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L. S. Lima K. P. Gramacho A. S. Gesteira U. V. Lopes F. A. Gaiotto H. A. Zaidan J. L. Pires J. C. M. Cascardo F. Micheli 《Molecular breeding : new strategies in plant improvement》2008,22(2):315-318
Theobroma cacao L.–Moniliophthora perniciosa expressed sequence tags (ESTs) were converted into useful satellite markers for population analysis and genetic mapping.
Forty-nine flanking primer pairs from TSH1188 (a resistant genotype) and Catongo (a susceptible genotype) ESTs were designed
and screened for polymorphism analysis. Eleven were polymorphic, with an average of 3.81 alleles per locus and a total of
42 alleles. The satellite markers were tested on 21 cacao accessions and two bulked DNAs generated from 6 resistant and 6
susceptible plants from a segregating F2 (SCA6 × ICS1) population for witches’ broom resistance. These results show that EST-derived microsatellites (short sequence
repeats, SSRs) in Theobroma cacao have many potential applications in linkage mapping and the planning of crosses. 相似文献
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Lívia Santos Lima Karina Peres Gramacho José Luis Pires Didier Clement Uilson Vanderlei Lopes Nicolas Carels Abelmon da Silva Gesteira Fernanda Amato Gaiotto Júlio Cézar de Mattos Cascardo Fabienne Micheli 《Tree Genetics & Genomes》2010,6(5):663-676
In this study, we report results of the detection and analysis of SSR markers derived of cacao–Moniliophthora perniciosa expressed sequence tags (ESTs) in relation to cacao resistance to witches’ broom disease (WBD), and we compare the polymorphism
of those ESTs (EST-simple sequence repeat (SSR)) with classical neutral SSR markers. A total of 3,487 ESTs was used in this
investigation. SSRs were identified in 430 sequences: 277 from the resistant genotype TSH 1188 and 153 from the susceptible
one Catongo, totalizing 505 EST-SSRs with three types of motifs: dinucleotides (72.1%), trinucleotides (27.3%), and tetranucleotides
(0.6%). EST-SSRs were classified into 16 main categories; most of the EST-SSRs belonged to “Unknown function” and “No homology”
categories (45.82%). A high frequency of SSRs was found in the 5’UTR and in the ORF (about 27%) and a low frequency was observed
in the 3’UTR (about 8%). Forty-nine EST-SSR primers were designed and evaluated in 21 cacao accessions, 12 revealed polymorphism,
having 47 alleles in total, with an average of 3.92 alleles per locus. On the other hand, the 11 genomic SSR markers revealed
a total of 47 alleles, with an average of 5.22 alleles per locus. The association of EST-SSR with the genomic SSR enhanced
the analysis of genetic distance among the genotypes. Among the 12 polymorphic EST-SSR markers, two were mapped on the F2 Sca 6 × ICS 1 population reference for WBD resistance. 相似文献
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Mangrove oysters Crassostrea rhizophorae were sampled monthly in the estuary of Jaguaribe River, on the east coast of Ceará State, Brazil, between August, 2000 and December, 2001, making up 170 individuals. The water temperature varied from 26 to 30 degrees C and salinity from 21 to 42. The animals' size ranged from 3.4 to 7.2 cm height. Macroscopical and histopathological analyses were carried out in the oysters' tissues. The histological exams showed protozoans and metazoans of genera Nematopsis and Tylocephalum, respectively. Nematopsis prevalence varied from 60 to 100% and it was higher in the gills and mantle. The oocysts presented a mean size of 11.5 microm (+/-1.32) length and 9.1 microm (+/-1.06) width (n = 30), up to 3 oocysts/phagocyte having been observed. Several animals presented focal hemocitical reaction. The percentage of Tylocephalum was 1.7%. In spite of the high infection prevalence by Nematopsis, infected animals did not have their reproductive cycle impaired. 相似文献
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Analysis of the NAC transcription factor gene family in citrus reveals a novel member involved in multiple abiotic stress responses 总被引:1,自引:0,他引:1
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Gesteira AS Micheli F Carels N Da Silva AC Gramacho KP Schuster I Macêdo JN Pereira GA Cascardo JC 《Annals of botany》2007,100(1):129-140
BACKGROUND AND AIMS: Witches' broom disease is caused by the hemibiotrophic basidiomycete Moniliophthora perniciosa, and is one of the most important diseases of cacao in the western hemisphere. Because very little is known about the global process of such disease development, expressed sequence tags (ESTs) were used to identify genes expressed during the Theobroma cacao-Moniliophthora perniciosa interaction. METHODS: Two cDNA libraries corresponding to the resistant (RT) and susceptible (SP) cacao-M. perniciosa interactions were constructed from total RNA, using the DB SMART Creator cDNA library kit (Clontech). Clones were randomly selected, sequenced from the 5' end and analysed using bioinformatics tools including in silico analysis of the differential gene expression. KEY RESULTS: A total of 6884 ESTs were generated from the RT and SP cDNA libraries. These ESTs were composed of 2585 singlets and 341 contigs for a total of 2926 non-redundant sequences. The redundancy of the libraries was low and their specificity high when compared with the few other cacao libraries already published. Sequence analysis allowed the assignment of a putative functional category for 54 % of sequences, whereas approx. 22 % of sequences corresponded to unknown function and approx. 24 % of sequences did not show any significant similarity with other proteins present in the database. Despite the similar overall distribution of the sequences in functional categories between the two libraries, qualitative differences were observed. Genes involved during the defence response to pathogen infection or in programmed cell death were identified, such as pathogenesis related-proteins, trypsin inhibitor or oxalate oxidase, and some of them showed an in silico differential expression between the resistant and the susceptible interactions. CONCLUSIONS: As far as is known this is the first EST resource from the cacao-M. perniciosa interaction and it is believed that it will provide a significant contribution to the understanding of the molecular mechanisms of the resistance and susceptibility of cacao to M. perniciosa, to develop strategies to control witches' broom, and as a source of polymorphism for molecular marker development and marker-assisted selection. 相似文献
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Codon usage in the vertebrate hemoglobins and its implications 总被引:2,自引:0,他引:2
A study of codon usage in vertebrate hemoglobins revealed an evolutionary
trend toward elevated numbers of CpG codon boundary pairs in mammalian
hemoglobin alpha genes. Selection for CpG codon boundaries countering the
generally observed CpG suppression is strongly suggested by these data.
These observations parallel recently published experimental results that
indicate that constitutive expression of the human alpha-globin gene
appears to be determined by regulatory information encoded within the
structural gene. The possibility is raised that, in the absence of
selection, CpG decay can be used to date the evolutionary origin of a
mammalian alpha pseudogene from its active alpha gene.
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Coulson-Thomas VJ Coulson-Thomas YM Gesteira TF de Paula CA Mader AM Waisberg J Pinhal MA Friedl A Toma L Nader HB 《Cell and tissue research》2011,346(2):223-236
During cancer cell growth many tumors exhibit various grades of desmoplasia, unorganized production of fibrous or connective
tissue, composed mainly of collagen fibers and myofibroblasts. The accumulation of an extracellular matrix (ECM) surrounding
tumors directly affects cancer cell proliferation, migration and spread; therefore the study of desmoplasia is of vital importance.
Stromal fibroblasts surrounding tumors are activated to myofibroblasts and become the primary producers of ECM during desmoplasia.
The composition, density and organization of this ECM accumulation play a major role on the influence desmoplasia has upon
tumor cells. In this study, we analyzed desmoplasia in vivo in human colorectal carcinoma tissue, detecting an up-regulation of collagen I, collagen IV and collagen V in human colorectal
cancer desmoplastic reaction. These components were then analyzed in vitro co-cultivating colorectal cancer cells (Caco-2 and HCT116) and fibroblasts utilizing various co-culture techniques. Our findings
demonstrate that direct cell-cell contact between fibroblasts and colorectal cancer cells evokes an increase in ECM density,
composed of unorganized collagens (I, III, IV and V) and proteoglycans (biglycan, fibromodulin, perlecan and versican). The
desmoplastic collagen fibers were thick, with an altered orientation, as well as deposited as bundles. This increased ECM
density inhibited the migration and invasion of the colorectal tumor cells in both 2D and 3D co-culture systems. Therefore
this study sheds light on a possible restricting role desmoplasia could play in colorectal cancer invasion. 相似文献