Wheat 1BL/1RS translocations are widely planted in China as well as in most of the wheat producing area in the world for their good qualities of disease resistance and high yield. 1BL/1RS translocations are however po...Wheat 1BL/1RS translocations are widely planted in China as well as in most of the wheat producing area in the world for their good qualities of disease resistance and high yield. 1BL/1RS translocations are however poor in bread making, partially caused by a family of small monomeric proteins, ω-secalins, which are encoded by genes on 1RS. Based on published sequence of a rye ω-secalin gene we designed a pair of primers to cover the whole mature protein coding sequence. A major band could be amplified from 1BL/1RS translocations but not from euploid wheat. Using this primer set we conducted PCR amplification by using high fidelity Pfu polymerase on the genomic DNAs and cDNAs purified from a 1BL/1RS translocation Lankao 906. Sequencing analysis indicated that this gene family contains several mem- bers of 1150 bp, 1076 bp, 1075 bp, 1052 bp and 1004 bp genes, including two pseudogenes and three active genes. The gene transcripts were differentially expressed in developing seeds.展开更多
Wheat-rye T1BL·1RS translocation lines are widely used,especially in China,but their processing quality is generally poor.An interfering expression vector targeting theω-secalin genes was constructed with the 1B...Wheat-rye T1BL·1RS translocation lines are widely used,especially in China,but their processing quality is generally poor.An interfering expression vector targeting theω-secalin genes was constructed with the 1Bx7 seed-specific promoter.Biolistic-mediated genetic transformation of the wheat cultivar KN199 carrying the T1BL·1RS translocation generated 10 transgenic lines.Two representative transgenic lines,8-2 and 13-7,were selected for analysis.Compared with the control,the two transformants showed an up to 4.5-fold decrease in totalω-secalins and various levels of decrease inω-gliadins,γ-gliadins,and low-molecular-weight glutenins.A decrease in high molecular weight(HMW)glutenin 1Bx7 was detected only in 8-2,owing possibly to promoter methylation.Increased levels ofα-gliadins were observed in both transformants,but increased levels of HMW glutenins were observed only in 13-7.Line 13-7 showed increases in gluten index,Zeleny sedimentation value,stabilization time,and maximum resistance.Its bread volume was 849.6 mL,an 11.9%increase over that of the control.Line 8-2 showed decreases in these parameters,but its total cake-making quality score was 88,an 17.3%increase over that of the control.The study demonstrates that the same RNAi construct may produce different effects on wheat processing quality and highlights the influence of the vector promoter in RNA interference.展开更多
基金supported by the National Basic Research Program of China(973)(No.2004CB117200).
文摘Wheat 1BL/1RS translocations are widely planted in China as well as in most of the wheat producing area in the world for their good qualities of disease resistance and high yield. 1BL/1RS translocations are however poor in bread making, partially caused by a family of small monomeric proteins, ω-secalins, which are encoded by genes on 1RS. Based on published sequence of a rye ω-secalin gene we designed a pair of primers to cover the whole mature protein coding sequence. A major band could be amplified from 1BL/1RS translocations but not from euploid wheat. Using this primer set we conducted PCR amplification by using high fidelity Pfu polymerase on the genomic DNAs and cDNAs purified from a 1BL/1RS translocation Lankao 906. Sequencing analysis indicated that this gene family contains several mem- bers of 1150 bp, 1076 bp, 1075 bp, 1052 bp and 1004 bp genes, including two pseudogenes and three active genes. The gene transcripts were differentially expressed in developing seeds.
基金The financial support by Key Research and Development Plan of Hebei Province(20326348D)Hebei Modern Agricultural Science and Technology Innovation Project(2019-4-08-1)are gratefully acknowledged.
文摘Wheat-rye T1BL·1RS translocation lines are widely used,especially in China,but their processing quality is generally poor.An interfering expression vector targeting theω-secalin genes was constructed with the 1Bx7 seed-specific promoter.Biolistic-mediated genetic transformation of the wheat cultivar KN199 carrying the T1BL·1RS translocation generated 10 transgenic lines.Two representative transgenic lines,8-2 and 13-7,were selected for analysis.Compared with the control,the two transformants showed an up to 4.5-fold decrease in totalω-secalins and various levels of decrease inω-gliadins,γ-gliadins,and low-molecular-weight glutenins.A decrease in high molecular weight(HMW)glutenin 1Bx7 was detected only in 8-2,owing possibly to promoter methylation.Increased levels ofα-gliadins were observed in both transformants,but increased levels of HMW glutenins were observed only in 13-7.Line 13-7 showed increases in gluten index,Zeleny sedimentation value,stabilization time,and maximum resistance.Its bread volume was 849.6 mL,an 11.9%increase over that of the control.Line 8-2 showed decreases in these parameters,but its total cake-making quality score was 88,an 17.3%increase over that of the control.The study demonstrates that the same RNAi construct may produce different effects on wheat processing quality and highlights the influence of the vector promoter in RNA interference.
