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Study on Technology of Good Agricultural Practice of Gynostemma Pentaphyllum (Thunb.) Makino

Author: WangPeiXiang
Tutor: XiaoZuoPing
School: Shaanxi Normal University
Course: Botany
Keywords: Gynostemma pentaphyllum (Thunb.) Makino tissue culture Good Agricultural Practice (GAP) biotechnology
CLC: S567.239
Type: Master's thesis
Year: 2005
Downloads: 234
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Abstract

The plant Gynostemma pentaphyllum (Thunb.) Makino is a kind of perennial herb liane, which belongs to Cucurbitaceae. There is a long history in China that people have planted it for eating and treatment. Many traditional books such as "Textual Research on Reality and Titles of Plants", "Compendium of Materia Medica" etc. record that Gynostemma pentaphyllum has great medicinal function as behaving antioxidation, diminishing inflammation, detoxification, relieving a cough and dispelling phlegm, curing tracheitis and contagious hepatitis, promoting immunity, reducing fat in the blood etc. It has a wide market prospect.For a long time, the source of Gynostemma pentaphyllum mainly depends on the wild resources and several farmers’ planting here and there. With the demand increasing, the resource reduces gradually. In addition, the government has released a policy for protecting the wild resources. So, pursuing some study on good agricultural practice of Gynostema pentaphyllum is extremely urgent. In 2001, Pingli, one of the main planting regions of Gynostemma pentaphyllum, was named as Chinese traditional medicine demonstration area by Department of Science and Technology of Shaanxi Province. GAP of Gynostemma pentaphyllum, an important project of Chinese traditional medicine modernization, has been examined successfully by the experts of the subject in May, 2004 and by Chinese Administration Bureau of the National Food and Medicines in November, 2004.This paper mainly studies the resources of Gynostemma pentaphyllum in Pingli, Shaanxi province, through investigation and studies on its biological feature and tissue culture. We compared the main active components of it based on different habitats, different varieties, different picking time and different cultivating measure by high performance liquid chromatography (HPLC). The paper makes clearly the proper picking time, which provides theory foundation for GAP of Gynostemma pentaphyllum. The main results could be concluded as following:1) Through the investigation of the source of Gynostemma pentaphyllum in Pingli, five good varieties are chosen on the basis of the biological features of branches, leaves and roots combined with its saponin A content and its yield.2) As for leaves of different varieties, the paper describes the number of stomatal on leaves and the morphological features of trichomata by using scanning electron microscopy (SEM).3) The proper culture condition of callus, adventitious buds, proliferation of buds and rooting is screened after the tissue culture and regeneration technique are studied. We also established the regeneration system of Gynostemma pentaphyllum.When the basic medium is MS supplemented with 2, 4-D l.Omg/L, 6-BA 0.5mg/L, the callus induction rate of the stem and leaf could reach 100%. The medium of MS+KT 2.0 mg/L + 6-BA 2.0mg/L + NAA 0.01 mg/L was used to propagate callus. The callus was 3.9 times increased after three subcultured on this medium.The ratio of induction of shoots, roots and callus is studied by single factor and orthogonal test. The result shows that low concentrated NAA and high concentrated 6-BA or KT have better effect on the formation rate of adventitious bud than 6-BA or KT respectively.Low concentration of inorganic salt in medium is the major factor to produce root. Root will be greatly induced if adding suitable content of NAA or IAA. The effect of IAA is superior to that of NAA. The effect of using leaf as explant to induce root is better than that of using stem as explant. When we use stem as explant to induce root, we find that stem with some leaves are better than pure stem.4) The paper studies the effect of ultrasound on Gynostemma pentaphyllum grown in vitro. The buds can be proliferated 10 times if we use ultrasonic treatment with 8-10 min.5) The paper also studies the effect of fertilization, sun light and water on Gynostemma pentaphyllum. We discover that fertilization and enough water can improve the growth of Gynostemma pentaphyllum. When the plant grows under 65% of full light, saponin A content can reach 7.2%. It is most suitable for Gynostemma pentaphyllum to grow under the soil humidity of 30%, while proper drought threatening is benefitable for the accumulation of saponin A.6) A good method to determine the content of saponin A by HPLC is established. The method should be carried out under the condition of Methanol-water-phosphoric acid (68: 32: 0.2) and the detecting wavelength at 205 ran. The method can be repeated and is a good detecting method for saponin A of Gynostemma pentaphyllum.7) The content of saponin A of Gynostemma pentaphyllum is picked at the different time during a year. We determine that the optimum picking time is from late September to early November by considering the content of saponin A and the yield of individual.

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