| M14 beet is a monosomic addition line of Beta corolliflora Zoss which constitutes of the normal 18 B. vulgaris chromosomes plus Beta corolliflora chromosome 9. Through the investigation we found the transmission frequencies of M14 up to 96% in four generations. Our studies revealed that the reproduction mode of M14 is allium-odorum type diplospory, autonomous endospermgenesis but pollen stimulation is needed. We believed the gene(s) controlling apomixis may be located on the alien chromosome from Beta corolliflora Zoss.In order to clone the putative gene(s) controlling apomixes of M14, we constructed a plant-transformation-competent binary BAC library from the genomic DNA of M14, the library consists of 49,920 clones with an average insert size of 127kb, representing 7.5xhaploid genome equivalents and providing a grater than 99% probability of isolating single copy DNA sequences from this library. We used pBC1054 probe from the alien chromosome Beta corolliflora genome-specific dispersed repetitive DNA sequences to isolate BAC clones derived from the alien chromosome in the library. In result a total of 2.377 positive clones were obtained and rearrayed into a sublibary specific for Beta corolliflora chromosome 9, these clones represent 3.8xhaploid plus Beta corolliflora chromosome 9 equivalents.We invented a series new methods to isolated BAC plasmid and convalent DNA-glass chemistries attach to an unmodified glass surface. Make use of these technology we invented the BAC chip which 2,377 BAC DNA from Beta corolliflora chromosome 9 in the array. It can testified that the false-positive rate was very low through the positive probe and randomprimes detected. Our microarray system is fundamentally different from most current microarray technologies in that activated DNA is printed on nature glass surfaces, a strategy that invariably introduces hybridization backgrounds. Finally we make the simple express analysis between Ml4 beet and B.vulgaris through the BAC chip. |
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