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Development of Sequence Characterized Amplified Region (SCAR) Primers for the Detection of Resistance to Sporisorium reiliana in Maize 被引量:4
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作者 SHI Hong-liang LI Xin-hai +5 位作者 ZHANG De-gui XIE Chuan-xiao HAO Zhuan-fang LI Ming-shun PAN Guang-tang ZHANG Sbi-huang 《Agricultural Sciences in China》 CAS CSCD 2009年第8期910-919,共10页
Head smut of maize (Zea mays L.), which was caused by Sporisorium reiliana, occurred in most of the maize growing areas of the world. The purpose of this study was to develop SCAR markers for map-based cloning of re... Head smut of maize (Zea mays L.), which was caused by Sporisorium reiliana, occurred in most of the maize growing areas of the world. The purpose of this study was to develop SCAR markers for map-based cloning of resistance genes and MAS. Two sets of BC3 progenies, one (BC3Q) derived from the cross Qi319 (resistance)×Huangzao 4 (susceptible), the other (BC3M) from Mol7 (resistance)× Huangzao 4 (susceptible), were generated. Huangzao 4 was the recurrent parent in both progenies. A combination of BSA (bulked segregant analysis) with AFLP (amplified fragment length polymorphism) method was applied to map the genes involving the resistance to S. reiliana, and corresponding resistant and susceptible bulks and their parental lines were used for screening polymorphic AFLP primer pairs. One fragment of PI3M61-152 was converted into SCAR (sequence charactered amplified fragment) marker S130. The marker was mapped at chromosome bin 2.09, the interval of a major QTL region previously reported to contribute to S. reiliana resistance. Furthermore, S130 was highly and facilitate map-based cloni associated with resistance to S. reiliana, and could be useful for marker-assisted selection ng of resistance genes. 展开更多
关键词 maize (Zea mays L.) Sporisorium reiliana bulked segregant analysis amplified fragment length polymorphism sequence characterized amplified region
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Genetic Mapping of Laminaria japonica and L. Iongissima Using Amplified Fragment Length Polymorphism Markers in a "Two-Way Pseudo- Testcross" Strategy 被引量:8
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作者 Yuhui Li Yingxia Yang +3 位作者 Jidong Liu Xiuliang Wang Tianxiang Gao Delin Duan 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2007年第3期392-400,共9页
With a "two-way pseudo-testcross" mapping strategy, we applied the amplified fragment length polymorphism (AFLP) markers to construct two moderate density genetic linkage maps for Laminaria. The linkage maps were ... With a "two-way pseudo-testcross" mapping strategy, we applied the amplified fragment length polymorphism (AFLP) markers to construct two moderate density genetic linkage maps for Laminaria. The linkage maps were generated from the 60 progenies of the F1 cross family (Laminaria iongissima Aresch. × L. Japonica Miyabe) with twenty pairs of primer combinations. Of the 333 polymorphic loci scored in 60 progenies, 173 segregated in a 1:1 ratio, corresponding to DNA polymorphisms heterozygous in a single parent, and the other 58 loci existing in both parents followed a 3:1 Mendelian segregation ratio. Among the loci with 1:1 segregating ratios, 79 loci were ordered in 14 linkage groups (648.6 cM) of the paternal map, and 72 loci were ordered in 14 linkage groups (601.9 cM) of the maternal map. The average density of loci was approximately 1 per 8 cM. To Investigate the homologies between two parental maps, we used 58 loci segregated 3:1 for further analysis, and deduced one homologous linkage group. The linkage data developed in these maps will be useful for detecting loci-controlling commercially important traits for Laminaria. 展开更多
关键词 amplified fragment length polymorphism makers genetic linkage map Laminaria japonica Laminaria longissima segregation ratio two-way pseudo-testcross strategy
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Discussion on AFLP Molecular Markers in Piper methysticum and Pepper 被引量:2
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作者 施江 辛莉 +1 位作者 杨彦 郑学勤 《Agricultural Science & Technology》 CAS 2007年第3期5-10,19,共7页
The aim of the research was to discuss the genetic relationships between Piper methysticum, Pepper and other wild species in Pepper genus. DNA was extracted from leaves which belonged to 28 germplasms including 6 mate... The aim of the research was to discuss the genetic relationships between Piper methysticum, Pepper and other wild species in Pepper genus. DNA was extracted from leaves which belonged to 28 germplasms including 6 materials of P. methysticum, 21 maerials of cultivated and wild Pepper, 1 material of Peperomia pellucida belonged to different genus. Premiers with good band-type and high polymorphism and resolution were selected from 64 pairs of primers for AFLP amplification and the clustering analysis was conducted with MVSP3.13f software. 191 bands were amplified by 4 pairs of premiers, 189 of which had polymorphism, being 98.6%. 28 germplasms were classified into 6 different groups at the genetic similarity coefficient of 0.52 by silver staining AFLP, in which 6 materials of Piper methysticum were clustered into a single group, indicating that P. methysticum belonged to Pepper family of Pepper genus but were distantly related to the others. The research provided the basis for selecting rootstocks for P. methysticum graft, molecular identification of P. methysticum and the fingerprint construction of P. methysticum. 展开更多
关键词 Piper methysticum PEPPER amplified fragment length polymorphism
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Discriminating ability of molecular markers and morphological characterization in the establishment of genetic relationships in cultivated genotypes of almond and related wild species 被引量:2
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作者 Karim Sorkheh Behrouz Shiran +7 位作者 Soghra Kiani Nazanin Amirbakhtiar Sadegh Mousavi Vahid Rouhi Shahram Mohammady-D Thomas M. Gradziel Lyudmyla V. Malysheva-Otto Pedro Martinez-Gomez 《Journal of Forestry Research》 SCIE CAS CSCD 2009年第3期183-194,共12页
A total 23 morphological traits, 19 AFLP-primer combinations, 80 RAPD primers and 32 SSR primer pair were used to compare the informativeness and efficiency of random amplified polymorphic DNA (RAPD), amplified frag... A total 23 morphological traits, 19 AFLP-primer combinations, 80 RAPD primers and 32 SSR primer pair were used to compare the informativeness and efficiency of random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers in establishing genetic relationships among 29 almond cultivars and three related wild species. SSRs presented a high level of polymorphism and greater information content, as assessed by the expected hetrozygosity, compared to AFLPs and RAPDs. The lowest values of expected hetrozygosity were obtained for AFLPs; however AFLPs showed the highest efficiency, owing to their capacity to reveal large numbers of bands per reaction, which led to high values for various types of indices of diversity. All the three techniques discriminated almond genotypes very effectively, except that SSRs failed to discriminate between 'Monagha' and 'Sefied' almond genotypes. The correlation coefficients of similarity were statistically significant for all the three marker systems, but were lower for the SSR data than for RAPDs and AFLPs. For all the markers, high similarity in dendrogram topologies was obtained, although some differences were observed. All the dendrograms, including that obtained by the combined use of all the marker data, reflect relationships for most of cultivars according to their geographic diffusion. AMOVA detected more variation among cultivated and related wild species of almond within each geographic group. Bootstrap analysis revealed that the number of markers used was sufficient for reliable estimation of genetic similarity and for meaningful comparisons of marker types. 展开更多
关键词 amplified fragment length polymorphisms (AFLPs) Random amplified Potymorphic DNA (RAPDs) Simple-SequenceRepeats (SSRs) germplasm genetic relationships breeding prunus dulcis
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Discriminating ability of molecular markers and morphological characterization in the establishment of genetic relationships in cultivated genotypes of almond and related wild species
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作者 Karim Sorkheh Behrouz Shiran +7 位作者 Soghra Kiani Nazanin Amirbakhtiar Sadegh Mousavi Vahid Rouhi Shahram Mohammady-D Thomas M.Gradziel Lyudmyla V.Malysheva-Otto Pedro Martínez-Gómez 《Journal of Forestry Research》 SCIE CAS CSCD 2009年第A3期183-194,285,共13页
A total 23 morphological traits, 19 AFLP-primer combinations, 80 RAPD primers and 32 SSR primer pair were used to compare the informativeness and efficiency of random amplified polymorphic DNA (RAPD), amplified fragme... A total 23 morphological traits, 19 AFLP-primer combinations, 80 RAPD primers and 32 SSR primer pair were used to compare the informativeness and efficiency of random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers in establishing genetic relationships among 29 almond cultivars and three related wild species. SSRs presented a high level of polymorphism and greater information content, as assessed by the expected hetrozygosity, compared to AFLPs and RAPDs. The lowest values of expected hetrozygosity were obtained for AFLPs; however AFLPs showed the highest efficiency, owing to their capacity to reveal large numbers of bands per reaction, which led to high values for various types of indices of diversity. All the three techniques discriminated almond genotypes very effectively, except that SSRs failed to discriminate between ‘Monagha’ and ‘Sefied’ almond genotypes. The correlation coefficients of similarity were statistically significant for all the three marker systems, but were lower for the SSR data than for RAPDs and AFLPs. For all the markers, high similarity in dendrogram topologies was obtained, although some differences were observed. All the dendrograms, including that obtained by the combined use of all the marker data, reflect relationships for most of cultivars according to their geographic diffusion. AMOVA detected more variation among cultivated and related wild species of almond within each geographic group. Bootstrap analysis revealed that the number of markers used was sufficient for reliable estimation of genetic similarity and for meaningful comparisons of marker types. 展开更多
关键词 amplified fragment length polymorphisms (AFLPs) Random amplified Polymorphic DNA (RAPDs) Simple-Sequence Repeats (SSRs) germplasm genetic relationships breeding prunus dulcis
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Isolation and characterization of the AGAMOUS homologous gene NTAG in Chinese narcissus (Narcissus tazetta var. chinensis Roem) 被引量:7
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作者 Wang Zheng-ke Gao Jian +1 位作者 Li Lu-bin Peng Zhen-hua 《Forestry Studies in China》 CAS 2006年第1期21-26,共6页
Amaryllidaceae, a monocot plant family, consists of many important ornamental bulb flower species. Chinese narcissus (Narcissus tazetta var. chinensis Roem), its flowers developed at high temperatures and bloomed at... Amaryllidaceae, a monocot plant family, consists of many important ornamental bulb flower species. Chinese narcissus (Narcissus tazetta var. chinensis Roem), its flowers developed at high temperatures and bloomed at lower temperatures during the Chinese Spring Festival, is a traditional Chinese flower with high economic and ornamental value. To study its flower development, a full length cDNA containing MADS box domain from narcissus was isolated by a reverse transcription polymerase chain reaction (RT-PCR) with degenerate oligo-nucleotide primers derived from a conserved MADS- and K-box domain sequence. The 5' and the 3' regions of the gene were amplified using the PCR protocol for the rapid amplification of cDNA ends (RACE). The 690 bp open reading frame encodes 230 amino acid residues. A comparison of the deduced amino acid sequence of NTAG with the sequence of other MADS box proteins showed 91.3% amino acid identities with HAG (Hyacinthus orientalis). Sequence analysis and alignment showed significant similarity with other AG homologues. RNA blot analysis indicated that the narcissus NTAG gene was expressed only in reproductive organs, especially in stamens and carpels. These results indicated that the NTAG gene was an AG homologue and that the AG genes appeared to be structurally and functionally conserved between dicots and monocots. 展开更多
关键词 NTAG (Narcissus tazetta var. chinensis Roem AGAMOUS) cDNA amplified fragment length polymorphism Chinese narcissus
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Genetic Relationships among Prunus mume var. pendula Using AFLP Markers 被引量:3
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作者 Ming Jun Zhang Qixiang +3 位作者 Ru Guangxin Mao Qingshan Yan Xiaolan Lan Yanping 《Forestry Studies in China》 CAS 2003年第4期26-30,共5页
Genetic relationships among Prunus mume var. pendula were studied by using AFLP markers. 18 accessions representing 14 cultivars of Prunus mume var. pendula were selected from the germplasm collection at the Research ... Genetic relationships among Prunus mume var. pendula were studied by using AFLP markers. 18 accessions representing 14 cultivars of Prunus mume var. pendula were selected from the germplasm collection at the Research Center of China Mei Flower. Seven Mse I-EcoR I AFLP primer combinations revealed 450 legible bands, and 269 of which were polymorphic markers. A similarity matrix was prepared using the simple matching coefficient of similarity and Neis (72) distance coefficient. A UPGMA dendrogram demonstrated the genetic relationships of the cultivars. The information given by AFLP markers was basically consistent with the morphological classification and the evolutionary history of the morphotypes, and roughly supported the new revised classification system for Chinese Mei Cultivars. But there were still several exceptions: 1) the Guhong Chuizhi inserted between the Tiaoxue Chuizhi and the Danfen Chuizhi; 2) the Wufu Chuizhi kept off the Pink Pendant Form, and the Moshan Chuizhi was removed from Viridiflora Pendant Form; 3) the Danbi Chuizhi and the Shuangbi Chuizhi of Viridiflora Pendant Form got together well but fell within the Pink Pendant Form. 展开更多
关键词 Prunus mume Sieb. et Zucc. amplified fragment length polymorphism (AFLP) Prunus mume var. pendula (Pendulous Mei Group) genetic relationship
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Development of a SCAR marker for male gametophyte of Gracilariopsis lemaneiformis based on AFLP technique 被引量:1
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作者 周伟 丁弘叶 +2 位作者 隋正红 王钟霞 王津果 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2014年第3期522-526,共5页
The red alga Gracilariopsis lemaneiformis(Bory) is an economically valuable macroalgae. As a means to identify the sex of immature Gracilariopsis lemaneiformis, the amplifi ed fragment length polymorphism(AFLP) techni... The red alga Gracilariopsis lemaneiformis(Bory) is an economically valuable macroalgae. As a means to identify the sex of immature Gracilariopsis lemaneiformis, the amplifi ed fragment length polymorphism(AFLP) technique was used to search for possible sex- or phase-related markers in male gametophytes, female gametophytes, and tetrasporophytes, respectively. Seven AFLP selective amplifi cation primers were used in this study. The primer combination E-TG/M-CCA detected a specifi c band linked to male gametophytes. The DNA fragment was recovered and a 402-bp fragment was sequenced. However, no DNA sequence match was found in public databases. Sequence characterized amplifi ed region(SCAR) primers were designed from the sequence to test the repeatability of the relationship to the sex, using 69 male gametophytes, 139 female gametophytes, and 47 tetrasporophytes. The test results demonstrate a good linkage and repeatability of the SCAR marker to sex. The SCAR primers developed in this study could reduce the time required for sex identifi cation of Gracilariopsis lemaneiformis by four to six months. This can reduce both the time investment and number of specimens required in breeding experiments. 展开更多
关键词 Gracilariopsis lemaneiformis amplified fragment length polymorphism (AFLP) male gametophyte sequence characterized amplified region (SCAR) marker
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Range-wide genetic diversity in natural populations of Larix principis-rupprechtii Mayr. 被引量:1
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作者 Xiaoyan Di Xiangxiang Meng Mengben Wang 《Journal of Forestry Research》 SCIE CAS CSCD 2021年第1期319-327,共9页
Prince Rupprecht’s larch(Larix principis-rupprechtii Mayr.),a deciduous conifer,widely grows in middle and high elevations of Northern China.Its natural distribution has sharply decreased and has become fragmented,wh... Prince Rupprecht’s larch(Larix principis-rupprechtii Mayr.),a deciduous conifer,widely grows in middle and high elevations of Northern China.Its natural distribution has sharply decreased and has become fragmented,which may have resulted in the loss of genetic variation.In this study,ten natural populations across the entire range of this species were analyzed using amplifi ed fragment length polymorphism markers.A total of 309 loci were detected from 225 individuals of these populations,of which 261(84.5%)were polymorphic.At the species level,the genetic diversity was high(average of the Nei’s genetic diversity H e=0.2602,and Shannon’s information index I=0.3967).The results of molecular variance analysis showed that 90.71%of the genetic diversity occurred within populations.The genetic diff erentiation among populations was moderate as a whole(F ST=0.0929,G ST=0.1510),which is consistent with the moderate level of gene fl ow among populations(N m=2.8116).Based on the unweighted pair group method with arithmetic mean and STRU CTU RE analysis,these populations were grouped into three genetically distinct clusters.The degree of inter-population diff erentiation(G ST=0.1338)for the south group was larger than that for the north group(G ST=0.0915).There was a signifi cant correlation between genetic distance and geographic distance across the species range(r=0.316,P<0.05).Genetic diversity was signifi-cantly associated with longitude but not elevation or climatic factors.The populations with high genetic diversity from each cluster are therefore recommended for future conservation and management of this species. 展开更多
关键词 Larix principis-rupprechtii Amplifi ed fragment length polymorphism markers(AFLP) Genetic diversity Environmental factors
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Analysis of the Relationship between Genome Diversity and Adult Survive Rate of Botryllus Schlosseri by AFLP
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作者 FENGXiao-rong ZHUJian-zhong +2 位作者 DENGFeng-jiao JacobDouek BaruchRinkevich 《Journal of Nanjing Medical University》 2004年第2期67-77,共11页
Objective: The self cross colonial prochordate, Botryllus schlosseri ( B.schlosseri ) occupy a key phylogenetic position in the evolution of vertebrates. To clarify the relationship of genome diversity and survive rat... Objective: The self cross colonial prochordate, Botryllus schlosseri ( B.schlosseri ) occupy a key phylogenetic position in the evolution of vertebrates. To clarify the relationship of genome diversity and survive rate, five generations of B. schlosseri was investigated by amplified fragment length polymorphism (AFLP). Methods: AFLP markers are extremely sensitive to even small sequence variation, using PCR and high resolution electrophoresis to examine restriction fragments. Results: AFLP polymorphism was high in the parent and lower in its F1, F2, F3 and F4. Each primer combination generated from 80 to more than 120 bands, of which average 25.85% polymorphic loci in parent, 15.79% polymorphic among F1, 9.16% and 5.58% in F2, F3. The AFLP markers were transmitted from F1 to F2, F3 and F4 and inherited, segregated in expected Mendelian ratio. However, some of the markers were lost in F2, F3 and F4 while it disappeared in their mother. In addition, gene mutation new loci and lost loci among F1, F2, F3 and F4 were observed. These special fragments were cloned and sequenced. Then, the genomic DNA was analyzed by Southern hybridization with the probes from these specific fragments and the mechanism of gene mutation was clarified. Conclusion: These results suggest that there are high frequency of polymorphic loci and mutation in genome of B. schlosseri. Gene deletion or low diversity may be the reason for high rate of death of the offspring of inbred laboratory reared strains. 展开更多
关键词 Botryllus schlosseri genome diversity MUTATION amplified fragment length polymorphism POLYMORPHISM survive rate
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Impact of Crop Rotation on Pathotype and Genetic Structure of Phythophthora sojae in Fields
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作者 Zhao Li-ming Li Shuang +6 位作者 Sui Zhe Huang Jing Chen Qiu-ming Suo Bing Ding Jun-jie Liu Wei-ting Wen Jing-zhi 《Journal of Northeast Agricultural University(English Edition)》 CAS 2016年第2期1-11,共11页
To estimate the impact of crop rotation on the pathotype and genetic structure of Phythophthora sojae in fields, 372 isolates of P. sojae were obtained from long-term localisation experimental fields in Heilongjiang P... To estimate the impact of crop rotation on the pathotype and genetic structure of Phythophthora sojae in fields, 372 isolates of P. sojae were obtained from long-term localisation experimental fields in Heilongjiang Province of China. The hypocotyl inoculation method was used to characterize the virulence of P. sojae on 13 differential cultivars, and the amplified fragment length polymorphism(AFLP) technique was used to analyze difference in the genetic structure of P. sojae. The results indicated that an abundant diversity of genetic structures and pathotypes of P. sojae, a more uniform distribution of pathotypes and less dominance of pathotypes occurred in corn-soybean and wheat-soybean rotation fields than in a continuous soybean mono-cropping field. These findings suggested that P. sojae did not easily become the dominant race in rotation fields, which maintain disease resistance in soybean varieties. Therefore, the results of this study suggested that Phytophthora stem and root rot of soybeans could be effectively controlled by rotating soybeans with non-host crops of corn and wheat. 展开更多
关键词 Phytophthora sojae pathotype composition genetic structure crop rotation amplified fragment length polymorphism(AFLP)
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Molecular Study of the Genetic Variability of Pumpkins Landraces from Brazilian Amazon
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作者 Lúcia Helena Pinheiro Martins Maria Teresa Gomes Lopes +4 位作者 Hiroshi Noda Pedro de Queiroz Costa Neto Ayrton Luiz Urizzi Martins Fabíola Viana de Almeida Liane Cristine Reboucas Demosthenes 《Agricultural Sciences》 2018年第4期511-523,共13页
The Cucurbita maxima Duchesne is a vegetable crop plant cultivated and maintained by traditional Amazon communities, Brazil. The situation is worsened by the possibility of disappearance of local populations and genet... The Cucurbita maxima Duchesne is a vegetable crop plant cultivated and maintained by traditional Amazon communities, Brazil. The situation is worsened by the possibility of disappearance of local populations and genetic variability of this specie, taking into account the today changes promoted in family farming. The aim of this study was to estimate the current levels of genetic variability of local cultivars through the use of molecular markers (Amplified Fragment Length Polymorphism—AFLP). We chose to collect in two distinct micro regions in order to identify possible influences of geographic isolation and different levels of market requirements in the conservation of the genetic variability of the C. maxima. For the molecular analysis, bulk samples of fresh leaves of 15 plants/half-sibling family were collected in paper bags. There were 34 samples from the half-sib families. The analysis of the results half-sib obtained by methods of estimation of genetic variation by molecular markers shows that the forms of cultivation and management adopted by family farmers maintain the identities of the local/landraces (native cultivars) and, at the same time, the levels of diversity for the assurance of adaptability macro-environmental. 展开更多
关键词 Crop Genetic Variability Molecular Marker amplified fragment length Polymorphism Cucurbita maxima Duchesne
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Use of the Metabolomics Approach to Characterize Chinese Medicinal Material Huangqi 被引量:19
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作者 Li-Xin Duan Tian-Lu Chen +7 位作者 Ming Li Min Chen Ye-Qing Zhou Guang-Hong Cui Ai-Hua Zhao Wei Jia Lu-Qi Huang Xiaoquan Qi 《Molecular Plant》 SCIE CAS CSCD 2012年第2期376-386,共11页
Integration of the genetic and metabolic fingerprinting can provide a new approach to differentiate similar Traditional Chinese Medical (TCM) materials. Two leguminous plants, Mojia Huangqi and Menggu Huangqi, are i... Integration of the genetic and metabolic fingerprinting can provide a new approach to differentiate similar Traditional Chinese Medical (TCM) materials. Two leguminous plants, Mojia Huangqi and Menggu Huangqi, are important medical herbs and share great similarities in morphology, chemical constituent, and genomic DNA sequence. The taxonomy of Mojia Huangqi and Menggu Huangqi has been debated for more than 50 years and discrimination of TCM materials directly affects the pharmacological and clinical effects. AFLP based genetic fingerprinting and GC-TOF/MS-based meta- bolic fingerprinting were used to successfully discriminate the two species. The results of AFLP supported the opinion that Menggu Huangqi was a variant of Mojia Huangqi. The metabolic fingerprinting showed growth locations have greater impacts on the metabolite composition and quantity than the genotypes (cultivated versus wild) in Menggu Huangqi. The difference of some soluble sugars, fatty acids, proline, and polyamine reflected plant adaptation to different growth environments. Using multivariate and univariate statistical analysis, three AFLP markers and eight metabolites were identified as candidate DNA and metabolic markers to distinguish the two herb materials. The correlation network between AFLP markers and metabolites revealed a complex correlation network, which indicated the special metabolic pathways and the regulation networks of Huangqi. 展开更多
关键词 Astragalus membranaceus metabolic fingerprinting amplified fragment length polymorphisms gas chromatography time-of-flight mass spectrometry.
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A High-throughput Genomic Tool: Diversity Array Technology Complementary for Rice Genotyping 被引量:4
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作者 Yong Xie Kenneth McNally +2 位作者 Cheng-Yun Li Hei Leung You-Yong Zhu 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2006年第9期1069-1076,共8页
Diversity array technology (DART^TM) was a genotyping tool characterized gel-independent and high throughput. The main purpose of present study is to validate DArT for rice (Oryza sativa L.)genotyping in a high th... Diversity array technology (DART^TM) was a genotyping tool characterized gel-independent and high throughput. The main purpose of present study is to validate DArT for rice (Oryza sativa L.)genotyping in a high throughput manner. Technically, the main objective was to generate a rice general purpose gene pool, and optimize this genomic tool in order to evaluate rice germplasm genetic diversity. To achieve this, firstly, a generalpurpose DArT array was developed. Ten representatives from 24 varieties were hybridized with the general-purpose array to determine the informativeness of the clones printed on the array. The informative 1 152 clones were re-arrayed on a slide and used to fingerprint 17 of 24 germplasms. Hybridizing targets prepared from the germplasm to be assayed to the DNA array gave DNA fingerprints of germplasms. Raw data were normalized and transformed into binary data, which were then analyzed by using NTSYSpc (Numerical taxonomy system for cluster and ordination analysis, v. 2.02j) software package. The graphically displayed dendrogram derived from the array experimental data was matched with simple sequence repeats genotyping outline and varieties' pedigree deviation of the different varieties. Considering DArT is a sequence-independent genotyping approach, it will be applied in studies of the genetic diversity and the gene mapping of diverse of organisms, especially for those crops with less-developed molecular markers. 展开更多
关键词 amplified fragment length polymorphisms diversity array technology genome complexity reduction microarray rice genotyping
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Loss of Genetic Diversity of Domesticated Panax notoginseng F H Chen as Evidenced by ITS Sequence and AFLP Polymorphism: A Comparative Study with P. stipuleanatus H T Tsai et K M Feng 被引量:15
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作者 Shi-LiangZHOU Gao-MingXIONG +1 位作者 Zhong-YiLI JunWEN 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2005年第1期107-115,共9页
In the present study, we evaluated the genetic diversity of Panax notoginseng F H Chen, a domesticated species, and P. stipuleanatus H T Tsai et K M Feng, an endangered wild species in southeastern Yunnan and adjacent... In the present study, we evaluated the genetic diversity of Panax notoginseng F H Chen, a domesticated species, and P. stipuleanatus H T Tsai et K M Feng, an endangered wild species in southeastern Yunnan and adjacent areas in Vietnam, using sequences of the internal transcribed spacer (ITS) regions of nuclear ribosomal DNA and amplified fragment length polymorphism (AFLP) markers. Twenty-four accessions from three plantations of P. notoginseng and 51 samples from eight populations of P. stipuleanatus were assayed. A total of 694 bp of partial sequences of 18S, ITS 1, 5.8S, ITS2, and partial sequences of 26S were obtained. No sequence variation was detected within P. notoginseng and nine sites (1.30%) were variable in P. stipuleanatus. Two-thirds of the variable sites were found between Langqiao and other populations. In P. notoginseng, four pairs of AFLP primer combinations generated 312 bands, of which 240 (76.9%) were polymorphic and 60.15% of the polymorphisms were harbored within plantations. Approximately 41.0% and 66.9% of bands were polymorphic in population D7 and 5589, respectively. In P.stipuleanatus, the same four primer combinations produced 346 bands, of which 334 (96.5%) were polymorphic and approximately 62.14% of polymorphisms were maintained within populations. Considerable variations were observed. The percentage of polymorphic bands ranged from 50.2% to 84.9% and the average over populations was 70.9%. Cluster analysis did not show correlation of genetic differentiation with the distinctive leaf morphology of P. stipuleanatus (i.e. one form with bipinnatifid leaflets and the other with undivided leaflets). Because over 40% of genetic variations were maintained among populations and because of the very restricted distribution of P. stipuleanatus, all natural populations of this species should be conserved in situ. Considering that there are variations in P. notoginseng within and among plantations, we suggest establishing a genetic resource conservation garden or reintroducing P. notoginseng into its native habitats in southwestern China. Such reintroduction should be carefully executed after large-scale screening of genetic variation within the species. 展开更多
关键词 amplified fragment length polymorphism ARALIACEAE conservation genetics internal transc-ribed spacer Panax notoginseng F H Chen P.stipuleanatus H T Tsaiet KM Feng.
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Drug-resistant gene based genotyping for Acinetobacter baumannii in tracing epidemiological events and for clinical treatment within nosocomial settings 被引量:18
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作者 JIN Hui XU Xiao-min +2 位作者 MI Zu-huang MOU Yi LIU Pei 《Chinese Medical Journal》 SCIE CAS CSCD 2009年第3期301-306,共6页
Background Acinetobacter baumannfi has emerged as an important pathogen related to serious infections and nosocomial outbreaks around the world. However, of the frequently used methods, pulsed-field gel electrophores... Background Acinetobacter baumannfi has emerged as an important pathogen related to serious infections and nosocomial outbreaks around the world. However, of the frequently used methods, pulsed-field gel electrophoresis (PFGE) and amplified fragment length polymorphism (AFLP) in Acinetobacter baumannfi genotyping lack the direct molecular proof of drug resistance. This study was conducted to establish a typing method based on drug resistant gene identification in contrast to traditional PFGE and AFLP in the period of nosocomial epidemic or outbreak. Methods From January 2005 to October 2005, twenty-seven strains of Acinetobacter species from Intensive Care Units, the Second Affiliated Hospital in Ningbo were isolated, including both epidemic and sporadic events. Susceptibility test, PFGE, AFLP and drug resistance gene typing (DRGT) were carried out to confirm the drug resistance and analyze the genotyping, respectively. PFGE was used as a reference to evaluate the typeability of DRGT and AFLP. Results Twenty-seven strains of Acinetobacter displayed multiple antibiotic resistance and drug resistant genes, and β-1actamase genes were detected in 85.2% strains. The result of DRGT was comparable to PFGE in Acinetobacter strains with different drug resistance though a little difference existed, and even suggested a molecular evolution course of different drug-resistant strains. AFLP showed great polymorphism between strains and had weak ability in distinguishing the drug resistance. Conclusion Compared to AFLP and PFGE, DRGT is useful to analyze localized molecular epidemiology of nosocomial infections and outbreaks, which would benefit clinical diagnosis and therapy. 展开更多
关键词 Acinetobacter baumannii drug-resistance gene typing pulsed-field gel electrophoresis amplified fragment length polymorphism fingerprinting
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Cloning and Characterization of the Microspore Development-Related Gene BcMF2 in Chinese Cabbage Pak-Choi (Brassica campestris L. ssp. chinensis Makino) 被引量:9
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作者 Yong-QinWANG Wan-ZhiYE +3 位作者 Jia-ShuCAO Xiao-LinYU XunXIANG GangLU 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2005年第7期863-872,共10页
: For the sake of providing some important information relevant to the study of the molecular mechanism of genic male sterility in plants, gene differential expression in flower buds at different developmental stages,... : For the sake of providing some important information relevant to the study of the molecular mechanism of genic male sterility in plants, gene differential expression in flower buds at different developmental stages, as well as in rosette leaves, florescence leaves, and scapes was analyzed using cDNA amplified fragment length polymorphism (cDNA-AFLP) in the genic male sterile A and fertile B line of Chinese cabbage pak-choi. Following amplification of 125 pairs of primer combinations, 11 differential fragments were obtained, of which eight were from the B line and the other three were from the A line. Of 11 differential fragments, four were verified by Northern hybridization that were expressed preferentially in fertile flower buds. Results of GenBank BLAST showed that one fragment was with unknown function, whereas the other fragments have strong nucleotide sequence similarities with the polygalacturonase (PG) gene, the pectinesterase (PE) gene, and the polygalacturonase inhibitory protein (PGIP4) gene. Only full-length cDNA from the differential fragment BcMF-A18T16-1 was amplified by rapid amplification of cDNA ends (RACE) and Northern analysis showed that this fragment was expressed only in medium and large-sized flower buds of the B line. The full-length cDNA, designated as BcMF2 (Brassica campestris Male Fertile 2), was 1 485 bp long and was composed of a 1 263-bp open reading frame, which had 83% nucleotide similarity to a PG gene from Arabidopsis encoding polygalacturonase. Analysis of the basic structure of the protein revealed that it had one polygalacturonase active site (RVTCGPGHGLSVGS) at 256th site of amino acids and was classified as being a member of family 28 of the glycosyl hydrolases. The role of the BcMF2 gene on microspore development is discussed in the present paper. 展开更多
关键词 BcMF2 (Brassica campestris Male Fertile 2) gene cDNA amplified fragment length polymorphism Chinese cabbage pak-choi genic male sterility
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Analysis of genetic diversity in banana cultivars(Musa cvs.) from the South of Oman using AFLP markers and classification by phylogenetic,hierarchical clustering and principal component analyses 被引量:2
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作者 Umezuruike Linus OPARA Dan JACOBSON Nadiya Abubakar AL-SAADY 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2010年第5期332-341,共10页
Banana is an important crop grown in Oman and there is a dearth of information on its genetic diversity to assist in crop breeding and improvement programs.This study employed amplified fragment length polymorphism(AF... Banana is an important crop grown in Oman and there is a dearth of information on its genetic diversity to assist in crop breeding and improvement programs.This study employed amplified fragment length polymorphism(AFLP) to investigate the genetic variation in local banana cultivars from the southern region of Oman.Using 12 primer combinations,a total of 1094 bands were scored,of which 1012 were polymorphic.Eighty-two unique markers were identified,which revealed the distinct separation of the seven cultivars.The results obtained show that AFLP can be used to differentiate the banana cultivars.Further classification by phylogenetic,hierarchical clustering and principal component analyses showed significant differences between the clusters found with molecular markers and those clusters created by previous studies using morphological analysis.Based on the analytical results,a consensus dendrogram of the banana cultivars is presented. 展开更多
关键词 MUSA Genetic diversity amplified fragment length polymorphism(AFLP) PHYLOGENETICS Principal component analysis(PCA) Hierarchical clustering analysis(HCA) Oman
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Assessment of Genetic Stability Among In Vitro Plants of Arachis retusa Using RAPD and AFLP Markers for Germplasm Preservation 被引量:1
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作者 Rachel Fatima Gagliardi Luiz Ricardo Hanai +5 位作者 Georgia Pacheco Carlos Alberto Oliveira Leonardo Alves Carneiro José Francisco Montenegro Valls Elisabeth Mansur Maria Lucia Carneiro Vieira 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2007年第3期307-312,共6页
Arachis retusa Krapov. et W. C. Gregory et Valls is endemic in the West-central region of Brazil, occurring In areas endangered by human actions. The establishment of in vitro preservation methods for wild species of ... Arachis retusa Krapov. et W. C. Gregory et Valls is endemic in the West-central region of Brazil, occurring In areas endangered by human actions. The establishment of in vitro preservation methods for wild species of Arachis is an alternative to seed banks for germplasm storage, multiplication and distribution. The risk of genetic changes Induced by tissue culture and the monitoring of the genetic stability of the biological material before, during and after storage must be considered In the context of conservation. Random amplified polymorphlc ONA (RAPO) and amplified fragment length polymorphism (AFLP) fingerprinting were used to evaluate the genetic stability of in vitro plants originated from cotyledons and embryo axes of A. retusa. Cotyledons originated shoots through direct organogenesls and embryo axes displayed muItishoot formation Induced by 110 mmol/L and 8.8 mmol/L BAP, respectively. Ninety genomlc regions (loci) generated from RAPO and 372 from AFLP analyses were evaluated. All amplified fragments detected by both techniques in plants derived from the two explant types were monomorphic. The results Indicate that the recovered shoots are genetically stable at the assessed genomic regions. 展开更多
关键词 amplified fragment length polymorphism Arachis retusa in vitro preservation MICROPROPAGATION random amplifiedpolymorphic DNA somaclonal variation
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Effects of Logging on the Genetic Diversity of Quercus tiaoloshanica Chun et Ko in a Tropical Montane Forest of Hainan Island, Southern China 被引量:1
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作者 Jian-Wei ZHENG Shu-Qing AN Lin CHEN Xin LENG Zhong-Sheng WANG Hua-Jun XIANG 《Journal of Integrative Plant Biology》 SCIE CAS CSCD 2005年第10期1184-1192,共9页
Quercus tiaoloshanica Chun et Ko, which has a narrow range of distribution, is one of the important endemic species of the tropical montane rain forest on Hainan Island, southern China. Long-term logging and habitat d... Quercus tiaoloshanica Chun et Ko, which has a narrow range of distribution, is one of the important endemic species of the tropical montane rain forest on Hainan Island, southern China. Long-term logging and habitat destruction have resulted in population decline and distribution retreat of Q. tiaoloshanica. To determine the impact of logging on the genetic diversity of Q. tiaoloshanica, the authors investigated the genetic structures using amplified fragment length polymorphism markers in four regenerated stands after logging and in one unlogged stand. Compared with the unlogged stand, the effective number of alleles per locus dropped by 1.0% in selective logging stands and by 2.0% in clear logging stands, corresponding to reductions of 3.8% and 5.2%, respectively, in mean Nei's gene diversity and 2.9% and 3.5%, respectively, in mean Shannon diversity index. No substantial genetic erosion was detected in any of the regenerated stands owing to the high tree density and high heterogeneity of the Q. tiaoloshanica stands investigated. Meanwhile, there was no natural regeneration of the species observed in a Dacrydium pierrei Hickel plantation 700 m away from the regenerated stands, suggesting the limited ability of seed dispersal of Q. tiaoloshanica. Clear logging should be undertaken cautiously because the total number of plant species dropped by 15.2% in the clear-logged stands compared with the unlogged stand. To conserve the genetic diversity of this species, as well as the plant biodiversity of tropical forests, the habitats of Q. tiaoloshanica should be protected against exploitation in terms of agricultural or other forms of land use, and some mature trees should be preserved as seed sources to maintain an adequate regeneration base for this species in the management of logging. 展开更多
关键词 amplified fragment length polymorphism genetic erosion LOGGING Quercus tiaoloshanica tropical forest.
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