RNA silencing of Waxy gene results in low levels of amylose in the seeds of transgenic wheat (Triticum aestivum L.).
In this study, the level of amylose was reduced in wheat seeds by RNAi strategy. Because the synthesis of amylose is catalyzed by the granule-bound starch synthase I (GBSSI or WAXY protein), the Waxy gene of wheat was isolated from wheat seeds by using RT-PCR. Southern analysis confirmed that there were three Waxy genes in wheat genome. Northern hybridization showed that Waxy mRNA accumulated in seeds following pollination. By RNAi strategy,the 683 bp sense and antisense fragments in reverse orientation separated by a 150 bp intron were cloned into pCAMBIA 3300 just downstream of the maize ub/1 promoter. By Agrobacteriurn-mediated wheat transformation method, four transgenic plants (Cultivar Yangmai 10) were identified by PCR, RT-PCR and leaf painting assay. The level of amylose in the endosperm were significantly reduced in transgenic seeds as checked by iodine staining and analysis of amylose content. The results indicated that RNA silencing of Waxy gene resulted in low level of amylose in the seeds of transgenic wheat.
Li
Jia-Rui
JR
The Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Taian 271018, China.
Zhao
Wei
W
Li
Quan-Zi
QZ
Ye
Xing-Guo
XG
An
Bao-Yan
BY
Li
Xiang
X
Zhang
Xian-Sheng
XS
D000688:Amylose / Q000378:metabolism*; D015152:Blotting, Northern; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012231:Rhizobium / Q000235:genetics; D012639:Seeds / Q000378:metabolism; D013214:Starch Synthase / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic; D014908:Triticum / Q000235:genetics* / Q000378:metabolism
D010940:Plant Proteins; D012333:RNA, Messenger; C060756:waxy protein, plant; D000688:Amylose; D013214:Starch Synthase