In this study,intelligent,pH-responsive colorimetric films were prepared by encapsulating anthocyanins in nanocomplexes prepared from glutenin and carboxymethyl chitosan.These nanocomplexes were added to a corn starch...In this study,intelligent,pH-responsive colorimetric films were prepared by encapsulating anthocyanins in nanocomplexes prepared from glutenin and carboxymethyl chitosan.These nanocomplexes were added to a corn starch matrix and used in the freshness monitoring of chilled pork.The effects of anthocyanin-loaded nanocomplexes on the physical,structural,and functional characteristics of the films were investigated.The addition of anthocyanin-loaded nanocomplexes increased the tensile strength,elongation at break,hydrophobicity,and light transmittance of the films while decreasing their water vapor permeability.This is because new hydrogen bonds are formed between the film components,resulting in a more homogeneous and dense structure.The colorimetric film has a significant color response to pH changes.These films were used in experiments on the freshness of chilled pork,in which the pH changes with changing freshness states.The results show that the colorimetric film can monitor changes in the freshness of chilled pork in real time,where orange,pink,and green represent the fresh,secondary fresh,and putrefied states of pork,respectively.Therefore,the intelligent colorimetric film developed in this study has good application potential in the food industry.展开更多
Sedimentation values, flour glutenin macropolymer (GMP) contents, composition and contents of high-molecular-weight (HMW) glutenin subunits (GS) of 233 flour samples were determined. Our data indicated that subunit 1 ...Sedimentation values, flour glutenin macropolymer (GMP) contents, composition and contents of high-molecular-weight (HMW) glutenin subunits (GS) of 233 flour samples were determined. Our data indicated that subunit 1 occurred more frequently at Glu-A1 , subunit pair 7 + 8 at Glu-B1 and 2 + 12 at Glu-D1. The significant relationships between Glu-1 quality score and total HMW glutenin content, sedimentation value and GMP content suggested that the composition of HMW-GS affects wheat quality strongly. Moreover, the total content of HMW-GS was correlated with certain quality parameters more significantly. Relationship between subunit 5 + 10 content and breadmaking quality was better than others, but 2 + 12, 7 + 8, 7 + 9 and 4 + 12 also correlated with certain quality parameters significantly. The contents of total HMW-glutenin, x-type subunits and y-type subunits related with sedimentation value, flour GMP content, and Glu-1 quality score more strongly than that of individual subunit or subunit pair. The flour GMP content, with excellent correlation to sedimentation value, total contents of HMW glutenin, x- and y-type subunits and many other quality parameters, could be an ideal indicator of breadmaking quality at earlier generations for breeding purpose for its simple procedure and small scale.展开更多
The high-molecular-weight (HMW) glutenin subunits and their coding genes from Aegilops umbellulata Zhuk. (UU, 2n = 2x = 14) were characterized using SDS-PAGE analysis and molecular approaches. SDS-PAGE analysis showed...The high-molecular-weight (HMW) glutenin subunits and their coding genes from Aegilops umbellulata Zhuk. (UU, 2n = 2x = 14) were characterized using SDS-PAGE analysis and molecular approaches. SDS-PAGE analysis showed that the 1Ux subunits from four different accessions possessed electrophoretic mobilities close to, or slower than, that displayed by the 1Dx2.2 subunit of common wheat. The electrophoretic mobilities of the 1Uy subunits were generally similar to those shown by the 1Dy subunits of common wheat. The complete open reading frames of the 1Ux and 1Uy genes were amplified by PCR and subsequently cloned and sequenced. Amino acid sequence comparisons suggested that the primary structure of the 1Ux and 1Uy subunits were identical to that of published HMW glutenin subunits from related species, Phylogenetic analysis indicated that the HMW glutenin subunits of Ae. umbellulata were most closely related to those encoded by the D genome of Triticeae.展开更多
In common wheat, insoluble glutenin （IG） is an important fraction of flour glutenin macropolymers, and insoluble glutenin content （IGC） is positively associated with key end-use quality parameters. Here, we presen...In common wheat, insoluble glutenin （IG） is an important fraction of flour glutenin macropolymers, and insoluble glutenin content （IGC） is positively associated with key end-use quality parameters. Here, we present a genetic analysis of the chromosomal loci affecting IGC with the data collected from 90 common wheat varieties cultivated in four environments. Statistical analysis showed that IGC was controlled mainly genetically and influenced by the environment. Among the major genetic components known to affect end-use quality, 1BL/1RS translocation had a significantly negative effect on IGC across all four environments. As to the different alleles of Glu-A1, -B1 and -D1 loci, Glu-Ala, Glu-Blb and Glu-Dld exhibited relatively strong positive effects on IGC in all environments. To identify new loci affecting IGC, association mapping with 1355 DArT markers was conducted. A total of 133 markers were found associated with IGC in two or more environments （P 〈 0.05）, ten of which consistently affected IGC in all four environments. The phenotypic variance explained by the ten markers varied from 4.66% to 8.03%, and their elite alleles performed significantly better than the inferior counterparts in enhancing IGC. Among the ten markers, wPt-3743 and wPt-733835 reflected the action of Glu-D1, and wPt-664972 probably indicated the effect of GIu-A1. The other seven markers, forming three clusters on 2AL. 3BL or 7BL chromosome arms, represented newly identified genetic determinants of IGC. Our work provided novel insights into the genetic control of IGC, which may facilitate wheat end- use quality improvement through molecular breeding in the future.展开更多
The biosynthetic time and accumulations of A-, B-, and C-type glutenin subunits in 7 winter wheat cultivars with different quality （strong, medium, weak gluten） were analyzed by SDS-PAGE. The results showed that no ...The biosynthetic time and accumulations of A-, B-, and C-type glutenin subunits in 7 winter wheat cultivars with different quality （strong, medium, weak gluten） were analyzed by SDS-PAGE. The results showed that no glutenin subunit was observed within 8 d after anthesis. Parts or all A-, B-, and C-type subunits appeared around day 12 in different cultivars. Other A-, B-, and C-type subunits appeared gradually. The accumulation of A-, B-, and C-type subunits fluctuated before maturity. The results of analysis of correlation between the ratios of A/T （total content of glutenin subunits）, A/C, AJ （B＋C）, （A＋B）/C, and （A＋B）/T and SDS-sedimentation value suggested that they were more significant. The negative correlation between the ratio of （B＋C）/T and SDS-sedimentation value was more significant, and the correlations between the ratio C/T and the SDS-sedimentation value were significantly negative.展开更多
Nitrogen(N),a macronutrient essential for plant growth and development,is needed for biosynthesis of protein and starch,which affect grain yield and quality.Application of high-N fertilizer increases plant growth,grai...Nitrogen(N),a macronutrient essential for plant growth and development,is needed for biosynthesis of protein and starch,which affect grain yield and quality.Application of high-N fertilizer increases plant growth,grain yield,and flour quality.In this study,we performed the first comparative analysis of gliadin and glutenin subproteomes during kernel development in the elite Chinese wheat cultivar Zhongmai 175 under high-N conditions by reversed-phase ultra-performance liquid chromatography and twodimensional difference gel electrophoresis(2D-DIGE).Application of high-N fertilizer led to significant increases in gluten macropolymer content,total gliadin and glutenin content,and the accumulation of individual storage protein components.Of 126 differentially accumulated proteins(DAPs)induced by high-N conditions,24 gliadins,12 high-molecularweight glutenins,and 27 low-molecular-weight glutenins were significantly upregulated.DAPs during five kernel developmental stages displayed multiple patterns of accumulation.In particular,gliadins and glutenins showed respectively five and six accumulation patterns.The accumulation of storage proteins under high-N conditions may lead to improved dough properties and bread quality.展开更多
Using degenerate oligonucleotide primers and genomic PCR reactions,the complete coding region sequences of two 1Ay high molecular weight(HMW)glutenin subunit genes were amplified from Triticum urartu accessions that s...Using degenerate oligonucleotide primers and genomic PCR reactions,the complete coding region sequences of two 1Ay high molecular weight(HMW)glutenin subunit genes were amplified from Triticum urartu accessions that showed differential expression of the 1Ay HMW glutenin subunits in their seeds.The coding sequence amplified from the accession that expressed the 1Ay gene(Tu1Ay-e)was highly homologous to that of known y type HMW glutenin subunit genes.Consequently,the primary structure of the protein translated from the coding sequence of Tu/Ay-e was identical to that of previously published y type subunits.Bacterial expression of the coding sequence of Tu/Ay-e produced a polypeptide identical to the 1Ay subunit extracted from seeds,indicating that the cloned sequence was an accurate representation of the original coding region of Tu/Ay-e.In contrast,the coding region sequence amplified from the accession that did not express 1Ay subunit contained three in-frame premature stop codons.Based on past findings on silenced HMW glutenin subunit genes,we conclude that the presence ofin-frame premature stop codon(s)is an important feature of the silenced 1Ay gene(Tu1Ay-s)in T.urartuThe potential value of the active 1Ay gene in improving the end use quality of common wheat and the mechanism underlying 1Ay gene silencing are discussed.展开更多
Parental varieties Suneca and Cook have contrasting alleles at each of the five glutenin subunit loci (Glu-B1, Glu-D1 Glu-A3, Glu-B3, and Glu-D3), a set of 60 lines homozygous at these loci from the F4 progeny populat...Parental varieties Suneca and Cook have contrasting alleles at each of the five glutenin subunit loci (Glu-B1, Glu-D1 Glu-A3, Glu-B3, and Glu-D3), a set of 60 lines homozygous at these loci from the F4 progeny population of Suneca X Cook was chosen to analyze the variation of the size distribution of glutenin polymeric protein (measured by SE-HPLC) and flour mixing properties of these lines and to study relationship between the size distribution of glutenin polymeric protein and wheat flour mixing properties. The results showed that there were very significant differences among the relative size distributions of glutenin polymeric protein (i.e. percentage of unextracTable polymeric protein in the total polymeric protein, or UPP%) and dough development times (i.e. peak time of mixograph, or PTM) of different homozygous lines, respectively. Flour mixograph shape was closely related to UPP% value. The results also indicated that UPP% was very strongly positive correlation with PTM and negative correlation with peak height of mixograph (PHM). Comparing with flour protein content (FP%), UPP% gave greater effect on PTM and PHM, i.e. flour mixing properties, and it can be considered as one of criteria for quality selecting from early generation of breeding program.展开更多
Allelic variation and genetic diversity at HMW glutenin subunits loci, Glu-A1, Glu-B1and Glu-D1 were investigated in 64 accessions of three unique wheats of western Chinausing sodium dodecyl sulphate polyacrylamide ge...Allelic variation and genetic diversity at HMW glutenin subunits loci, Glu-A1, Glu-B1and Glu-D1 were investigated in 64 accessions of three unique wheats of western Chinausing sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Two HMWglutenin patterns (i.e., null, 7+8, 2+12 and null, 7, 2+12) in 34 Yunnan wheataccessions, 3 HMW glutenin patterns (i.e., null, 7+8, 2+12; null, 6+8, 2+12 andnull, 7+8, 2) in 24 Tibetan accessions and 1 HMW glutenin pattern (null, 7, 2+12) in6 Xinjiang wheat accessions were found. The Tibetan accession TB18 was found to be witha rare subunit 2 encoded by Glu-D1. A total of 4 (i.e., Glu-A1c, Glu-B1a, Glu-B1b andGlu-D1a), 5 (i.e., Glu-A1c, Glu-B1d, Glu-B1b, Glu-D1a and Glu-D1) and 3 alleles (i.e.,Glu-A1c, Glu-B1a and Glu-D1a) at Glu-1 locus were identified among Yunnan, Tibetan andXinjiang unique wheat accessions, respectively. For Yunnan wheat, Tibetan wheat andXinjiang wheat, the Neis mean genetic variation indexes were 0.1574, 0.1366 and 0,respectively, which might indicate the higher genetic diversity at HMW glutenin subunitsloci of Yunnan and Tibetan wheat accessions as compared to that of Xinjiang wheataccessions. Among the three genomes of hexaploid wheats of western China, the highestNeis genetic variation index was appeared in B genome with the mean value of 0.2674,while the indexes for genomes A and D were 0 and 0.0270, respectively. It might bereasonable to indicate that Glu-B1 showed the highest, Glu-D1 the intermediate and Glu-A1 always the lowest genetic diversity.展开更多
The objective of the present study was to characterize the high molecular glutenin subunits （HMW-GS） composition and the presence of 1B/1R translocation in newly developed wheat （Triticum aestivum L.） germplasm, w...The objective of the present study was to characterize the high molecular glutenin subunits （HMW-GS） composition and the presence of 1B/1R translocation in newly developed wheat （Triticum aestivum L.） germplasm, which have one or more traits that are useful in wheat improvement. Sodium dodecyl sulphate polyacrylamide-gel electrophoresis （SDS-PAGE） and acid polyacrylamide-gel electrophoresis （A-PAGE） were used to detect HMW-GS composition and the presence of 1B/1R wheat-rye （Secale cereale L.） chromosome translocation in the wheat germplasm. Bread-making quality scores of these lines were determined. A high level of variations in HMW-GS encoded by Glu-1 locus was observed. Sixteen major HMW-GS, with 30 combinations, were detected. The percentage of cultivars with more than two desirable subunits was 38.7%. Thirteen cultivars had bread-making quality scores of 10 in combination with one or two desirable agronomical traits, such as high-yield potential, dwarfing stem, resistance to diseases, and/or tolerance to abiotic stress. Sixty-eight （36.6%） cultivars possessed 1B/1R translocation. The newly developed germplasm with HMW-GS for good quality can be promising resources for improving bread-making quality of wheat.展开更多
The swelling index of glutenin (SIG) and the protein fraction of 25 Chinese wheat varieties were studied with new protein fractions extracting method. The protein fractions compose of monomeric protein, soluble glut...The swelling index of glutenin (SIG) and the protein fraction of 25 Chinese wheat varieties were studied with new protein fractions extracting method. The protein fractions compose of monomeric protein, soluble glutenin and insoluble glutenin. The relations between other protein index, dough character, and fresh noodle quality were also discussed. The SIG results at different time is positively and highly significantly related to the insoluble glutenin content (r= 0.808 -0.867, P< 0.01). The SIG result can reflect the insoluble glutenin content. The protein content, gluten index, farinograph stability time, extensibility length and extensigram energy were positively and significantly correlated with SIG5 and SIG20 (r= 0.516 - 0.734, P<0.05, 0.01).SIG proved to be applicable in Chinese wheat dough evaluation. Fresh noodle making quality parameters were evaluated by fresh noodle length, thickness, maximum resistance to extension, extension area and extension distance, while cooked noodle texture was determined by cutting firmness, compression recovery, surface firmness and TPA by using a texture analyzer of TA.XT2i. The noodle cooking quality was significantly correlated with SIG value. The surface firmness and cutting firmness were more desirable for predicating the quality difference than TPA test and compression. Cooking loss and water absorption were negatively related to SIG value and insoluble content (r = -0.556 - - 0.787, P < 0.05, 0.01). The results showed that SIG test was also suitable in evaluating noodle making and cooking quality in very small sample size, which was very important in wheat breeding programs. Therefore, SIG test, as a small scale test, is suitable to evaluate dough rheological properties for Chinese wheat varieties, and will be helpful in cereal research and wheat breeding program, especially, in early generations.展开更多
Size distribution of glutenin macropolymer （GMP） particles significantly affects the quality of final products in wheat. Six wheat varieties in eastern China were used for investigating the GMP content, particle siz...Size distribution of glutenin macropolymer （GMP） particles significantly affects the quality of final products in wheat. Six wheat varieties in eastern China were used for investigating the GMP content, particle size distribution and their relation. The results showed that the particle diameter of GMP changed from 0.38 μm to 269 μm. A typical bimodal curve of volume distribution of particles with peak values ranged from 4.44 μm to 5.36 μm and from 45.76 μm to 116.30 μm, respectively. Number distribution of particles showed the typical population with peak values in the range of 0.57-1.00 p.m. Proportions of granules 〈12μm, 12-80 μm and 〉80 μm were in the range of 17.1%-47.8%, 32.1%-50.3% and 10.2%-38.1% of total volume, respectively. The content of GMP was negatively correlated with the volume of GMP particles 〈12μm, but positively correlated with the volume of particles 〉80μm. The results suggested that the greater percentage of larger GMP particles is, the more GMP content there is in wheat grain.展开更多
As one of the most important cereals,wheat(Triticum aestivum)has high nutritional value and is widely cultivated in China.However,wheat can cause severe allergic reactions,and a growing number of people are developing...As one of the most important cereals,wheat(Triticum aestivum)has high nutritional value and is widely cultivated in China.However,wheat can cause severe allergic reactions,and a growing number of people are developing allergies to Chinese wheat.Low molecular weight glutenin(LMW-GS),an important allergen in susceptible populations,is responsible for celiac disease and wheat contacts dermatitis.In this study,LMW-GS was highly purified from Chinese wheat(Xiaoyan 6)and further identified and characterized.In addition,8 peptides were predicted efficiently by 5 immunological tools,among which 5 peptides showed significant immunoglobulin E binding abilities.Two specific epitopes were found to be in the non-conserved region of the amino acid sequence of LMW-GS,which was speculated to be the specific epitope of Chinese wheat.This systematic research of LMW-GS may provide new insights into the prevention of wheat allergy and development of hypoallergenic wheat products.展开更多
The low molecular weight （LMW） glutenln subunlts account for 40% of wheat gluten protein content by mass and these proteins are considered to significantly affect dough quality characteristics. Five new full-length ...The low molecular weight （LMW） glutenln subunlts account for 40% of wheat gluten protein content by mass and these proteins are considered to significantly affect dough quality characteristics. Five new full-length LMW glutenln genes （designated LMW-5, LMW-7, LMW-42, LMW-58, and LMW-34） were isolated from the Chinese elite wheat cultivar ＂Xlaoyan 54＂ by PCR amplification of genomlc DNA using a pair of degenerate primers designed from the conserved sequences of the N- and C-terminal regions of published LMW glutenln genes. Deduced amino acid sequence analysis showed that LMW-5 belongs to the LMW-i type genes and that the other four belong to LMW-m type genes. Sequence comparisons revealed that point mutations occasionally occurred in signal peptide and N-terminus domains and often existed in domain III and domain V. Small insertions and deletions are represented in the repetitive domain. There is a stop codon after amino acid position 110 In the repetitive domain of LMW.34, indicating that It is a pseudogene. The other four genes have complete open reading frames and the putative mature regions of these genes were subcloned Into pET-30a expression vector and successfully expressed in Escherlchla coll. Protein sodium dodecyl sulfate-polyacrylamlde gel electro- phoresls analysis showed that all proteins expressed in E. coil by the four genes could be related to B-group LMW glutenln subunits of wheat.展开更多
Aegilops tauschii is the donor of the D genome of common wheat (Triticum aestivum). Genetic variation of HMW glutenin subunits encoded by the Glu-1Dt locus of Ae. tauschii has been found to be higher than that specifi...Aegilops tauschii is the donor of the D genome of common wheat (Triticum aestivum). Genetic variation of HMW glutenin subunits encoded by the Glu-1Dt locus of Ae. tauschii has been found to be higher than that specified by the Glu-1D locus in common wheat. In the present note, we report the identification of a novel HMW glutenin subunit, Dy13t, from Ae. tauschii. The newly identified subunit pos-sessed an electrophoretic mobility that was faster than that of the Dy12 subunit of common wheat. The complete ORF of encoding the Dy13t subunit contained 624 codons (excluding the stop codons). The amino acid sequence deduced from the Dy13t gene ORF was the shortest among those of the previ-ously reported subunits derived by the D genome. A further comparison of Dy13t amino acid sequence with those of the subunits characterized from the A, B, D, R genomes of Triticeae showed that the smaller size of the Dy13t subunit was associated with a reduction in the size of its repetitive domain.展开更多
The Aegilops genus contains species closely related to wheat. In common with wheat, Aegilops species accumulate high molecular weight (HMW) glutenin subunits in their endospermic tissue. In this study, we investigated...The Aegilops genus contains species closely related to wheat. In common with wheat, Aegilops species accumulate high molecular weight (HMW) glutenin subunits in their endospermic tissue. In this study, we investigated the composition of HMW glutenin subunits in four multiploid Aegilops species using SDS-PAGE analysis. Furthermore, by working with Ae. ventricosa, we established an efficient ge-nomic PCR condition for simultaneous amplification of DNA sequences coding for either x- or y-type HMW glutenin sub-units from polyploid Aegilops species. Using the genomic PCR condition, we amplified and subsequently cloned two DNA fragments that may code for HMW glutenin subunits in Ae. ventricosa. Based on an analysis of the deduced amino acid sequences, we concluded that the two cloned sequences encode one x- and one y-type of HMW glutenin subunit, respectively.展开更多
基金funded by the Hainan Provincial Natural Science Foundation of China[Grant Number 2019RC031]National Natural Science Foundation of China[Grant Number 31460407].
文摘In this study,intelligent,pH-responsive colorimetric films were prepared by encapsulating anthocyanins in nanocomplexes prepared from glutenin and carboxymethyl chitosan.These nanocomplexes were added to a corn starch matrix and used in the freshness monitoring of chilled pork.The effects of anthocyanin-loaded nanocomplexes on the physical,structural,and functional characteristics of the films were investigated.The addition of anthocyanin-loaded nanocomplexes increased the tensile strength,elongation at break,hydrophobicity,and light transmittance of the films while decreasing their water vapor permeability.This is because new hydrogen bonds are formed between the film components,resulting in a more homogeneous and dense structure.The colorimetric film has a significant color response to pH changes.These films were used in experiments on the freshness of chilled pork,in which the pH changes with changing freshness states.The results show that the colorimetric film can monitor changes in the freshness of chilled pork in real time,where orange,pink,and green represent the fresh,secondary fresh,and putrefied states of pork,respectively.Therefore,the intelligent colorimetric film developed in this study has good application potential in the food industry.
基金the National Natural Science Foundation of China(No.39970456 ,39930110).
文摘Sedimentation values, flour glutenin macropolymer (GMP) contents, composition and contents of high-molecular-weight (HMW) glutenin subunits (GS) of 233 flour samples were determined. Our data indicated that subunit 1 occurred more frequently at Glu-A1 , subunit pair 7 + 8 at Glu-B1 and 2 + 12 at Glu-D1. The significant relationships between Glu-1 quality score and total HMW glutenin content, sedimentation value and GMP content suggested that the composition of HMW-GS affects wheat quality strongly. Moreover, the total content of HMW-GS was correlated with certain quality parameters more significantly. Relationship between subunit 5 + 10 content and breadmaking quality was better than others, but 2 + 12, 7 + 8, 7 + 9 and 4 + 12 also correlated with certain quality parameters significantly. The contents of total HMW-glutenin, x-type subunits and y-type subunits related with sedimentation value, flour GMP content, and Glu-1 quality score more strongly than that of individual subunit or subunit pair. The flour GMP content, with excellent correlation to sedimentation value, total contents of HMW glutenin, x- and y-type subunits and many other quality parameters, could be an ideal indicator of breadmaking quality at earlier generations for breeding purpose for its simple procedure and small scale.
文摘The high-molecular-weight (HMW) glutenin subunits and their coding genes from Aegilops umbellulata Zhuk. (UU, 2n = 2x = 14) were characterized using SDS-PAGE analysis and molecular approaches. SDS-PAGE analysis showed that the 1Ux subunits from four different accessions possessed electrophoretic mobilities close to, or slower than, that displayed by the 1Dx2.2 subunit of common wheat. The electrophoretic mobilities of the 1Uy subunits were generally similar to those shown by the 1Dy subunits of common wheat. The complete open reading frames of the 1Ux and 1Uy genes were amplified by PCR and subsequently cloned and sequenced. Amino acid sequence comparisons suggested that the primary structure of the 1Ux and 1Uy subunits were identical to that of published HMW glutenin subunits from related species, Phylogenetic analysis indicated that the HMW glutenin subunits of Ae. umbellulata were most closely related to those encoded by the D genome of Triticeae.
基金supported by the Ministry of Science and Technology of China (Grant No. 2014CB138104)Chinese Academy of Sciences (Grant No. XDA08010302)the National Science Foundation of China (Grant No. 31371611)
文摘In common wheat, insoluble glutenin （IG） is an important fraction of flour glutenin macropolymers, and insoluble glutenin content （IGC） is positively associated with key end-use quality parameters. Here, we present a genetic analysis of the chromosomal loci affecting IGC with the data collected from 90 common wheat varieties cultivated in four environments. Statistical analysis showed that IGC was controlled mainly genetically and influenced by the environment. Among the major genetic components known to affect end-use quality, 1BL/1RS translocation had a significantly negative effect on IGC across all four environments. As to the different alleles of Glu-A1, -B1 and -D1 loci, Glu-Ala, Glu-Blb and Glu-Dld exhibited relatively strong positive effects on IGC in all environments. To identify new loci affecting IGC, association mapping with 1355 DArT markers was conducted. A total of 133 markers were found associated with IGC in two or more environments （P 〈 0.05）, ten of which consistently affected IGC in all four environments. The phenotypic variance explained by the ten markers varied from 4.66% to 8.03%, and their elite alleles performed significantly better than the inferior counterparts in enhancing IGC. Among the ten markers, wPt-3743 and wPt-733835 reflected the action of Glu-D1, and wPt-664972 probably indicated the effect of GIu-A1. The other seven markers, forming three clusters on 2AL. 3BL or 7BL chromosome arms, represented newly identified genetic determinants of IGC. Our work provided novel insights into the genetic control of IGC, which may facilitate wheat end- use quality improvement through molecular breeding in the future.
文摘The biosynthetic time and accumulations of A-, B-, and C-type glutenin subunits in 7 winter wheat cultivars with different quality （strong, medium, weak gluten） were analyzed by SDS-PAGE. The results showed that no glutenin subunit was observed within 8 d after anthesis. Parts or all A-, B-, and C-type subunits appeared around day 12 in different cultivars. Other A-, B-, and C-type subunits appeared gradually. The accumulation of A-, B-, and C-type subunits fluctuated before maturity. The results of analysis of correlation between the ratios of A/T （total content of glutenin subunits）, A/C, AJ （B＋C）, （A＋B）/C, and （A＋B）/T and SDS-sedimentation value suggested that they were more significant. The negative correlation between the ratio of （B＋C）/T and SDS-sedimentation value was more significant, and the correlations between the ratio C/T and the SDS-sedimentation value were significantly negative.
基金financially supported by the National Key Research and Development Program of China(2016YFD0100502)the National Natural Science Foundation of China(31171773)
文摘Nitrogen(N),a macronutrient essential for plant growth and development,is needed for biosynthesis of protein and starch,which affect grain yield and quality.Application of high-N fertilizer increases plant growth,grain yield,and flour quality.In this study,we performed the first comparative analysis of gliadin and glutenin subproteomes during kernel development in the elite Chinese wheat cultivar Zhongmai 175 under high-N conditions by reversed-phase ultra-performance liquid chromatography and twodimensional difference gel electrophoresis(2D-DIGE).Application of high-N fertilizer led to significant increases in gluten macropolymer content,total gliadin and glutenin content,and the accumulation of individual storage protein components.Of 126 differentially accumulated proteins(DAPs)induced by high-N conditions,24 gliadins,12 high-molecularweight glutenins,and 27 low-molecular-weight glutenins were significantly upregulated.DAPs during five kernel developmental stages displayed multiple patterns of accumulation.In particular,gliadins and glutenins showed respectively five and six accumulation patterns.The accumulation of storage proteins under high-N conditions may lead to improved dough properties and bread quality.
文摘Using degenerate oligonucleotide primers and genomic PCR reactions,the complete coding region sequences of two 1Ay high molecular weight(HMW)glutenin subunit genes were amplified from Triticum urartu accessions that showed differential expression of the 1Ay HMW glutenin subunits in their seeds.The coding sequence amplified from the accession that expressed the 1Ay gene(Tu1Ay-e)was highly homologous to that of known y type HMW glutenin subunit genes.Consequently,the primary structure of the protein translated from the coding sequence of Tu/Ay-e was identical to that of previously published y type subunits.Bacterial expression of the coding sequence of Tu/Ay-e produced a polypeptide identical to the 1Ay subunit extracted from seeds,indicating that the cloned sequence was an accurate representation of the original coding region of Tu/Ay-e.In contrast,the coding region sequence amplified from the accession that did not express 1Ay subunit contained three in-frame premature stop codons.Based on past findings on silenced HMW glutenin subunit genes,we conclude that the presence ofin-frame premature stop codon(s)is an important feature of the silenced 1Ay gene(Tu1Ay-s)in T.urartuThe potential value of the active 1Ay gene in improving the end use quality of common wheat and the mechanism underlying 1Ay gene silencing are discussed.
文摘Parental varieties Suneca and Cook have contrasting alleles at each of the five glutenin subunit loci (Glu-B1, Glu-D1 Glu-A3, Glu-B3, and Glu-D3), a set of 60 lines homozygous at these loci from the F4 progeny population of Suneca X Cook was chosen to analyze the variation of the size distribution of glutenin polymeric protein (measured by SE-HPLC) and flour mixing properties of these lines and to study relationship between the size distribution of glutenin polymeric protein and wheat flour mixing properties. The results showed that there were very significant differences among the relative size distributions of glutenin polymeric protein (i.e. percentage of unextracTable polymeric protein in the total polymeric protein, or UPP%) and dough development times (i.e. peak time of mixograph, or PTM) of different homozygous lines, respectively. Flour mixograph shape was closely related to UPP% value. The results also indicated that UPP% was very strongly positive correlation with PTM and negative correlation with peak height of mixograph (PHM). Comparing with flour protein content (FP%), UPP% gave greater effect on PTM and PHM, i.e. flour mixing properties, and it can be considered as one of criteria for quality selecting from early generation of breeding program.
文摘Allelic variation and genetic diversity at HMW glutenin subunits loci, Glu-A1, Glu-B1and Glu-D1 were investigated in 64 accessions of three unique wheats of western Chinausing sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Two HMWglutenin patterns (i.e., null, 7+8, 2+12 and null, 7, 2+12) in 34 Yunnan wheataccessions, 3 HMW glutenin patterns (i.e., null, 7+8, 2+12; null, 6+8, 2+12 andnull, 7+8, 2) in 24 Tibetan accessions and 1 HMW glutenin pattern (null, 7, 2+12) in6 Xinjiang wheat accessions were found. The Tibetan accession TB18 was found to be witha rare subunit 2 encoded by Glu-D1. A total of 4 (i.e., Glu-A1c, Glu-B1a, Glu-B1b andGlu-D1a), 5 (i.e., Glu-A1c, Glu-B1d, Glu-B1b, Glu-D1a and Glu-D1) and 3 alleles (i.e.,Glu-A1c, Glu-B1a and Glu-D1a) at Glu-1 locus were identified among Yunnan, Tibetan andXinjiang unique wheat accessions, respectively. For Yunnan wheat, Tibetan wheat andXinjiang wheat, the Neis mean genetic variation indexes were 0.1574, 0.1366 and 0,respectively, which might indicate the higher genetic diversity at HMW glutenin subunitsloci of Yunnan and Tibetan wheat accessions as compared to that of Xinjiang wheataccessions. Among the three genomes of hexaploid wheats of western China, the highestNeis genetic variation index was appeared in B genome with the mean value of 0.2674,while the indexes for genomes A and D were 0 and 0.0270, respectively. It might bereasonable to indicate that Glu-B1 showed the highest, Glu-D1 the intermediate and Glu-A1 always the lowest genetic diversity.
文摘The objective of the present study was to characterize the high molecular glutenin subunits （HMW-GS） composition and the presence of 1B/1R translocation in newly developed wheat （Triticum aestivum L.） germplasm, which have one or more traits that are useful in wheat improvement. Sodium dodecyl sulphate polyacrylamide-gel electrophoresis （SDS-PAGE） and acid polyacrylamide-gel electrophoresis （A-PAGE） were used to detect HMW-GS composition and the presence of 1B/1R wheat-rye （Secale cereale L.） chromosome translocation in the wheat germplasm. Bread-making quality scores of these lines were determined. A high level of variations in HMW-GS encoded by Glu-1 locus was observed. Sixteen major HMW-GS, with 30 combinations, were detected. The percentage of cultivars with more than two desirable subunits was 38.7%. Thirteen cultivars had bread-making quality scores of 10 in combination with one or two desirable agronomical traits, such as high-yield potential, dwarfing stem, resistance to diseases, and/or tolerance to abiotic stress. Sixty-eight （36.6%） cultivars possessed 1B/1R translocation. The newly developed germplasm with HMW-GS for good quality can be promising resources for improving bread-making quality of wheat.
文摘The swelling index of glutenin (SIG) and the protein fraction of 25 Chinese wheat varieties were studied with new protein fractions extracting method. The protein fractions compose of monomeric protein, soluble glutenin and insoluble glutenin. The relations between other protein index, dough character, and fresh noodle quality were also discussed. The SIG results at different time is positively and highly significantly related to the insoluble glutenin content (r= 0.808 -0.867, P< 0.01). The SIG result can reflect the insoluble glutenin content. The protein content, gluten index, farinograph stability time, extensibility length and extensigram energy were positively and significantly correlated with SIG5 and SIG20 (r= 0.516 - 0.734, P<0.05, 0.01).SIG proved to be applicable in Chinese wheat dough evaluation. Fresh noodle making quality parameters were evaluated by fresh noodle length, thickness, maximum resistance to extension, extension area and extension distance, while cooked noodle texture was determined by cutting firmness, compression recovery, surface firmness and TPA by using a texture analyzer of TA.XT2i. The noodle cooking quality was significantly correlated with SIG value. The surface firmness and cutting firmness were more desirable for predicating the quality difference than TPA test and compression. Cooking loss and water absorption were negatively related to SIG value and insoluble content (r = -0.556 - - 0.787, P < 0.05, 0.01). The results showed that SIG test was also suitable in evaluating noodle making and cooking quality in very small sample size, which was very important in wheat breeding programs. Therefore, SIG test, as a small scale test, is suitable to evaluate dough rheological properties for Chinese wheat varieties, and will be helpful in cereal research and wheat breeding program, especially, in early generations.
基金Supported by the National Key Research and Development Program of China(2016YFD0300408)the Anhui Provincial Natural Science Foundation(1408085MC48)
文摘Size distribution of glutenin macropolymer （GMP） particles significantly affects the quality of final products in wheat. Six wheat varieties in eastern China were used for investigating the GMP content, particle size distribution and their relation. The results showed that the particle diameter of GMP changed from 0.38 μm to 269 μm. A typical bimodal curve of volume distribution of particles with peak values ranged from 4.44 μm to 5.36 μm and from 45.76 μm to 116.30 μm, respectively. Number distribution of particles showed the typical population with peak values in the range of 0.57-1.00 p.m. Proportions of granules 〈12μm, 12-80 μm and 〉80 μm were in the range of 17.1%-47.8%, 32.1%-50.3% and 10.2%-38.1% of total volume, respectively. The content of GMP was negatively correlated with the volume of GMP particles 〈12μm, but positively correlated with the volume of particles 〉80μm. The results suggested that the greater percentage of larger GMP particles is, the more GMP content there is in wheat grain.
基金financially supported by the State Key Research and Development Plan(2019YFC1605002)。
文摘As one of the most important cereals,wheat(Triticum aestivum)has high nutritional value and is widely cultivated in China.However,wheat can cause severe allergic reactions,and a growing number of people are developing allergies to Chinese wheat.Low molecular weight glutenin(LMW-GS),an important allergen in susceptible populations,is responsible for celiac disease and wheat contacts dermatitis.In this study,LMW-GS was highly purified from Chinese wheat(Xiaoyan 6)and further identified and characterized.In addition,8 peptides were predicted efficiently by 5 immunological tools,among which 5 peptides showed significant immunoglobulin E binding abilities.Two specific epitopes were found to be in the non-conserved region of the amino acid sequence of LMW-GS,which was speculated to be the specific epitope of Chinese wheat.This systematic research of LMW-GS may provide new insights into the prevention of wheat allergy and development of hypoallergenic wheat products.
基金Supported by the National Natural Science Foundation of China （30571153）.
文摘The low molecular weight （LMW） glutenln subunlts account for 40% of wheat gluten protein content by mass and these proteins are considered to significantly affect dough quality characteristics. Five new full-length LMW glutenln genes （designated LMW-5, LMW-7, LMW-42, LMW-58, and LMW-34） were isolated from the Chinese elite wheat cultivar ＂Xlaoyan 54＂ by PCR amplification of genomlc DNA using a pair of degenerate primers designed from the conserved sequences of the N- and C-terminal regions of published LMW glutenln genes. Deduced amino acid sequence analysis showed that LMW-5 belongs to the LMW-i type genes and that the other four belong to LMW-m type genes. Sequence comparisons revealed that point mutations occasionally occurred in signal peptide and N-terminus domains and often existed in domain III and domain V. Small insertions and deletions are represented in the repetitive domain. There is a stop codon after amino acid position 110 In the repetitive domain of LMW.34, indicating that It is a pseudogene. The other four genes have complete open reading frames and the putative mature regions of these genes were subcloned Into pET-30a expression vector and successfully expressed in Escherlchla coll. Protein sodium dodecyl sulfate-polyacrylamlde gel electro- phoresls analysis showed that all proteins expressed in E. coil by the four genes could be related to B-group LMW glutenln subunits of wheat.
基金This work was supported by the "973" Key Basic Research and Development Program (Grant No. G1998010208) the special fund for biotechnology research from the Chinese Academy of Sciences.
文摘Aegilops tauschii is the donor of the D genome of common wheat (Triticum aestivum). Genetic variation of HMW glutenin subunits encoded by the Glu-1Dt locus of Ae. tauschii has been found to be higher than that specified by the Glu-1D locus in common wheat. In the present note, we report the identification of a novel HMW glutenin subunit, Dy13t, from Ae. tauschii. The newly identified subunit pos-sessed an electrophoretic mobility that was faster than that of the Dy12 subunit of common wheat. The complete ORF of encoding the Dy13t subunit contained 624 codons (excluding the stop codons). The amino acid sequence deduced from the Dy13t gene ORF was the shortest among those of the previ-ously reported subunits derived by the D genome. A further comparison of Dy13t amino acid sequence with those of the subunits characterized from the A, B, D, R genomes of Triticeae showed that the smaller size of the Dy13t subunit was associated with a reduction in the size of its repetitive domain.
基金a special fund for biotechnology research from the Chinese Academy of Sciences (Grant No. STZ-3-11). The nucleotide sequences for the Aeveny1.9 and Aevenx2.5 subunit have been deposited in the EMBO database with the accession numbers AF226698 and AF22669
文摘The Aegilops genus contains species closely related to wheat. In common with wheat, Aegilops species accumulate high molecular weight (HMW) glutenin subunits in their endospermic tissue. In this study, we investigated the composition of HMW glutenin subunits in four multiploid Aegilops species using SDS-PAGE analysis. Furthermore, by working with Ae. ventricosa, we established an efficient ge-nomic PCR condition for simultaneous amplification of DNA sequences coding for either x- or y-type HMW glutenin sub-units from polyploid Aegilops species. Using the genomic PCR condition, we amplified and subsequently cloned two DNA fragments that may code for HMW glutenin subunits in Ae. ventricosa. Based on an analysis of the deduced amino acid sequences, we concluded that the two cloned sequences encode one x- and one y-type of HMW glutenin subunit, respectively.
