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Despite the fact that R. oryzae s. s. and R. delemar can be resolved by multi-gene phylogenetic analyses and physiological differences in acid production, morphological examinations and molecular analyses of more conservative loci indicate that these two cryptic species are very close relatives.
We performed a suite of phylogenetic and population genetic analyses of the mcy genes using three "conservative" loci within the mcy gene cluster as markers (designated " mcyMLST" by analogy with MLST).
Our phylogenetic and population genetic analyses of multiple conservative loci within the microcystin synthetase (mcy) gene cluster suggested that mcy genes of M. aeruginosa are subdivided into two cryptic clades, which have been primarily generated and maintained as a result of homology-dependent recombination and neutral genetic drift.
With the use of universal primers that recognize highly conservative loci such as the 16S rDNA encoding gene, species-specific sequences can be amplified directly from diseased host tissues and compared with a reference-sequence database to infer phylogenetic relationships (3, 4 ).
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The ability of the more conservative locus rbcLa to provide such a highly resolved tree is due in large part to the very low ratio of species to genera in the Luquillo forest (see above).
Complementing a rapidly evolving locus such as the trnH-psbA spacer with a more conservative locus (such as the coding locus rbcL) can minimize type I errors (such that sequences are robustly assigned to the correct genus at least) and type II errors (higher rates of sequence divergence can discriminate among closely allied species in highly speciose genera).
Compared to (iHSVoight) [ 9] our method is more conservative for loci with low minor allele frequencies.
These were used to construct a phylogenetic tree we termed "conservative" because loci with any missing data were excluded (Table S12).
Single locus methods, such as classical interval mapping (Lander and Botstein 1989), are simple and computationally efficient, but in cases where the sample size is small, investigated markers are numerous, and significance testing is conservative, single locus effects may suffer from severe overestimation (denoted the "Beavis effect") (Beavis 1994; Slate 2013).
The overall PID-sib is the upper limit of the possible ranges of PID in a population, and thus, provides the most conservative number of loci required to resolve all individuals, including relatives.
The overall PID among siblings is the upper limit of the possible ranges of PID in a population, thus providing the most conservative number of loci required to resolve all individuals, including relatives.
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