ISHS Acta Horticulturae 763: XXVII International Horticultural Congress - IHC2006: International Symposium on Structural and Functional Genomics of Horticultural Plants
APPLICATION OF SUPPRESSIVE SUBTRACTION HYBRIDIZATION IN THE STUDY OF EMBRYO ABORTION RELATED TO GENE EXPRESSION IN LITCHI (LITCHI CHINENSIS SONN.) |
| Authors: | Xiang Xu, Zhang YiShun, Huang ShangZhi, Ou Liangxi, Qiu Yanping, Li Zhiqiang, Chen Jiezhen |
| Keywords: | virtual northern blot, S-adenosylmethionine synthetase gene (SAM gene) |
| Abstract:
To study the molecular mechanisms associated with embryo abortion in seed-shriveled litchi varieties, suppressive subtraction hybridization (SSH) was used to investigate differential gene expression of normal-seeded and seed-shriveled embryos during stages of I to II of fruit-set. The differential expression cDNA library, representing the specific cDNA for aborting embryos of 'GuiWei' litchi, was built using the embryos from different fruit-set stages, with the normal-seeded embryo as the driver and the seed-shriveled embryo as the tester. After testing by Virtual Northern, 3 positive clones were obtained. We isolated 3 cDNAs from the 3 clones, which were 342, 599 and 391 nucleotides long and were designated Lce1, Lce2 and Lce3, respectively. The Lce1 amino acid sequence showed 97% homology with ubiquitin-conjugated protein of Capsicum and Solanum, Lce2 showed 94% homology with a Brassica S-adenosylmethionine synthetase and Lce3 showed 98% homology with a Zea mays methionine adenosyltransferase. These are the first reported sequences in litchi. The full-length sequence was obtained by PCR amplification and a complete cDNA for SAM synthetase gene was obtained. The sequence was 1515 base pairs (bp) with an opening reading frame (ORF) encoding 393 amino acids (GenBank accession NO: AY259227). This SAM synthetase gene in embryos of 'Guiwei' litchi may be associated with abortion. | |
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