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Published on: March 28, 2018
A bacterial artificial chromosome library for sugarcane.
J P Tomkins1, Y Yu, H Miller-Smith
1Clemson University Genomics Institute Room 100 Jordan Hall Clemson, SC 29634, USA, JO.
Summary
A new bacterial artificial chromosome (BAC) library was created for sugarcane (Saccharum officinarum) to aid in cloning genes, like the rust resistance locus. This comprehensive library enables efficient map-based cloning and comparative genomics studies in sugarcane.
Area of Science:
- Plant genetics
- Genomics
- Molecular biology
Background:
- Sugarcane (Saccharum officinarum) is a complex polyploid, making gene cloning challenging.
- A rust resistance locus with monogenic inheritance presents an opportunity for genetic study.
- A bacterial artificial chromosome (BAC) library is crucial for isolating specific genes in complex genomes.
Purpose of the Study:
- To construct a high-quality BAC library for sugarcane cultivar R570.
- To facilitate map-based cloning of the rust resistance locus.
- To enable comparative genomics with other monocot species.
Main Methods:
- A BAC library was created with 103,296 clones, representing 4.5 genome equivalents.
- Average insert size was 130 kb, ensuring high probability of sequence recovery.
- Robotic high-density filter gridding and screening with RFLP probes were employed.
Main Results:
- The library demonstrated low chloroplast DNA content (<1%).
- Screening with mapping probes identified 8-60 positive signals per probe.
- BAC clones were fingerprinted to assemble contigs and identify candidate clones for the rust locus.
Conclusions:
- The constructed BAC library is a valuable resource for sugarcane genetic research.
- The library supports map-based cloning of important agronomic traits, such as rust resistance.
- Comparative analysis with rice and sorghum BAC libraries aids in understanding monocot genome evolution.
