Study On The Creat Induction Autoteraploid,Identification And Determination Of Effective Componnents Content Of Andrographis Paniculata (Burm. F.) Nees

ObjectiveAndrographis paniculata (Burm. f.) Nees. belongs to Acathaceous plants. It also known as andrographis paniculata, cutting snake grass, eel grass, olive lotus, with medicinal materials in Guangdong Province, is originated from dried aerial parts of Andrographis paniculata (Burm. f.) Nees., bitter to taste, cold in mature. It has significant effect in clearing away and relieving internal heat or fever, colding blood, redu-cing swelling, so it is used to the cold and fevers, sore throat, sore tongue, pertussis, overstrain cough, diarrhoea, dysentery, pain in heat strangury, welling-abscess swellings and sores, bites caused by insects or snakes. Wild resources of our country are ectremely rare, more than 99% of commercial drugs belongs to cultivated variety. However, because of the difference of geographi cal environment around, water and soil, natural climate, administration of manufactur ing and the artificial cultivated for a long time lack of scientific management syste m, the phenomena of variety degeneration, production drawdown, effective constituen t decreased the content becomes more and more serious. This makes the medicinal materials of Andrographis paniculata (Burm. f.) Nees. Uneven. Understanding the germplasm resources of Andrographis paniculata (Burm. f.) Nees., to carry out the cultivation of high yield, high content of medicinal active ingredients and good qual ity in new germplasm is extremely urgent, also is the demand of the market. Medic inal pl-ant polyploid plants, due to chromosome doubling in the cell, it often manife sts itself in nutritional organs such as roots, stems,leaves the giant, medicinal effecti ve component content increased,ecological adaptability and resistance to strengthen. As a result, to cultivate chromosome of Andrographis paniculata (Burm. f.) Nees., is one of the effective way to solve the variety degeneration, production drawdown, effective xonstituent decreased the content and so onMethodsThis experiment investigates 0.2% mercuric chloride sterilization time(1、3、5、7、 9 min), period of harvest(harvest for half a year, harvest for one year), maturational stage(green, full ripe), nutrient medium(pure AGAR,1/2 MS, MS, MS+2 mg/L6-B A), pretreament ways(0.5% potassium nitrating 2 h,0.1 mol/L salicylic acid soaking 2 h,30% hydrogen peroxide solution soak for 2 h,40% of sulfuric acid soaking 2 h, blank control), light condition(illumination, darkness)on the influence of seed ger mination rate using single factor method.It also inspects the impact on the growth rate of fresh weiht about growth regulator(6-BA、KT、NAA), flocculus banyan aeri-al root solution soncentration(0、20、40、60、80、100%).The experiment explores the induction effect of different experimental materials(stem tip, stem, root), soaking method and Agar group. Inspecting the influence of colchi cine’s concentration (0,200,400,600,800,1000 mg/L), co-Ichicine’s effecting time(7,14,21,28 d), DMSO’s concentration (0,10,20,40 g/L) and KT’s conc entration (0,0.1,0.2,0.4 mg/L) on polyploid induction rate using the orthogonal de-sign method. Comparing the difference of tetraploid and diploid plant morphology, cell, chromosome and effective components of andrographolied total lactone conten, to clear creat different chromosome ploidy type and medicinal materials production and creat the correlation of total lactone content.RusultsUse half a year’s paniculta fully mature seeds harvested, soaking with 40% sulfuric acid pretreatment of 2 h,0.2% mercuric chloride disinfection time for 5 min, vaccin ation in 1/2 MS basic culture medium, light condition.1.0 mg/L of KT is more adv antageous to creat proliferation.KT concentration of 1.0 mg/L, NAA concentration of 0.05 mg/L, creat organization fresh weight growth rate of 1000%, maximum.20% of the aerial root solution to creat most affect reproductive growth rate of fresh wei ght,reached 737%,100% of the aerial toot solution minimum impact on the growth rate of fresh weight of andrographolide breeding, only 261%. 2000 mg/L of colchicine induced 6 times of morphological variation rate,tetraploid variation strains, tetraploid double rate were significantly increased the induction rate was 8.7%. To creat stem section(with interaodes)as material handling,inducing rate can reach 12.5%.AGAR method is adopted to MS+10 mg/L KT+0.01 mg/L NAA as the medium, add 200 mg/L colchicine concentration,10 mg/LDMSO, dealing with 28 d, creat polyploid induction rate can reach 20%, higher than the above 16 induct ion rate of experiments, the induced effect is good. The strains of tetraploid plants salience shorter than kiploid,tetraploid of stem diameter, leaf length, leaf width, leaf area, fruit length, seed diameter significantly greater than the diploid, polyploid plant small, stem diameter, leaf thickness, leaf area, fruit length, seed charcateristics, and the color is diploid thick green leaf. The cell morphological identification, tetraploi d of leaf epidermal cells under the stomata length, width, density and chloroplast nu-mber were diploid were 130%,107%,130% and 107%. The chromosome identifica tion, the tetraploid number (100)is the diploid chromosome number(about 50)2 times, the planat can be determined to creat tetraploid plants.ConclusionsOf this study was carried out by creat sterile seeds germination and breeding resear ch,that made a successful andrographis propagation system, the polyploidy induction and identification of andrographis,successful polyploid plants, is a sollution to creat varieties degradation, the decrease of the quality of the production and a variety of problems, such as one of the effective ways to lay a foundation for further research on paniculta breeding,etc.