| 1The study of The RTNS of Artemisia annua L1.1Selecting RTNS sequence from the EST database of Artemisia annua The Artemisia annua is the dry aerial parts of Compositae Artemisia annua L. it has the affect of releasing the hotness and eliminating the steam and malaria. It is designed to relieve hotness and humid, smelling fragrant, being the critical medicine for wet plague Pandora. Its main medicinal ingredient is Artemisinin (sesquiterpene lactones). Modern pharmacological studies have shown the the annuasignificant antimalarial, phytopathogenic microorganisms, antipyretic and analgesic effects. For the important medicinal value, Artemisia annua is the most abundant species of medicinal plants in the genetic data. Currently, in GenBank, the nucleic acidsequence of Artemisia annua is more than237,000, its number is after the model plant Arabidopsis (more than298,000), the EST sequences is more than94,000, and the Artemisia annua genome project are being conducted.In the evolution of the genome, turn (Transportableelements TEs) has an important position, in our human genome, nearly50%is constituted by TEs. In the evolution of gene regulatory networks, TEs also play an important role; TEs can be divided into two categories: retrotransposons (Retrotransposons, RTNs) and DNA transposons. The former will make the genome via RNA replication transposition, increasing the genome.For RTNs largely existing in a wide range of single-cell algae, bryophytes, gymnosperms and angiosperms, so tapping the massive accumulation of Artemisia annua EST data and establishing analysis system of the EST-the RTNs sequence of Artemisia annua have great value in exploring the homology in the different tissues of Artemisia annua gene, studying the complex physiological and pathological processes, and then understanding the evolutionary relationship between the universality of the biological mechanisms and genetic background in the analysis of species differences and species, comparative mapping and other aspects of research with application value.However, the research of the the RTNs of Artemisia annua is still in the blank stage. In the nucleic acid sequence, only our laboratory get a sequences of ty3-gypsy with the mehod of cloning the homologous. Due to the the RTNs sequence changes, it has weak conservative, so we try t clone o the RTNs sequence of Artemisia annua with a data mining method.The results are as follows:The length of Artemisia annua EST data is50～1074bp, the average length is620bp, arranging the Artemisia annua EST data with est_trimmer Perllanguage, removing sequence shorter than100bp to get93874, compared with the carrier database, found9EST sequences vector contaminated after removing,93865clean, higher-quality Artemisia annua EST sequences was got. RTNs sequence characteristics of data sets with the Bioperl Auxiliary mining, the source is the NCBI plant nucleic acid sequences, with total of39documents, the amount of data is8GB, defining gene with retrotransposon, copia, gypsy, reverse genetic and selecting the field parallels of keywords (KEYWORDS). Conversing the sequence data format, filtering out RTNs sequences from GenBank format conversion for the fastaformat for the CAP3clustering,636RTNs seed sequence is obtained, the type including:knownRTNs covering the types of flowering plants, set the RTNs seed sequence Blast of EST data to search Artemisia annua, the result maybe RTNs of Artemisia annua, extracting EST with a Perl program. Extracted EST after CAP3clusteringmosaic,196contigs are obtained, using Blast to search non-redundant protein database, and filter out recognized the RTNs the annua ESTsplicing sequence22sequence composed of the following table, select the length of the large a900bp of contig1pair of PCR primers, experiments verify that the expected length of the annuaEST data drawn from the analysis of the nucleotide sequence of the Artemisia annua retrotransposons, both consistentdescription of forecast sequence present in the Artemisia annuagenome data mining is practicable.1.2The conditions optimization for the Artemisia annua iPBSIPBS is based on retrotransposons, molecular markers,compared with other retrotransposons marker technology, the advantage of the conserved regions is able to use the sequenceof the entire PBS, it not only can be intuitively seen between individual genetic normality quickly clone LTR fragment from genomic DNA, but also can be a computer database for the query of the fragments, which makes the the iPBS method become a powerful DNA fingerprinting technology, and does not require previous sequence knowledge. Germplasm resources is particularly important due to significant differences in the content of artemisinin in Artemisia region, so we will experiment to explore the conditions for optimization of Artemisia annua iPBS.The experimental results are as follows:Artemisia annua DNA material, established the IPBS molecular marker system based on retrotransposons, preferably selecting aprimer (2378) for Artemisia annua, ptimizing the content of each component whichis more appropriate for Artemisia annua experimental (50μl system) in this group:The concentration of template DNA is15～36.9ng,,5μ’s10×buffer (Mg2+free) buffer,MgCL22.0mmol/L, dNTPs,0.2mmol/L of Taq enzyme,3U,primers (2378)2.0μmol/L,; amplification program:initial denaturation at5min; With this basic system optimization results, you can make the IPBS better used in molecular markers of Artemisia annua, lay a good foundation for future Artemisia annua germplasm research.2The leaf anatomical structure of Polygonum cuspidatumThe medicinal properties of Polygonum cuspidatum are recorded in "Herbal Supplements" and "Herbal Chen", it is bitter, slightly astringent, slightly cold. It is included in liver. It can calming liver, relieving expectorant. It is used for the liver, high blood pressure, dizziness,. the anatomical structure of Polygonum cuspidatum is in blank, this article the use bio-microscopy combined with mathematical knowledge, paraffin sections of Polygonum leaves, torn them into piece by hands, and at the same time, powder the Polygonum cuspidatum leavesfor microscopic observation, measure of cell thickness,calculate the fence percentage. Providing amicroscopic basis for the using of Polygonum cuspidatum leaves as medicine, the resistance mechanism of Polygonum cuspidatum and Polygonum cuspidatum leaf ontogeny.Polygonum cuspidatum leaves are composed of epidermis, mesophyll, veins,it is the typical surface leaves. View from the surface, epidermal cells are regularly arranged on Polygonum cuspidatum, nearly equal in diameter, the lower epidermal cells o fPolygonum cuspidatum epidermis neat arrangement is less than the pores, it is distributed only with the lower epidermis, stomatal type is infinitive, the anticlinal walls are vertical. View from the cross-section, Polygonum cuspidatum leaves the upper and lower epidermis are composed of two layers of cells on epidermal cells, nearly rectangular,thickness of66.3microns, small epidermal cells is small, nearly square,, with a thickness of38microns. Mesophyll, palisade tissue is below epidermidis, arranged within a layer, spongy tissue is located between the palisade parenchyma and lower epidermis, palisade tissue cells were cylindrical, arranged in neat rows and close, the longest is95microns and with width of28.5microns, fence percentage is1to2, the spongy tissue is connected with the lowerepidermis phase, compose by some of the nearly circular or irregular parenchyma cells, the cellgap is loosely arranged, and thin-walled cells have druse, Polygonum cuspidatum leaves are in stockwork shape, has developed main veins down processing, compose by the main vein vascular and basic organization. It have a big and a small vasculars, Including the outer xylem, phloem, xylem in the adaxial, phloemabaxial. The vascular diameter is218.5microns, the vascular tissue is composed by the xylem, cambium, and phloem. The xylem is developed, has multiple catheters, fiber and textured catheter, the formation of the layer is not obvious, the phloem cells are arranged closely in neat rows. |
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