Search Results - (Author, Cooperation:D. Labonte)

2014-11-22

American Association for the Advancement of Science (AAAS)

0036-8075

1095-9203

Biology

Chemistry and Pharmacology

Computer Science

Medicine

Natural Sciences in General

Physics

Antibodies, Viral/blood/*immunology ; Antigenic Variation/genetics/immunology ; Evolution, Molecular ; Humans ; Influenza A Virus, H3N2 Subtype/genetics/*immunology ; Influenza Vaccines/*immunology ; Influenza, Human/blood/*immunology/prevention & control ; *Vaccination

1432-203X

KeywordsTy1-copia-like reverse transcriptase sequences ; Sweetpotato ; Clonal ; Ipomoea species ; Convolvulaceae ; AbbreviationsLTR: Long terminal repeats ; PCR: Polymerase chain reaction ; RT: Reverse transcriptase ; URL: Universal Resource Locator ; VT: Virus tested

Springer Online Journal Archives 1860-2000

Biology

Abstract Ty1-copia -like sequences were PCR amplified from sweetpotato [Ipomoea batatas (L.) Poir.] L87-95 genomic DNA samples by using Ty1-copia reverse transcriptase-specific primers. PCR fragments within the expected size range were isolated, cloned, and sequenced. Inferred amino acid sequences of two randomly selected cloned fragments were found to be highly homologous to Ty1-copia-like reverse transcriptase sequences in the GenBank database. Subsequent sequencing of an additional 22 cloned fragments revealed a high level of reverse transcriptase sequence diversity (sequence divergence ranged from 2% to 73%). Southern blot hybridization analysis indicated that these sequences are present in the genome of I. batatas and taxonomic relatives in high copy numbers. PCR amplification from leaf cDNA obtained from a sweetpotato clone using Ty1-copia reverse transcriptase-specific primers yielded a Ty1-copia-like fragment. This is the first known report of the presence of genomic, and putatively expressed, Ty1-copia-like reverse transcriptase sequences in I. batatas.

Electronic Resource

1432-2242

Key words Microsatellite DNA ; Polyploid ; Repeat instability ; Sweetpotato ; Simple sequence repeats

Springer Online Journal Archives 1860-2000

Biology

Abstract  The objectives of the present study were to evaluate the inheritance and nucleotide sequence profiles of microsatellite genetic markers in hexaploid sweetpotato [Ipomoea batatas (L.) Lam.] and its putative tetraploid and diploid ancestors, and to test possible microsatellite mutation mechanisms in polyploids by direct sequencing of alleles. Sixty three microsatellite loci were isolated from genomic libraries of I. batatas and sequenced. PCR primers were designed and used to characterize microsatellite loci in two hexaploid I. batatas populations, a tetraploid Ipomoea trifida population, and a diploid I. trifida population. Nine out of the sixty three primer pairs tested yielded a clearly discernible, heritable banding pattern; five showed Mendelian segregation. All other primer pairs produced either smeared banding patterns, which could not be scored, or no bands at all in I. batatas. All of the primers which produced discernible banding patterns from I. batatas also amplified products of similar size in tetraploid and diploid I. trifida accessions. The sequence analysis of several alleles in the three species showed differences due to mutations in the repeat regions consistent with small differences in the repeat number. However, in some cases insertions/deletions and base substitutions in the microsatellite flanking regions were responsible for polymorphisms in both polyploid and diploid species. These results provide strong empirical evidence that complex genetic mechanisms are responsible for SSR allelic variation in Ipomoea. Four I. batatas microsatellite loci showed polysomic segregation fitting tetraploid segregation ratios. To our knowledge this is the first report of segregation ratios for microsatellites markers in polyploids.

Electronic Resource