Effect of low temperature on cell synchronization division in suspension culture cells of Pinellia ternata / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of low temperature on cell synchronization division in suspension culture cells of Pinellia ternata, in order to find a proper condition for cell synchronization.</p><p><b>METHOD</b>The loosened calli of P. ternata were used as materials. The effect of low temperature on cell synchronization division was studied and researched in suspension culture cells by the method of orthogonal experiment, the obtained experimental data was compared and analyzed through the analysis of range and analysis of variance.</p><p><b>RESULT</b>After the low temperature treatment, the cell division index was increased in various degrees, among the total results, when the temperature was set at 8 degrees C for 24 hours and then recovered for 36 hours, the effect was the best, the mitotic index was significantly increased to 11.3%.</p><p><b>CONCLUSION</b>The low temperature could obviously influence the cell synchronization division in suspension culture cells of P. ternata, and the mitotic index was efficiently increased.</p>

Differential display of mRNA from microtubers of Pinellia ternata in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To study mRNA differential expression from microtubes of Pinellia ternata in vitro, and give more information of the molecular mechanism in the formation of microtubers.</p><p><b>METHOD</b>DDRT-PCR was used to identify the expressed gene fragments related to the microtubers development of P. ternata in vitro.</p><p><b>RESULTS AND CONCLUSION</b>Fifteen cDNA fragments differentially expressed in the induction of microtubers of P. ternata in vitro were identified and characterized by using mRNA differential display DDRT-PCR. The deduced amino-acid sequences of six fragments of cDNA showed no significant homology with ESTs and genes in the Genbank databases, and they could be new cDNA fragments. However, the remaining three showed significant homologies with sequences encoding components of eukaryotic translation initiation factor 3 subunit H, MADS-box protein and ethylene signal transcription factor, respectively. Their differential expression patterns were confirmed by semi-quantitative RT-PCR analysis. And the expression level of the induction of microtubers of P. ternata in vitro was different.</p>

Study on preservation of Fritillaria anhuiensis by vitrification in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To establish cryopreservation system of shoot-tips from Fritillaria anhuiensis.</p><p><b>METHOD</b>Taking vitrification as system of cryopreservation, the shoot tips with length 2-3 mm were precultured in MS medium enriched with 0.4 mol x L(-1) sucrose for 3 d. They were treated for 20 min with 60% PVS2 at 25 degrees C, and then subjected to ice-cooled vitrification solution for 60 min and transferred to 2 mL cryotubes with fresh vitrification solution (PVS2) and plunged into liquid nitrogen. After rapid thawing in 40 degrees C water bath for 1 min, shoot-tips were expelled into MS medium containing 1.2 mol x L(-1) sucrose for 20 min. Further recovery and growth took place on regeneration medium in the dark at 25 degrees C for 2 weeks, and then with light/dark cycle of 12/12 h. The genetic integrity of cryopreserved shoot-tips was identified through products of PCR with arbitrary primers.</p><p><b>RESULT AND CONCLUSION</b>The highest survival rates of shoot-tips reached 79.9% by vitrification, and the regeneration rates were 52.3%. No changes were found between treated materials and untreated materials in genomic DNA.</p>

Study on hydroxyproline tolerance of different cultivars of Chrysanthemum morifolium in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate hydroxyproline tolerance of Chrysanthemum morifolium plantlets included " Boju", "Huaiju", "Chuju", "Gongju" and "Hangju",and provide references basis for excellent cultivar and breeding of Ch. morifolium.</p><p><b>METHOD</b>Plantlets in vitro from five kinds of Ch. morifolium were inoculated on medium added with different concentrations of hydroxyproline. Free proline in leaves from plantlets was determined, then the damage index and survival rate were compared.</p><p><b>RESULT AND CONCLUSION</b>The results showed that hydroxyproline tolerance of " Boju" and "Huaiju" were superior, the survival rates and free proline of them were higher, but the damage index was inferior. The hydroxyproline tolerance of "Hangju" was the worst, and the survival rate was minimum. The survival rate of "Chuju" and "Gongju" was between "Boju" and " Hangju", and the hydroxyproline tolerance of them was also medium.</p>

Changes of total RNA and mRNA differential expressions in leaves of Pinellia ternata under high temperature stress / 中国中药杂志

<p><b>OBJECTIVE</b>To study the total RNA and mRNA differential expression in leaves of Pinellia ternata under high temperature, provide more information of the molecular mechanism of the sprout tumble.</p><p><b>METHOD</b>The total RNA and mRNA differential expression in leaves of P. ternata at different stress time was analyzed.</p><p><b>RESULT</b>The results showed that the trend of total RNA content was divided into three descending stages and two ascending stages, the total RNA content was the highest at 0, 6 h, but it was the lowest at and 42 h, as well as when the sprout tumbled. The differential display showed that the polymorphism and type of bands of the sample at 6 h were similar to those at 0 h. But the bands numbers at other time were far less than those at 0, 6 h. And there were some different mRNA differential expression bands between the different samples.</p><p><b>CONCLUSION</b>In the process of the sprout tumble caused by high temperature stress, the RNA and mRNA differential expression in leaves of P. ternata changed.</p>

Variation of endogenous hormones in formation of microtuber of Dioscorea opposite in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>Through analysis of variation and function of 5 main endogenous hormones in the formation of microtuber of Dioscorea opposite in vitro to explore the physiological and biochemical mechanism of microtuber development.</p><p><b>METHOD</b>When microtubers were induced on MS + 6-BA 1.5 mg x L(-1) + NAA 1.5 mg x L(-1) + sucrose 5% medium, the endogenous hormones were isolated during different formation stages of microtubers, then purified and detected with enzyme-linked immunosorbent assays (ELISA).</p><p><b>RESULT</b>The results showed that GA3 slightly decreased in initial period, rose suddenly 20 days later, and than decreased. IAA showed a dropping tendency in the total course, ABA and ZR increased in a long period, dropped at last. JA continuously rose and never dropped, GA3 and ABA and the ratio of GA3 and JA varied obviously.</p><p><b>CONCLUSION</b>IAA, ABA, JA , ZR and GA3 play an important role in controlling formation of microtubers in D. opposite in vitro.</p>