ABSTRACT
<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>
Subject(s)
Agrobacterium tumefaciens , Genetics , DNA, Plant , Genetics , Genetic Engineering , Methods , Glucuronidase , Genetics , Metabolism , Heat-Shock Proteins, Small , Genetics , Pinellia , Genetics , Metabolism , Plant Leaves , Genetics , Metabolism , Plants, Genetically Modified , Polymerase Chain Reaction , Reproducibility of Results , Tissue Culture Techniques , Methods , Transformation, GeneticABSTRACT
<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>
Subject(s)
Benzyl Compounds , Dose-Response Relationship, Drug , Kinetin , Pharmacology , Naphthaleneacetic Acids , Pharmacology , Plant Growth Regulators , Pharmacology , Plant Leaves , Plant Roots , Purines , Rehmannia , Seedlings , Sucrose , Pharmacology , Tissue Culture Techniques , MethodsABSTRACT
<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>
Subject(s)
China , DNA Primers , Genetics , DNA, Plant , Genetics , Electrophoresis , Magnesium , Metabolism , Nucleic Acid Amplification Techniques , Methods , Nucleotides , Genetics , Pinellia , Genetics , Polymerase Chain Reaction , Methods , Reproducibility of Results , Taq Polymerase , Metabolism , Templates, GeneticABSTRACT
<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>