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Microdissection of Single Chromosome and Construction of Its DNA Library in Sunflower (Helianthus annuus L.)
Sun Ruifen1, Yan Suli1,2, An Yulin1*
1Biotechnology Research Center, Inner Mongolia Academy of Agriculture and Animal Husbandry Sciences, Huhhot 010031, China; 2Institute of Agronomy, Inner Mongolia Agriculture University, Huhhot 010019, China
Abstract: The satellite chromosome of triple cross and single cross of “Inner Mongolia Hibrid oil sunflower 3” were isolated by glass needle method with the help of micromanipulator. After two rounds of LA-PCR amplification， the PCR products ranging from 250 to 1 500 bp were obtained. Southern blot with genomic DNA revealed hybridization signal， indicating that DNAs from the satellite chromosome had been successfully amplified.The second-round PCR products were microcloned into plasmid vector， and their DNA libraries were constructed.About 2.26×105 and 2.57×105 recombinant clones were obtained from triple cross and single cross of sunflower respectively. 30 clones were randomly selected for size evaluation. The results demonstrated that the size of the DNA fragmenrts from triple hybrid sunflower varied approximately from 200 to 700 bp with an average of 535 bp， and the DNA fragmenrts from single cross hybrid sunflower varied approximately from 200 to 500 bp with an average of 480 bp. The microdissection and microcloning technology has been applied for sunflower chromosome analysis.
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