Studies On The Rooting Of Branch Cuttings And The Accumulation Of Active Ingredients In The Roots, Stems And Leaves Of Morinda Officinalis How

For the development and utilization of the medicial plant of Morinda officinalis How, we had studied the rooting of branch cutting and the accumulation of rubiadin, monotropein and polysaccharide in the roots, stems and leaves of Morinda officinalis How for the following goals:(1)To optimize the cutting propagation technique and investigate the rooting mechanism of branch cuttings of Morinda officinalis How to provide a scientific basis for the rapid propagation of its seedlings in industrial production.(2)To analyze the distribution and accumulation of rubiadin, monotropein and polysaccharide in the roots, stems and leaves of Morinda officinalis How to provide a scientific basis for its GAP cultivation, harvest and the comprehensive utilization of the stems and leaves resources.1.Study on the cutting propagation: Taking the cutting rooting rate, root number, length of the longest root as examining index,the effects and the mechanism of IBA treatment on rooting of branch cuttings collected from 1~3- year-old mother trees or kept different nodes were studied. The results showed that there were best effects on rooting of cuttings with three nodes collected from 3-year-old mother tree when treated with 50 mg/L IBA. And the average rooting rate, the average root number and the average length of the longest root were respectively 93.3%, 23.18, 2.21 cm. During the rooting process, POD activity showed a tendency of rise to decline. PPO activity showed a trend of rise to decline and then to rise. The activity of IAAO showed a tendency of decline to rise The POD and PPO activity of IBA treatment group cuttings were higher than the control group, while IAAO activity was lower than the control group. Therefore, IBA treatment affected the rooting of cuttings by changing the activity of POD, PPO and IAAO.2.Study on the distribution and accumulation of rubiadin: Use the HPLC to determine the contents of rubiadin in the roots, stems, leaves of Morinda officinalis How collected from different areas at different growing years. HPLC condition was that Agilent TC-C18(4.6mm×250mm, 5μm) column was employed as analytic column; the mobile phase was acetonitrile-water(52:48); the velocity of flow was 1.0ml/min; the detection wavelength was at 278nm; the column temperature was 25℃; and the injection volume was 20μl. The results showed that the contents of rubiadin in the roots, stems, leaves were at the rang of 1.41~9.98μg/g, 0.69~3.87μg/g, 0.11~1.02μg/g, respectively. The contents of rubiadin in Morinda officinalis How were higher in the roots, and less in the stems and leaves. During the planting year of 2-4, the contents of rubiadin in the roots of Morinda officinalis How were fromed 2.16μg/g increased to 9.98μg/g, and it began to decline at the plangting years of 5-6.3.Study on the distribution and accumulation of monotropein: 1.TLC method was adopted for qualitative identification of monotropein, with silica gel G plates, chloroform:methanol:water:acetic acid(8:2:0.5:0.5)as developing system, and Trim-Hill solution as chromogenic reagent, at daylight viewing. 2.Using HPLC to determine the content of monotropein in the root, stem, leaf of Morinda officinalis How collected from different areas at different growing years. The HPLC condition was that Waters C8(4.6mm×150mm, 3.5um) column was employed as analytic column; the mobile phase was methanol-0.1% phosphate solution(5:95); the velocity of flow was 0.8ml/min; the detection wavelength wae at 235nm; the column temperature was 25℃; and the injection volume was 10μl.The TLC results showed that the root, stem and leaf sample had corresponding spots with monotropein reference in the TLC chromatogram. The results showed that the contents of monotropein in the roots, stems, leaves were at the rang of 8.54~16.64mg/g 、8.48~19.95mg/g、12.06~25.23mg/g. In Guangdong and Guangxi producing areas, the monotropein mainly distributed in leaves, while the content of monotropein was highest in the stems of Morinda officinalis How in the Fujan producing areas. During the planting years of 2-6, the contents of monotropein in the root of Morinda officinalis How were fromed 9.17mg/g increased to 16.47mg/g.4.Study on the distribution and accumulation of polysaccharide: Use sulfuric acid-phenol method to determine the contents of polysaccharide in the roots, stems, leaves of Morinda officinalis How collected from different areas at different growing years. Take the glucose as reference, the sample wae measured at the wavelength of 489 nm, and the contents of polysaccharide in the sample were calculated by the conversion factor. The results showed that the contents of polysaccharide in the roots, stems, leaves were at the rang of 7.68~24.97%、4.44~12.28%、6.11%~16.79%. The contents of polysaccharide were higher in the roots and leaves of Morinda officinalis How, and less in the stems. During the planting years of 2-5, the content of polysaccharide in the roots of Morinda officinalis How were fromed 7.68% increased to 24.97%, and it began to decline at the plangting year of six.Conclusion:(1) By studing the the effect of IBA treatment on rooting of branch cuttings collected from different years old mother trees or kept different nodes. There was best effects on rooting of cuttings with three nodes collected from 3-year-old mother tree, when treated with 50 mg/L IBA, the rooting rate of cutting was 93.3%.(2) All of the roots, stems and leaves contained rubiadin, monotropein and polysaccharide. The content of rubiadin and polysaccharide were highest in the roots, and the content of monotropein was highest in the leaves. During the planting years of 2-5, the content of rubiadin and polysaccharide in roots of Morinda officinalis How showed the trend of rise to decline, while the content of monotropein continued to accumulate.