Analysis of shading on DNA methylation by MSAP in Pinellia ternata / 中国中药杂志

Pinellia ternata is a medicinal herb of Araceae, and its tubers are used as medicines. It is a common Chinese herbal medicine in China and has a large market demand. When exposing to strong light intensity and high temperature during the growth process, P. ternata withers in a phenomenon known as "sprout tumble", which largely limits tuber production. Shade can effectively delay sprout tumble formation and increase its yield, however the relevant regulation mechanism is unclear. DNA methylation, as a self-modifying response to environmental changes, is often involved in the regulation of plant growth and development. In this study, P. ternata grown under natural light and 90% shading were selected as the control group and the experimental group for genomic DNA methylation analysis by using methylate sensitive amplification polymorphism(MSAP). The results showed that a total of 617 loci were detected with 20 pairs of primers, of which 311 were in the natural light group and 306 in the shading group. The methylation sites in the light and shading groups accounted for 58.2% and 71.57%, respectively, and the methylation ratios in the methylation sites were 27.65% and 29.41%, respectively, indicating that shading significantly induced the genome DNA methylation of P. ternata. Compared to the natural light group, shading promoted 32.51% of the genes methylation, while inducing 16.25% gene demethylation. This study reveals the DNA methylation variation of P. ternata under shading conditions, which lays a preliminary theoretical foundation for further analysis of the mechanism of shading regulation of P. ternata growth from epigenetic level.

Genetic transformation of Pinellia ternata with Agrobacterium tumefaciens-mediated sHSP genes / 中国中药杂志

<p><b>OBJECTIVE</b>To establish an efficient genetic transformation system of Pinellia ternata.</p><p><b>METHOD</b>With petioles from test-tube seedlings of P. ternata as explants, Agrobacterium tumefaciens mediation method was adopted to explore the effect of phenolic substances, A. tumefaciens's concentration, infection time, pre-incubation time and co-cultivation time on genetic transformation efficiency of P. ternata.</p><p><b>RESULT AND CONCLUSION</b>The genetic transformation efficiency could be effectively enhanced by infecting in A. tumefaciens culture containing AS 40 mg x L(-1) for 15 min for three days. The petioles were put into the differentiation medium containing 150 mg x L(-1) Kan and 350 mg x L(-1) Carb to screening and cultivation. After around 30 days, microtubers could be observed at both sides of the petioles. Gus staining and PCR verification on the regenerated plants showed that the exogenous gene sHSP had been integrated into genome of P. ternata.</p>

Study on optimization of induction system of test-tube tuberous roots from leaves of Rehmannia glutinosa / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of sucrose and plant growth substances of different concentrations on the induction of test-tube tuberous roots of Rehmannia glutinosa, in order to establish an efficient system for the induction of test-tube tuberous roots from leaves of R. glutinosa.</p><p><b>METHOD</b>Leaves from test-tube seedlings of 85-5 R. glutinosa were used as explants. After rooting induction, they were transferred to medium with orthogonal design for inducing test-tube tuberous roots of R. glutinosa.</p><p><b>RESULT AND CONCLUSION</b>NAA played a significant role in induction of test-tube tuberous roots of R. glutinosa, followed by sucrose and 6-BA. With leaves from test-tube seedlings as the explants, the optimal medium for inducing test-tube tuberous roots of R. glutinosa was MS + BA 3.0 mg x L(-1) + NAA 0.1 mg x L(-1) + sucrose 7%. The study provides an efficient induction system for studies on artificial seeds and secondary metabolism with test-tube tuberous roots of R. glutinosa.</p>

Study on optimization of SRAP-PCR reaction system for Pinellia ternata in Suzhou / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the optimization system of SRAP-PCR molecular marker technology in the analysis on Pinellia ternata.</p><p><b>METHOD</b>SRAP-PCR reaction system for P. ternata was optimized by L16 (5(4)) orthogonal design with five elements (dNTPs, Mg2+, the template DNA, primers, Taq enzyme) and four standards.</p><p><b>RESULT</b>The most suitable forward primer for SRAP for Pinellia ternata was 5'-TGAGTCCAAACCGGAAG-3', while the reverse primer was 5'-GACTGCGTACGAATTACG-3'. The optimized reaction system contained 70 ng DNA template, 0.9 micromol x L(-1) primer, 0.20 mmol x L(-1) dNTP s, 1.5 - 2.0 mmol x L(-1) Mg2+, and 2 U Taq enzyme.</p><p><b>CONCLUSION</b>SRAP-PCR system for P. ternata is established to lay a foundation for future construction of SRAP genetic map of P. ternata.</p>

Effects of different factors on direct induction of microtubers from explants in Pinellia ternate / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effects of different factors on direct induction of microtubers. These factors included plant growth substances, casein hydrolysate (CH), active carbon (AC), polyethylene glycol (PEG 4000), sucrose and glucose.</p><p><b>METHOD</b>Using the orthogonal design method.</p><p><b>RESULT AND CONCLUSION</b>The optimal media to directly induce microtubers from leaves were MS + MET 0.5 mg x L(-1) + 6-BA 0.5 mg x L(-1) + sucrose 3% and MS+ 6-BA 0.5 mg x L(-1) + IAA 0.5 mg x L(-1) + sucrose 3%. Optimal media to directly induce microtubers from tubers were MS + 6-BA 1.0 mg x L(-1) + PEG 5% + sucrose 5% and MS+ CH 500 mg x L(-1) + MET 0.5 mg x L(-1) + 6-BA 0.5 mg x L(-1) + AC 0.5% + sucrose 5%. Media suitable for plantlet growth were MS + 6-BA 0.5 mg x L(-1) + IAA 0.5 mg x L(-1) + AC 0.5% + sucrose 5% and MS + MET 0.5 mg x L(-1) + 6-BA 0.5 mg x L(-1) + sucrose 3%.</p>