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標題: 馬鈴薯冷馴化相關基因之選殖與表現分析
Molecular Cloning and Characterization of Cold-Acclimation Genes in Potato (Solanum commersonii Dun.)
作者: 陳俊麟
Jiun-Lin Chen
Menq-Jiau Tseng
關鍵字: 馬鈴薯;Cold-acclimation gene;基因選殖;冷馴化基因;耐寒性;Frost resistance;Potato;Molecular cloning
Project: Horticulture NCHU, Volume 27, Issue 3, Page(s) 49-64.
興大園藝, Volume 27, Issue 3, Page(s) 49-64.
Low temperature stress restricts the distribution of plants and causes crop loss and reduced yield. Tolerance of crops to low temperature stress is thus a critical element in which can affect the balance of food supplement and the price of commodities. The objectives of the research are to isolate cold acclimation genes from S. commersonii, to examine the temporal-specific expression of selected genes, and to study the composition of these genes.
A cDNA library was constructed from polyA-RNA of S. commersonii leaves after 14 days of acclimation at 4℃. Two hundred and forty-one plagues that hybridized with 32P-labeled single-strand cDNA of 14 days of acclimated leaves but not for controls, were preliminary selected. Followed in vitro excision of selected phages and repeating DNA dot blot hybridization , four genes (ca101, ca106, ca127, ca153) were selected.
The results of northern blot hybridization showed that ca106 and ca153 genes were induced, ca101 and ca127 genes were increased in expression by cold acclimation. There are two patterns of expression in these four genes in regard to the amount of expression during 14 days of cold acclimation periods. Rapid increases in expression of ca101 and ca106 were found in S. commersonii leaves after one day of acclimation, followed by little decline, and then increases on day 10. Whereas, ca127 and ca153 didn't exhibit rapid increase in expression until day 3, then levels fall off during further acclimation. The expression of these 4 genes were decreased to the level approximate to those of controls after one day of deacclimation at 20℃ . According to the nucleotides sequencing, the selected 4 genes are different genes. The fragments of ca101, ca106, ca127, ca153 are 0.56, 1.5, 1.7, 2 kb, respectively.

本試驗以冷馴化處理14天之馬鈴薯的polyA-RNA構築成cDNA library。利用差異性篩選法篩選100,000個cDNA clones中獲得4個冷馴化處理所誘導之基因(ca101, ca106, ca127, ca153)。其片段大小分別為556 bp,1.5 kb,1.7 kb與2 kb。調查四個基因在冷馴化處理不同時期之表現,顯示ca101,ca106,於冷馴化1天後即急遽增加,隨後表現量稍降,於第10天再小幅度上昇,隨後再下降。然而ca127與153的表現要到冷馴化第3天才急遽增加,但隨後繼續冷馴化,其表現量遽減。經過14天冷馴化後,再去馴化處理一天,所選殖的四個基因的表現量皆有下降回復至與對照組相似之量的趨勢。
ISSN: 0255-5921
Appears in Collections:第27卷 第03期

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