A majority of commercial rose varieties bloom repeatedly throughout the year, as
compared to most rose species, other woody ornamentals, and fruit crops that bloom once a
year. This recurrent flowering feature of the commercial roses resulted from a flowering
mutation named everblooming (evb). The mutation is recessive to once blooming and is
found in the rose species Rosa chinensis. Although several molecular maps have been
developed for rose, little is known about the evb gene, except for its classic genetics. The
purpose of this study was to develop a large-insert bacterial artificial chromosome (BAC)
library as a starting tool for molecular cloning and analysis of the evb gene by map-based
cloning. To construct the large-insert BAC library, nuclear megabase-size DNA was
isolated from the recurrent blooming diploid species, Rosa chinensis cv. Old Blush. The
DNA was then partially digested with BamHI and separated on agarose gels by multi-phase
pulsed-field gel electrophoresis. Size selected fragments estimated between 100 kb and 150
kb in size were cloned into the pECBAC1 BAC vector and the clones having rose DNA
inserts were arrayed in 80 384-well microplates individually, with each clone being barcoded.
The library contains 30,720 clones, has an average insert size of 108 kb and covers
roughly 5.9x genome equivalents, with a >99% probability of isolating a single-copy clone from the library. The library is now available to be screened with the genes cloned from
other species that control vernalization and floral development and will be used in mapbased
cloning of the evb gene using a Rosa wichuraiana (âÂÂBasyeâÂÂs ThornlessâÂÂ) x âÂÂOld
Blushâ backcross population.
| Identifer | oai:union.ndltd.org:tamu.edu/oai:repository.tamu.edu:1969.1/4318 |
| Date | 30 October 2006 |
| Creators | Hess, Gregory |
| Contributors | Zhang, Hongbin |
| Publisher | Texas A&M University |
| Source Sets | Texas A and M University |
| Language | en_US |
| Detected Language | English |
| Type | Book, Thesis, Electronic Thesis, text |
| Format | 415130 bytes, electronic, application/pdf, born digital |
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