Protoplasts isolation, purification and plant regeneration of Pinellia cordata / 中国中药杂志

The main factors which affected the isolation, purification and cultivation of Pinellia cordata protoplasts from leaves were studied. The results indicated that the optimum enzyme solution for P. cordata leaves was 13% CPW + 1.0% Cellulose +0.1% Pectolase, at pH 6.0, temperature (25-28 degrees C ) for 4 h. The sucrose density gradient centrifugation was adopted to purificate the protoplasts collected, when 25% sucrose was used as mediator, centrifugating at 500 rpm for 10 min. When the protoplasts were shallow liquid and liquid-solid double layer cultured on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA + 13% mannitol at the density of 2.5 x 104 protoplasts/mL, or fed and nursed cultured at the density of 100-500 protoplasts/mL, cell division could be observed for 3 days; granular calli appeared for 30 days. Calli was proliferated on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA solidified by 0.55% agar, and differentiated and regenerated after 5-6 months. Plant generation of P. cordata is successfully established.

Study function of endophytic fungus in parasitism process of mistletoe / 中国中药杂志

<p><b>OBJECTIVE</b>To research the function of endophytes of mistletoe in parasitism process of mistletoe in Pterocarya stenoptera.</p><p><b>METHOD</b>Endophytes from eight different parts of the mistletoe were separated by explant culture, and further screened by different CMC plates culture and DNS method to get cellulase high productive strains. The distribution of the endophytic fungus parasitized in mistletoe were prepared and stained to demonstrate by histological section of the intumescentia part of the P. stenoptera.</p><p><b>RESULT</b>The histological section indicated that aboundent of hyphasma were distributed around the haustorium of the mistletoe. Eighty three strains of endophytic fungus were separated, 38 of them were able to degrade cellulose, 19 strains showed high cellulase activity and 10 of which were separated from the parasitic position.</p><p><b>CONCLUSION</b>Endophytic fungus of mistletoe can secrete cellulase and assist the haustorium of mistletoe to breakthrough the cell walls as well as intercellular space tissues of the P. stenoptera, thus, the endophytic fungus plays an important role in the parasitism process of mistletoe in P. stenoptera.</p>

Application of SCAR molecular marker technology in identification of Monascus strains / 中国中药杂志

<p><b>OBJECTIVE</b>To establish an effective way for rapid identification of Monascus strains based on DNA molecular marker.</p><p><b>METHOD</b>A random amplified polymorphic DNA (RAPD) marker named F421 in genomic DNA of Monascus F strain was observed during a comparison of DNA fingerprints derived from 10 cultivated strains of Monascus. F421 was cloned and sequenced. Comparing the sequence of F421 (GenBank accession number EF063107) with other relative sequences in the GenBank databases, no distinct comparability was found. A pair of sequence characterized amplified region (SCAR) primers were designed based on the sequence of the cloned fragment and tested for the specific detection of Monascus F.</p><p><b>RESULT</b>The results of polymerase chain reaction showed that only a 421bp segment of Monascus F strain was amplified compared with other 9 cultivated strains of Monascus. And the acquired SCAR marker of strain F could be used as a specific DNA fingerprint to identify Monascus strain F within one day.</p><p><b>CONCLUSION</b>SCAR molecular marker technology is an effective new way to identify Monascus strains more rapidly. And also is an assistant tool to identify Monascus strains more accurately when disagreements come out using traditional classification. It could be applied widely to the protection of germ plasm resources, classification and identification distinguishing false strains of pharmaceutical fungi.</p>