Carbohydrate profiling in seeds and seedlings of transgenic triticale modified in the expression of sucrose:sucrose-1-fructosyltransferase (1-SST) and sucrose:fructan-6-fructosyltransferase (6-SFT).
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.
Agriculture and Agri-Food Canada Research Centre, 5403 1st Avenue South, Lethbridge, Alberta T1J 4B1, Canada.
Calliste
C
Diedhiou
Denis
D
Gaudet
Yehong
Y
Liang
Jinyue
J
Sun
Zhen-Xing
ZX
Lu
François
F
Eudes
André
A
Laroche
D002241:Carbohydrates / Q000032:analysis*; D003080:Cold Temperature; D002523:Edible Grain / Q000737:chemistry* / Q000235:genetics* / Q000502:physiology; D005630:Fructans / Q000032:analysis; D018525:Germination; D006602:Hexosyltransferases / Q000235:genetics / Q000378:metabolism; D030781:Organisms, Genetically Modified / Q000235:genetics / Q000502:physiology; D012639:Seeds / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins
D002241:Carbohydrates; D005630:Fructans; D006602:Hexosyltransferases; C097181:sucrose - fructan 6-fructosyltransferase; C021451:inulosucrase; D001426:Bacterial Proteins