Abstract
BackgroundChromosome microdissection is one of the most important techniques in molecular cytogenetic research. Cotton (Gossypium Linnaeus, 1753) is the main natural fiber crop in the world. The resistance gene analog (RGA) cloning after its single chromosome microdissection can greatly promote cotton genome research and breeding.ResultsUsing the linker adaptor PCR (LA-PCR) with the primers of rice disease-resistance homologues, three nucleotide sequences PS016 (KU051681), PS054 (KU051682), and PS157 (KU051680) were obtained from the chromosome Ah01 of upland cotton (cv. TM-1). The Blast results showed that the three sequences are the nucleotide binding site-leucine rich repeat (NBS-LRR) type RGAs. Clustering results indicated that they are homologous to these published RGAs. Thus, the three RGAs can definitely be confirmed as NBS-LRR class of RGAs in upland cotton.ConclusionsUsing single chromosome microdissection technique, DNA libraries containing cotton RGAs were obtained. This technique can promote cotton gene cloning, marker development and even the improvement of cotton genome research and breeding.
Highlights
Chromosome microdissection is one of the most important techniques in molecular cytogenetic research
The positive control produced a weak initial band (Figure 2, lane 4) and an obvious smearing pattern after the second linker adaptor PCR (LA-PCR). These results indicated that the Ah01 chromosome was amplified successfully
We successfully developed a technique to separate a single chromosome from upland cotton pollen mother cells (PMC) with the laser cutting
Summary
Chromosome microdissection is one of the most important techniques in molecular cytogenetic research. The resistance gene analog (RGA) cloning after its single chromosome microdissection can greatly promote cotton genome research and breeding. Research focusing on a single chromosome or a chromosomal subsection can greatly reduces subsequent work, such as identifying, screening and minimizing the whole genome screening. This technique has been widely used in Drosophila, humans and many other animals since its Plants have developed defensive mechanisms to protect themselves from pathogen infection through a number of evolutionary processes. Many R genes have been cloned from different host plants using positional cloning and transposon tagging methods. Considering the large number of physiological races of pathogens, transposon tagging and positional cloning methods are clearly inefficient.
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