Rice storage proteins(RSPs)are plant proteins with high nutritional quality.As the second largest type of storage substance in rice,it is the main source of protein intake for people who consume rice as a staple food....Rice storage proteins(RSPs)are plant proteins with high nutritional quality.As the second largest type of storage substance in rice,it is the main source of protein intake for people who consume rice as a staple food.The content and type of RSPs affect the appearance,processing quality and eating quality of rice.These effects involve the distribution of RSPs in rice grains as well as the interactions of RSPs with other components such as starch in rice grains.In the past two decades,some progress has been made in the genetic improvement of RSPs.However,the determination mechanism of protein content and composition in rice is still unclear,and the mechanism of the effect of RSPs on rice quality has not been elucidated.In this review,the composition,biosynthesis and distribution of RSPs,and quantitative trait loci mapping and cloning of RSP genes are summarized,the research progress of the influence of RSPs and their components on rice quality are reviewed,and the research directions in the future are proposed.展开更多
One hundred and fifteen varieties （including cultivars and lines） with different ecotypes in japonica rice （Oryza sativa L.） were analyzed for endosperm storage proteins by sodium dodecyl sulfate polyacrylamide ge...One hundred and fifteen varieties （including cultivars and lines） with different ecotypes in japonica rice （Oryza sativa L.） were analyzed for endosperm storage proteins by sodium dodecyl sulfate polyacrylamide gel electrophoresis （SDS-PAGE） to estimate their genetic diversity for the purpose of genetic improving and variety identification. Nineteen types of profile were identified according to 1） presence/absence of 65 kDa bands, 2） staining intensity of 70, 60, 57, 37-39, 22-23, 13 and 10 kDa bands, 3） migration velocity of 35 kDa （α-4） and 4） band number at 57 kDa location. An unweighted-pair group average method with arithmetic mean （UPGMA） dendregram based on the cluster analysis of genetic similarity of the protein bands showed a small genetic variation among the tested materials, with the similarity coefficients varying between 0.75 and 1.00. Three distinct groups were identified from the cluster analysis of the rice varieties studied at the similarity coefficient level of 0.894. The first group included eight varieties with high amylose content, the second group contained fifteen varieties with high protein content, and the third group had the remaining ninety-two varieties, which accounted for 80% of the total materials. Clear relationship between ecotypes distinguished by maturity and groups revealed by cluster analysis was not found in this study. Only the group of high amylose linked with medium-maturity medium japonica ecotype. The bands of 70 kDa and 65 kDa can be used as protein markers to identify F1 seed purity of japonica hybrid rice Liuyanyou 422.展开更多
Rice(Oryza sativa L.)is the most important food crop for at least half of the world’s population.Due to improved living standards,the cultivation of high-quality rice for different purposes and markets has become a m...Rice(Oryza sativa L.)is the most important food crop for at least half of the world’s population.Due to improved living standards,the cultivation of high-quality rice for different purposes and markets has become a major goal.Rice quality is determined by the presence of many nutritional components,including seed storage proteins(SSPs),which are the second most abundant nutrient components of rice grains after starch.Rice SSP biosynthesis requires the participation of multiple organelles and is influenced by the external environment,making it challenging to understand the molecular details of SSP biosynthesis and improve rice protein quality.In this review,we highlight the current knowledge of rice SSP biosynthesis,including a detailed description of the key molecules involved in rice SSP biosynthetic processes and the major environmental factors affecting SSP biosynthesis.The effects of these factors on SSP accumulation and their contribution to rice quality are also discussed based on recent findings.This recent knowledge suggests not only new research directions for exploring rice SSP biosynthesis but also innovative strategies for breeding high-quality rice varieties.展开更多
In order to Identify appropriate plant materials for studying the gene expression and biological function of vegetative storage proteins （VSPs） in woody plants, the VSPs in the seedlings of Swietenla rnacrophylla Ki...In order to Identify appropriate plant materials for studying the gene expression and biological function of vegetative storage proteins （VSPs） in woody plants, the VSPs in the seedlings of Swietenla rnacrophylla King were investigated by using light microscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE） and Western.blotting. The seed of S. macrophylla was rich in storage proteins that accumulated In the vacuoles of cotyledon parenchyma cells in appearance of compact spherical grains. The growth and development of S. macrophylla seedlings were characterized by an obvious growth rhythm. The storage proteins In seeds disappeared during seedling growth while VSPs appeared in the stem 2 weeks after seedling leaves matured. Thereafter, the VSPs In the seedling stem almost exhausted during new shoot growth, and when the leaves of new shoot Just matured, both the stem beneath the new shoot of seedlings and the stem of new shoot started to accumulate VSPs. Nitrogen application dramatically Increased the level of VSPs, but had little influence on the dynamics of VSP consumption and accumulation in seedling stem. Together with these data, the fluctuation of VSPs in seedlings was very similar to that in the branches of the adult trees. In addition, seedlings are easy to be treated due to their small size. Our results suggested that S. rnacrophylla seedlings were suitable for Investigating the biological roles of VSPs and the mechanism of nitrogen storage in trees.展开更多
The occurrence of twin-arginine motifs （-R-R-） in the amino acid sequences of animal pro-proteins frequently defines the cleavage site（s） for their structural/functional maturation. No information is available on ...The occurrence of twin-arginine motifs （-R-R-） in the amino acid sequences of animal pro-proteins frequently defines the cleavage site（s） for their structural/functional maturation. No information is available on the presence and possible biological meaning of these motifs in the seed storage proteins. In this work, a novel endopeptidase activity with cleavage specificity to twin-arginine pairs has been detected in mature dry Lupinus albus seeds. The endopeptidase was tested with a number of endogenous and exogenous protein substrates, which were selected according to the pres- ence of one or more twin-arginine residue motifs in their amino acid sequences. The observed hydrolysis patterns were limited and highly specific. Partial proteolysis led to stable polypeptide fragments that were characterized by 1- and 2-D electrophoresis. Selected polypeptides were submitted to N-terminal amino acid sequencing and mass spectrometry anal- yses, These approaches, supported by bioinformatic analysis of the available sequences, allowed the conclusion that the polypeptide cleavage events had occurred at the peptide bonds comprised between twin-arginine residue pairs with all tested protein substrates. The endopeptidase activity was inhibited by 4-（2-AminoEthyl）Benzene-Sulphonyl Fluoride hy- drochloride （AEBSF）, leupeptin, and serine proteinase protein inhibitors, while it was not affected by pepstatin, trans- EpoxysuccinyI-L-leucylamido（4-guanidino）butane （E64）, and ethylenediaminetetraacetic acid （EDTA）, thus qualifying the Arg-Arg cleaving enzyme as a serine endopeptidase. The structural features of storage proteins from lupin and other legume seeds strongly support the hypothesis that the occurrence of an endopeptidase activity cleaving -R-R- bonds may be functional to facilitate their degradation at germination and possibly generate polypeptide fragments with specific biological activity.展开更多
The content and composition of wheat storage proteins are the major determinants of dough rheological properties and breadmaking quality and are influenced by cultivation conditions.This study aimed to investigate the...The content and composition of wheat storage proteins are the major determinants of dough rheological properties and breadmaking quality and are influenced by cultivation conditions.This study aimed to investigate the effects of water deficit and high N-fertilizer application on wheat storage protein synthesis,gluten secondary structure,and breadmaking quality.Reverse-phase ultrahigh-performance liquid chromatography analysis showed that storage protein and gluten macropolymer accumulation was promoted under both independent applications and a combination of water-deficit and high N-fertilizer treatments.Fourier-transform infrared spectroscopy showed that water deficit and high N-fertilizer treatments generally improved protein secondary structure formation and lipid accumulation,and reduced flour moisture.In particular,high N-fertilizer application increasedβ-sheet content by 10.4%and the combination of water-deficit and high N-fertilizer treatments increased random coil content by 7.6%.These changes in gluten content and secondary structure led to improved dough rheological properties and breadmaking quality,including superior loaf internal structure,volume,and score.Our results demonstrate that moderately high N-fertilizer application under drought conditions can improve gluten accumulation,gluten secondary structure formation,and baking quality.展开更多
The ultrastructural characteristics and the subcellular localization of vegetative storage proteins(VSPs)of Ginkgo biloba L.were systematically studied under the electron microscope.Results indicated that the VSPs of ...The ultrastructural characteristics and the subcellular localization of vegetative storage proteins(VSPs)of Ginkgo biloba L.were systematically studied under the electron microscope.Results indicated that the VSPs of G.biloba were mostly distributed in the small vacuoles of the phloem parenchyma cells.The VSPs of phloem parenchyma cells were produced in cytoplasm,then separated by the inflated cisternae of endoplasmic reticulum(ER),plasmalemma invagination or Golgi body vesicles,resulting in the formation of vacuole filled with proteins.Three kinds of VSPs were detected:granular,floccular and massive VSPs,which were distributed in different tissue cells and different cells of the same tissue or vacuoles of the same cells.VSPs accumulated in autumn and kept at high level throughout the winter.In the following spring,the bud started growing and VSPs were completely mobilized.With new shoots growth,VSPs resumed accumulating in late summer and early autumn.展开更多
Wide hybridization is an effective approach for enhancing the resistance of bread wheat （Triticum aestivum L.） to biotic and abiotic stresses by introducing favorable alien genes （Sepsi et al., 2008）. Wheatgrass, ...Wide hybridization is an effective approach for enhancing the resistance of bread wheat （Triticum aestivum L.） to biotic and abiotic stresses by introducing favorable alien genes （Sepsi et al., 2008）. Wheatgrass, Thinopyrum intermedium （Host） Barkworth ＆ D.R. Dewey or Agropyron intermedium （Host） Beauvoir （2n = 42; genome formula JJjSjSstst）, is a perennial species in the tribe Triticeae and an important source of wheat improvement for biotic and abiotic stress resistance and quality-related traits, such as high grain protein concentration （Chen et al., 1998; 2001; 2003; Han et al., 2004; Li and Wang, 2009）. In addition, the ready crossing ability of wheatgrass with various Triticum species has made it popular in germ- plasm development.展开更多
Soybean seed storage protein is one of the most important plant vegetable proteins, and β subunit is of great significance to enhance soybean protein quality and processing property. F2 segregated population and resi...Soybean seed storage protein is one of the most important plant vegetable proteins, and β subunit is of great significance to enhance soybean protein quality and processing property. F2 segregated population and residual heterozygous lines（RHL） derived from the cross between Yangyandou（low level of β subunit） and Zhonghuang 13（normal level of β subunit） were used for mapping of β subunit content. Our results showed that β subunit content was controlled by a single dominant locus, qBSC-1（β subunit content）, which was mapped to a region of 11.9 cM on chromosome 20 in F2 population of 85 individuals. This region was narrowed down to 2.5 cM between BARCSOYSSR_20_0997 and BARCSOYSSR_20_0910 in RHL with a larger population size of 246 individuals. There were 48 predicted genes within qBSC-1 region based on the reference genome（Glyma 1.0, Williams 82）, including the two copies of β subunit coding gene CG4. An InDel marker developed from a thymine（TT） insertion in one copy of CG4 promoter region in Yangyandou cosegregrated with BARCSOYSSR_20_0975 within qBSC-1 region, suggesting that this InDel marker maybe useful for marker-assisted selection（MAS）.展开更多
The characteristics of 48 rice varieties from Uttarakhand Himalaya, India were detected by morphological and biochemical markers. The grains of the selected rice varieties varied in their morphological （grain length,...The characteristics of 48 rice varieties from Uttarakhand Himalaya, India were detected by morphological and biochemical markers. The grains of the selected rice varieties varied in their morphological （grain length, grain width and grain weight） and biochemical characters （sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE）. Based on the presence of 70, 65, 60, 57, 37-39, 22-23, 13 and 10 kDa protein bands in the 48 rice varieties, seven types of profiles were identified. An unweighted pair group average method with arithmetic mean （UPGMA） dendrogram based on cluster analysis of genetic similarity of the protein bands showed two distinct groups with 1%-78% similarity coefficients. The presence of characteristic bands in selected varieties is a useful parameter for identification of rice germplasm.展开更多
Veronica erciyasdagi （M. A. Fischer） C. Vural comb. ＆ stat. nov., previously regarded as a variety, is proposed as a new combination, based on the morphological characters and seed storage protein polymorphism pres...Veronica erciyasdagi （M. A. Fischer） C. Vural comb. ＆ stat. nov., previously regarded as a variety, is proposed as a new combination, based on the morphological characters and seed storage protein polymorphism presented in this study. In addition, information about the ecology and conservation status of Veronica erciyasdagi was reported. This taxon is endemic to central Anatolia, Turkey and is critically endangered.展开更多
The profile of polypeptides separated by SDS-PAGE from seed of major crop species such as pea (Pisum sativum) is complex, resulting from cleavage (processing) of precursors expressed from multiple copies of genes enco...The profile of polypeptides separated by SDS-PAGE from seed of major crop species such as pea (Pisum sativum) is complex, resulting from cleavage (processing) of precursors expressed from multiple copies of genes encoding vicilin and legumin, the major storage globulins. Translation in vitro of mRNAs hybrid-selected from mid-maturation pea seed RNAs by denned vicilin and legumin cDNA clones provided precursor molecules that were cleaved in vitro by a cell-free protease extract obtained from similar stage seed; the derived polypep tides were of comparable sizes to those observed in vivo. The feasibility of transcribing mENA in vitro from a cDNA clone and cleavage in vitro of the derived translation products was established for a legumin clone, providing a method for determining polypeptide products of an expressed sequence. This approach will also be useful for characterising cleavage site requirements since modifications an readily be introduced at the DNA level.展开更多
Dwarfing is useful to reduce plant height,when breeding high-yielding and non-lodging crops.In this study,a set of natural storage protein subunit-null dwarf mutants of soybean was reported that showed strongly reduce...Dwarfing is useful to reduce plant height,when breeding high-yielding and non-lodging crops.In this study,a set of natural storage protein subunit-null dwarf mutants of soybean was reported that showed strongly reduced plant stature and deficiency in various 7S and 11S subunits,designated as snd1 mutants.Under normal growth conditions,the snd1 mutants showed a severe dwarf phenotype,with plant height of about 25 cm.Compared with wild-type DN47,the mutant snd1 exhibited no obvious morphological differences at the early stage of development.All the snd1 mutants examined had fewer nodes and shorter than normal internodes;the leaves were similar in shape to normal parents,but were dark-green at the mature stage.The flower size was similar to DN47;however,the flowering period was shorter than in the wild-type.Significant variation was noted for protein content,oil content of the seeds and size of seeds(weight of 100 seeds)among 17 snd1 dwarf lines.Genetic analysis indicated that the dwarfism of snd1 was controlled by a single recessive gene.The snd1 dwarf mutant had markedly different dynamic levels of the endogenous hormones gibberellin(GA),brassinosteroid,indole-3-acetic acid and abscisic acid,at the seedling stage.Exogenous GA3 treatment led to recovery of the plant height phenotype of the snd1 mutant;GA3 at 0.1 mm had the largest effect on enhancing plant height.Using molecular markers,snd1 gene was approximately mapped in an interval of 603 kb between markers Satt166 and Satt561 on chromosome 19.Snd1 mutant provided valuable material for hypoallergenic soybean breeding and the snd1 gene might be a novel gene related to plant height in soybean.展开更多
High molecular weight glutenin subunits(HMW-GS),major components of seed storage proteins in wheat,have large effects on processing quality.GLU-1 genes encode HMW-GS and their expression is mainly controlled at the tr...High molecular weight glutenin subunits(HMW-GS),major components of seed storage proteins in wheat,have large effects on processing quality.GLU-1 genes encode HMW-GS and their expression is mainly controlled at the transcriptional level by interactions between cis-regulatory elements and transcription factors.We previously identified an Aux/IAA transcription factor TaIAA10-6D that bound to a conserved cis-regulatory module CCRM1-1,the most essential conserved cis-regulatory module in GLU-1.Here,we confirmed the binding of TaIAA10-6D to CCRM1-1 using yeast one hybrid and dualluciferase reporter assays.The enhanced expression of TaIAA10-6D suppressed glutenin accumulation and increased gliadin content.Dynamic transcriptome analyses revealed that TaIAA10-6D overexpression down-regulated glutenin and gliadin genes during an early stage of grain filling,but up-regulated gliadin genes during a late stage probably by endoplasmic reticulum stress,accounting for its effect on the tradeoff between glutenin and gliadin.Rheological property and processing quality assays showed that TaIAA10-6D overproduction reduced stabilization time and bread quality,but enhanced cookie quality.Overexpression of TaIAA10-6D also reduced plant height,leaf size,kernel number and grain yield.We identified two major haplotypes of TaIAA10-6D,Hap I and Hap II,and developed a breeding-friendly diagnostic marker.Hap I conferred higher expression of TaIAA10-6D and concomitantly reduced plant height and kernel number,but had little effect on grain yield,contributing to lodging resistance without yield penalty.Hap I was subjected to positive selection in breeding.The findings provide a useful gene for wheat improvement and broaden insights into the regulatory machinery underpinning auxin-mediated quality formation,plant morphogenesis and yield gain.展开更多
The effects of protein characteristics and the proportion of gluten on end-use quality in 13 Korean wheat cultivars for three years were verified in this study. Year, cultivar, and the interaction between the year and...The effects of protein characteristics and the proportion of gluten on end-use quality in 13 Korean wheat cultivars for three years were verified in this study. Year, cultivar, and the interaction between the year and the cultivar influenced protein characteristics, the proportion of gluten except for γ-and ω-gliadin using RP-HPLC(reversed-phase high-performance liquid chromatography), and end-use quality. Protein characteristics and the proportion of gluten in Korean wheat cultivars were between those of Australian standard white(ASW) and hard wheat(AH). Korean wheat cultivars exhibited a higher average α+β gliadin proportion than imported wheat, a γ-gliadin proportion similar to that of dark northern spring wheat, and the same ω-gliadin proportion as AH. They showed a bread-loaf volume intermediate between those of ASW and AH and a texture of cooked noodles similar to that of soft white wheat, but less springiness than imported wheat. The cookie diameter of Korean wheat cultivars was similar to that of hard red winter wheat. There was a correlation between bread-loaf volume and protein characteristics, except for the protein content in Korean wheat cultivars. Springiness and cohesiveness of cooked noodles were not correlated with protein characteristics, while hardness was correlated with the protein content and water absorption of a mixograph. Cookie diameter was negatively correlated with the sodium dodecyl sulfate(SDS) sedimentation volume and water absorption of a mixograph. The end-use quality was not correlated with any proportion of gluten composition. Principal component analysis(PCA) showed that the proportion of gluten was not related to the quality of the bread(both PCs, 81.3%), noodle(77.7%), and cookie(82.4%). PCA explained that Keumkang is suitable for superior bread, while Uri is good for cooked noodles and cookies.展开更多
ω-Secalin was an important factor influencing processing quality of wheat 1BL/1RS translocations. On the basis of four ω-secalin gene sequences cloned from Lankao 906 （a wheat cultivar with 1BL/1RS translocation） ...ω-Secalin was an important factor influencing processing quality of wheat 1BL/1RS translocations. On the basis of four ω-secalin gene sequences cloned from Lankao 906 （a wheat cultivar with 1BL/1RS translocation） with putative transcription activity, a pair of primers with suitable restriction endonucleases added at their 5＂ ends were designed to amplify the mature protein-coding regions of the four genes. After digestion with restriction endonucleases, the amplified products were ligated into the prokaryotic expression vector pET30a（＋）. The prokaryotically expressed recombinant proteins and gliadin isolated from the Lankao 906 seed were analyzed on the same acid polyacrylamide gel electrophoresis. All four prokaryotically expressed secalin bands had corresponding seed-expressed gliadin bands. The four corresponding gliadin bands were confirmed to be the expression products of the four ω-secalin genes by liquid chromatography tandem mass spectrometry （LC-MS/MS）. This conclusion was further confirmed because the four ω-secalin bands could be detected in all 14 wheat 1BL/1 RS translocation cultivars used in the study, although there was some interference for the detection of one ω-secalin band from nearby wheat gliadin bands. The sequence information of ω-secalin genes with expression activity will be helpful for improving the processing quality of wheat with 1BL/1RS translocations by using RNA interference method to silence the expression of the ω-secalin genes.展开更多
Insects enter in diapause in response to diverse environmental cues. During diapause, insects arrest their development and many genes are down-regulated while a small number of genes uniquely expressed at this time. T...Insects enter in diapause in response to diverse environmental cues. During diapause, insects arrest their development and many genes are down-regulated while a small number of genes uniquely expressed at this time. This review aims to present available data regarding the regulation of diapause in the moth Sesamia nonagrioides （Lepidoptera： Noctuidae）. Studying the transcriptional regulation of several genes （five heat shock proteins, two storage proteins and one juvenile hormone esterase） showed that these genes may play various roles in the diapause programming. The results show that SnoHsp19.5 gene was consistently expressed, while SnoHsp20.8 was down-regulated in deep diapause and was up-regulated at the termination of diapause. SnoHsc70 may play important roles in assisting protein conformation during specific stages of diapause. SnoHsp83 displays a similar pattern to SnoHsc70 under diapause conditions, when extra larval moults occur, indicating that could be involved in the developmental process that occurs between two moults. Expression of two SnoSP1 and SnoSP2 hexamerin genes was also observed throughout diapause. And the results lead us to the conclusion that larval diapause of S. nonagrioides is associated with continuous synthesis and accumulation of storage proteins. In addition, the transcript level of the carboxylesterase SnoJHER was higher in non-diapausing larvae than in diapausing ones. During the fifth instar of the non-diapausing and the ninth instar of the diapausing larvae, SnoJHER mRNAs reached higher expression levels on the days close to each larval molt.展开更多
Many antimicrobial peptides (AMPs) have been identified in plants. These peptides are highly divergent at the primary sequence level and vary in their hierarchical structures. Some common biochemical features include ...Many antimicrobial peptides (AMPs) have been identified in plants. These peptides are highly divergent at the primary sequence level and vary in their hierarchical structures. Some common biochemical features include the ability to form disulfide bonds, tandemly repeated amino acid sequences and a net charge at pH 7. Unusual Cysteine containing repeats has been identified in several plant seed storage proteins that may act as AMPs. We identified a Cys repeat within a vicilin (seed storage protein) of a wild legume, Centrosema virginianum. Cleavage of the vicilin protein during germination would generate a vicilin derived Cys peptide (VDCP). We investigated the antimicrobial properties of this VDCP and compared its efficacy as an antimicrobial agent to VDCPs from other species. We developed transgenic tobacco plants that expressed cloned sequences encoding the Cysteine repeat unit from C. virginianum, Theobroma cacao and Gossypium hirsutum. Extracts from fully expanded leaves were tested for antimicrobial activity against a fungal pathogen, Botrytis cinerea. The Cys motif from C. virginianum was also expressed in two E. coli cell lines (reducing or oxidizing cytoplasm) and peptide fusion protein fractions were tested for antimicrobial activity against a battery of fungal strains. The unique Cysteine repeat single unit from C. virginianum exhibited antimicrobial properties greater than or equal to the antimicrobial activity associated with expression of the multiple Cys-repeat VDCPs from G. hirsutum or T. cacao in transgenic tobacco. When expressed in bacteria, a C. virginianum VDCP fusion protein exhibited antifungal activity against 3 of the 4 fungi tested. Although the primary role of seed storage proteins is to provide a pool of amino acids and nitrogen for germinating seeds and developing plantlets, it is likely that seed storage protein proteolytic products also provide beneficial antimicrobial properties during germination and young plantlet development.展开更多
基金supported by the Scientific Research Foundation of Hunan Provincial Education Department,China(Grant Nos.19A532,19B594 and 19B597)National Key Research and Development Program of China(Grant No.2022YFD2101303)+1 种基金Hunan Key Research and Development Plan Project,China(Grant No.2022NK2032)the Program for Science&Technology Innovation Platform of Hunan Province in China(Grant Nos.2019TP1028 and 2019TP1029).
文摘Rice storage proteins(RSPs)are plant proteins with high nutritional quality.As the second largest type of storage substance in rice,it is the main source of protein intake for people who consume rice as a staple food.The content and type of RSPs affect the appearance,processing quality and eating quality of rice.These effects involve the distribution of RSPs in rice grains as well as the interactions of RSPs with other components such as starch in rice grains.In the past two decades,some progress has been made in the genetic improvement of RSPs.However,the determination mechanism of protein content and composition in rice is still unclear,and the mechanism of the effect of RSPs on rice quality has not been elucidated.In this review,the composition,biosynthesis and distribution of RSPs,and quantitative trait loci mapping and cloning of RSP genes are summarized,the research progress of the influence of RSPs and their components on rice quality are reviewed,and the research directions in the future are proposed.
文摘One hundred and fifteen varieties （including cultivars and lines） with different ecotypes in japonica rice （Oryza sativa L.） were analyzed for endosperm storage proteins by sodium dodecyl sulfate polyacrylamide gel electrophoresis （SDS-PAGE） to estimate their genetic diversity for the purpose of genetic improving and variety identification. Nineteen types of profile were identified according to 1） presence/absence of 65 kDa bands, 2） staining intensity of 70, 60, 57, 37-39, 22-23, 13 and 10 kDa bands, 3） migration velocity of 35 kDa （α-4） and 4） band number at 57 kDa location. An unweighted-pair group average method with arithmetic mean （UPGMA） dendregram based on the cluster analysis of genetic similarity of the protein bands showed a small genetic variation among the tested materials, with the similarity coefficients varying between 0.75 and 1.00. Three distinct groups were identified from the cluster analysis of the rice varieties studied at the similarity coefficient level of 0.894. The first group included eight varieties with high amylose content, the second group contained fifteen varieties with high protein content, and the third group had the remaining ninety-two varieties, which accounted for 80% of the total materials. Clear relationship between ecotypes distinguished by maturity and groups revealed by cluster analysis was not found in this study. Only the group of high amylose linked with medium-maturity medium japonica ecotype. The bands of 70 kDa and 65 kDa can be used as protein markers to identify F1 seed purity of japonica hybrid rice Liuyanyou 422.
基金supported by grants from the National Natural Science Foundation of China(NSFC31871396,32000208,32070769)the Science and Technology Innovation Program of Hunan Province(2020WK2014)+2 种基金the Changsha Municipal Natural Science Foundation(kq2014039)the Scientific Innovation Fund for Post-graduates of Central South University of Forestry and Technology(CX202101026)the Postgraduate Scientific Research Innovation Project of Hunan Province(CX20210844)。
文摘Rice(Oryza sativa L.)is the most important food crop for at least half of the world’s population.Due to improved living standards,the cultivation of high-quality rice for different purposes and markets has become a major goal.Rice quality is determined by the presence of many nutritional components,including seed storage proteins(SSPs),which are the second most abundant nutrient components of rice grains after starch.Rice SSP biosynthesis requires the participation of multiple organelles and is influenced by the external environment,making it challenging to understand the molecular details of SSP biosynthesis and improve rice protein quality.In this review,we highlight the current knowledge of rice SSP biosynthesis,including a detailed description of the key molecules involved in rice SSP biosynthetic processes and the major environmental factors affecting SSP biosynthesis.The effects of these factors on SSP accumulation and their contribution to rice quality are also discussed based on recent findings.This recent knowledge suggests not only new research directions for exploring rice SSP biosynthesis but also innovative strategies for breeding high-quality rice varieties.
基金Supported by the National Natural Science Foundation of China （30460107）
文摘In order to Identify appropriate plant materials for studying the gene expression and biological function of vegetative storage proteins （VSPs） in woody plants, the VSPs in the seedlings of Swietenla rnacrophylla King were investigated by using light microscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis （SDS-PAGE） and Western.blotting. The seed of S. macrophylla was rich in storage proteins that accumulated In the vacuoles of cotyledon parenchyma cells in appearance of compact spherical grains. The growth and development of S. macrophylla seedlings were characterized by an obvious growth rhythm. The storage proteins In seeds disappeared during seedling growth while VSPs appeared in the stem 2 weeks after seedling leaves matured. Thereafter, the VSPs In the seedling stem almost exhausted during new shoot growth, and when the leaves of new shoot Just matured, both the stem beneath the new shoot of seedlings and the stem of new shoot started to accumulate VSPs. Nitrogen application dramatically Increased the level of VSPs, but had little influence on the dynamics of VSP consumption and accumulation in seedling stem. Together with these data, the fluctuation of VSPs in seedlings was very similar to that in the branches of the adult trees. In addition, seedlings are easy to be treated due to their small size. Our results suggested that S. rnacrophylla seedlings were suitable for Investigating the biological roles of VSPs and the mechanism of nitrogen storage in trees.
文摘The occurrence of twin-arginine motifs （-R-R-） in the amino acid sequences of animal pro-proteins frequently defines the cleavage site（s） for their structural/functional maturation. No information is available on the presence and possible biological meaning of these motifs in the seed storage proteins. In this work, a novel endopeptidase activity with cleavage specificity to twin-arginine pairs has been detected in mature dry Lupinus albus seeds. The endopeptidase was tested with a number of endogenous and exogenous protein substrates, which were selected according to the pres- ence of one or more twin-arginine residue motifs in their amino acid sequences. The observed hydrolysis patterns were limited and highly specific. Partial proteolysis led to stable polypeptide fragments that were characterized by 1- and 2-D electrophoresis. Selected polypeptides were submitted to N-terminal amino acid sequencing and mass spectrometry anal- yses, These approaches, supported by bioinformatic analysis of the available sequences, allowed the conclusion that the polypeptide cleavage events had occurred at the peptide bonds comprised between twin-arginine residue pairs with all tested protein substrates. The endopeptidase activity was inhibited by 4-（2-AminoEthyl）Benzene-Sulphonyl Fluoride hy- drochloride （AEBSF）, leupeptin, and serine proteinase protein inhibitors, while it was not affected by pepstatin, trans- EpoxysuccinyI-L-leucylamido（4-guanidino）butane （E64）, and ethylenediaminetetraacetic acid （EDTA）, thus qualifying the Arg-Arg cleaving enzyme as a serine endopeptidase. The structural features of storage proteins from lupin and other legume seeds strongly support the hypothesis that the occurrence of an endopeptidase activity cleaving -R-R- bonds may be functional to facilitate their degradation at germination and possibly generate polypeptide fragments with specific biological activity.
基金financially supported by the National Natural Science Foundation of China(31771773 and 31971931)the National Key Research and Development Program of China(2016YFD0100502)。
文摘The content and composition of wheat storage proteins are the major determinants of dough rheological properties and breadmaking quality and are influenced by cultivation conditions.This study aimed to investigate the effects of water deficit and high N-fertilizer application on wheat storage protein synthesis,gluten secondary structure,and breadmaking quality.Reverse-phase ultrahigh-performance liquid chromatography analysis showed that storage protein and gluten macropolymer accumulation was promoted under both independent applications and a combination of water-deficit and high N-fertilizer treatments.Fourier-transform infrared spectroscopy showed that water deficit and high N-fertilizer treatments generally improved protein secondary structure formation and lipid accumulation,and reduced flour moisture.In particular,high N-fertilizer application increasedβ-sheet content by 10.4%and the combination of water-deficit and high N-fertilizer treatments increased random coil content by 7.6%.These changes in gluten content and secondary structure led to improved dough rheological properties and breadmaking quality,including superior loaf internal structure,volume,and score.Our results demonstrate that moderately high N-fertilizer application under drought conditions can improve gluten accumulation,gluten secondary structure formation,and baking quality.
文摘The ultrastructural characteristics and the subcellular localization of vegetative storage proteins(VSPs)of Ginkgo biloba L.were systematically studied under the electron microscope.Results indicated that the VSPs of G.biloba were mostly distributed in the small vacuoles of the phloem parenchyma cells.The VSPs of phloem parenchyma cells were produced in cytoplasm,then separated by the inflated cisternae of endoplasmic reticulum(ER),plasmalemma invagination or Golgi body vesicles,resulting in the formation of vacuole filled with proteins.Three kinds of VSPs were detected:granular,floccular and massive VSPs,which were distributed in different tissue cells and different cells of the same tissue or vacuoles of the same cells.VSPs accumulated in autumn and kept at high level throughout the winter.In the following spring,the bud started growing and VSPs were completely mobilized.With new shoots growth,VSPs resumed accumulating in late summer and early autumn.
基金supported by the Provincial Prize Fund for Distinguished Young and Middle-aged Scientists of Shandong Province(No.BS2011SW053)State Key Laboratory of Plant Cell and Chromosome Engineering(No.PCCE-KF-2014-01)State Key Laboratory of Crop Biology(No.2015KF06)
文摘Wide hybridization is an effective approach for enhancing the resistance of bread wheat （Triticum aestivum L.） to biotic and abiotic stresses by introducing favorable alien genes （Sepsi et al., 2008）. Wheatgrass, Thinopyrum intermedium （Host） Barkworth ＆ D.R. Dewey or Agropyron intermedium （Host） Beauvoir （2n = 42; genome formula JJjSjSstst）, is a perennial species in the tribe Triticeae and an important source of wheat improvement for biotic and abiotic stress resistance and quality-related traits, such as high grain protein concentration （Chen et al., 1998; 2001; 2003; Han et al., 2004; Li and Wang, 2009）. In addition, the ready crossing ability of wheatgrass with various Triticum species has made it popular in germ- plasm development.
基金funded by the National High-Tech R&D Program of China(2012AA101106)the National Basic Research Program of China(2009CB118404)+1 种基金the Key Technologies R&D Program of China during the 12th Five-Year Plan period(2011BAD35B06)the National Transgenic Major Program,China(2008ZX08009-003)
文摘Soybean seed storage protein is one of the most important plant vegetable proteins, and β subunit is of great significance to enhance soybean protein quality and processing property. F2 segregated population and residual heterozygous lines（RHL） derived from the cross between Yangyandou（low level of β subunit） and Zhonghuang 13（normal level of β subunit） were used for mapping of β subunit content. Our results showed that β subunit content was controlled by a single dominant locus, qBSC-1（β subunit content）, which was mapped to a region of 11.9 cM on chromosome 20 in F2 population of 85 individuals. This region was narrowed down to 2.5 cM between BARCSOYSSR_20_0997 and BARCSOYSSR_20_0910 in RHL with a larger population size of 246 individuals. There were 48 predicted genes within qBSC-1 region based on the reference genome（Glyma 1.0, Williams 82）, including the two copies of β subunit coding gene CG4. An InDel marker developed from a thymine（TT） insertion in one copy of CG4 promoter region in Yangyandou cosegregrated with BARCSOYSSR_20_0975 within qBSC-1 region, suggesting that this InDel marker maybe useful for marker-assisted selection（MAS）.
文摘The characteristics of 48 rice varieties from Uttarakhand Himalaya, India were detected by morphological and biochemical markers. The grains of the selected rice varieties varied in their morphological （grain length, grain width and grain weight） and biochemical characters （sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE）. Based on the presence of 70, 65, 60, 57, 37-39, 22-23, 13 and 10 kDa protein bands in the 48 rice varieties, seven types of profiles were identified. An unweighted pair group average method with arithmetic mean （UPGMA） dendrogram based on cluster analysis of genetic similarity of the protein bands showed two distinct groups with 1%-78% similarity coefficients. The presence of characteristic bands in selected varieties is a useful parameter for identification of rice germplasm.
基金The present study is part of the project "EUBAP-FBT.04.46" supported by Erciyes University Research Fund
文摘Veronica erciyasdagi （M. A. Fischer） C. Vural comb. ＆ stat. nov., previously regarded as a variety, is proposed as a new combination, based on the morphological characters and seed storage protein polymorphism presented in this study. In addition, information about the ecology and conservation status of Veronica erciyasdagi was reported. This taxon is endemic to central Anatolia, Turkey and is critically endangered.
文摘The profile of polypeptides separated by SDS-PAGE from seed of major crop species such as pea (Pisum sativum) is complex, resulting from cleavage (processing) of precursors expressed from multiple copies of genes encoding vicilin and legumin, the major storage globulins. Translation in vitro of mRNAs hybrid-selected from mid-maturation pea seed RNAs by denned vicilin and legumin cDNA clones provided precursor molecules that were cleaved in vitro by a cell-free protease extract obtained from similar stage seed; the derived polypep tides were of comparable sizes to those observed in vivo. The feasibility of transcribing mENA in vitro from a cDNA clone and cleavage in vitro of the derived translation products was established for a legumin clone, providing a method for determining polypeptide products of an expressed sequence. This approach will also be useful for characterising cleavage site requirements since modifications an readily be introduced at the DNA level.
基金Supported by the Ministry of Science and Technology of China(2016YFD0100500)Funding from Harbin Science and Technology Bureau(2016RQYXJ018,2017RAQXJ104)+4 种基金the Key Laboratory of Soybean Biology in the Chinese Ministry of Education,Northeast Agricultural University(SB17A01)National Natural Science Foundation of China(31801386)Heilongjiang Natural Science Foundation(LC2018008)Heilongjiang General Young Innovative Talents Training Plan(UNPYSCT-2018158)Certificate of China Postdoctoral Science Foundation Grant(2018M641839)
文摘Dwarfing is useful to reduce plant height,when breeding high-yielding and non-lodging crops.In this study,a set of natural storage protein subunit-null dwarf mutants of soybean was reported that showed strongly reduced plant stature and deficiency in various 7S and 11S subunits,designated as snd1 mutants.Under normal growth conditions,the snd1 mutants showed a severe dwarf phenotype,with plant height of about 25 cm.Compared with wild-type DN47,the mutant snd1 exhibited no obvious morphological differences at the early stage of development.All the snd1 mutants examined had fewer nodes and shorter than normal internodes;the leaves were similar in shape to normal parents,but were dark-green at the mature stage.The flower size was similar to DN47;however,the flowering period was shorter than in the wild-type.Significant variation was noted for protein content,oil content of the seeds and size of seeds(weight of 100 seeds)among 17 snd1 dwarf lines.Genetic analysis indicated that the dwarfism of snd1 was controlled by a single recessive gene.The snd1 dwarf mutant had markedly different dynamic levels of the endogenous hormones gibberellin(GA),brassinosteroid,indole-3-acetic acid and abscisic acid,at the seedling stage.Exogenous GA3 treatment led to recovery of the plant height phenotype of the snd1 mutant;GA3 at 0.1 mm had the largest effect on enhancing plant height.Using molecular markers,snd1 gene was approximately mapped in an interval of 603 kb between markers Satt166 and Satt561 on chromosome 19.Snd1 mutant provided valuable material for hypoallergenic soybean breeding and the snd1 gene might be a novel gene related to plant height in soybean.
基金supported by the STI 2030-Major Projects(2023ZD0406903)the National Natural and Science Foundation of China(32272182)+1 种基金the Postdoctoral Fellowship Program of CPSF(GZC20241955)the Science and Technology Innovation Program of Chinese Academy of Agricultural Sciences(CAAS).
文摘High molecular weight glutenin subunits(HMW-GS),major components of seed storage proteins in wheat,have large effects on processing quality.GLU-1 genes encode HMW-GS and their expression is mainly controlled at the transcriptional level by interactions between cis-regulatory elements and transcription factors.We previously identified an Aux/IAA transcription factor TaIAA10-6D that bound to a conserved cis-regulatory module CCRM1-1,the most essential conserved cis-regulatory module in GLU-1.Here,we confirmed the binding of TaIAA10-6D to CCRM1-1 using yeast one hybrid and dualluciferase reporter assays.The enhanced expression of TaIAA10-6D suppressed glutenin accumulation and increased gliadin content.Dynamic transcriptome analyses revealed that TaIAA10-6D overexpression down-regulated glutenin and gliadin genes during an early stage of grain filling,but up-regulated gliadin genes during a late stage probably by endoplasmic reticulum stress,accounting for its effect on the tradeoff between glutenin and gliadin.Rheological property and processing quality assays showed that TaIAA10-6D overproduction reduced stabilization time and bread quality,but enhanced cookie quality.Overexpression of TaIAA10-6D also reduced plant height,leaf size,kernel number and grain yield.We identified two major haplotypes of TaIAA10-6D,Hap I and Hap II,and developed a breeding-friendly diagnostic marker.Hap I conferred higher expression of TaIAA10-6D and concomitantly reduced plant height and kernel number,but had little effect on grain yield,contributing to lodging resistance without yield penalty.Hap I was subjected to positive selection in breeding.The findings provide a useful gene for wheat improvement and broaden insights into the regulatory machinery underpinning auxin-mediated quality formation,plant morphogenesis and yield gain.
基金the support of Cooperative Research Program for Agriculture Science & Technology Development (Project title: Establishment of quality criteria for high uniformity in end-use of Korean wheat cultivars, PJ011009), Rural Development Administration, Republic of Korea
文摘The effects of protein characteristics and the proportion of gluten on end-use quality in 13 Korean wheat cultivars for three years were verified in this study. Year, cultivar, and the interaction between the year and the cultivar influenced protein characteristics, the proportion of gluten except for γ-and ω-gliadin using RP-HPLC(reversed-phase high-performance liquid chromatography), and end-use quality. Protein characteristics and the proportion of gluten in Korean wheat cultivars were between those of Australian standard white(ASW) and hard wheat(AH). Korean wheat cultivars exhibited a higher average α+β gliadin proportion than imported wheat, a γ-gliadin proportion similar to that of dark northern spring wheat, and the same ω-gliadin proportion as AH. They showed a bread-loaf volume intermediate between those of ASW and AH and a texture of cooked noodles similar to that of soft white wheat, but less springiness than imported wheat. The cookie diameter of Korean wheat cultivars was similar to that of hard red winter wheat. There was a correlation between bread-loaf volume and protein characteristics, except for the protein content in Korean wheat cultivars. Springiness and cohesiveness of cooked noodles were not correlated with protein characteristics, while hardness was correlated with the protein content and water absorption of a mixograph. Cookie diameter was negatively correlated with the sodium dodecyl sulfate(SDS) sedimentation volume and water absorption of a mixograph. The end-use quality was not correlated with any proportion of gluten composition. Principal component analysis(PCA) showed that the proportion of gluten was not related to the quality of the bread(both PCs, 81.3%), noodle(77.7%), and cookie(82.4%). PCA explained that Keumkang is suitable for superior bread, while Uri is good for cooked noodles and cookies.
基金supported by the Natural Science Foundation of Hebei Province,China (C2014301005)
文摘ω-Secalin was an important factor influencing processing quality of wheat 1BL/1RS translocations. On the basis of four ω-secalin gene sequences cloned from Lankao 906 （a wheat cultivar with 1BL/1RS translocation） with putative transcription activity, a pair of primers with suitable restriction endonucleases added at their 5＂ ends were designed to amplify the mature protein-coding regions of the four genes. After digestion with restriction endonucleases, the amplified products were ligated into the prokaryotic expression vector pET30a（＋）. The prokaryotically expressed recombinant proteins and gliadin isolated from the Lankao 906 seed were analyzed on the same acid polyacrylamide gel electrophoresis. All four prokaryotically expressed secalin bands had corresponding seed-expressed gliadin bands. The four corresponding gliadin bands were confirmed to be the expression products of the four ω-secalin genes by liquid chromatography tandem mass spectrometry （LC-MS/MS）. This conclusion was further confirmed because the four ω-secalin bands could be detected in all 14 wheat 1BL/1 RS translocation cultivars used in the study, although there was some interference for the detection of one ω-secalin band from nearby wheat gliadin bands. The sequence information of ω-secalin genes with expression activity will be helpful for improving the processing quality of wheat with 1BL/1RS translocations by using RNA interference method to silence the expression of the ω-secalin genes.
文摘Insects enter in diapause in response to diverse environmental cues. During diapause, insects arrest their development and many genes are down-regulated while a small number of genes uniquely expressed at this time. This review aims to present available data regarding the regulation of diapause in the moth Sesamia nonagrioides （Lepidoptera： Noctuidae）. Studying the transcriptional regulation of several genes （five heat shock proteins, two storage proteins and one juvenile hormone esterase） showed that these genes may play various roles in the diapause programming. The results show that SnoHsp19.5 gene was consistently expressed, while SnoHsp20.8 was down-regulated in deep diapause and was up-regulated at the termination of diapause. SnoHsc70 may play important roles in assisting protein conformation during specific stages of diapause. SnoHsp83 displays a similar pattern to SnoHsc70 under diapause conditions, when extra larval moults occur, indicating that could be involved in the developmental process that occurs between two moults. Expression of two SnoSP1 and SnoSP2 hexamerin genes was also observed throughout diapause. And the results lead us to the conclusion that larval diapause of S. nonagrioides is associated with continuous synthesis and accumulation of storage proteins. In addition, the transcript level of the carboxylesterase SnoJHER was higher in non-diapausing larvae than in diapausing ones. During the fifth instar of the non-diapausing and the ninth instar of the diapausing larvae, SnoJHER mRNAs reached higher expression levels on the days close to each larval molt.
文摘Many antimicrobial peptides (AMPs) have been identified in plants. These peptides are highly divergent at the primary sequence level and vary in their hierarchical structures. Some common biochemical features include the ability to form disulfide bonds, tandemly repeated amino acid sequences and a net charge at pH 7. Unusual Cysteine containing repeats has been identified in several plant seed storage proteins that may act as AMPs. We identified a Cys repeat within a vicilin (seed storage protein) of a wild legume, Centrosema virginianum. Cleavage of the vicilin protein during germination would generate a vicilin derived Cys peptide (VDCP). We investigated the antimicrobial properties of this VDCP and compared its efficacy as an antimicrobial agent to VDCPs from other species. We developed transgenic tobacco plants that expressed cloned sequences encoding the Cysteine repeat unit from C. virginianum, Theobroma cacao and Gossypium hirsutum. Extracts from fully expanded leaves were tested for antimicrobial activity against a fungal pathogen, Botrytis cinerea. The Cys motif from C. virginianum was also expressed in two E. coli cell lines (reducing or oxidizing cytoplasm) and peptide fusion protein fractions were tested for antimicrobial activity against a battery of fungal strains. The unique Cysteine repeat single unit from C. virginianum exhibited antimicrobial properties greater than or equal to the antimicrobial activity associated with expression of the multiple Cys-repeat VDCPs from G. hirsutum or T. cacao in transgenic tobacco. When expressed in bacteria, a C. virginianum VDCP fusion protein exhibited antifungal activity against 3 of the 4 fungi tested. Although the primary role of seed storage proteins is to provide a pool of amino acids and nitrogen for germinating seeds and developing plantlets, it is likely that seed storage protein proteolytic products also provide beneficial antimicrobial properties during germination and young plantlet development.
