4.Physiological characters of Catharanthus roseus mutant cells
Xiusheng ZHANG ; Lili NIE ; Beibei XIANG ; Shufang WANG ; Yerong ZHU ; Yong WANG
Chinese Traditional and Herbal Drugs 1994;0(11):-
Objective To investigate the characteristics of growth, accumulation of indole alkaloid, and the optical concentration of the nutrition components in the Catharanthus roseus mutant cells induced by the colchicine, looking forward to getting the ideal material producing alkaloids which was suited for industrialized cell cultivation. Methods Mutant cells cultivated on the MS medium were harvested according to experiment designs,and the fresh weight was got.The indole alkaloid as extracted by organic solvent. The ajmalicine and catharanthine extracted from the samples were analyzed by RP-HPLC. Results The growth rate was faster and the indole alkaloid accumulation was more when the mutant cells were cultivated generations 30 and the both were sharply decreased at generations 45, while the contents of medical compounds amounted to the peak at the generations 20. The content of medical components and alkaloid accumulation were evidently prompted by the tryptophan added in the media, especially at the concentration of Ca2+ 1 760 and Zn2+ 12.6 mg/L, respectively. Conclusion C.roseus mutant cells probably is an ideal material for industrialized cell cultivation.
5.Methods of ultra-dry and pre-humidification in Scutellaria baicalensis seeds
Jue WANG ; Khumanghlyva ULARGUL ; Jianhua WANG
Chinese Traditional and Herbal Drugs 1994;0(11):-
Objective To optimize the suitable method of ultra-try and pre-humidity for Scutellaria baicalensis seeds. Methods Taking S. baicalensis seeds as experimental materials, the seeds were dried by silica gel in a certain proportion on the constant temperature 30 ℃ in the desicator to obtain the seeds of S. baicalensis with various ultra dried moisture The proper moisture of ultra dry for S. baicalensis seeds was evaluated by measuring the germination percentage, germination energy, etc.The optimized pre-humidity was studied by six kinds of pre-humidifications. Results The optimum ultra-dry moisture of S. baicalensis seeds was 3.6%. The viability of S. baicalensis seeds decreased significantly while the ultra-dry moisture was 2.6%. The viability of S. baicalensis seeds damaged by ultra-dry was improved by PEC 30% pre-humidification. Conclusion The S. baicalensis seeds can be kept in a good quality by reducing the moisture with the ultra-dry method.
6.Inducing and culturing technology of callus in Cynanchum komarovii
Chinese Traditional and Herbal Drugs 1994;0(11):-
Objective To provide the initiative experimental reference for resource utilization of Cynanchum komarovii by biological technology. Methods Using root, stem, and leaf of C. komarovii as explants, the proper media for inducing of callus were optimized by orthogonal test. In culturing process for proliferation of callus, the fresh and dry weights were determined at various times and callus growth curve was drawn. Meanwhile, division test of callus was taken. Results Root callus test showed the best effect in successful inducing percentage of 90%-100%. The best media combination of stem and leaf were MS+2,4-D 1.0 mg/L+6-BA 2.0 mg/L+NAA 0.5 mg/L and MS+2,4-D 0.5 mg/L+6-BA 0.5 mg/L+NAA 0.5 mg/L. The stem segament was more easily induced than leaf. The callus induced by various treatments is in loose character and fresh yellow colour with no brown phenomenon. At day 10 the growth entered the exponent period of callus offspring, the maximal fresh weight to 4.961 g and the dry weight to 0.496 g. The division rate of callus was rather low and not easy to shoot. However, the nail root was commonly seen in the test. Conclusion The root is proved to be the best explants in the test of inducing callus, which shows the best medium of stem and leaf for forming the callus. The growth of callus is bigger, but the division is not easy.
7.Tissue culture system of Cynanchum otophylum in genetic transformation
Lu QIU ; Bo WANG ; Shuguo FAN ; Chunmei LUO ; Yanhua LI ; Kai CHEN ; Yinyan ZHOU ; Qiguo YANG ; Fusuo YANG
Chinese Traditional and Herbal Drugs 1994;0(11):-
Objective To establish the tissue culture system of Cynanchum otophylum in genetic transformation. Methods Asepsis seedling was set up by using different explant parts, disinfectors, and disinfecting time. The callus was induced, breeded, and differentiated by using different media, different hormone categories and combinations, and different hormone concentrations of 2,4-D and KT. Results It was the best method that asepsis seeding was built up by using seed as the explant and using 10% NaClO to treat the seed for 20—30 min or using 0.2% HgCl2 to treat the stem for 3 min. The seed, root, stem, and leaf of C. otophylum can form callus easily. The 2,4-D was important in forming callus. C. otophylum was more sensitivity to KT. The effect was better when KT was used to induce callus or adventitious buds. MS+2,4-D 1.0 mg/L+KT 1.0 mg/L was the better medium to induce callus to form. The callus cant be induced to form adventitous buds without reference to using seed or root, stem, and leaf as the explant, without reference to using hormone 6-BA or KT, ZT, 2ip. Gentamicin 100 ?g/mL is the optimum pressure in genetic transformation of C. otophylum. Conclusion The seed or stem is the ideal material bombarded by particle gun that is induced to form callus for 14 d in the media MS+2, 4-D 1.0 mg/L+KT 1.0 mg/L and without forming adventitious buds.
8.AFLP Analysis on diversity of germplasm resource in cultivated and wild Cistanche deserticola
Rong XU ; Jun CHEN ; Shilin CHEN ; Tongning LIU ; Ren NA
Chinese Traditional and Herbal Drugs 1994;0(11):-
Objective To determine the genetic diversity of germplasm resource in cultivated and wild Cistanche deserticola. Methods Fifty-eight samples from three populations of cultivated and wild C. deserticola were analyzed by amplified fragment length polymorphism (AFLP) DNA markers, and the gene- tic diversity was evaluated by PopGen32. Results The average percentage of polymorphic loci (PPL) of cultivated C. deserticola is 79.16%. The PPL of wild population is 89.53%. Average Neis gene diversity index (He) from four populations was 0.193 8, Shannons genetic diversity index (I) was 0.300 4, and genetic differentiation index (Gst) was 0.097 9. Conclusion The diversities of cultivated and wild C. deserticola are both higher and theres no differentiation between them. It shows that genetic diversity of inner-species is higher, which is not the reason for endangerment. Therefore, wild nursery and artificial cultivating are the best measures for the conservation and sustainable utilization in C.deserticola.
10.Effect of Qinglongyi polysaccharide on sialic acid level, ATPase activity, and membrane potential in erythrocytes of S_(180) mice
Chenfeng JI ; Feng XIAO ; Yubin JI
Chinese Traditional and Herbal Drugs 1994;0(11):-
Objective To study the effects of Qinglongyi polysaccharide on sialic acid (SA) content, Na+, K+-ATP and Ca2+, Mg2+-ATPase activity, and membrane potential in erythrocytes of S180 mice. Methods Using Kit to determine the SA content and Na+, K+-ATPase, Ca2+, Mg2+-ATPase activity, using flow cytometry to determine the membrane potential of erythrocytes of S180 mice. Results In the treated groups with three doses, it showed Qinglongyi polysaccharide could make the SA content increased (P