Stress responsive NAC transcription factors involve in plant abiotic stress tolerance. Overexpression of SNAC1 significantly enhances drought, cold and salinity resistance in transgenic rice(Oryza sativa). In this study, TaSNAC1 was obtained from common wheat (Triticum aestivum) by homology-based cloning, its sub-celluar localization was analyzed, and its expression patterns in different tissues and under PEG or salt stress were investigated by quantitative RT-PCR. The cDNA of amplified TaSNAC1 including complete CDS was 1 076 bp in size, and the gDNA was 1 222 bp including a 146 bp intron (GenBank accession No. JN621240). TaSNAC1 encoded a protein with 329 amino acids, which showed 97.3%, 86.3%, 81.1%, 79.1% and 79.2% identity with SNAC1 of barley(Hordeum vulgare), false brome(Brachupodium distachyon), rice, maize(Zea mays) and sorghum(Sorghum bicolor), respectively. Results of phylogenetic analysis showed that TaSNAC1 was different from other wheat NAC transcription factors, it was clustered into a separate clade with other grass stress-responsive NAC. Structure prediction showed that TaSNAC1 might form a dimer, including a untypical nuclear localization signal (NLS) and a typical no apical meristem (NAM) domain. The core motif "WKATGXDK100-107" was located in a β sheet, which formed a concave surface and confered the ability of DNA binding. Based on transient expression assay using Arabidopsis thaliana mesophyll protoplasts, we found TaSNAC1 localized in the nucleus specifically. The expression levels of TaSNAC1 in both leaf and root were increased significantly in similar pattern during the application of high salt, and the increase in root was more dramatic (upto ~60 folds in root and ~10 folds in leaf ). Under PEG stress, the transcripts of TaSNAC1 were elevated quickly and sharply in root, but the change in leaf was delayed and the amplitude was decreased (about 15 folds in root and 6 folds in leaf). These data suggest that TaSNAC1 plays a vital role in abiotic stress response, and it possesses potential utility in improving drought and salinity tolerance in wheat.

To explore more genes for the improvement of drought-tolerance and water use efficiency in crops, the differential gene expression during re-watering after serious drought stress in common wheat (Triticum aestivum L.) was investigated by constructing SSH-cDNA library with the seedlings of Shaan 229 as materials. A total of 59 positive clones with insertions more than 400 bp were randomly picked from the library and sequenced. Then redundancy sequences were removed, finally 32 high quality EST (GenBank accessions of ES466767~ES466798) were identified. Homologous comparison of those EST sequences indicated that there were some similar responses of wheat to re-watering after serious drought stress with that to the other abiotic stresses. Among the 32 EST identified, 17 EST sequences were similar to known genes encoding functional proteins, which involved in the signal transduction, energy metabolism and transcriptional regulation, etc., the others were new EST in wheat. Based on an EST sequence highly similar to ERS gene in the SSH-cDNA library, four cDNA sequences encoding ethylene receptor in wheat were isolated using RT-PCR and RACE approaches. All the deduced proteins of these four cDNA sequences had the typical domains of ERS (three transmembrane segments, GAF domain, HisKA and HATPase-c domain). The full-length cDNA sequence of TaERS genes in wheat were 2 090, 2 271, 2 216 and 1 886 bp, and designated as TaERS1 (HM347272), TaERS2 (HM601437), TaERS3(HM601438) and TaERS4 (HQ111523), respectively. Multiple sequence alignment analysis based on the amino acids encoded by the ERS genes from wheat (T. aestivum L.), rice (Oryza sabiva), corn (Zay mays), sugarcane (Saccharum officinarum), Arabidopsis (Arabidopsis thaliana), tobacco (Nicotiana tabacum) and tomato (Lycopersicoum esculentum) indicated that TaERS had the highest similarity with that of rice (93%). The sequence alignment analysis revealed 23 SNPs among the four cDNA sequences of TaERS genes in wheat. Real-time PCR analysis indicated that TaERS was up-regulated in wheat by water stress and re-watering, and suggested that TaERS genes might be involved in responses of wheat to water stress. Those results may be beneficial for understanding the gene expression network of wheat in response to re-watering after drought stress and further characterization of the function of TaERS genes in improving the drought tolerance and water use efficiency in wheat.

Ear traits closely related to grain yield are important agronomic characters of wheat. In this study QTLs of traits related spike, spike length, grains per spike and fertile spikelet number, were analyzed using simple sequence repeat (SSR) markers in a population which consisted of 99 F8 recombinant inbred lines (RILs) derived from the cross of Poland wheat (Triticum polonicum L.)×common wheat (T. aestivum L.) line Zhong 13. The analysis of variance showed that there was highly significant difference between Poland wheat and Zhong 13 in spike length and grains per spike in 2010 and 2011, and significant difference was observed between Poland wheat and Zhong 13 in fertile spikelet number in 2010. It was clear that the hereditary basis of two parents about spike length, grains per spike and fertile spikelet number were very different. Highly significant difference among RILs was found in all three traits in two years. Of 355 pairs of polymorphic SSR marker between parents, 115 SSR markers were located to the 14 linkage groups including genome A and B. On average, there were 8.2 marker loci each chromosome. Chromosome 3B, with the most marker loci, had 13 marker loci and there were only 5 marker loci on chromosome 1A, 5B and 6B. The total length of 14 linkage groups was 822.9 cM, and average genetic distance between markers was 7.16 cM. With the method of composite interval mapping, the QTLs of traits related spike were detected using the field data in two environments. 6 QTLs for spike length, 4 located on chromosome 3A and 1 located on chromosomes 2A and 5B, respectively, were found in two years. The single of those QTLs for spike length could explain phenotypic variance from 9.21% to 22.94%. Two QTLs for fertile spikelet number located on chromosome 3B accounted for phenotypic variance 11.48% and 13.01% , respectively. Five QTLs for grains per spike were detected in 2010 and 2011. 4 QTLs were targeted to chromosome 3A and 1 to chromosome 7B. The single of those QTLs for grains per spike could explain phenotypic variance about 9.18%~19.71%. A major QTL for grains per spike was detected in the interval of Xbarc12~Xbarc310 on chromosome 3A in both 2010 and 2011. It indicated that this QTL was a stable and reliable grain per spike,which was less susceptible to environment conditions. This QTL was 0.01cM away from the nearest marker. So it can be used in molecular marker- assisted breeding for the improvement of grains per spike in wheat.

Recent studies have already indicated that vasoactice intestinal peptide (VIP) is closely related to the habit of birds' nesting. In this study, we first cloned VIP receptor gene and analyzed its characteristics of expression in different tissues of muscovy duck. Reversed transcript PCR was used to clone vasoactice intestinal peptide receptor-1 (VIPR-1) gene from the hypothalamus of black muscovy ducks(Cairina moschata), the cDNA sequence of this gene consisted of 1 125 nucleotides, which coded 335 amino acids(GenBank accession No. JN625215). The sequence of VIPR-1 was highly conserved in avian species. The muscovy duck VIPR-1 shared 96% amino acid identity to that of chicken (Gallus gallus), turkey(Meleagris gallopavo), and quail(Coturnix japonica), while it shared only 70%~72% amino acid identing to that of some mammalians like human(Homo sapiens), mice(Mus musculus), pig(Sus scrofa) and cattle(Bos taurus). The significant differences were detected in levels of VIPR-1 expressed in the stage of egg-laying, incubating and out-of-lay (P<0.05) by Real-time PCR, and the highest expression level was in the stage of incubating, while the lowest was in out-of-lay. This finding indicated that expression level VIPR-1 was closely correlated with the reproduction stages. In addition, VIPR-1 mRNA was found to be expressed in the hypothalamus, pituitary and ovary of black muscovy ducks. The highest expression level of VIPR-1 was found in hypothalamus, while the lowest level was in ovaries (P<0.01). In summary, VIPR-1 gene is highly conservative, and its tissue-specific expression patterns imply that VIPR-1 gene is involved in the role of hypothalamus-pituitary-ovary axis in regulating the incubation of muscovy ducks.

Abstract Establishment and preservation of livestock fibroblasts play a important role in protecting of animal genetic resources. The fetal fibroblast cell line of Jinhua pig was successfully established from 40 d's embryos by direct explant technique, and the cell morphology, growth dynamic, vitality, chromosome, isoenzymes, microbial inspection, cell apoptosis, as well as the transfection of fluorescent protein report plasmid(pEGFP-N3) had been studied in the line. The results showed that the primary cells proliferated rapidly, and overgrew on the bottom of culture flast in 2~3 days after explanted with a good situation, the growth curve had an obviously typical S shape and the population doubling time(PDT) was 36 h. The viabilities before freezing and after recovery were 95.2% and 92.8%, respectively. The frequency of chromosome number(2n=38) was 91% in metaphase, and it had met the establishment standard of fibroblast cell line. The analysis of lactic dehydrogenase(LDH) and malic dehydrogenase(MDH) isoenzymes showed no cross-contamination among the cells. No bacteria, fungi and mycoplasm were found, and there were without the microbial pollution in fibroblasts. The cell apoptosis rate of fibroblasts in the 15th generation was 2.0%, and without large-scale cell apoptosis. The plasmid transfection efficiency peaked at 32.4%, when the dose of cationic liposome(lipofectamin 2000) was 0.3 μL and fluorescent protein report plasmid(pEGFP-N3) was 0.5 μg. The individual item of the cell line had met the standard quality controls of American Type Culture Collection(ATCC). Establishment of Jinhua fetal fibroblast cell line not only preserves genetic resources of Jinhua pig but also provides a necessary experimental material for bioresearch, embryonic cloning and transgenosis animal. Moreover, the establishment of Jinhua fetal fibroblast cell line can provides both technical and theoretical support for storing the genetic resources of other animals and poultry at the cell level.

Bone morphogenetic protein 4 (BMP-4) plays an important role in animal bone growth and organism development. In this study, the methylation pattern of CpG island of bone morphogenetic protein 4(BMP4) gene promoter was detected using methylation-specific PCR(MSP), and compared and analyzed with carcass traits in four rabbit population of 80, New Zealand(NZ) rabbit(New Zealand Oryctolagus cuniculus), Fujian Yellow(FY) rabbit(Fujian Yellow O. cuniculus) and reciprocal cross between NZ and FY, 20 of each population. The results showed that cross populations on slaughter traits were higher than that of the average of FY rabbit and NZ rabbit, and they were lower than that of NZ rabbit but no significant(P>0.05) . It was very significant(P<0.01) between cross populations and FY rabbit on slaughter traits. Carcass traits were different in different methylation level in each group, but it showed no significant(P>0.05). Methylation level of NZ rabbit was significant compared with FY Rabbit and FY rabbit×NZ rabbit (P<0.05) and it showed no significant from one to each other rabbit populations(P>0.05) . Slaughter traits were different under different methylation level, but they did not match very well with methylation level and this might be cross by the methylation state of specific site. This study suggest a theoretical reference for explaining the relationship between methylation of BMP4 promoter region and heterosis and whether methylation of BMP4 promoter region can be a molecular marker or not.

The application of microsatellite markers for parentage determination has been approved in aquaculture which not only can maintain the parentage information, but also greatly reduce the cost in space and man-power and greatly improve breeding selection effect in communal environment, especially in the process of selective breeding. In this study, two selected cross-breeding families (the 49 full-sib families: 7 of Cyprinus carpio var. mirror♀×7 of C. carpio var. wuyuanensis♂and the 24 full-sib families: 3 of C. carpio var. wuyuanensis♀×8 of C. carpio var. mirror♂) were evaluated for genetic diversity and inheritance characteristics by 21 microsatellites loci, and examined several properties of microsatellites which could be considered to affect the accuracy of loci in parentage assignment. Finally, 13 suitability loci were obtained for parentage assignment, and performance of them showed that: (1) These 13 screened loci were high polymorphic. Mean number of alleles (K), mean expected heterozygosity (He), mean observed heterozygosity (Ho) and mean polymorphic information content (PIC) were respectively 0.505, 0.648, 0.791 and 0.5889 in the 49 full-sib families and 0.548, 0.670, 0.819 and 0.6138 in the 24 full-sib families. (2) 13 screened loci were of high discriminatory power. The combined exclusion probability (CPE) was 0.999997978 in the 49 full-sib families and 0.999999583 in the 24 full-sib families. (3) High efficiency of the loci subset was obtained. The CPE of 4 core microsatellite loci (MFW29, HLJ392, HLJ044 and HLJ855) could be accurate to more than 99% in the two cross-breeding groups.(4) Combined non-exclusion probabilities when the first parent was available (NE-1P), the second parent was available (NE-2P) and parent pairs were available (NE-PP) were respectively 1.34E-02, 3.91E-04 and 2.02E-06 in the 49 full-sib families, and were respectively 1.34E-02, 3.91E-04 and 2.02E-06 in the 24 full-sib families. (5) The simulations with 99% confidence indicated that 100% offspring of 49 full-sib families could be assigned to a parent pair using the 11 most polymorphic loci or more loci, 99% offspring of 24 full-sib families could be assigned to a parent pair using the best 9 or more loci. (6) The distance-based method of cluster analysis showed that: the set of 13 loci could clearly distinguished between the offspring of cross-breeding and the unrelated family in the polyculture. Based on these results, a suitable microsatellite-based tool for parentage assignment in cross-breeding carp is obtained, and displays a reliability and high efficacy alternative to physical tagging in subsequently selecting program.

Polysaccharides effects on immune regulation, anti-tumor and other activities. This experiment studied the effects of different dose Apostichopus japonicus intestinal polysaccharides on the immune function and anti-tumor of the mice(Mus musculus) by the method of stomach perfusion. Results showed that the intestinal polysaccharides had the dose-dependent inhibition for H22 tumor. The best anti-tumor effect was the high-dose(400 mg/kg/d) group, and its inhibition rate was up to 77.2%. The middle-dose(200 mg/kg/d) group's inhibition rate reached 62.6%, also significant higher than that of tumor control group. In the thymus index, the high-dose tumor group was greater than that of blank control group(P<0.05). Although no significant difference between other groups, from low-dose(100 mg/kg/d) to high-dose(400 mg/kg/d) the thymus index showed a rising trend in tumor groups. The spleen index in the tumor group increased first and then decreased following with the dose rising, the highest values appeared in the middle-dose group. While the trend of the blank groups always increased, but it was no significant difference among them. In the thymus index and the spleen index, tumor groups were always higher than that of blank groups in the same dose, this phenomenon may be related with the immune response. High-dose polysaccharides increased the IL-2 level significantly(P<0.05) of the tumor mice than that of the blank mice. The IL-2 level increased with the dose rising, to a certain extent it had concentration-dependent. The TNF-α level for the tumor mice high-dose group up to 441.685 pg/mL had significant difference than that of control group(P<0.05), the level rising followed the dose from control group to high-dose group. For the blank mice there was not such a regular pattern like that, the TNF-α level reached 60.518 pg/mL at middle-dose group and then down to 42.855 pg/mL at high-dose group. Such as the thymus index and the spleen index, on the IL-2 level and TNF-α level, tumor groups were higher than blank groups in the same dose. The tumor-bearing mice's NK cells' activity was less than that of the blank mice's. And following with the dose increasing, the blank mice's NK cell's activity showed a rising trend. In tumor group NK cell's activity increased first, in middle-dose the tumor group's NK cell's activity overtook the that of blank group's, and then the activity decreased. The polysaccharides could return the NK cell's activity, and its dose was not the higher the better. The experiment proved that Apostichopus japonicus intestinal polysaccharides can promote the immune function, and have the concentration-dependent inhibition in a certain dose on the effect to the tumor. This polysaccharides effect on immune function and anti-tumor activity can not unlimited increasing with its dose rising, there may be an optimal dose between middle-dose(200 mg/kg/d) and high-dose(400 mg/kg/d).

UGPase(UDP-glucose pyrophosphorylase) is one of the enzymes which mainly involves in plant carbohydrate metabolism, and plays an important role in the process of plant growth and development. The plant expression vector was constructed by ligating the cDNA fragment of UGPase from Saccharum Officinarum into expression vector pBI121, then the recombinant plasmid pBI-UGP was confirmed by restriction enzyme digestion analysis with BamHⅠ/SacⅠ, and further verified by DNA sequencing. The expression vector was introduced into Arabidopsis thaliana by floral-dip method, with Agrobacterium tumefaciens as media. Five T0 transgenic plants were selected by combining kanamycin screening with PCR detection. After selecting by kanamycin screening, T1 transgenic plants were identified by PCR and Southern blot, the results showed that the target gene was transformed into Arabidopsis thaliana successfully, and different transgenic plants contained various copy numbers of UGPase from Saccharum officinarum. T2 transgenic plants were identified by PCR and RT-PCR, the results indicated that the gene not only inherited stably in self-progeny, but also expressed at the RNA level. Meanwhile, content of soluble sugar, sucrose and starch were determinated in T2 transgenic plants and wild plants. Compared to wild plants, the soluble sugar content of transgenic plants had little change, however, sucrose content of transgenic plants improved obviously, increased by 50.85%~96.99%. Starch content of transgenic plants was lower than that of wild plants, decreased by 9.69%~36.76%. The research proved that UGPase plays a critical role in the process of converting sugar and starch, and the direction of its catalysis influneces distribution of these products in plant organization. It will lay foundation for further study of UGPase.

S-locus receptor kinase(SRK) and S-locus cysteine rich protein(SCR) are determinant factors of male and female in self-incompatibility, they are receptor-ligand relation and act with each other in allele-specific manner. To establish a technology system suitable for analysis of the allele-specific recognition and interaction mechanism between SRK and SCR, Brassica oleracea L. inbred lines E and F were used as plant materials. Firstly, the compatible index were analyzed by pollination assay, the result showed that E and F were all self-incompatible lines, they belonged to different haplotypes and their cross-pollination were compatible. Secondly, eSRKE, eSRKF, SCRE and SCRF were cloned from genomic DNA(gDNA) of E and F, nucleotide Blast revealed that E was S28 haplotype, and F was S7 haplotype. Thirdly, the interactions among the SCRE, SCRF, eSRKE and eSRKF were detected by MatchmakerTM Gold yeast two-hybrid system. The results indicated that SCRE could specific interact with eSRKE, SCRF could specific interact with eSRKF, whereas SCRF could not interact with eSRKE and SCRE could not interact with eSRKF, which were in entirely agreement with the result of pollination assay. The analysis of the interaction SCRE, SCRF, eSRKE and eSRKF provides a technology system for further study on recognition and interaction mechanism between SRK and SCR.

Real-time quantitative PCR(qPCR) has been widely used in gene expression analysis to date, and selection of suitable reference genes for qPCR according to specific experimental materials or conditions is very important for accurate normalization of target gene expression. However, only a few studies on reference genes have been done in plants. The present research here was conducted to identify appropriate reference gene(s) for normalization in future expression studies on tree peony (Paeonia suffruticosa Andr.). In this study, expression of five commonly used housekeeping genes such as β-tubulin, ubiquitin, glyceraldehyde- 3-phosphate dehydrogenase(GAPDH), Actin and 18S ribsomal RNA(18S rRNA) were assessed by qPCR in various tissues(roots, stems, leaves and petals) of tree peony and the petals of this cut flower during different opening phases or under different treatments(with ethylene or glucose). All the candidate reference genes could amplify specifically with high efficiency in qPCR reaction. To determine the expression stability of these genes, qPCR data were determined by two commonly used applets(geNorm and NormFinder), which produced highly comparable results depending on the experimental parameters. According to the analysis of geNorm program, the widely used reference genes ubiquitin and GAPDH were top ranked under all above conditions, which demonstrated they were both in the highest stability, whereas others including β-tubulin, actin and 18S rRNA were poorly ranked. Similarly, the results from NormFinder showed that ubiquitin was the most stable gene in the first two experimental conditions, while GAPDH was the most stable one in the petals of cut flower. As a result, for various tissues, as well as petals at different opening stages of tree peony cut flower, ubiquitin could be chosen as the proper reference gene, while compared to differences of gene expression in petals of this cut flower among diverse treatments, GAPDH was considered as the most appropriate reference gene. Besides, given using a normalization factor that was the combination of multiple housekeeping genes, the analysis recommended two most suitable reference genes (ubiquitin and GAPDH) were enough to achieve more accurate and reliable results among all investigated experimental conditions. The present study has provided an important reference for analysis the expression of critical genes in tree peony.

Abstract Mediator is a large, modular protein complex, and plays the important roles in regulating transcription in eukaryotes. Growth and development of a plant are controlled by programmed expression of suits of genes at the appropriate time, tissue and abundance. Although mediator has been studied rapidly in recent years in Arabidopsis thaliana, but the expression profiling patterns of mediator subunits remains largely unknown. In this study, using the coding sequences (CDS) of mediator subunits in Arabidopsis thaliana as searching sequences, a total of 24 mediator subunits CDS were found in rice(Oryza sativa) by BLAST with nucleonic acid data base of NCBI. Med19 had two copies, all the rest had only one copy in rice among the obtained 24 genes. The expression profiles of 23 subunit genes based on 59 Affymetrix GeneChip samples of japonica rice were obtained by clustering method. In these subunit genes, OsMed12 had the highest specificity expression, the fellowing was OsMed11, all the rest 21 genes had differential expression even close to constitutive expression. Analysis of OsMed8 promoter activity by GUS expression showed that OsMed8 highly expressed in meristems, early stages of vegetative and reproductive development, which consistented with the expression profile of OsMed8 in clustering analysis. Results of subcellular locations of mediator subunits in Nicotiana benthamiana leaves indicated that OsMed6 was localized in nucleus, and OsMed8 was localized in both the nucleus and the plasma membrane. The results of the expression profiling patterns and subcellular locations of mediator subunits can provide useful informations for studying of their biological roles in rice.

A reverse genetic technology platform of Porcine reproductive and respiratory syndrome virus (PRRSV) with foreign gene is established, which can be used to develop a DIVA (differentiating infected from vaccinated animals) marker vaccine. In this study, a molecular marker of immunodominant B-cell epitope (49 amino acids) of Newcastle disease virus (NDV) nucleoprotein (NP) was inserted into nonessential region of nsp2 by the method of mutant PCR, basing on the infectious clone of HuN4-F112 vaccine strain. The marker full-length cDNA clone (psk-HuN4-F112-Δ52+NP49) was assembled by cloning and splicing of the gene fragments. The completely assembled full-length cDNA clone was confirmed by sequence and Swa Ι enzyme digestion. Capped RNA was transcribed in vitro from a full-length cDNA clone of the viral genome and transfected into baby hamster kidney cells(BHK-21) by liposome to acquire the rescued virus. The rescued recombinant virus was passaged on a highly permissive subclone of African green monkey kidney epithelial cell line(MARC-145). The successfully rescued virus was tested by RT-PCR, digestion, and genome sequence. The results showed that the rescued virus could be distinguished from the parental with the mutant genetic marker (MluⅠenzyme site of genome at 14667nt) and the inserted nsp2 gene region. The results of indirect immunofluorescence assay(IFA) showed that the inserted foreign marker could be expressed by the recombinant virus and the inserted foreign gene was stable during the virus serial passage. The results of plaque assay and viral growth curve showed that the recovery virus possessed similar characters of viral growth to those of the parental virus. The results suggest that this stable infectious clone can be used as an important tool for development of novel vaccine against PRRSV.

Oomycetes is a class of diploid eukaryotic microorganisms in the kingdom of Chromista, including many of important plant pathogens. The principal impediment to progress in the molecular analysis of oomycetes has been the lack of efficient genetic and molecular biological tools. To evoke researchers to better understand the molecular genetics of oomycets and find out efficient tools for functional analysis of genes, three kinds of methods (disturbing gene expression, disrupting specific genes and spurring gene expression) used for the gene function analysis in oomycete study are reviewed in this paper. The review places an emphasis on the kind of disturbing gene expression. The advantages and disadvantages of the different techniques in three kinds were discussed. Future research trends of molecular genetics tools were discussed as well.

Tartary buckwheat (Fagopyrum tartaricum) seeds contain abundant protein with balanced amino acids and functional flavonoids. It is one typical crop for food and medical functions. Seed-filling development period is the crucial stage for the accumulation of those functional substances, so revealing the gene expression profile of this period is important for further regulation of functional component and improving the seed quality. For this reason, A full-length cDNA library was constructed by swithching mechanism at 5'end of RNA transcript(SMART) technology from filling stage Tartary buckwheat seeds and partial expressed sequence tags(EST) were analyzed. The results indicated that the titer of primary library and amplified library were 4.86×105 pfu/mL and 6.2×109 pfu/mL, respectively. The blue-white screening showed that it had a recombination rate of 99.58%, and the length of cDNA inserts was range from 500 to 3 500 bp. Some ESTs of the cDNA library (GenBank accession No. GO477569 and GO496285~GO496321) were obtained by sequencing of the random circularized plasmid clones. Bioinformatics analysis indicated that 54% ESTs showed high homology with the functional known genes, 28% ESTs had high homology with unknown protein or hypothetical protein, and 5% ESTs had no-hits. Among all known genes, ESTs of storage protein accounted for 17.93%, including 10, 16 and 22 kD storage protein, 11S and 13S globulin, vicilin and oleosin，etc. In addition, two genesrelated to the secondary metabolism named flavonoid 3'-hydroxylase and phenylalanine ammonia-lyase were obtained. All those results can lay a foundation for further isolation the genes related with the metabolism of seed storage protein and secondary metabolites.

Two new wheat cutivars Nongda 211 and Nongda 212 with high identities of agronomic traits(plant height, plant type, growth period, disease resistance, maturation appearance, etc.) except the seed coat color, were selected from the wheat(Triticum aestivum) cultivar Nongda3291 population by College of Agronomy and Biotechnology, China Agricultural University These two cultivars were officially approved by Beijjing Crop Variety Approval Committee in 2007 and 2010, respectively. In this paper, the cultivar selection process, features and extention situation of Nongda211 and Nongda212 were described, and experiences of wheat cultivar selection in China Agricultural University were discussed.