甘肃小檗属植物分类检索表

经查阅有关小檗属分类的文献及标本 ,结合多年野外调查采集的资料 ,初步整理出甘肃小檗属植物 41种 4变种 ,并以检索表的形式对其进行了分组和分种鉴定 ,同时记载了种的分布。

The Application of RAPD Markers in Diversity Detection and Variety Identification of Porphyra

RAPD (random amplified polymorphic DNA) markers were generated from filaments of 15 Porphyra lines representing four important groups, P. yezoensis, P. haitanensis, P. katadai var. hemiphylla and P. oligospermatangia . Among the total 69 fragments generated by 6 selected primers (among 50 primers), 67 appeared to be polymorphic (97.1%). Cluster analysis based on the RAPD results was performed. The 15 Porphyra lines were divided into 3 groups. This result was consistent with that from taxonomy analysis. A DNA fingerprinting based on 8 bands amplified with OPN_02 and OPJ_18 was constructed and might be used in Porphyra variety identification. Five specific RAPD fragments of 5 Porphyra lines were isolated and cloned into pGEM_T easy vector. These five RAPD fragments may be useful in germplasm identification and property protection of Porphyra .

Molecular Identification and Cultivar Fingerprints of Prunus persica (L.) Batsch Germplasms

[Objective] The aim was to study the molecular identification and cultivar fingerprints of Prunus persica (L.) Batsch germplasms.[Method] Sixty peach genotypes,representing China common local cultivars and European samples were screened by microsatellites (simple sequence repeats,SSRs) and Inter-Simple Sequence Repeat (ISSR) markers.[Result] 26 reproducible bands were amplified by Nine SSR primers,and 24 of which were polymorphic; 236 bands were amplified by 30 ISSR primers,and 113 of which were polymorphic.31 genotypes were discriminated with 1-3 distinct polymorphic bands generated from the primers ISSR and SSR.Seven cultivar-specific ISSR fragments and two SSR unique alleles obtained from this study were available to be converted into Sequence Characterized Amplified Region (SCAR) markers.The genetic similarity coefficient (GS) estimated from these molecular data averaged were 0.939 (ranged from 0.856 to 0.983) for ISSR and 0.646 (ranged from 0.240 to 1.000) for SSR,respectively.The combined grouping association indicated that most local Chinese peach cultivars and exotic accessions were clustered together.This could be related to the mode of introduction and maintenance of the peach cultivars involving limited foundation germplasm,exchange of cultivars between plantations,and periodic development of new recombinant cultivars following sexual reproduction.[Conclusion] The results obtained in this work would help to improve the conservation,molecular identification and management of peach germplasm in breeding.

Identification of Five Sea Cucumber Species Through PCR-RFLP Analysis

Sea cucumbers are traditional marine food and Chinese medicine in Asia. The rapid expansion of sea cucumber market has resulted in various problems, such as commercial fraud and mislabeling. Conventionally, sea cucumber species could be distinguished by their morphological and anatomical characteristics; however, their identification becomes difficult when they are processed. The aim of this study was to develop a new convenient method of identifying and distinguishing sea cucumber species. Polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) analysis of mitochondrial cytochrome oxidase I gene(COI) was used to identifing five sea cucumber species(Apostichopus japonicus, Cucumaria frondosa, Thelenota ananas, Parastichopus californicus and Actinopyga lecanora). A 692 bp fragment of COI was searched for BamHI, KpnI, PstI, XbaI and Eco31 I restriction sites with DNAMAN 6.0, which were then used to PCR-RFLP analysis. These five sea cucumber species can be discriminated from mixed sea cucumbers. The developed PCR-RFLP assay will facilitate the identification of sea cucumbers, making their source tracing and quality controlling feasible.

Species identification and phylogenetic analysis of genus Nassarius(Nassariidae)based on mitochondrial genes

Genus Nassarius contains many subgenera, such as Zeuxis, Telasco, Niotha, Varicinassa, Plicarcularia, Nassarius s. str. and Reticunassa. On the basis of morphological characteristics of the shell and radula and sequences of mitochondrial cytochrome oxidase subunit I (COI) and 16S rRNA genes, Nassarius specimens collected from the South China Sea were identified and phylogenetically analyzed. Although Nassarius sp. and Nassarius (Varicinassa) variciferus were morphologically different in their shells, few variations were found among their radular teeth and sequences of mtCOI and mt16S RNA genes. Therefore, Nassarius sp. should be classified as N. (Varicinassa) variciferus. Nassarius (Zeuxis) sp. has only a subtle difference from Nassarius (Zeuxis) algidus on the shell, but it shows obvious differences in radular teeth and DNA sequence, indicating that they are two distinct species. Sequence divergence of mtCOI and mt16S RNA genes within Nassarius species was much lower than that between species, suggesting that these two genes are suitable for Nassarius species identification. Phylogenetic analysis (neighbor-joining and maximum parsimony) based on mtCOI and mt16S rRNA genes revealed the presence of two groups in genus Nassarius and a closest relationship between subgenera Zeuxis and Telasco. Species of subgenus Plicarcularia did not form a single clade. The molecular phylogeny was not congruent with the previous morphological phylogeny. The subgeneric divisions of genus Nassarius appear to be uncertain and unreliable.

Preliminary Studies and Antimicrobial Evaluation of the Aerial Parts of Genista numidica ssp. numidica

Genista numidica Spach ssp numidica （Spach） Batt. is a northeast Algerian endemic plant; the aim of this work is about the phytochemical screening, the microscopic analysis and the antimicrobial activity of the aerial parts of this species （stems, flowers, seeds and pods）; there is a wealth in polyphenolic compounds： flavonoids （method of Charaux, Paris, 1964） and saponins （method of Braune et al, 1969）, tannins, terpenes, sterols and fatty acids. The microscopic analysis doesn＇t allow a differential identification of the species. The results of aromatogram by the incorporation method of Muller-Hinton on solid medium, showed a significant antimicrobial activity （method Duraffourd,1987） of the extracts, infusion, and the active substances towards bacterial strains, essentially those recognized as antibiotic resistant （Staphylococcus, Acinetobacter, Pseudomonas, Enterobacter, Citrobacter）; the results obtained indicate that the most efficient antibacterial activity was obtained with the pod infusion, the seed extract and the flavonoids. The raw extract has an important antifungal activity on Candida albicans. This could open opportunities for using this species in the treatment of urinary, respiratory, intestinals and cutaneous infections, and as a disinfecting additive on nosocomial area.

Molecular identification based on ITS sequences for Kappaphycus and Eucheuma cultivated in China

The systematic classification of the Eucheumatoideae is difficult because of their variable morphology and interpretation of reproductive structures. Kappaphycus and Eucheuma specimens cultivated on the Hainan and Fujian coast of China were introduced from Vietnam, the Philippines and Indonesia. Combined with morphological characteristics, all Kappaphycus and Eucheuma cultivated strains were identified by internal transcribed spacer (ITS) sequences. The phylogenetic tree was constructed using neighbor-joining and maximum likelihood methods. The results indicate that different ITS sequence lengths occurred in the different genera and species. An obvious difference in morphology could be found in the protuberance shape between Kappaphycus and Eucheuma. The protuberance in Eucheuma was thorn-like and in Kappaphycus was wartlike or papillate. Their ITS sequence lengths differed significantly in nucleotide variation rates up to 58.55%-63.90%. All nucleotide variations occurred in the ITS1 and ITS2 regions except for five nucleotide transversions in the 5.8S rDNA region. In addition, the difference was at the branches among congeneric species. Kappaphycus sp. had branches with small buds, while K. alvarezii did not have such a feature. The nucleotide variation rates varied from 7.02% to 7.48% among species; within the same species of the clades it was <1.20%. Eucheumatoideae algae cultivated in China consisted of three clades, K. alvarezii, Kappaphycus sp., and E. denticulatum. The results indicate that ITS sequence analysis was an effective way for identification of interspecies and intraspecies phylogenetic relationships and might provide a clue for molecular identification of algal Eucheumatoideae.

沙门氏菌MALDI-TOF MS分型方法建立

沙门菌（Salmonella）是一种重要的食源性致病菌,其抗原结构复杂,根据Kauffmann-White抗原表中O抗原和H抗原分型,沙门菌具有2 500多个血清型。沙门菌在不同地区分布不同的菌型,地区分布差异大,目前全球已知有2 489个不同血清型,分属于46个O群。截止2000年底,中国只检出292个血清型,分属于35个O群[1]。

Evaluation of Landsat 7 ETM＋ Data for Spectral Discrimination and Classification of Sugarcane Varieties in Colombia

The identification of sugarcane varieties through remote sensing is studied to reduce the time taken to identify in the field, also is useful to identify non-certified varieties and to monitor the adoption of new varieties. The purpose of this study is to evaluate the Landsat 7 ETM＋ images to discriminate varieties CC85-92 and CC84-75 in the Cauca river valley in Colombia. The method used to measure the spectral separability between varieties was Jeffries-Matusita. The results indicated that the only period where a clear discrimination of the varieties is between 4th and 5th months, with a global precision of 80.8% and kappa index 0.62. The proposed methodology and preliminary results show that remote sensing is a useful tool for monitoring and identification of varieties and could be used for identification of varieties already registered and planted in other countries without the consent of their true creators.

Molecular markers for identifying a new selected variety of Pacific white shrimp Litopenaeus vannamei

Selective breeding of the Pacific white shrimp Litopenaeus vannamei during the last decade has produced new varieties exhibiting high growth rates and disease resistance.However,the identification of new varieties of shrimps from their phenotypic characters is difficult.This study introduces a new approach for identifying varieties of shrimps using molecular markers of microsatellites and mitochondrial control region sequences.The method was employed to identify a new selected variety,Kehai No.1(KH-1),from three representative stocks(control group):Zhengda;Tongwei;and a stock collected from Fujian Province,which is now cultured in China's Mainland.By pooled genotyping of KH-1 and the control group,five microsatellites showing differences between KH-1 and the control group were screened out.Individual genotyping data confirmed the results from pooled genotyping.The genotyping data for the five microsatellites were applied to the assignment analysis of the KH-1 group and the control group using the partial Bayesian assignment method in GENECLASS2.By sequencing the mitochondrial control regions of individuals from the KH-1 and control group,four haplotypes were observed in the KH-1 group,whereas14 haplotypes were obtained in the control group.By combining the microsatellite assignment analysis with mitochondrial control region analysis,the average accuracy of identification of individuals in the KH-1group and control group reached 89%.The five selected microsatellite loci and mitochondrial control region sequences were highly polymorphic and could be used to distinguish new selected varieties of L.vannamei from other populations cultured in China.

Study on Complete Mitochondrial Genome of Oula Sheep(Ovis aries)

Mitochondrial genome has been widely used in species identification and gene conservation.In the present study,the complete mitochondrial genome of Oula sheep(Ovis aries)was determined using next-generation sequencing.This genome was16 618 bp(NCBI accession number:KU575248)and contained 13 protein coding genes,22 transfer RNA genes,two ribosomal RNA genes,and a typical control region.The overall nucleotide composition was 33.7%A,27.4%T,25.8%C,and 13.1%G,with a total A+T content of 61.1%.The phylogenetic analysis of selected sheep breeds showed that Oula sheep were clustered within branch A and originated from approximately 6 ka.This mitochondrial genome will provide valuable information for molecular genetic research of Oula sheep.

Identification of Essential Oil Composition of Four Picea Mill, （Pinaceae） Species from Canada

The aim of this study was to determine the volatile composition of essential oil of four Picea Mill. species （Piceapungens Engelm., Picea mariana （Mill.） Britton, Picea glauca （Moench） Voss., Picea rubens Sarg.） needles. The volatile components extracted from these four species needles were analyzed by using headspace solid phase microextraction （HS-SPME）/gas chromatography-mass spectrum （GC-MS） and 31, 34, 27 and 24 compounds were identified representing 91.77%, 92.70% 92.38% and 94.06% of the total oil, respectively. The major constituents were found to be bornylacetate （29.40%）, camphor （26.43%）, 13-myrcene （7.47%） and camphene （7.01%） in P. pungens; camphene （22.03%）, bornylacetate （21.64%）, α-pinene （16.62%） and borneol （7.79%） in P. mariana; bornylacetate （31.25%）, limonene （17.27%）, α-pinene （15.85%）; camphene （13.65%） in P. glauca and borneol （12.38%）, α-pinene （10.36%）, germacrene D （9.86%） and δ-cadinene （8.25%） in P. rubens. This study sought to detecte new phytochemical data on the Picea genus to help chemotaxonomy and usable of studied species.

Screening of Drought Resistance Identification Indexes for Rice(Oryza sativa L.) Cultivars in Ningxia

The morphological characters, physiological characters, yield traits and yield per plant of total 49 rice cultivars from Ningxia were investigated under conditions of water stress and non-water stress so as to determine the relationship between each trait and yield per plant under water stress and the relationship between each relative character and drought resistance coefficient under water stress and non-water stress. The correlation, grey correlation, stepwise regression and path analyses showed that the tiller number per plant, plant height, grain density, effective panicle number per plant and grain number per panicle, total 5 traits, were significantly correlated with the drought resistance of rice, and they could be used to identify the drought resistance of rice in Ningxia. In addition, the drought resistance of rice was graded qualitatively according to the subordinate function value of corresponding drought resistance coefficient. The results showed that among the 49 rice cultivars from Ningxia,6 rice cultivars were highly drought resistant, and 9 rice cultivars were moderately drought resistant, suggesting that the evaluation method was feasible and effective.

Fast species identification of mycobacterium by rpoB gene chip technology

Based on rpoB gene micro array as target gene, we are going to use gene chip technology to test 24 mycobacterium standard specimens, 8 non-mycobacterium specimens and 86 mycobacterium clinical isolated specimens. As a result, after mycobacterium and non-mycobacterium standard specimens were duplicated by PCR, mycobacterium standard specimens reproduced 360bp DNA fragments; on the other hand, non-mycobacterium specimens did not reproduce any fragments except for hemolytic streptococcus and corynebacterium pseudodiphtheriticum which had the same results as mycobacterium standard specimens. Sensitive test is able to detect lpg tuberculosis mycobacterium DNA. The probe test showed that, among 21 oligonucleotide probes, probe-M. fortuitum and M. marinum were cross-hybrid; the other probes were specific. We used the new method to identify 126 mycobacterium clinical isolated specimens. The test results of this new method matched with conventional method. In conclusion, compared to the traditional method, the use of rpob gene chip technology to identify mycobacterium species will be faster, more accurate and higher value in application.

Identification of the novel recessive gene pi55(t) conferring resistance to Magnaporthe oryzae

The elite rice cultivar Yuejingsimiao 2 （YJ2） is characterized by a high level of grain quality and yield, and resistance against Magnaporthe oryzae. YJ2 showed 100% resistance to four fungal populations collected from Guangdong, Sichuan, Liaoning, and Heilongjiang Provinces, which is a higher frequency than that shown by the well-known resistance （R） gene donor culti- vats such as Sanhuangzhan 2 and 28zhan. Segregation analysis for resistance with F2 and F4 populations indicated the re- sistance of YJ2 was controlled by multiple genes that are dominant or recessive. The putative R genes of YJ2 were roughly tagged by SSR markers, located on chromosomes 2, 6, 8, and 12, in a hulked-segregant analysis using genome-wide selected SSR markers with F4 lines that segregated into 3 resistant （R）： 1 susceptible （S） or 1R：3S. The recessive R gene on chromosome 8 was further mapped to an interval ~1.9 cM/152 kb in length by linkage analysis with genomic position-ready markers in the mapping population derived from an F4 line that segregated into 1R：3S. Given that no major R gene was mapped to this inter- val, the novel R gene was designated as pi55（t）. Out of 26 candidate genes predicted in the region based on the reference genomic sequence of the cultivar Nipponbare, two genes that encode a leucine-rich repeat-containing protein and heavy-metalassociated domain-containing protein, respectively, were suggested as the most likely candidates for pi55（t）.

Local multifractal detrended fluctuation analysis for tea breeds identification

In recent years, the popular multifractal detrended fluctuation analysis （MF-DFA） is extended to two-dimensional （2D） version, which has been applied in some field of image processing. In this paper, based on the 2D MF-DFA, a novel multifractal estimation method for images, which we called the local multifractal detrended fluctuation analysis （LMF-DFA）, is proposed to recognize and distinguish 20 types of tea breeds. A set of new multifractal descriptors, namely the local multifractal fluctuation exponents is defined to portray the local scaling properties of a surface. After collecting 10 tea leaves for each breed and photographing them to standard images, the LMF-DFA method is used to extract characteristic parameters for the images. Our analysis finds that there are significant differences among the different tea breeds＇ characteristic parameters by analysis of variance. Both the proposed LMF-DFA exponents and another classic parameter, namely the exponent based on capacity measure method have been used as features to distinguish the 20 tea breeds. The comparison results illustrate that the LMF-DFA estimation can differentiate the tea breeds more effectively and provide more satisfactory accuracy.