesis,sucrose synthesis,transfer and maturation.In addition to RNA polymerase ? subunit,the remaining protein expression quantity was down-regulated,the different protein expression of Sparganii Rhizoma under oxyfluorfen stress provided an important basis for revealing the molecular mechanisms between oxyfluorfen and Sparganii Rhizoma.Meanwhile the determination method of oxyfluorfen was established to analyze its residues in Sparganii Rhizoma and soil,the results showed that oxyfluorfen was degraded.It provided the basis for further study on the mutually effects between oxyfluorfen and Sparganii Rhizoma.5.When it was too late to scrape the skin in harvesting of Sparganii Rhizoma,we sprayed metsulfuron-methyl,so we determined the metsulfuron-methyl residual quantity in the tubers and skin of Sparganii Rhizoma by HPLC,the contents of seven phenolic compounds including rhydroxybenzaldehde,vanillic acid,rcoumaric acid,ferulic acid,rutin,kaempferol and formononetin and total polyphenol were determined by the validated procedure of HPLC.Metsulfuron-methyl had a different influence on the diverse Phenolic component in Sparganii Rhizoma.The Metsulfuron-methyl degraded in different degree on the skin and tubers of Sparganii Rhizoma.The result showed the residual quantity of metsulfuron-methyl in Sparganii Rhizoma exceeded the EU maximum residue limits in other plant and animal food.It was Proposed that metsulfuron-methyl should be abolished to the harvesting and processing for Sparganii Rhizoma.6.Based on multi-index evaluation technology,Sparganii Rhizoma was processed using five drying methods including drying in the shade,drying in the sun,drying in the oven,microwave drying and infrared drying,the contents of the functional ingredients such as seven phenolic compounds including r-hydroxybenzaldehde,vanillic acid,r-coumaric acid,ferulic acid,rutin,kaempferol and formononetin,mannitol,polysaccharide,total saponins and total polyphenol were determined by the validated procedure of HPLC and ultraviolet spectrophotometry.The results showed that different drying techniques have significant influence on the quality of Sparganii Rhizoma,the contents of seven phenolic compounds,polysaccharide and total polyphenol were the highest using microwave drying,when Rhizoma Sparganii was processed by drying in the shade the contents of the mannitol was the hightest over other four drying techniques,while the contents of total saponins was the highest by drying in the oven.The different drying techniques had significant influence on the quality of Sparganii Rhizoma.