Coupe Simon A, Sinclair Ben K, Somerfield Sheryl D, Hurst Paul L
New Zealand Institute for Crop and Food Research Ltd, Private Bag 11 600, Palmerston North, New Zealand. Current address: Department of Animal and Plant Sciences, University of Sheffield, Sheffield S10 2TN, UK.Corresponding author; email:
New Zealand Institute for Crop and Food Research Ltd, Private Bag 11 600, Palmerston North, New Zealand.
Funct Plant Biol. 2002 Aug;29(9):1045-1053. doi: 10.1071/PP01237.
A cDNA clone encoding malate synthase (MS; EC 4.1.3.2) was isolated from a 48-h postharvest asparagus (Asparagus officinalis L.) spear cDNA library using a MS clone from Brassica napus. The asparagus MS (AoMS1) cDNA hybridized to a 1.9-kb transcript that increased in abundance preferentially in spear-tip tissue during postharvest storage. The AoMS1 transcript also accumulated during natural foliar senescence of asparagus fern. The cDNA consists of 1960 nucleotides with an open reading frame of 1665 nucleotides or 555 amino acids, and encodes a deduced protein with a predicted Mr of 63 kDa and a pI of 8.1. The deduced amino acid sequence of AoMS1 showed high identity with the B. napus MS clone (77.2%) used to isolate it, and with MS from cucumber (77%). Genomic Southern analysis suggests that a single gene in asparagus encodes AoMS1. Controlled- atmosphere treatments aimed at reducing deterioration of harvested asparagus spears reduced the expression of AoMS1. The reduction was correlated with the reduced oxygen level, and reduced MS enzyme activity was also observed. Asparagus cell cultures were used to test the role of sugar status in regulating AoMS1 gene expression. In cultures without sucrose there was an accumulation of AoMS1 transcript that was absent in cultures containing sucrose.
利用来自甘蓝型油菜的苹果酸合酶(MS;EC 4.1.3.2)克隆,从采后48小时的芦笋(Asparagus officinalis L.)嫩茎cDNA文库中分离出一个编码苹果酸合酶的cDNA克隆。芦笋MS(AoMS1)cDNA与一个1.9 kb的转录本杂交,该转录本在采后贮藏期间优先在嫩茎尖组织中丰度增加。AoMS1转录本在芦笋蕨自然叶片衰老过程中也会积累。该cDNA由1960个核苷酸组成,开放阅读框为1665个核苷酸或555个氨基酸,编码一个推导蛋白,预测分子量为63 kDa,pI为8.1。AoMS1的推导氨基酸序列与用于分离它的甘蓝型油菜MS克隆具有高度同一性(77.2%),与黄瓜的MS也具有高度同一性(77%)。基因组Southern分析表明,芦笋中的一个单基因编码AoMS1。旨在减少采后芦笋嫩茎变质的气调处理降低了AoMS1的表达。这种降低与氧气水平的降低相关,并且还观察到MS酶活性降低。利用芦笋细胞培养物来测试糖状态在调节AoMS1基因表达中的作用。在没有蔗糖的培养物中,AoMS1转录本会积累,而在含有蔗糖的培养物中则不存在这种积累。