Bachem C W, Kuyt S, Horvath B M, Claassens M M, Vreugdenhil D, Visser R G
Laboratory of Plant Breeding, Graduate School of Experimental Plant Sciences, Wageningen Agricultural University, The Netherlands.
Plant Mol Biol. 2000 Jul;43(4):473-82. doi: 10.1023/a:1006492205788.
Using the cDNA-AFLP method, we have isolated a transcript-derived fragment (TDF) which shows a differential expression pattern during tuber organogenesis of Solanum tuberosum L. The TDF was used to isolate a cDNA clone carrying a 1.5 kb insert and potentially coding for a 32.5 kDa peptide which, by homology, represents a potato homologue of an alpha-snap gene and has been designated Stsnap. Northern analysis showed that the Stsnap gene is expressed in actively dividing tissues throughout the potato plant. Analysis of genomic DNA from potato revealed that the Stsnap gene is likely to be a single-copy gene. The expression of antisense Stsnap cDNA under the control of the CaMV 35S promoter results in plants with an altered morphology such as curled leaves. Several of these transgenic lines also display cellular and developmental abnormalities with distinct changes in assimilate transport including accumulation of starch and soluble sugars in source leaves. We argue that these findings are consistent with the hypothetical function of the StSNAP gene product in vesicle targeting and fusion during plant development.
利用cDNA-AFLP方法，我们分离出一个转录本衍生片段（TDF），该片段在马铃薯块茎器官发生过程中呈现差异表达模式。该TDF用于分离一个携带1.5 kb插入片段的cDNA克隆，该克隆可能编码一个32.5 kDa的肽，通过同源性分析，它代表α-SNAP基因的马铃薯同源物，已被命名为Stsnap。Northern分析表明，Stsnap基因在整个马铃薯植株的活跃分裂组织中表达。对马铃薯基因组DNA的分析表明，Stsnap基因可能是单拷贝基因。在CaMV 35S启动子控制下反义Stsnap cDNA的表达导致植株形态改变，如叶片卷曲。其中一些转基因株系还表现出细胞和发育异常，同化物运输发生明显变化，包括源叶中淀粉和可溶性糖的积累。我们认为这些发现与StSNAP基因产物在植物发育过程中囊泡靶向和融合的假设功能一致。
