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. olig...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 .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
Randomly amplified polymorphic DNA (RAPD) technique was applied to assess the genetic variations and phylogenetic relationships in 4 species ofPorphyra. The samples were collected from the coast of Canada, Vietnam, Zh...Randomly amplified polymorphic DNA (RAPD) technique was applied to assess the genetic variations and phylogenetic relationships in 4 species ofPorphyra. The samples were collected from the coast of Canada, Vietnam, Zhoushan, Fujian and Qingdao in China. Amplifications with 20 primers were carried out under predetermined optimal reaction conditions (samples were first heated at 94°C for 5 min. and followed by 45 cycles of 1 min at 94°C, 1 min at 36°C, and 2 min at 72°C, then held at 72°C for 10 min). The amplified products were scored as present (1) or absent (0) for each DNA sample and an index of genetic similarity (F) was calculated by using Nei & Li’s matching coefficient method (1979). The value of (1?F) was used to quantify the genetic distances between species and construct a phylogenetic tree. The relationship indicated by the UPGMA and NJ cluster analysis on the values of the genetic distance is in good overall agreement with classical taxonomy. The obvious differences between natural and cultivated population ofP. haitanensi suggest that variation or hybridization with other species occurred during the culture.展开更多
Germplasm collection is important to preserve and maximize genetic diversity for germplasm conservation.Tsuga dumosa(D.Don)Eichler in Engler & Prantl.and T.chinensis var.forrestii(Downie)Silba germplasm was collec...Germplasm collection is important to preserve and maximize genetic diversity for germplasm conservation.Tsuga dumosa(D.Don)Eichler in Engler & Prantl.and T.chinensis var.forrestii(Downie)Silba germplasm was collected from three localities in China:Mt.Yulong,Wenfeng Temple and Mt.Dishiergu,Yunnan Province.Accessions were identified based on morphological characters and RAPD markers.The shapes of the apices and margins of needles were examined,and the length and width of needles,cones and seeds from accessions of mature plants were used to compare the morphological differences and to identify the germplasm.Molecular markers generated by randomly amplified polymorphic DNA(RAPD)were also used to characterize the taxa.Although the clustering based on RAPD markers was inconsistent with the morphological characters of the needles,based on the overall morphological characters and on RAPD markers,the accessions from Mt.Yulong and Wenfeng Temple were identified as T.chinensis var.forrestii,and those from Mt.Dishiergu identified as T.dumosa.Taxonomic identification of the accessions was made based on morphology and by RAPD markers concurred.The results indicate that the shapes of the apices and margins of needles particularly from young plants could not be used as a possible key to identify T.dumosa and T.chinensis var.forrestii.Fig 6,Tab 3,Ref展开更多
Eighteen gametophytes includingL. japonica, L. ochotensisandL. longissima, were verified with random amplified polymorphic DNA (RAPD) technique. Eighteen ten-base primers were chosen from 100 primers selected for fina...Eighteen gametophytes includingL. japonica, L. ochotensisandL. longissima, were verified with random amplified polymorphic DNA (RAPD) technique. Eighteen ten-base primers were chosen from 100 primers selected for final amplification test. Among the total of 205 bands amplified, 181 (88.3%) were polymorphic. The genetic distance among different strains ranged from 0.072 to 0.391. The dendrogram constructed by unweighted pair-group method with arithmetic (UPGMA) method showed that the female and male gametophytes of the same cell lines could be grouped in pairs respectively. It indicated that RAPD analysis could be used not only to distinguish different strains ofLaminaria, but also to distinguish male and female gametophyte within the same cell lines. There is ambiguous systematic relationship if judged merely by the present data. It seems that the use of RAPD marker is limited to elucidation of the phylogenetic relationship among the species ofLaminaria.展开更多
Objective： To identify the plants in Vitex including Vitex negundo L. , V. negundo var. cannabifolia （Sieb. et Zuee. ） Hand. -Mazz. , V. trifolia L. and V. trifolia L. var. simplicifolia Cham.. Methods： Both intra...Objective： To identify the plants in Vitex including Vitex negundo L. , V. negundo var. cannabifolia （Sieb. et Zuee. ） Hand. -Mazz. , V. trifolia L. and V. trifolia L. var. simplicifolia Cham.. Methods： Both intra- and inter-species relationships among these plants were analyzed by RAPD marker. Twenty-one samples collected from different locations in China were tested using 25 RAPD arbitrary 10- mer primers which were screened from 32 primers. Cluster analysis was conducted by Ward＇s minimumvariance method of SAS software. Results： A total of 224 bands were produced and 128 bands showed polymorphism among 21 samples. The level of polymorphism within species was 51.7%. The dendrogram constructed based on RAPD analysis showed that 21 samples can be placed in 2 groups at the level of SPRS value of 0. 3636. The first group included V. trifolia var. simplicifolia and V. trifolia. The other consisted of V. negundo and V. negundo var. cannabifolia.. At the level of SPRS value of 0. 1, 21 samples can be obviously divided into 3 groups. V. trifolia and V. trifolia var. simplicifolia were clustered into one group, V. negundo and V. negundo var. cannabifolia were separately clustered to 2 groups. Conclusion： RAPD analysis is in good agreement with the traditional plant taxonomic classification, and fundamentally identical with their origins and morphologie characteristics, which indicates that the genetic relationship among samples is related to their center of origin. These medicinal species in Vitex have their specific bands which are available to identify. These specific bands can be used as species-specific molecular markers of Fruetus Vitieis for the application of germplasm identification and classification.展开更多
限制性位点扩增多态性（restriction site amplification polymorphism，RSAP）是根据基因组内普遍存在的酶切位点来设计特殊的引物，通过简单的梯度PCR反应来产生多态性。本研究首次将RSAP标记技术成功应用于紫菜遗传多样性检测和种质...限制性位点扩增多态性（restriction site amplification polymorphism，RSAP）是根据基因组内普遍存在的酶切位点来设计特殊的引物，通过简单的梯度PCR反应来产生多态性。本研究首次将RSAP标记技术成功应用于紫菜遗传多样性检测和种质鉴定。利用所建立的适合于紫菜RSAP分析的PCR反应体系和反应条件，使用30对引物对15个紫菜系DNA进行了PCR扩增，经过3次重复验证，有12对引物能够扩增出清晰且稳定的条带。这12对引物共扩增出413条带，其中408条为多态性条带，多态性比例96％，平均每对引物产生34条多态性条带，片段大小在50-500bp之间。利用这413条带进行聚类分析，产生了这15个紫菜系的进化树。15个紫菜系在0.69相似系数水平上分为两大类：一类包括坛紫菜；另一类包括条斑紫菜和少精紫菜。选2对引物R1／R6和R3／R4所扩增的10条稳定且重复性好的条带，构建了这15个紫菜系的RSAP-DNA指纹图谱。在该指纹图谱中，每个紫菜系都有其独一无二的指纹模式，能够很容易地与其它紫菜系相区分。展开更多
文摘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 .
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Randomly amplified polymorphic DNA (RAPD) technique was applied to assess the genetic variations and phylogenetic relationships in 4 species ofPorphyra. The samples were collected from the coast of Canada, Vietnam, Zhoushan, Fujian and Qingdao in China. Amplifications with 20 primers were carried out under predetermined optimal reaction conditions (samples were first heated at 94°C for 5 min. and followed by 45 cycles of 1 min at 94°C, 1 min at 36°C, and 2 min at 72°C, then held at 72°C for 10 min). The amplified products were scored as present (1) or absent (0) for each DNA sample and an index of genetic similarity (F) was calculated by using Nei & Li’s matching coefficient method (1979). The value of (1?F) was used to quantify the genetic distances between species and construct a phylogenetic tree. The relationship indicated by the UPGMA and NJ cluster analysis on the values of the genetic distance is in good overall agreement with classical taxonomy. The obvious differences between natural and cultivated population ofP. haitanensi suggest that variation or hybridization with other species occurred during the culture.
文摘Germplasm collection is important to preserve and maximize genetic diversity for germplasm conservation.Tsuga dumosa(D.Don)Eichler in Engler & Prantl.and T.chinensis var.forrestii(Downie)Silba germplasm was collected from three localities in China:Mt.Yulong,Wenfeng Temple and Mt.Dishiergu,Yunnan Province.Accessions were identified based on morphological characters and RAPD markers.The shapes of the apices and margins of needles were examined,and the length and width of needles,cones and seeds from accessions of mature plants were used to compare the morphological differences and to identify the germplasm.Molecular markers generated by randomly amplified polymorphic DNA(RAPD)were also used to characterize the taxa.Although the clustering based on RAPD markers was inconsistent with the morphological characters of the needles,based on the overall morphological characters and on RAPD markers,the accessions from Mt.Yulong and Wenfeng Temple were identified as T.chinensis var.forrestii,and those from Mt.Dishiergu identified as T.dumosa.Taxonomic identification of the accessions was made based on morphology and by RAPD markers concurred.The results indicate that the shapes of the apices and margins of needles particularly from young plants could not be used as a possible key to identify T.dumosa and T.chinensis var.forrestii.Fig 6,Tab 3,Ref
文摘Eighteen gametophytes includingL. japonica, L. ochotensisandL. longissima, were verified with random amplified polymorphic DNA (RAPD) technique. Eighteen ten-base primers were chosen from 100 primers selected for final amplification test. Among the total of 205 bands amplified, 181 (88.3%) were polymorphic. The genetic distance among different strains ranged from 0.072 to 0.391. The dendrogram constructed by unweighted pair-group method with arithmetic (UPGMA) method showed that the female and male gametophytes of the same cell lines could be grouped in pairs respectively. It indicated that RAPD analysis could be used not only to distinguish different strains ofLaminaria, but also to distinguish male and female gametophyte within the same cell lines. There is ambiguous systematic relationship if judged merely by the present data. It seems that the use of RAPD marker is limited to elucidation of the phylogenetic relationship among the species ofLaminaria.
基金Supported by specific TCM modernization fund.Shanghai Commission of Sci &.Tech (No.04D19810)
文摘Objective： To identify the plants in Vitex including Vitex negundo L. , V. negundo var. cannabifolia （Sieb. et Zuee. ） Hand. -Mazz. , V. trifolia L. and V. trifolia L. var. simplicifolia Cham.. Methods： Both intra- and inter-species relationships among these plants were analyzed by RAPD marker. Twenty-one samples collected from different locations in China were tested using 25 RAPD arbitrary 10- mer primers which were screened from 32 primers. Cluster analysis was conducted by Ward＇s minimumvariance method of SAS software. Results： A total of 224 bands were produced and 128 bands showed polymorphism among 21 samples. The level of polymorphism within species was 51.7%. The dendrogram constructed based on RAPD analysis showed that 21 samples can be placed in 2 groups at the level of SPRS value of 0. 3636. The first group included V. trifolia var. simplicifolia and V. trifolia. The other consisted of V. negundo and V. negundo var. cannabifolia.. At the level of SPRS value of 0. 1, 21 samples can be obviously divided into 3 groups. V. trifolia and V. trifolia var. simplicifolia were clustered into one group, V. negundo and V. negundo var. cannabifolia were separately clustered to 2 groups. Conclusion： RAPD analysis is in good agreement with the traditional plant taxonomic classification, and fundamentally identical with their origins and morphologie characteristics, which indicates that the genetic relationship among samples is related to their center of origin. These medicinal species in Vitex have their specific bands which are available to identify. These specific bands can be used as species-specific molecular markers of Fruetus Vitieis for the application of germplasm identification and classification.
文摘限制性位点扩增多态性（restriction site amplification polymorphism，RSAP）是根据基因组内普遍存在的酶切位点来设计特殊的引物，通过简单的梯度PCR反应来产生多态性。本研究首次将RSAP标记技术成功应用于紫菜遗传多样性检测和种质鉴定。利用所建立的适合于紫菜RSAP分析的PCR反应体系和反应条件，使用30对引物对15个紫菜系DNA进行了PCR扩增，经过3次重复验证，有12对引物能够扩增出清晰且稳定的条带。这12对引物共扩增出413条带，其中408条为多态性条带，多态性比例96％，平均每对引物产生34条多态性条带，片段大小在50-500bp之间。利用这413条带进行聚类分析，产生了这15个紫菜系的进化树。15个紫菜系在0.69相似系数水平上分为两大类：一类包括坛紫菜；另一类包括条斑紫菜和少精紫菜。选2对引物R1／R6和R3／R4所扩增的10条稳定且重复性好的条带，构建了这15个紫菜系的RSAP-DNA指纹图谱。在该指纹图谱中，每个紫菜系都有其独一无二的指纹模式，能够很容易地与其它紫菜系相区分。
