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Canavanine is a natural non-protein amino acids, and is very similar to the molecular structure of arginine, because it could be instead of arginine, arginine tRNA synthetase mistakenly believe that arginine synthesis to protein, leaving the The physiological function of canavanine-containing proteins to be defective, and thus has a good anti-tumor properties. Canavanine the Papilionoideae (Papilionnoidea) and legumes (Leguminosae) seeds in soluble nitrogen storage library, which is also considered a protected plant pests and diseases infested plants are natural times The metabolites, which showed some toxic effects of certain pests because canavanine, be considered a potential insecticide because of its toxicity and its performance advantages of low toxicity, non-polluting nature, judge from its mechanism of action, because it does not damage the parasitic natural enemies of the pest, so it has to protect the pest natural enemies of insect characteristics. The main source of canavanine synthetic and extracted from plants two ways. Plant extracts can be obtained by tissue culture techniques of secondary metabolites and generally low cost, high charged, less environmental pollution, has not been reported on the Con callus, this study is the use of concanavalin seed callus induction, cell fresh weight, dry weight, the cells relative growth rate and the canavanine content as index Multilevel optimization experiments, analysis of the physiological and biochemical indexes in liquid suspension culture process, which canavanine content was measured by the paper is to develop a new simple, rapid determination of canavanine The results show that: (1) The 2,4 - Dinitrofluorobenzene concanavalin ammonia acid in pH 9.5 borate buffer solution, the optimum reaction conditions for the establishment of a new, quick and easy method of measuring the canavanine, i.e. the use of a spectrophotometric determination of the absorbance of the complex at 571 nm at. The results showed that: 50 ° C water bath for 30 min, canavanine and 2,4 - dinitrofluorobenzene occurred color reaction, forming a green 1:1 stable complex. Its maximum absorption peak at 571nm, the apparent molar absorption coefficient e = 1.086 × 104 L / (mol · cm), obeys Beer's law in 13.75 to 220.00μg/mL within the scope of, the corresponding linear regression equation: y = 0.0051 0.0682; related The coefficient: R2 = 0.9939. Recoveries ranged from 96.60 to 101.56%, relative standard deviation of 1.768%, thereby establishing a canavanine is simple, rapid determination of the new method. (2) In concanavalin seed material induced by concanavalin A callus, concanavalin callus system, and further culturing and proliferation, measured canavanine content and type of different colors callus growth period callus canavanine content. The results showed that: light green sword bean callus can be induced in MS medium and concanavalin callus growth showed the characteristics of an \The study found that green callus canavanine the highest content 43.462μg/gFW (fresh weight) of cells, followed by light green, followed by white is a callus 21.626μg/gFW browning Minimum content . Canavanine content increased and then decreased trend in the callus tissue growth. Period with the highest content of culture 24d, canavanine content 30.675μg/gFW. (3) the use of orthogonal design, fresh weight, dry weight, the cells relative growth rate and canavanine content as an indicator, Multilevel Con callus to inspect. The results showed that: sucrose is the most significant impact on callus growth. Also significant canavanine content. If the callus growth target: the best medium for of MS of 3.0 mg/L6-BA 0.8 mg / L NAA 40g / L sucrose, 7.8 g / L agar, the best medium conditions callus relative growth rate of 0.1016 dry weight 1.1318g bottle -1; canavanine content callus goal: the best medium for MS 3.0 BA 0.2 mg / L NAA 20g / L sucrose, 7.8 g / L agar best medium conditions canavanine content 63.2432μg / L FW and 617.5320μg / L DW. (4) selection of loose texture, uniform growth state, good dispersion concanavalin callus suspension culture as inoculation material, a comparative analysis of a variety of physiological and biochemical indexes during suspension culture and canavanine content changes. Experimental results show that: Con callus suspension culture was \Intracellular and extracellular soluble protein content and cell viability was first increased and then decreased, while the culture solution pH changes and extracellular peroxidase content was first decreased and then increased; canavanine content - a slight decrease in and then rise again, and finally a downward trend, indicating that the culture broth of pH value and extracellular peroxidase activity correlated with canavanine content.
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