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A Map-based Cloning Approach for the Identification of a Low Temperature Sensitive Gene sy-2 in Chilli pepper (Capsicum chinense)
- 언어ENG
- URLhttps://db.koreascholar.com/Article/Detail/298122
sy-2 (Seychelles-2) is a temperature sensitive mutant of Capsicum chinense and native to Seychelles Island in Africa. Previously we showed that sy-2 leaves were irregularly shaped and defective in chlorophyll development at temperatures lower than 24℃. A segregation test revealed that the sy-2 gene is controlled by a single recessive gene. To identify the sy-2 gene, we performed a map-based cloning approach using a total of 1,010 F2 plants derived from crossing sy-2 and the wild type C. chinense ‘No.3341’. sy-2 gene is located on chromosome 1, 0.3 cM and 0.1cM away from cosII markers C2_At4g29120 and C2_At1g09070, respectively. The tomato genome sequence between those two markers was compared with pepper genome sequence. We found three of pepper scaffold sequences in this region. We developed seven ingle nucleotide polymorphism (SNP) markers on the pepper scaffold sequences, among which five SNP markers were co-segregated with sy-2. To fill the gap between the scaffolds which contains co-segregating markers, we screened a bacterial artificial chromosome (BAC) library, and end-sequences of total of 22 AC clones were i. We found that five clones were overlapped to cover the gap. We fully sequenced four AC clones and found that the physical distance between C2_At4g29120 and C2_At1g09070 is 343kb. This region contains 70 putative genes such as HSP90-like ATPase family proteins, lipid-transfer proteins, calmodulin-domain protein kinases, and zinc finger proteins (ZFPs). To identify the sy-2 gene, we performed RT-PCR and found that a ZFP-like gene is differentially expressed between WT and sy-2 leaves. This result suggests that the ZFP-like gene is a strong candidate for the sy-2 gene. We are currently characterizing this candidate gene.
