DNA fingerprinting among members of the Chinese drug Pu Gong Ying(Taraxacum mongolicum Hand,-Mazz.)and six adulterants of Tu Gong Ying were demonstrated with random-primed polymerase chain reaction(PCR)including arbit...DNA fingerprinting among members of the Chinese drug Pu Gong Ying(Taraxacum mongolicum Hand,-Mazz.)and six adulterants of Tu Gong Ying were demonstrated with random-primed polymerase chain reaction(PCR)including arbitrarily primed polymerase chain reaction(AP-PCR)and random amplified polymorphic DNA(RAPD).Distinctive,reproducible genomic fingerprints from DNA from 7 species belonged to Compositae were generated with two long(20 and 24 mer)and one short(10 mer)randomly chosen primers.The Pu Gong Ying can be differentiated from six species of Tu Gong Ying according to the banding pattems of their amplified DNA on agarose gels.The results showed that AP-PCR and RAPD methods can be used for identifying Chinese drugs.Moreover,the Similarity Indexes of the genomic DNA fingerprints showed that Pu Gong Ying and its adulterants are unrelated.Therefore,AP-PCR and RAPD methods can be used for identifying Chinese drugs.展开更多
RAPD (Randomly Amplified Polymorphic DNA) analysis was performed with filaments of 15 Porphyra lines representing four important groups (P. yezoensis, P. haitanensis, P. katadai var. Hemiphylla and P. digospermatangia...RAPD (Randomly Amplified Polymorphic DNA) analysis was performed with filaments of 15 Porphyra lines representing four important groups (P. yezoensis, P. haitanensis, P. katadai var. Hemiphylla and P. digospermatangia). Eight stable and repeatable RAPD bands amplified with two primers, OPN-02 and OPJ-18, were selected for the construction of DNA fingerprinting. The RAPD results were scored based on the presence or absence of each of the 8 bands and then converted to computer language expressed with two digitals, 1 and 0, which represented the presence (numbered as 1) or absence (numbered as 0) of each band, respectively. Based on these results, a model DNA fingerprint and a computerized DNA fingerprint were constructed. In the constructed DNA fingerprint, each Porphyra line has its unique fingerprinting pattern and can be easily distinguished from each other. Later, a software, named as PhGI, was designed based on this DNA fingerprinting. It can be used in practical Porphyra line identification.展开更多
We developed a computerized seed identification system. Fifteen rice varietiesthat were widely used in China were analyzed by AFLP fingerprinting. 12 primerpairs were screened, In order to simplify the procedure and c...We developed a computerized seed identification system. Fifteen rice varietiesthat were widely used in China were analyzed by AFLP fingerprinting. 12 primerpairs were screened, In order to simplify the procedure and cut down the cost inseed identification. the least number of primer pairs for practical seed identifi-cation should be seleeled. In this study. 3 primer pairs were selected. They展开更多
Rapid and reliable identification of olive plants using DNA markers has been attempted in the past but the selection of polymorphic regions for discrimination at varietal level remained obscure. Recent sequencing of p...Rapid and reliable identification of olive plants using DNA markers has been attempted in the past but the selection of polymorphic regions for discrimination at varietal level remained obscure. Recent sequencing of plastid genome of the olive flaunts high resolution Cp markers for olive DNA fingerprinting. Using this information, we designed a combination of chloroplast markers to amplify genes recruited in photosynthesis, ribosomal and NADH energy metabolism for varietal identification of olive plants. Concatenated DNA sequences of more than 100 unknown and 10 reference plants samples were analyzed using various bioinformatics and phylogenetic tools. Conserved blocks of nucleotide sequences were detected in multiple alignments. Phylogenetic reconstruction differentiated the unknown plants into various clusters with known varieties. Further narrowing down of the samples through UPGMA tree clearly separated the plants into Arbosana, Frantoio and Koroneiki as the major varieties. Multiple alignments of these clusters revealed important variety specific SNPs including G and T nucleotides at specific positions. Sequence identifying at intra cultivar level was more than 98.79% while it dropped to 97%, and even to 96% at inter varietal level. Furthermore, a neighbor net network analysis separated these three clusters, thus validating the results of UPGMA tree. Over all, out of 100 plants samples, 49 plants were identified that fall into 10 varieties including Arbosana, Carolea, Chetoui, Coratina, Domat, Frantoio, Gemlik, Koroneiki,Leccino and Moraiolo. The maximum number of known plants belongs to Frantoio and Gemlik (8 each). The least number of samples was identified from Carolea, Domat and Moraiolo with 2 samples each. However, 51 plants could not be identified, as plants were not clustered with any of reference control. Our results have implications in on-farm conservation of olive germplasm and provision of genuine material for multiplication of authentic varieties. This strategy can be extended to varietal identification of other plant species.展开更多
应用‘Myo’小卫星 DNA 探针,Southern 印迹杂交技术,对血斑、精斑、同一个体不同组织进行 DNA 指纹图分析,均获得清晰的图谱。同一个体的血斑与血液、精斑与精液以及不同的组织其 DNA 指纹图谱完全相同。可以根据斑痕或组织与嫌疑个体...应用‘Myo’小卫星 DNA 探针,Southern 印迹杂交技术,对血斑、精斑、同一个体不同组织进行 DNA 指纹图分析,均获得清晰的图谱。同一个体的血斑与血液、精斑与精液以及不同的组织其 DNA 指纹图谱完全相同。可以根据斑痕或组织与嫌疑个体的血液或某一组织 DNA 的指纹图谱比对以做出同一认定。50μl 血液量的血斑、5μl 精液量的精斑可以获得清晰易辨的指纹图谱。五年的精斑、两年的血斑亦可做出与同源个体新鲜精液、血液完全一致的 DNA 指纹图谱。对杀人、强奸杀人、碎尸等不同案件的血痕、精斑、不同组织碎块进行了 DNA 指纹图检验,均做出了正确的个体认定。本方法的应用为我国法医物证检验提供了新的分析手段,使个体认定得以实现。展开更多
从木材识别的角度,对木材DNA提取方法(CTAB、SDS、PTB、DNeasy Plant Mini Kit等)和DNA条形码及DNA指纹图谱等基于DNA的木材树种识别方法进行了综合评述。其中,针对DNA条形码方法,重点阐述了基因组DNA中可用的特征序列来源叶绿体基因(ch...从木材识别的角度,对木材DNA提取方法(CTAB、SDS、PTB、DNeasy Plant Mini Kit等)和DNA条形码及DNA指纹图谱等基于DNA的木材树种识别方法进行了综合评述。其中,针对DNA条形码方法,重点阐述了基因组DNA中可用的特征序列来源叶绿体基因(chloroplast DNA,cpDNA)rbcL、matK、trnH-psbA间隔区序列,核糖体基因(ribosomal DNA,rDNA)ITS序列,以及变异位点的识别和系统进化树的应用等问题;针对DNA指纹图谱方法,重点阐述了(RAPD、ISSR、SSR、SNP)4种DNA分子标记方法在DNA指纹图谱中的研究和应用现状。笔者认为,以DNA特征序列为依据的木材树种识别理论上虽然是可行的,但要应用于实际还需开展更多的研究。展开更多
文摘DNA fingerprinting among members of the Chinese drug Pu Gong Ying(Taraxacum mongolicum Hand,-Mazz.)and six adulterants of Tu Gong Ying were demonstrated with random-primed polymerase chain reaction(PCR)including arbitrarily primed polymerase chain reaction(AP-PCR)and random amplified polymorphic DNA(RAPD).Distinctive,reproducible genomic fingerprints from DNA from 7 species belonged to Compositae were generated with two long(20 and 24 mer)and one short(10 mer)randomly chosen primers.The Pu Gong Ying can be differentiated from six species of Tu Gong Ying according to the banding pattems of their amplified DNA on agarose gels.The results showed that AP-PCR and RAPD methods can be used for identifying Chinese drugs.Moreover,the Similarity Indexes of the genomic DNA fingerprints showed that Pu Gong Ying and its adulterants are unrelated.Therefore,AP-PCR and RAPD methods can be used for identifying Chinese drugs.
基金This study was supported by China Ocean "863" ProjectNational Natural Science Foundation of China.
文摘RAPD (Randomly Amplified Polymorphic DNA) analysis was performed with filaments of 15 Porphyra lines representing four important groups (P. yezoensis, P. haitanensis, P. katadai var. Hemiphylla and P. digospermatangia). Eight stable and repeatable RAPD bands amplified with two primers, OPN-02 and OPJ-18, were selected for the construction of DNA fingerprinting. The RAPD results were scored based on the presence or absence of each of the 8 bands and then converted to computer language expressed with two digitals, 1 and 0, which represented the presence (numbered as 1) or absence (numbered as 0) of each band, respectively. Based on these results, a model DNA fingerprint and a computerized DNA fingerprint were constructed. In the constructed DNA fingerprint, each Porphyra line has its unique fingerprinting pattern and can be easily distinguished from each other. Later, a software, named as PhGI, was designed based on this DNA fingerprinting. It can be used in practical Porphyra line identification.
文摘We developed a computerized seed identification system. Fifteen rice varietiesthat were widely used in China were analyzed by AFLP fingerprinting. 12 primerpairs were screened, In order to simplify the procedure and cut down the cost inseed identification. the least number of primer pairs for practical seed identifi-cation should be seleeled. In this study. 3 primer pairs were selected. They
文摘Rapid and reliable identification of olive plants using DNA markers has been attempted in the past but the selection of polymorphic regions for discrimination at varietal level remained obscure. Recent sequencing of plastid genome of the olive flaunts high resolution Cp markers for olive DNA fingerprinting. Using this information, we designed a combination of chloroplast markers to amplify genes recruited in photosynthesis, ribosomal and NADH energy metabolism for varietal identification of olive plants. Concatenated DNA sequences of more than 100 unknown and 10 reference plants samples were analyzed using various bioinformatics and phylogenetic tools. Conserved blocks of nucleotide sequences were detected in multiple alignments. Phylogenetic reconstruction differentiated the unknown plants into various clusters with known varieties. Further narrowing down of the samples through UPGMA tree clearly separated the plants into Arbosana, Frantoio and Koroneiki as the major varieties. Multiple alignments of these clusters revealed important variety specific SNPs including G and T nucleotides at specific positions. Sequence identifying at intra cultivar level was more than 98.79% while it dropped to 97%, and even to 96% at inter varietal level. Furthermore, a neighbor net network analysis separated these three clusters, thus validating the results of UPGMA tree. Over all, out of 100 plants samples, 49 plants were identified that fall into 10 varieties including Arbosana, Carolea, Chetoui, Coratina, Domat, Frantoio, Gemlik, Koroneiki,Leccino and Moraiolo. The maximum number of known plants belongs to Frantoio and Gemlik (8 each). The least number of samples was identified from Carolea, Domat and Moraiolo with 2 samples each. However, 51 plants could not be identified, as plants were not clustered with any of reference control. Our results have implications in on-farm conservation of olive germplasm and provision of genuine material for multiplication of authentic varieties. This strategy can be extended to varietal identification of other plant species.
文摘应用‘Myo’小卫星 DNA 探针,Southern 印迹杂交技术,对血斑、精斑、同一个体不同组织进行 DNA 指纹图分析,均获得清晰的图谱。同一个体的血斑与血液、精斑与精液以及不同的组织其 DNA 指纹图谱完全相同。可以根据斑痕或组织与嫌疑个体的血液或某一组织 DNA 的指纹图谱比对以做出同一认定。50μl 血液量的血斑、5μl 精液量的精斑可以获得清晰易辨的指纹图谱。五年的精斑、两年的血斑亦可做出与同源个体新鲜精液、血液完全一致的 DNA 指纹图谱。对杀人、强奸杀人、碎尸等不同案件的血痕、精斑、不同组织碎块进行了 DNA 指纹图检验,均做出了正确的个体认定。本方法的应用为我国法医物证检验提供了新的分析手段,使个体认定得以实现。
