Establishment Of Callus Clones Derived From Anther Of Panax Quinquefolium L. And Selection Of High Ginsenoside-yielding Callus Clones

Panax quinquefolium L. is also known by its Chinese name Guangdong ginseng and Huaqishen.It’s a herbaceous perennial plant which belongs to genus Panax in the Araliaceae family. Panaxquinquefolium L., Panax ginseng L. and Panax notoginseng L. belong to the same genus but differentspecies. They are all precious herbal plants.Anther of Panax quinquefolium L.was used as explant in this paper. Selection of callus inductionmedium and subculture medium established callus clones; Selection of differentiation medium was usedto test the callus redifferentiation; Studies on growth condition and content of ginsenosides developed astable high ginsenoside-yielding callus clone. The main results are as follows:1. The effects of factors were investigated in anther induction from anther culture of Panaxquinquefolium L., such as medium type, hormone constitutions and sucrose concentrations. The resultshowed that2,4-D by itself promoted callus formation. Callus induction experiments demonstrated thehighest induction rate, up to37.53%, could be obtained by the combinations of2,4-D2mg/L and6-BA0.5mg/L.60g/L sucrose treatment had the highest induction rate. With the increasing concentration,sucrose did not contribute to promote the induction of callus derived from anther of Panaxquinquefolium L. MS medium was better than PⅡ medium in anther culture. The optimum mediumhad been screened out through callus induction experiments, which was MS medium supplemented with2,4-D2.0mg/L,6-BA0.5mg/L, sucrose60g/L, agar5.5g/L and LH0.5g/L.2. The effects of different combinations of2,4-D and6-BA were tested to promote callussubculture. It indicated that the concentration range of6-BA from0.5mg/L to2mg/L promoted growthof callus with2mg/L fixed concentration of2,4-D.This study preliminarily established callus clonesand conserved115callus resources.3. The effects of different hormone proportions on callus differentiation were explored. The workhad screened out11differentiation media. The findings showed that callus cultured on F6and F9medium induced eight green primordiums. However, the effect factors of callus differentiation have notdeeply researched yet.4. Twenty callus resources were used to investigate the situation of callus growth. Increasing rateof fresh callus is taken as index to check the situation of callus growth. The results indicated that allcallus resources had kept stable growth during continuous cultivation for four times. Increasing rate ofsix callus resources had been over200%, increasing rate of five callus resources had been between150~200%, increasing rate of five callus resources had been between100~150%and increasing rate offive callus resources had been below100%during continuous cultivation for four times.5. Content of ginsenosides in callus was determined by ultra performance liquid chromatograph(UPLC). The results showed that the content of total ginsenosides in callus8#、9#、14#、25#、47#wasmore than1%, the content of total ginsenosides in callus4#、5#、48#、69#was between0.5~1%, thecontent of other fourteen callus resources was below0.5%.25#had the highest total ginsenosides, morethan3%.This content is more close to the total ginsenosides in root of Panax quinquefolium L. at nature. Rg1、Re、Rb1were the chief component of the total ginsenosides in20callus resources.