Effects of different carbon sources on growth and active component contents in Salvia miltiorrhiza and S. castanea f. tomentosa hairy roots / 中国中药杂志

Salvia miltiorrhiza(Sm) and Salvia castanea f. tomentosa(Sc) hairy roots were used as experimental materials to study the effects of six different carbon sources, galactose, fructose, lactose, glucose, arabinose and sucrose(control), on fresh weight, dry weight, contents and yields of salvianolic acids and tanshinones. The results showed that galactose was most beneficial to the growth of two kinds of hairy roots, while lactose and arabinose were not conducive to their growth. As for Sm hairy roots, fructose significantly promoted the accumulation of salvianolic acid B, and the content increased by 5.801 times and 10.151 times compared with the control group, respectively. Glucose significantly promoted the accumulation of salvianolic acids. The content and yield of rosmarinic acid were 7.674 times and 9.260 times of that of the control group, and the content and yield of salvianolic acid B were 5.532 times and 6.675 times of the control group. For the hairy roots of Sc, galactose significantly increased the content and yield of rosmarinic acid, reaching 7.820 times and 9.944 times of the control group, respectively. Fructose promoted the increase of the content and yield of cryptotanshinone, reaching 9.242 times and 6.609 times of the control group, respectively. The study confirmed the optimal carbon source for the hairy root culture of Sm and Sc, and provided theoretical guidance for large-scale production of Sm drug-derived components and the utilization of Sc.

Mass Production of Pinellia ternate by Tissue Culture and the Selection for High-yield Individuals / 中国生物工程杂志

Using the tissue culture technology to achieve the fast propagation of P.ternata and through the photosynthetic characters and yield comparison,two high-yield indivaduals were obtained. The result showed that the coefficent of propagation can get to 1∶6.23 per generation, through 3 month culture the average diameter of the in vitro tuber can get 0.88 cm. P.ternata was a typical shade-plant, of which LCP was only 700?mol?photons/s?m2. Photo-inhibition was observed among all the three leaf-types of P.ternata, and the willow leaf type showed the most obvious photo-inhibition. Both of the light response Pn curve and single tuber weight were significant different among leaf styles. Result for studying the chlorophyll fluorescence parameters showed that all the difference of NPQ, qP and qN among three leaf-types of P.ternata were significant and the difference of potential-photosynthetic productivity among them was significant. The willow leaf style had the strongest light-tolerance capacity.The individuals of T2(peach-leaf type) had the highest Pn and single tuber weight among 11 superior individuals and the individuals of L2(willow-leaf type) showed the strongest light-tolerance capacity. Through 6 month propagation ten thousands in virto tuber of T2 and L2 each were gotten. And after growing in Wenzhou experimental base for one year, the total gross weight of the high-yield individual tuber get to 100kg each.

Expression of the gene coding for a thermostable alpha-amylase from Pyrococcus furious in Chiamydomonas reinhardtii chloroplast / 生物工程学报

Thermostable alpha-amylase from Pyrococcus furious is an important industrial enzyme in brewing and alcohol production. Eexpression of the thermostable a-amylase in plants can reduce greatly costs in the production of alcohol using crop plants. A chloroplast expression vector, p64A, containing the thermostable alpha-amylase gene from Pyrococcus furious, was constructed with clpP-trnL-petB-chlL-rp123-rpl2 as Chlamydomonas reinhardtii plastid homologous recombinant fragments and spetinomycin-resistant aadA gene as select marker. The plasmid p64A was transferred into the chloroplast genome of C. reinhardtii by the biolistic method. Nine independently transformed lines were obtained by 100 mg/L spectinomycin selection. PCR amplification, Southern blot analysis of the transgene and cultivation in the dark all showed that the a-amylase gene had been integrated into chloroplast genome of C. reinhardtii. The activity of amylase expressed in the chloroplast of C. reinhardtii was detected by amylase activity assay and found to be as high as 77.5 u/g fresh weight of cells. These experimental results demonstrated the possibility of using transgenic chloroplasts of plant as bioreactors for production of industrial enzymes.