Agriculture and Agri-Food Canada Research Centre, 5403 1st Avenue South, Lethbridge, Alberta T1J 4B1, Canada.
J Biosci Bioeng. 2012 Oct;114(4):371-8. doi: 10.1016/j.jbiosc.2012.05.008. Epub 2012 Jun 13.
Constructs with sucrose-sucrose 1-fructosyltransferase (1-SST) from rye and or sucrose-fructan 6-fructosyltransferase (6-SFT) from wheat were placed under the control of wheat aleurone-specific promoter and expressed in triticale using biolistic and microspore transformation. Transgenic lines expressing one or both the 1-SST and the 6-SFT accumulated 50% less starch and 10-20 times more fructan, particularly 6-kestose, in the dry seed compared to the untransformed wild-type (WT) triticale; other fructans ranged in size from DP 4 to DP 15. During germination from 1 to 4 days after imbibition (dai), fructans were rapidly metabolized and only in transgenic lines expressing both 1-SST and 6-SFT were fructan contents significantly higher than in the untransformed controls after 4 days. In situ hybridization confirmed expression of 6-SFT in the aleurone layer in imbibed seeds of transformed plants. When transgenic lines were subjected to a cold stress of 4°C for 2 days, synthesis of fructan increased compared to untransformed controls during low-temperature germination. The increase of fructan in dry seed and germinating seedling was generally associated with transcript expression levels in transformed plants but total gene expression was not necessarily correlated with the time course accumulation of fructan during germination. This is the first report of transgenic modification of cereals to achieve production of fructans in cereal seeds and during seed germination.
构建物的蔗糖-蔗糖 1-果糖基转移酶(1-SST)来自黑麦和/或来自小麦的蔗糖-果聚糖 6-果糖基转移酶(6-SFT)在黑小麦中受到小麦糊粉层特异性启动子的控制,并通过弹道和小孢子转化进行表达。与未转化的野生型(WT)黑小麦相比,表达 1-SST 和 6-SFT 之一或两者的转基因系在干燥种子中积累的淀粉减少了 50%,而果聚糖增加了 10-20 倍,特别是 6-蔗果三糖;其他果聚糖的大小范围从 DP4 到 DP15。在吸胀后 1 至 4 天( dai)的萌发过程中,果聚糖迅速代谢,只有在同时表达 1-SST 和 6-SFT 的转基因系中,果聚糖含量在 4 天后才显著高于未转化的对照。原位杂交证实了转化植物吸胀种子的糊粉层中 6-SFT 的表达。当转基因系受到 4°C 的冷应激 2 天时,与未转化的对照相比,在低温萌发期间,果聚糖的合成增加。在干燥种子和萌发幼苗中,果聚糖的增加通常与转化植物中的转录表达水平相关,但总基因表达不一定与萌发过程中果聚糖的积累时间相关。这是首次报道对谷物进行转基因修饰以在谷物种子和种子萌发期间产生果聚糖。