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The use of dna sequencing (ITS and trn L‐ F ), AFLP, and fluorescent in situ hybridization to study allopolyploid Miscanthus (Poaceae)
Author(s) -
Hodkinson Trevor R.,
Chase Mark W.,
Takahashi Chigusa,
Leitch Ilia J.,
Bennett Michael D.,
Renvoize Stephen A.
Publication year - 2002
Publication title -
american journal of botany
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.218
H-Index - 151
eISSN - 1537-2197
pISSN - 0002-9122
DOI - 10.3732/ajb.89.2.279
Subject(s) - biology , amplified fragment length polymorphism , genetics , genome , internal transcribed spacer , dna sequencing , fluorescence in situ hybridization , ribosomal dna , genomic dna , introgression , chromosome , nuclear dna , dna , ribosomal rna , phylogenetics , gene , mitochondrial dna , genetic diversity , population , demography , sociology
Two clones of Miscanthus , grown under the names M. × giganteus and M. sacchariflorus , have been used in biomass trials in Europe, but neither the identity of these clones nor their origin has been established. DNA sequencing, amplified fragment length polymorphism (AFLP), and chromosome studies confirm that M. × giganteus is an allotriploid (2 n = 3 x = 57) combining genomes from M. sinensis (2 n = 2 x = 38) and M. sacchariflorus (2 n = 38 or 76). Two alleles of the internal transcribed spacer of 18S–25S nuclear ribosomal DNA (ITS) were discovered in polymerase chain reaction products of M. × giganteus . Cloning of these revealed that one matched M. sinensis and the other M. sacchariflorus . Plastid trnL intron and trnL‐F spacer sequences showed that the maternal lineage of M. × giganteus was M. sacchariflorus . Fluorescent in situ hybridization, FISH, was used to investigate genome organization in Miscanthus but was unable to differentiate between the different parental genomes present in M. × giganteus, indicating that two parental genomes are still extremely similar at the repetitive DNA level. This study is an example in which rDNA sequences and AFLP fingerprints permit identification of the parental genomes in a hybrid, but FISH methods, at the repetitive DNA level (including genomic in situ hybridization, GISH), were unable to do so because their sequences remain too similar.