| Pueraria lobata (Willd) Ohwi. originating in China is a perennial leguminous plant and is also an important wild plant resource. The main components of biological activity are isoflavonoids such as puerarin, daidzin and daidzein. In recent years, Isoflavonoids from Pueraria radix (RP), the root of Pueraria lotaba (Willd.) Ohwi , are reported to have many important biological activities , and they are synthesized predominantly in legumes to show a lot of important physiological functions. Isoflavonoids also act as screens from harmful UV-light and scavengers of reactive oxygen species. By far, there are a few reports on the antioxidantion of related monomer compounds of isoflavonoids from Pueraria lobata, therefore there is a need to study isolation, purification and the antioxidantion of related component of isoflavonoids from Pueraria lobata.In recent years, P. lobata cell suspension cultures and callus cultures had been successfully induced and cultivated by plant culture techniques in vitro. A preliminary Optimization of culture conditions of P. lobata cell suspension cultures and callus cultures had been achieved through the regulation and controlling on the addition of carbon source, nitrogen source, the pH value and fungal elicitor, which provided an important technology mean and good materials for the production of isoflavonoids drugs ,then market supply and demand contradiction of isoflavonoids can be mitigated. Quality of light as an important plant growth regulator can phaseoloide the same important regulation and control significance. in vitro culture of plant cells and the suspension callus. Effective quality of light through appropriate regulation and control can maintain the vitality and stability of long-term high-yield synthesis of isoflavonoids. Therefore, the study on different quality of light regulation and the effects on cell growth and isoflavonoids biosynthesis, also have practical significance for further use of modern biotechnology methods to product isoflavonoidsIn addition, there are not only significant differences in the content and components of isoflavonoids from P. lobata cell suspension cultures and Pueraria Radix, but also related literature for research on isoflavonoids components of P. lobata cell suspension cultures are less. Therefore, related extraction, isolation and purification, identification of isoflavonoids and detection of antioxidation methods for isoflavonoids components from P. lobata in this study provided the very important reference value , which can be used for studying the isoflavonoids composition and antioxidant activity in vitro culture of P. lobata cell suspension cultures and callus cultures. In view of this, isoflavonoids components of Anhui Shuchen Pueraria Radix was researched by isolation and purification of it, and the antioxidant activity of isoflavonoids crude extract and isolated monomer compounds of isoflavonoids were studied; and the effcts of different quality of light regulation on cell growth and isoflavonoids biosynthesis were researched through different Light quality dealing with Pueraria lobata cell suspension cultures. In this paper, the main results were summarized as follows.There were 11 monomer compounds extracted , isolated and purified from Anhui Shuchen Pueraria Radix, which seven compounds with high yield were detected by HPLC and their structures were determined mainly by means of NMR, ESI-MS techniques.The current identified four compounds: Compound 1 was determined as daidzein, compound 7 was determined as daidzin, compounds 8 was determined as ononin, compounds 10 was determined as genistin.crude extracts from Pueraria Radix isoflavonoids and 5 isoflavonoid monomer compounds include puerarin, daidzein, daidzin, ononin and genistin for test samples, Vc and tea polyphenols for control samples, were studied their DPPH ? free radical scavenging ability, which were as followings:â‘ In the range of 0.01 ~ 0.5mg/mL concentration, it showed a positive correlation between the scavenging DPPH ? free radical rate of crude isoflavonoid extracts and puerarin and their concentrations. Crude isoflavonoid extracts possessed similar effect to Vc and tea polyphenols, but showed much stronger effect than the monomer puerarin.â‘¡In 2 ~ 10 mmol / L concentration range,the scavenging DPPH ? free radical ability of 5 isoflavonoid monomer compounds include puerarin, daidzein, daidzin, ononin and genistin and their concentrations were also into a positive correlation . And the efficiency of removal of DPPH ? free radical was shown as daidzein> daidzin> genistin > puerarin> ononin.crude extracts from Pueraria Radix isoflavonoids and 5 isoflavonoid monomer compounds include puerarin, daidzein, daidzin, ononin and genistin for test samples, Vc and tea polyphenols for control samples, were studied their superoxide radical inhibiting ability, which were as followings:â‘ In the range of 0.2 ~ 1mg/mL concentration, it showed a positive correlation between the inhibiting superoxide radical rate of crude isoflavonoid extracts and their concentrations. Crude isoflavonoid extracts possessed better effect than tea polyphenols ,but less than Vc ,and also showed more obvious effect than the monomer puerarin and daidzein. Puerarin and daidzein can be discavered their inhibiton of superoxide radical when their concentration was equal or greater than 1mg/mL;â‘¡ Experiments showed that the low concentration of isoflavonoid monomer compounds possessed much little inhibitory rate, 4 mmol / L of daidzein up to the inhibitory rate of 46.58 percent, which makes choosing a 4 mmol / L as the concentration of the samples in comparative experiment. And the efficiency of inhibiton of uperoxide radicall was shown as daidzein puerarin> daidzin> genistin > ononin. The elationships between their activity of antioxidant and radical scavenging and the molecular structures are also discussed.The red, yellow, blue, green and white colors of the five kinds of different color films and dark control were selected to process P. lobata cell suspension cultures, dealt with the same light intensity, it came to the conclusion as followings:â‘ yellow, white and red color films used to process P. lobata cell suspension cultures can promote the role of growth (higher than the dark control), with the promoting effect of yellow membrane> albuginea membrane > red membrane, but the yellow membrane with little difference between white membrane, white membrane and red membrane is not very different. But the Green film, blue film processing are not conducive to cell growth (below the dark control);â‘¡A variety of color film processing P. lobata cell suspension cultures can promote the role of extracellular and intracellular synthesis of isoflavonoids (higher than the dark control), and the blue membrane showed the best promoting role, the effect of the worst films are red, yellow and white film showed slightly higher role than the green , but little difference between the three. A variety of color films to deal with P. lobata cell suspension cultures can play the promoting role in synthesis of isoflavononids content in the total (higher than the dark control), with the promoting effect of yellow membrane> white membrane > blue membrane > green membrane > red membrane (the difference between albuginea membrane and yellow membrane was not obvious, and the red film showed little difference between the green membrane).
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