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In situ amplification techniques are designed to increase the mass of DNA in a fixed target, either whole cells or tissue sections. When combined with fluorescently labeled nucleotides, they can be used for locus detection. They also can be used to increase target mass for subsequent operations, such as cellular or chromosomal isolation by microdissection. When combined with chromosome microdissection, these techniques allow libraries to be made from single copies of chromosomes, chromosome fragments, or even bacteria.

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Characterization of Apomictic BC₇ and BC₈ Pearl Millet: Meiotic Chromosome Behavior and Construction of an ASGR‐carrier Chromosome‐specific Library

Singh

Conner

Zeng

et al. 2010

Crop Science

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Apospory in Pennisetum squamulatum Fresen is inherited as a single‐locus dominant trait. Molecular analysis has identified the apospory‐specific genomic region (ASGR), a large segment of one chromosome that is necessary and sufficient to confer apomixis. To transfer apospory from P. squamulatum to pearl millet [P. glaucum (L.) R. Br.], advanced backcrosses were generated. Here we report the characterization of apomictic BC7 and BC8 generations. A BC7–derived genotype has two P. squamulatum chromosomes, while BC8–derived apomictic lines inherited only the ASGR chromosome that confers apospory to pearl millet. Morphologically, no significant differences were observed between backcross generations for plant height, leaf length, leaf width, and first internode and inflorescence lengths. However, the BC7 and BC8 lines differed significantly with regard to pollen viability, flowering time, and seed set. Pollen viability increased to 81% in the BC8 lines compared with 37% in BC7. The frequency of aposporous embryo sacs was lower in BC8 (79%) lines compared to BC7 (93%). Meiotic behavior of chromosomes during metaphase I and anaphase I showed that BC8 typically had a single univalent, identified as the ASGR chromosome by FISH (fluorescence in situ hybridization) and GISH (genomic in situ hybridization). Utilizing its unique behavior during meiosis the ASGR chromosome was microdissected and a chromosome‐specific library was constructed and sequenced. Analysis of sequences identified both novel and previously known ASGR‐linked sequences.

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Cited by 3 publications

References 6 publications

The preparation of sex-chromosome-specific painting probes and construction of sex chromosome DNA library in half-smooth tongue sole (Cynoglossus semilaevis)

The preparation of sex-chromosome-specific painting probes and construction of sex chromosome DNA library in half-smooth tongue sole (Cynoglossus semilaevis)

Microdissection With PCR <i>In Situ</i>

Characterization of Apomictic BC₇ and BC₈ Pearl Millet: Meiotic Chromosome Behavior and Construction of an ASGR‐carrier Chromosome‐specific Library

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Cited by 3 publications

References 6 publications

The preparation of sex-chromosome-specific painting probes and construction of sex chromosome DNA library in half-smooth tongue sole (Cynoglossus semilaevis)

The preparation of sex-chromosome-specific painting probes and construction of sex chromosome DNA library in half-smooth tongue sole (Cynoglossus semilaevis)

Microdissection With PCR <i>In Situ</i>

Characterization of Apomictic BC7 and BC8 Pearl Millet: Meiotic Chromosome Behavior and Construction of an ASGR‐carrier Chromosome‐specific Library

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Characterization of Apomictic BC₇ and BC₈ Pearl Millet: Meiotic Chromosome Behavior and Construction of an ASGR‐carrier Chromosome‐specific Library