Journal of Food, Agriculture and Environment

cDNA cloning and SSCP analysis on Ss5MaT1 gene in Salvia splendens

Author(s):

Fenglan Li ¹, Feifei Qin ^{2, 3}, Yanzhong Feng ⁴, Hui-lian Xu ⁵, Baozhong Hu ^1*, Di Liu ¹, Guofu Hu ¹, Rongmei Liu ¹, Yongqing Xu ¹, Bingxiu Zhang ¹, Qi Wei ⁴, Shujun Ni ⁴, Ran Wang ⁶

Recieved Date: 2009-01-19, Accepted Date: 2009-04-05

Abstract:

In the present study, nested polymerase chain reaction-single-strand conformation polymorphism (nested PCR-SSCP) was used for detection of point mutations in flower color genes in Salvia splendens. Totally 916 bp of Ss5MaT1 cDNA segments were obtained in eight varieties of Salvia splendens flowers by RT-PCR. Four sets of specific primers were synthesized according to the obtained cDNA cloning for SSCP analysis. Results from assays of nested PCR-SSCP showed the presence of three kinds of polymorphic DNA bands using primers SS3. Six and four DNA bands were displayed in samples of Rose and Salmon flowers, respectively, compared to only two bands in other six varieties of Salvia splendens flowers. DNA sequencing showed particularly high homology in nucleotide sequences of partial Ss5MaT1 segment in eight varieties of Salvia splendens flowers. However, the point mutant appeared at size of 744 bp in Rose flower with a single base changed in nucleotide sequence from thymine (T) to cytosine (C) and at 678 bp in Salmon changed flower with from cytosine (C) to thymine (T). Consequently, these changes resulted in the amino acid mutations from isoleucine (Ile) to threonine (Thr) in Rose flower and from valine (Val) to alanine (Ala) in Salmon flower. In conclusion, the consistency of the result analyzed by DNA sequencing with that by nested PCR-SSCP suggests that PCR-SSCP could provide an accurate and rapid tool for detecting gene point mutations in ornamental flowers.

Keywords:

Cloning and sequencing, flower color gene, Salvia splendens, nested PCR, point mutation detection, SSCP analysis, Ss5MaT1

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Journal: Journal of Food, Agriculture and Environment
Year: 2009
Volume: 7
Issue: 2
Category: Agriculture
Pages: 581-589

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