Summary of the invention
The objective of the invention is for overcoming the deficiency of prior art; Provide a kind of and the artificial cultivation method raising irexis sonchifolia active constituent content that the active components in medicinal plant accumulation law is consistent, simple to operate, cost is low, output is high, article are of fine quality, good in economic efficiency.
A large amount of researchs show: healthy and strong high quality seeds is selected in (1), and through the atmospheric dielectric barrier discharge plasma treatment, afterwards with 0.05% gibberellin seed soaking, the survival rate that can significantly improve seedling is with robustness and shorten into the seedling time; (2) dregs of a decoction that produce in the Ixeris Sonchifolia Hance injection production process are processed fertilizer through fermenting and becoming thoroughly decomposed and are done base fertilizer, have improved soil K and P supply, and then improve the irexis sonchifolia content of effective.(3) waste liquid in the Ixeris Sonchifolia Hance injection production process (sowthistle-leaf ixeris seedling lime aqueous solution) not only contains the calcium ion of rich, and contains free OH
-, plant pathogenic microorganisms and plant insect there are prevention and killing action, can replace chemical pesticide, to avoid pollution to the irexis sonchifolia medicinal material; And be rich in amino acid, carbohydrate and various trace elements in the waste liquid, also be the ideal material that promotes plant growing.
The artificial cultivation method that the present invention improves the irexis sonchifolia active constituent content comprises: the pre-treatment of seed, whole ground, seed sowing, fertilising, the extermination of disease and insect pest, daily management and step such as gather; And seed pre-treatment, fertilizing method and the extermination of disease and insect pest are the key factors that influences the irexis sonchifolia active constituent content, and concrete steps are following:
1, seed pre-treatment
(1) seed collection: the irexis sonchifolia seed is tiny, and thousand kernel weight is 0.090-0.110g, when fruit still stretches out from fruit for green seed pappus, plucks usually; Be placed on the calm place airing that faces south after the withdrawal, behind cracking of fruit, rub out seed; Remove impurity, dry, cleanliness is not less than 95%;
(2) seed treatment: choose the healthy and strong high quality seed of irexis sonchifolia prior to seeding, handle through atmospheric dielectric barrier discharge plasma device (atmospheric dielectric barrier discharge plasma device, ceiling voltage are 30kV), electric field is set to 8kV; During processing, earlier the seed single flat in the broadcaster is laid on the conveyer belt of uniform motion, opens high voltage source then and begin discharge, handling grain weight is 1000/s, with 0.05% gibberellin seed soaking 24 hours, dries, and germination rate can reach more than 90%.
2, fertilizing method: the kind, consumption and the time that comprise fertilising:
(1) kind of fertilising: the sowthistle-leaf ixeris seedling dregs of a decoction become thoroughly decomposed and fertile mix through ploughing deeply with native soil surface 25cm table soil, the sowthistle-leaf ixeris seedling dregs of a decoction fertile consumption 2000kg/ mu that becomes thoroughly decomposed;
(2) fertilization time: spray with the 0.1% long-acting ammonium-potassium phosphate composite fertilizer aqueous solution, every square metre of 50ml-120ml, the fertilization time concrete steps in the planting process are following:
1. long during when the seedling basal leaf to 3-5 blade, carry out first time and apply fertilizer, with the aqueous solution sprinkling of 0.1% long-acting ammonium-potassium phosphate composite fertilizer, every square metre of 50ml-60ml;
2. carry out second time fertilising after 20 days, with the aqueous solution sprinkling of 0.1% long-acting ammonium-potassium phosphate composite fertilizer, every square metre of 70ml-80ml;
3. apply fertilizer for the third time after 45 days, the time is to take out the Roripa phase every square metre of 100ml-120ml plant;
3, the extermination of disease and insect pest comprises:
(1) when seedling length arrives 3-5 blade, carries out the extermination of disease and insect pest first time, irrigate or sprinkling with 1% sowthistle-leaf ixeris seedling lime aqueous solution;
After (2) 30 days, carry out the extermination of disease and insect pest second time, irrigate or sprinkling with 2% sowthistle-leaf ixeris seedling lime aqueous solution;
(3) extermination of disease and insect pest time is at bud in earlier stage for the third time, irrigates or sprinkling with 3% sowthistle-leaf ixeris seedling lime aqueous solution;
Above-mentioned percentage is weight percentage.
4, the sowthistle-leaf ixeris seedling dregs of a decoction fertilizer that becomes thoroughly decomposed is meant the dregs of a decoction in the Ixeris Sonchifolia Hance injection production process, through the natural microbial fermentation with become thoroughly decomposed and forming.
5, sowthistle-leaf ixeris seedling lime aqueous solution is meant the reacted aqueous solution of adding calcium oxide breast in the Ixeris Sonchifolia Hance injection production process.
6, active constituent content is relatively: carry out assay to the irexis sonchifolia of growing down by tame irexis sonchifolia of this experimental technique and carded sliver spare out of office, it is following to measure the result:
(1) general flavone: measure result such as table 1
Table 1 determination of total flavonoids result
(2) total phenolic acid: measure result such as table 2
The total phenolic content of table 2 is measured the result
(3) cyanidenon-7-0-β-D-glucuronide, cyanidenon-7-gentiobiose glycosides, Cichoric acid, the tartaric assay of caffeoyl
Chromatographic condition is filler with the octadecylsilane chemically bonded silica; Flowing phase: acetonitrile (A)-1.0% glacial acetic acid (B), gradient elution (0~10min:7%A~10%A; 10~20min:10%A~20%A; 20~30min:20%A~30%A; 30~40min:30%A~35%A; Be volume ratio); Flow velocity: 1.0mlmin
-1Detect wavelength: 327nm; Column temperature: room temperature (25 ℃); Sample size: 10 μ L, theoretical cam curve is calculated by cyanidenon 7-O-β-D glucuronide peak should be not less than 5000.
It is an amount of that cyanidenon-7-O-β-D-glucuronide, cyanidenon-7-gentiobiose glycosides, Cichoric acid, caffeoyl tartaric acid reference substance are got in the preparation of reference substance solution; Adding methyl alcohol respectively processes every 1ml and contains cyanidenon-7-O-β-D-glucuronide 0.1mg; Contain cyanidenon-7-gentiobiose glycosides 0.02mg; Contain Cichoric acid 0.05mg, contain caffeoyl tartaric acid 0.1mg.
Irexis sonchifolia medicinal material 20g is got in the preparation of need testing solution, pulverizes, and crosses 40 mesh sieves, mixing, and precision takes by weighing 2g, adds 50% methyl alcohol 100ml, and reflux 2h is condensed into 10ml afterwards, promptly gets;
Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject hplc determination respectively, promptly get.
Measure result such as following table 3,4,5,6
Table 3 cyanidenon-7-O-β-D-glucuronide assay result
Table 4 cyanidenon-7-gentiobiose glycosides assay result
Table 5 Cichoric acid assay result
Table 6 caffeoyl tartaric acid assay result
Can find out that by above determination data the inventive method has improved the content of Ixeris Sonchifolia Hance injection with traditional Chinese medicine irexis sonchifolia effective medicinal components through seed pre-treatment, fertilizing method are improved and the extermination of disease and insect pest; General flavone has improved 30%-40%, and total phenolic acid has improved 25%-35%, has accomplished real good quality and high output; But also the sowthistle-leaf ixeris seedling dregs of a decoction and sowthistle-leaf ixeris seedling lime aqueous solution have been fully utilized; Reduce waste disposal cost and cultivation cost, promoted economic benefit, realized the comprehensive utilization of sowthistle-leaf ixeris seedling; Make and medicinal material plantation and preparation production benign cycle realize the scientific and industrialization benign development of Ixeris Sonchifolia Hance injection.