ABSTRACT
To construct CRISPR/Cas9 vectors for the editing of SmPAL1 in the phenylpropane metabolic pathway of Salvia miltiorrhiza, CIRSPR/Cas9 target sites of SmPAL1 were designed by online software. Its target efficiencies were detected in vitro by enzyme digestion and sequences with highly efficiency were constructed into CRISPR/Cas9 vectors. Three possible CRISPR target sequences (SmPAL1-g1, SmPAL1-g2, SmPAL1-g3) were designed and the enzyme digestion efficiencies were 53.3%, 76.6% and 10.0%. SmPAL1-g1 and SmPAL1-g2 were constructed into vector VK005-03 named as VK005-03-g1 and VK005-03-g2. The results of sequencing showed that the two CRISPR/Cas target sequences were all constructed into VK005-03. Here we first laid the foundation for the study of SmPAL1 and provided an effective strategy for the screening of sgRNA.
Subject(s)
CRISPR-Cas Systems , Clustered Regularly Interspaced Short Palindromic Repeats , Metabolic Networks and Pathways , Genetics , Salvia miltiorrhiza , Chemistry , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To study the relationship between photosynthetic characteristics and environmental factors in leaves of P. lobata.</p><p><b>METHOD</b>Photosynthetic characteristics and environmental factors were measured by using CIRAS-2 portable photosynthesis system.</p><p><b>RESULT</b>The apparent quantum yield in leaves was 0.0173 micromol CO2 x micromol(-1) photon. The dark respiration rate was 2.9333 micromol x m(-2) x s(-1). The light compensation point of photosynthesis was 180 micromol x m(-2) x s(-1). The light saturation point was 1600 micromol x m(-2) x s(-1). The carboxylation efficiency was 0.0338 micromol x m(-2) x s(-1). The light respiration rate was 2.5 micromol x m(-2) x s(-1). The CO2 compensation point was 100 micromol x mol(-1), The CO2 saturation point was 1 600 micromol x mol(-1).</p><p><b>CONCLUSION</b>Photo flux density and air temperature are major environmental factors influencing diumal changes of net photosynthetic rate.</p>
Subject(s)
Photosynthesis , Physiology , Plant Leaves , Metabolism , Pueraria , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To purify salvianolic acids by macroreticular resin,then mensurate the contents of salvianolic acids and analyse the chromatogram with HPLC.</p><p><b>METHOD</b>Make salvianolic acids with macroreticular resin; mensurate the content of Salvianolic acids with UV spestrophotometry: the control compound is protocaechuic aldehyde, and the wavelength is 281 nm. Analysis the chromatogram with HPLC, and compare the chromatogram in different technics: zorbax ODS column (4.6 mm x 250 mm, 5 microm), mobilephase: 1% aceticacid-water and methanol in different proportions, the wavelength is 281 nm.</p><p><b>RESULT</b>The contents of salvianolic acids is 53.8%; HPLC chromatogram indicate that the method is reasonable to make salvianolic acids.</p><p><b>CONCLUSION</b>Determination of contents and HPLC chromatogram can control the quality of Salvianolic acids more accurately.</p>