Phosphorylation is one of the major posttranslational modifications to control plant growth and development.Opaque2(O2)represents a central hub for endosperm filling,which largely determines seed yield and nutrient st...Phosphorylation is one of the major posttranslational modifications to control plant growth and development.Opaque2(O2)represents a central hub for endosperm filling,which largely determines seed yield and nutrient storage in maize.However,it still remains unclear how O2 phosphorylation orchestrates endosperm filling and nutrient quality.Here,we systematically identified the phosphorylation sites of O2 during endosperm filling.A total of 18 phosphorylation sites were found in O2 and five sites were identified to apparently modulate its subcellular localization and transactivation capacity.In addition,a conserved protein kinase CK1 was confirmed to interact with and phosphorylate O2 at the residue Threonine(T)202 to promote O2-mediated transactivation and protein stability.Overexpression of CK1 resulted in increased kernel size,100-kernel weight and nutrient storage.Phosphorylation-mimic O2 seeds at T202 exhibited enhanced kernel dimension,test weight,vitreous endosperm area and nutrient accumulation,whereas the phosphorylation-deficient O2 seeds did not.Collectively,this study establishes a comprehensive phosphocode atlas of O2 during endosperm filling and highlights the importance of phosphorylation modification in O2 to precisely orchestrate maize yield and nutrient quality.展开更多
Inferior grains exhibit delayed developmental processes and reduced metabolic activities compared to superior grains,leading to unstable rice yield and quality.While significant advancements have been achieved in eluc...Inferior grains exhibit delayed developmental processes and reduced metabolic activities compared to superior grains,leading to unstable rice yield and quality.While significant advancements have been achieved in elucidating the physiology of endosperm filling in inferior grains,the role of the embryo remains underexplored and warrants comprehensive investigation.Two Wuyujing 3 mutants,DW024(relatively synchronous;syn-DW024)and DW179(significantly asynchronous;asyn-DW179),with different grain-filling patterns were used in this study.Samples of superior and inferior grains were collected at intervals from 5 to 60 d after fertilization and subsequently dissected into subsamples of the embryo and endosperm.Histochemical staining,biochemical analysis,and RNA sequencing(RNA-seq)were combined to systematically compare developmental and physiological traits between superior and inferior grains.Combining hierarchical clustering of mRNA datasets revealed three developmental phases of the endosperm and embryo:morphogenesis,endosperm filling/embryo enlargement,and maturation.In both syn-DW024 and asyn-DW179,the duration of the endosperm/embryo morphogenesis phase was identical in superior and inferior grains.The inferior grains of asyn-DW179 exhibited a 10-day prolongation in the endosperm filling phase and a 20-day extension in the embryo enlargement phase compared to the superior grains.The endosperm of inferior grains exhibited higher contents of sugars and free amino acids,along with slower accumulation of storage compounds,which was associated with the down-regulation of genes for starch synthesis and ABA signaling.In addition,transporters for nutrient exchanges between endosperm and embryo were down-regulated,suggesting a potential role of the embryo in adjusting the endosperm filling process.Collectively,our results reveal that the prolonged phases of endosperm filling and embryo enlargement may underlie the impaired development of inferior grains,offering a new perspective for breeding or cultivating rice with uniform grain quality.展开更多
The early development of the endosperm is crucial for balancing the allocation of maternal nutrients to offspring.This process is believed to be evolutionarily associated with genomic imprinting,resulting in parentall...The early development of the endosperm is crucial for balancing the allocation of maternal nutrients to offspring.This process is believed to be evolutionarily associated with genomic imprinting,resulting in parentally biased allelic gene expression.Beyond Fertilization Independent Seed(FIS)genes,the number of imprinted genes involved in early endosperm development and seed size determination remains limited.This study introduces early endosperm-expressed HAIKU(IKU)downstream Candidate F-box 1(ICF1)and ICF2 as maternally expressed imprinted genes(MEGs)in Arabidopsis thaliana.Although these genes are also demethylated by DEMETER(DME)in the central cell,their activation differs from the direct DME-mediated activation seen in classical MEGs such as the FIS genes.Instead,ICF maternal alleles carry pre-established hypomethylation in their promoters,priming them for activation by the WRKY10 transcription factor in the endosperm.On the contrary,paternal alleles are predominantly suppressed by CG methylation.Furthermore,we find that ICF genes partially contribute to the small seed size observed in iku mutants.Our discovery reveals a two-step regulatory mechanism that highlights the important role of conventional transcription factors in the activation of imprinted genes,which was previously not fully recognized.Therefore,the mechanism provides a new dimension to understand the transcriptional regulation of imprinting in plant reproduction and development.展开更多
Ultrastructural features of nucleus degradation during programmed cell death (PCD) of starchy endosperm cells in rice ( Oryza sativa L.) were observed using transmission electron microscopy. Several distinct morpho...Ultrastructural features of nucleus degradation during programmed cell death (PCD) of starchy endosperm cells in rice ( Oryza sativa L.) were observed using transmission electron microscopy. Several distinct morphological features of PCD have been found in the developing starchy endosperm cells, e.g. nucleus deformation, chromatin condensation, nuclear envelope disruption, and nuclear matrix leakage. DNA ladder displayed a smear of large DNA fragments from nucleus and evident bands of small DNA fragments (140-180 bp) from both nucleus and cytoplasm. In contrast with the rapid nucleus degradation, cell organelles in cytoplasm, such as rough endoplasmic reticulum, amyloplast, and mitochondrion, maintained their metabolic functions for a longer time. Seed reserves were continually synthesized and accumulated in the starchy endosperm cells despite the nucleus degradation during the PCD process. These results suggest that starchy endosperm cells remain active during reserve material synthesis and accumulation in the PCD process. The specific relationships between nucleus and cytoplasm in the developing endosperm cells and the morphological changes of nucleus in the endosperm PCD process were also discussed.展开更多
A statistical method for mapping quantitative trait loci （QTLs） underlying endosperm traits is proposed. The method is based on a genetic model containing both the direct effects and maternal effects of an endosperm...A statistical method for mapping quantitative trait loci （QTLs） underlying endosperm traits is proposed. The method is based on a genetic model containing both the direct effects and maternal effects of an endosperm QTL and on an experimental design termed two-stage hierarchical design, in which the trait information is obtained from F3 endosperms and molecular marker information is obtained from F2 plants and F3 embryos （plants）. Results of computer simulations indicate that the method can efficiently map endosperm QTLs and precisely estimate both the direct and maternal effects of endosperm QTLs when the sample size is sufficiently large.展开更多
[Objective] This research aimed to study the FTIR spectra of corn germs and endosperms so as to provide a scientific way for identifying corn of different types. [Method] The corn germs and endosperms of three types w...[Objective] This research aimed to study the FTIR spectra of corn germs and endosperms so as to provide a scientific way for identifying corn of different types. [Method] The corn germs and endosperms of three types were studied by using Fourier transform infrared spectroscopy（FTIR） technology, combined with cluster analysis. [Result] The overall characteristics of original FTIR spectra were basically similar within the range of 700-1 800 cm^-1. The FTIR spectra were mainly composed by the absorption peaks of polysaccharides, proteins and lipids. Within the wavelength range of 700-1 800 cm^-1, there were only tiny differences in original FTIR spectra among the corn germs and endosperms of three different types. The spectra were then processed by using first derivative and second derivative. The second derivative spectra were used for hierarchical cluster analysis（HCA）. The results showed that with the wavelength range of 700-1 800 cm^-1, the second derivative spectra of the 52 samples could be better clustered according to the tree types and corn germ and corn endosperm. The clustering correct rate reached 96.1%.[Conclusion] FTIR technology, combined with cluster analysis, can be used to identify different types of corn germs and endosperms, and it is characterized by convenience and rapidness.展开更多
Quantitative trait loci（QTL） for percentage of chalky grain,degree of chalkiness,and endosperm transparency were detected using 3 recombinant inbred line populations derived from crosses between parental lines of co...Quantitative trait loci（QTL） for percentage of chalky grain,degree of chalkiness,and endosperm transparency were detected using 3 recombinant inbred line populations derived from crosses between parental lines of commercial three-line hybrids of indica rice.Two of the populations showed great variations on heading date,and the other had a short range of heading date variation.A total of 40 QTLs were detected and fell into 15 regions of 10 chromosomes,of which 5 regions were detected for 1 or more same traits over different populations,2 were detected for different traits in different populations,3 were detected for 2 or all the 3 traits in a single population,and 5 were detected for a single trait in a single population.Most of these QTLs have been reported previously,but a region located on the long arm of chromosome 10 showing significant effects in all the 3 populations has not been reported before.It was shown that a number of gene cloned,including the Wx and Alk for the physiochemical property of rice grain,and GW2,GS3 and GW5 for grain weight and grain size,could have played important roles for the genetic control of grain chalkiness in rice,but there are many more QTLs exerting stable effects for rice chalkiness over different genetic backgrounds.It is worth paying more attentions to these regions which harbor QTL such as the qPCG5.2/qDC5.2/qET5.2 and qPCG10/qDC10/qET10 detected in our study.Our results also showed that the use of segregating populations having high-uniform heading date could greatly increase the efficiency of the identification of QTL responsible for traits that are subjected to great environmental influence.展开更多
This study provided visual evidence of a nitrogen effect on starch granules(SGs) in wheat endosperm. Winter wheat(Titicum aestivum L.) cultivar Xumai 30 was cultured under no nitrogen(control) and 240 kg ha-1of nitrog...This study provided visual evidence of a nitrogen effect on starch granules(SGs) in wheat endosperm. Winter wheat(Titicum aestivum L.) cultivar Xumai 30 was cultured under no nitrogen(control) and 240 kg ha-1of nitrogen applied at the booting stage. The number, morphology, and size of Aand B-type SGs in subaleurone of dorsal endosperm(SDE), center of dorsal endosperm(CDE), modified aleurone(MA), subaleurone of ventral endosperm(SVE), and center of ventral endosperm(CVE) were observed under light and electron microscopes.(1) The distribution of SGs in SDE was similar to that in SVE, the distributions of SGs in CDE and CVE were similar, but the distribution of SGs in MA was different from those in the other four endosperm regions. The number of SGs in the five endosperm regions was in the order SDE > CDE > SVE > CVE > MA.(2) Nitrogen increased the number of Aand B-type SGs in SDE and SVE. Nitrogen also increased the number of B-type SGs but decreased the number of A-type SGs in CDE and CVE. Nitrogen decreased the numbers of A-type and B-type SGs in MA. The results suggest that increased N fertilizer application mainly increased the numbers of small SGs and decreased the numbers of large SGs, but that the results varied in different regions of the wheat endosperm.展开更多
The translucent endosperm trait in a japonica rice variety ＇Kantou 194＇ is controlled by a Wx-mq gene which is allelic to Wx locus by genetic analysis and allelic test. The amylose content analysis showed that an in...The translucent endosperm trait in a japonica rice variety ＇Kantou 194＇ is controlled by a Wx-mq gene which is allelic to Wx locus by genetic analysis and allelic test. The amylose content analysis showed that an intermediate amylose content between those of glutinous and non-glutinous rice existed in endosperm of homozygous Wx-mq genotype. The slight changes of amylose content in different varieties and F1 grains with an identical Wx-mq genotype might be influenced by dissimilar genetic background. To identify the Wx-mq genotype simply and rapidly, a cleaved amplified polymorphic sequence （CAPS） marker was designed. The result from the molecular detection indicated that it could be used for marker-assisted selection for low amylose content varieties in rice breeding.展开更多
By using the laser scanning confocal microscope and plastic （Leica 7022 historesin embedding kit） semi-thin sectioning technique, comparative studies on the development of endosperm were carried out between autotetr...By using the laser scanning confocal microscope and plastic （Leica 7022 historesin embedding kit） semi-thin sectioning technique, comparative studies on the development of endosperm were carried out between autotetraploid and diploid rices. About one third of the ovaries in the autotetraploid showed normal endosperm development as those in the diploid. In these ovaries, one of the polar nuclei would fuse with the sperm nucleus, and the primary endosperm nucleus formed and underwent the first division in 4 hours after pollination; the anticlinal wall began to grow centripetally between the free nuclei starting from the wall ingrowths of the embryo sac near the micropylar end, and some of the phragmoplasts formed transformed into periclinal walls. In addition, some of the cell wall situated in the middle of the endosperm appeared to originate from phragmoplasts, whereas others seemed to develop randomly without the obvious formation of phragmoplasts. Cellulose began to accumulate in the wall of aleurone cell layer at 6 days after pollination. The cellulose wall of the cells of the aleurone cell layer appeared to have completely formed within 7 to 8 days after pollination. On the other hand, about two thirds of the ovaries in the autotetraploid showed abnormality in endosperm development with various types, such as non-fertilization, abnormal fertilization, endosperm development-delay and non-synchronization in the development of cellulose wall of cells of the aleurone layer. These abnormalities usually resulted in decreased seed setting in autotetraploid rice.展开更多
ZAG2 has been identified as a maternally expressed imprinted gene in maize endosperm.Our study revealed that paternally inherited ZAG2 alleles were imprinted in maize endosperm and embryo at 14 days after pollination(...ZAG2 has been identified as a maternally expressed imprinted gene in maize endosperm.Our study revealed that paternally inherited ZAG2 alleles were imprinted in maize endosperm and embryo at 14 days after pollination(DAP), and consistently imprinted in endosperm at 10, 12, 16, 18, 20, 22, 24, 26, and 28 DAP in reciprocal crosses between B73 and Mo17. ZAG2 alleles were also imprinted in reciprocal crosses between Zheng 58 and Chang7-2 and between Huang C and 178. ZAG2 alleles exhibited differential imprinting in hybrids of 178 × Huang C and B73 × Mo17, while in other hybrids ZAG2 alleles exhibited binary imprinting. The tissue-specific expression pattern of ZAG2 showed that ZAG2 was expressed at a high level in immature ears, suggesting that ZAG2 plays important roles in not only kernel but ear development.展开更多
To ascertain the effect mechanism of high temperature after anthesis on rice quality, the experiment was conducted with two rice lines, the heat-tolerant line 996 and heat-sensitive line 4628, with high temperature an...To ascertain the effect mechanism of high temperature after anthesis on rice quality, the experiment was conducted with two rice lines, the heat-tolerant line 996 and heat-sensitive line 4628, with high temperature and optimal temperature in the growth chamber to investigate the effect of high temperature stress after anthesis on rice appearance quality, milling quality, cooking and eating quality and starch granule structure of endo- sperm. The result showed that milled rice rate, head rice rate, amylose content and gel consistency of both lines decreased under high temperature stress after anthesis, while the ratio of grain length to width, chalky rate, chalkiness, protein content increased. Under high temperature treatments, gelatinization temperature, final viscosity, set back and peak time increased, breakdown decreased, Mg content and K content increased, Mg/K ra- tio decreased. Under same treatment, the extent of rice quality of heat tolerant line 996 affected by high temperature was lower than that of heat sensitive line 4628. Under high temperature stress after anthesis, starch granule arranged untightly, most of starch granules existed in the form of a single starch endosperm, refractive index decreased, transparency reduced, and led to the formation of chalk. Under high temperature stress af- ter anthesis, the increase of protein content, the decrease of Mg/K, the changes of rice RVA profile characteristics and starch granule structure of endosperm could be the main reason for the decrease of rice cooking and eating quality and appearance quality.展开更多
A high-yielding japonica rice variety, Wuyunjing 7, bred in Jiangsu Province, China as a female parent was crossed with a Japanese rice variety Kantou 194, which carries a rice stripe disease resistance gene Stv-b＇ a...A high-yielding japonica rice variety, Wuyunjing 7, bred in Jiangsu Province, China as a female parent was crossed with a Japanese rice variety Kantou 194, which carries a rice stripe disease resistance gene Stv-b＇ and a translucent endosperm mutant gene Wx-mq. From F2 generations, a sequence characterized amplified region （SCAR） marker tightly linked with Stv-b＇ and a cleaved amplified polymorphic sequence （CAPS） marker for Wx-mq were used for marker-assisted selection. Finally, a new japonica rice line, Ning 9108, with excellent agronomic traits was obtained by multi-generational selection on stripe disease resistance and endosperm appearance. The utilization of the markers from genes related to rice quality and disease resistance was helpful not only for establishing a marker-assisted selection system of high-quality and disease resistance for rice but also for providing important intermediate materials and rapid selection method for good quality, disease resistance and high yield in rice breeding.展开更多
Endosperm as the storage organ of starch and protein in cereal crops largely determines grain yield and quality.Despite the fact that several pentatricopeptide repeat(PPR)proteins required for endosperm development ha...Endosperm as the storage organ of starch and protein in cereal crops largely determines grain yield and quality.Despite the fact that several pentatricopeptide repeat(PPR)proteins required for endosperm development have been identified in rice,the molecular mechanisms of many P-type PPR proteins in endosperm development remains unclear.Here,we isolated a rice floury endosperm mutant ppr5 that developed small starch grains and an abnormal aleurone layer,accompanied by decreased starch,protein,and amylose contents.Map-based cloning combined with a complementation test demonstrated that PPR5 encodes a P-type PPR protein that is localized to the mitochondria.The mutation in PPR5 caused reduced splicing efficiency of mitochondrial NADH dehydrogenase 4(nad4)gene intron 3 and reduced complex I assembly and activity.Loss of PPR5 function greatly upregulated expression of alternative oxidases(AOXs),reduced ATP production,and affected mitochondrial morphology.We demonstrate that PPR5,as a P-type PPR protein,is required for mitochondrial function and endosperm development by controlling the cis-splicing of mitochondrial nad4 intron 3.展开更多
The major storage substance in rice endosperm is starch, which accounts for 80% of dry matter weight. In this study, rice mutant flo7, selected from the progeny of Nipponbare＇s tissue culture, displayed floury and op...The major storage substance in rice endosperm is starch, which accounts for 80% of dry matter weight. In this study, rice mutant flo7, selected from the progeny of Nipponbare＇s tissue culture, displayed floury and opaque endosperm. Compared with its corresponding wild type （WT） Nipponbare, the mutant rio7 produced longer, narrower, thinner and lighter grains. The levels of glucose, fructose and sucrose in the mutant flo7 endosperm were higher than those in the WT endosperm, whereas the protein content was not affected. With respect to both amylose content and gel consistency, the mutant flo7 was lower than WT, but its alkali value was higher. Scanning electron microscopic examinations showed that the endosperm of the mutant flo7 contained irregular, loosely packed and compound starch granules. Genetic analysis indicated that the mutant phenotype was determined by a single recessive nuclear gene The flo7 locus was mapped to a region on the long arm of chromosome 12, within a 95.1 kb interval defined by the markers C2-11 and C5-15. There are 13 open reading frames in the mapping interval. Transcription profiling of the developing grains showed that a number of genes involved in starch synthesis were affected differently in the mutant flo7.展开更多
Chalkiness characters affect not only the grain appearance,milling,eating and cooking qualities but also the grain nutritional quality in rice,thus it is one of the most important traits in rice. It is very important ...Chalkiness characters affect not only the grain appearance,milling,eating and cooking qualities but also the grain nutritional quality in rice,thus it is one of the most important traits in rice. It is very important for us to investigate the relation of the chalkiness formation and the development of endosperm structure and starch granule of different rice varieties. Here,we have investigated the chalkiness characters such as chalkiness rate,chalkiness degree and chalkiness area in 15 japonica rice varieties from southern Henan. Furthermore,the endosperm structure and starch granules of rice grain were also observed with scanning electron microscope. The results showed that the 15 japonica varieties have a significantly linear relationship between the chalkiness rate and chalkiness degree. Among the varieties,the biggest difference is the chalkiness rate,the second is the chalkiness area,and the last is the chalkiness degree. Moreover,there is a certain correlation between the distribution of starch granules,the arrangement of endosperm cells and the occurrence of grain chalkiness in the different rice varieties. For the same variety,the starch granules of chalky and non-chalky grains have obvious difference,while the starch granules from the transparent part of chalky rice and non-chalky rice do not have significant difference. The results would provide useful references for the improvement of grain quality in rice.展开更多
To reveal the suitability of using mature embryos as an explant source in wheat tissue culture, mature embryos from eight common wheat cultivars （Triticum aestivum L. cv.） were cultured with or without endosperm to ...To reveal the suitability of using mature embryos as an explant source in wheat tissue culture, mature embryos from eight common wheat cultivars （Triticum aestivum L. cv.） were cultured with or without endosperm to test their efficiency of callus induction and plant regeneration. When embryos were cultured together with endosperm （endosperm-supported culture, ES）, the percentage of callus induction was significantly lower than that when embryos were cultured in the absence of endosperm （non-endosperm-supported culture, NES）. This pattern was evident in most genotypes, regardless of whether 2 or 8 mg L^-1 2,4-D was added in the NES culture. However, in ES culture, more induced calli were differentiated into distinct green spots and they further developed into plantlets. Thus, more plants were regenerated in ES culture than in the NES treatment. Most of the eight tested genotypes showed a significant difference in callus induction rate and plantlet regeneration in both ES and NES cultures. In addition, the enzymatic activity of oxalate oxidase in the callus of ES culture condition was obviously higher than that in the callus of NES culture condition, suggesting that the activity of oxalate oxidase may be a parameter for selection of calli with potential for plantlet regeneration. These results indicate that wheat mature embryos are valuable explants for highly efficient callus induction and plant regeneration, if proper treatment and medium are used.展开更多
Morphological variations of the nucleus in starchy endosperm cell were observed by theelectron-transmisson microscope during endosperm development in rice. Along with thedevelopment of the starchy endosperm, the nucle...Morphological variations of the nucleus in starchy endosperm cell were observed by theelectron-transmisson microscope during endosperm development in rice. Along with thedevelopment of the starchy endosperm, the nuclei of the cells showed chromatin condensation,the typical feature of programmed cell death (PCD). The nuclei also showed nucleusdeformation, disruption of nuclear envelope, nucleoplasm leaking into the cytoplasm andnucleus disintegration resulting in nuclear residue formation. From the nucleus deformationto the nucleus disintegration, the morphological changes of the nucleus were orderlyprogressive. This indicated that the cell death of starchy endosperm in rice wasprogrammed cell death. Evans Blue staining observation showed that the cell death wasinitially detected in the central part of starchy endosperm in rice, then expandedoutward. The activities of superoxide dismutase (SOD) and catalase (CAT) in rice starchyendosperm both descended continuously as development progressed. The analysis of DNA ofrice starchy endosperm did not show the presence of DNA laddering. The above resultsshowed that the cell death of starchy endosperm in rice was a special form of PCD.展开更多
Endosperm mutants are critical to the studies on both starch synthesis and metabolism and genetic improvement of starch quality in maize.In the present study,a novel maize endosperm mutant A0178 of natural variation w...Endosperm mutants are critical to the studies on both starch synthesis and metabolism and genetic improvement of starch quality in maize.In the present study,a novel maize endosperm mutant A0178 of natural variation was used as the experimental material and identified and then characterized.Through phenotypic identification,genetic analysis,main ingredients measurement and embryo rescue,development of genetic mapping population from A0178,the endosperm mutant gene was located.The results showed that the mutant exhibited extremely low germination ability as attributed to the inhibited embryo development,and amounts of sugars were accumulated in the mutant seeds and more sugars content was detected at 23 days after pollination(DAP)in A0178 than B73.Employing genetic linkage analysis,the mutant trait was mapped in the bin 5.04 on chromosome 5.Sequence analysis showed that two sites of base transversion and insertion presented in the protein coding region and non-coding region of the mutant brittle-1(bt1),the adenylate translocator encoding gene involved in the starch synthesis.The single base insertion in the coding region cause frameshift mutation,early termination and lose of function of Brittle-1(BT1).All results suggested that bt1 is a novel allelic gene and the causal gene of this endosperm mutant,providing insights on the mechanism of endosperm formation in maize.展开更多
基金supported by Biological Breeding-National Science and Technology Major Project(2023ZD04069)Young Scientist Project(2023YFD1200008)+2 种基金the National Natural Science Foundation of China(32472122)Sichuan Provincial General Project(24NSFSC1704)the Open Project Program and Biological Breeding Program of State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China(SKL-ZY202211).
文摘Phosphorylation is one of the major posttranslational modifications to control plant growth and development.Opaque2(O2)represents a central hub for endosperm filling,which largely determines seed yield and nutrient storage in maize.However,it still remains unclear how O2 phosphorylation orchestrates endosperm filling and nutrient quality.Here,we systematically identified the phosphorylation sites of O2 during endosperm filling.A total of 18 phosphorylation sites were found in O2 and five sites were identified to apparently modulate its subcellular localization and transactivation capacity.In addition,a conserved protein kinase CK1 was confirmed to interact with and phosphorylate O2 at the residue Threonine(T)202 to promote O2-mediated transactivation and protein stability.Overexpression of CK1 resulted in increased kernel size,100-kernel weight and nutrient storage.Phosphorylation-mimic O2 seeds at T202 exhibited enhanced kernel dimension,test weight,vitreous endosperm area and nutrient accumulation,whereas the phosphorylation-deficient O2 seeds did not.Collectively,this study establishes a comprehensive phosphocode atlas of O2 during endosperm filling and highlights the importance of phosphorylation modification in O2 to precisely orchestrate maize yield and nutrient quality.
基金supported by the National Key Research and Development Program of China(2022YFD2300700)the National Natural Science Foundation of China(32201894)+1 种基金Hainan Provincial Natural Science Foundation of China(323QN193)Rothamsted Research receives strategic funding from the Biotechnological and Biological Sciences Research Council of the UK.Matthew J.Paul acknowledges funding from the Delivering Sustainable Wheat(BB/X011003/1)Strategic Program.
文摘Inferior grains exhibit delayed developmental processes and reduced metabolic activities compared to superior grains,leading to unstable rice yield and quality.While significant advancements have been achieved in elucidating the physiology of endosperm filling in inferior grains,the role of the embryo remains underexplored and warrants comprehensive investigation.Two Wuyujing 3 mutants,DW024(relatively synchronous;syn-DW024)and DW179(significantly asynchronous;asyn-DW179),with different grain-filling patterns were used in this study.Samples of superior and inferior grains were collected at intervals from 5 to 60 d after fertilization and subsequently dissected into subsamples of the embryo and endosperm.Histochemical staining,biochemical analysis,and RNA sequencing(RNA-seq)were combined to systematically compare developmental and physiological traits between superior and inferior grains.Combining hierarchical clustering of mRNA datasets revealed three developmental phases of the endosperm and embryo:morphogenesis,endosperm filling/embryo enlargement,and maturation.In both syn-DW024 and asyn-DW179,the duration of the endosperm/embryo morphogenesis phase was identical in superior and inferior grains.The inferior grains of asyn-DW179 exhibited a 10-day prolongation in the endosperm filling phase and a 20-day extension in the embryo enlargement phase compared to the superior grains.The endosperm of inferior grains exhibited higher contents of sugars and free amino acids,along with slower accumulation of storage compounds,which was associated with the down-regulation of genes for starch synthesis and ABA signaling.In addition,transporters for nutrient exchanges between endosperm and embryo were down-regulated,suggesting a potential role of the embryo in adjusting the endosperm filling process.Collectively,our results reveal that the prolonged phases of endosperm filling and embryo enlargement may underlie the impaired development of inferior grains,offering a new perspective for breeding or cultivating rice with uniform grain quality.
基金supported by the National Natural Science Foundation of China(31570322)。
文摘The early development of the endosperm is crucial for balancing the allocation of maternal nutrients to offspring.This process is believed to be evolutionarily associated with genomic imprinting,resulting in parentally biased allelic gene expression.Beyond Fertilization Independent Seed(FIS)genes,the number of imprinted genes involved in early endosperm development and seed size determination remains limited.This study introduces early endosperm-expressed HAIKU(IKU)downstream Candidate F-box 1(ICF1)and ICF2 as maternally expressed imprinted genes(MEGs)in Arabidopsis thaliana.Although these genes are also demethylated by DEMETER(DME)in the central cell,their activation differs from the direct DME-mediated activation seen in classical MEGs such as the FIS genes.Instead,ICF maternal alleles carry pre-established hypomethylation in their promoters,priming them for activation by the WRKY10 transcription factor in the endosperm.On the contrary,paternal alleles are predominantly suppressed by CG methylation.Furthermore,we find that ICF genes partially contribute to the small seed size observed in iku mutants.Our discovery reveals a two-step regulatory mechanism that highlights the important role of conventional transcription factors in the activation of imprinted genes,which was previously not fully recognized.Therefore,the mechanism provides a new dimension to understand the transcriptional regulation of imprinting in plant reproduction and development.
文摘Ultrastructural features of nucleus degradation during programmed cell death (PCD) of starchy endosperm cells in rice ( Oryza sativa L.) were observed using transmission electron microscopy. Several distinct morphological features of PCD have been found in the developing starchy endosperm cells, e.g. nucleus deformation, chromatin condensation, nuclear envelope disruption, and nuclear matrix leakage. DNA ladder displayed a smear of large DNA fragments from nucleus and evident bands of small DNA fragments (140-180 bp) from both nucleus and cytoplasm. In contrast with the rapid nucleus degradation, cell organelles in cytoplasm, such as rough endoplasmic reticulum, amyloplast, and mitochondrion, maintained their metabolic functions for a longer time. Seed reserves were continually synthesized and accumulated in the starchy endosperm cells despite the nucleus degradation during the PCD process. These results suggest that starchy endosperm cells remain active during reserve material synthesis and accumulation in the PCD process. The specific relationships between nucleus and cytoplasm in the developing endosperm cells and the morphological changes of nucleus in the endosperm PCD process were also discussed.
基金This work was supported by the Key Sci-Tech Project of Fujian Province (No. 2004NZ01-2) the Natural Science Foundation of Fujian Province of China (No. 2006J0300).
文摘A statistical method for mapping quantitative trait loci （QTLs） underlying endosperm traits is proposed. The method is based on a genetic model containing both the direct effects and maternal effects of an endosperm QTL and on an experimental design termed two-stage hierarchical design, in which the trait information is obtained from F3 endosperms and molecular marker information is obtained from F2 plants and F3 embryos （plants）. Results of computer simulations indicate that the method can efficiently map endosperm QTLs and precisely estimate both the direct and maternal effects of endosperm QTLs when the sample size is sufficiently large.
基金Supported by National Natural Science Foundation of China(30960179)Natural Science Foundation of Yunnan Province(2007A048M)~~
文摘[Objective] This research aimed to study the FTIR spectra of corn germs and endosperms so as to provide a scientific way for identifying corn of different types. [Method] The corn germs and endosperms of three types were studied by using Fourier transform infrared spectroscopy（FTIR） technology, combined with cluster analysis. [Result] The overall characteristics of original FTIR spectra were basically similar within the range of 700-1 800 cm^-1. The FTIR spectra were mainly composed by the absorption peaks of polysaccharides, proteins and lipids. Within the wavelength range of 700-1 800 cm^-1, there were only tiny differences in original FTIR spectra among the corn germs and endosperms of three different types. The spectra were then processed by using first derivative and second derivative. The second derivative spectra were used for hierarchical cluster analysis（HCA）. The results showed that with the wavelength range of 700-1 800 cm^-1, the second derivative spectra of the 52 samples could be better clustered according to the tree types and corn germ and corn endosperm. The clustering correct rate reached 96.1%.[Conclusion] FTIR technology, combined with cluster analysis, can be used to identify different types of corn germs and endosperms, and it is characterized by convenience and rapidness.
基金supported by the National 863 Program of China (2011AA10A101)the Chinese High-Yielding Transgenic Program (2011ZX08001-004)a project of the State Key Laboratory of Rice Biology,China(ZZKT201101)
文摘Quantitative trait loci（QTL） for percentage of chalky grain,degree of chalkiness,and endosperm transparency were detected using 3 recombinant inbred line populations derived from crosses between parental lines of commercial three-line hybrids of indica rice.Two of the populations showed great variations on heading date,and the other had a short range of heading date variation.A total of 40 QTLs were detected and fell into 15 regions of 10 chromosomes,of which 5 regions were detected for 1 or more same traits over different populations,2 were detected for different traits in different populations,3 were detected for 2 or all the 3 traits in a single population,and 5 were detected for a single trait in a single population.Most of these QTLs have been reported previously,but a region located on the long arm of chromosome 10 showing significant effects in all the 3 populations has not been reported before.It was shown that a number of gene cloned,including the Wx and Alk for the physiochemical property of rice grain,and GW2,GS3 and GW5 for grain weight and grain size,could have played important roles for the genetic control of grain chalkiness in rice,but there are many more QTLs exerting stable effects for rice chalkiness over different genetic backgrounds.It is worth paying more attentions to these regions which harbor QTL such as the qPCG5.2/qDC5.2/qET5.2 and qPCG10/qDC10/qET10 detected in our study.Our results also showed that the use of segregating populations having high-uniform heading date could greatly increase the efficiency of the identification of QTL responsible for traits that are subjected to great environmental influence.
基金supported by the National Natural Science Foundation (31171482)Jiangsu Natural Science Foundation (BK2011445)Jiangsu Graduate Innovation Project (CXLX12-0910)
文摘This study provided visual evidence of a nitrogen effect on starch granules(SGs) in wheat endosperm. Winter wheat(Titicum aestivum L.) cultivar Xumai 30 was cultured under no nitrogen(control) and 240 kg ha-1of nitrogen applied at the booting stage. The number, morphology, and size of Aand B-type SGs in subaleurone of dorsal endosperm(SDE), center of dorsal endosperm(CDE), modified aleurone(MA), subaleurone of ventral endosperm(SVE), and center of ventral endosperm(CVE) were observed under light and electron microscopes.(1) The distribution of SGs in SDE was similar to that in SVE, the distributions of SGs in CDE and CVE were similar, but the distribution of SGs in MA was different from those in the other four endosperm regions. The number of SGs in the five endosperm regions was in the order SDE > CDE > SVE > CVE > MA.(2) Nitrogen increased the number of Aand B-type SGs in SDE and SVE. Nitrogen also increased the number of B-type SGs but decreased the number of A-type SGs in CDE and CVE. Nitrogen decreased the numbers of A-type and B-type SGs in MA. The results suggest that increased N fertilizer application mainly increased the numbers of small SGs and decreased the numbers of large SGs, but that the results varied in different regions of the wheat endosperm.
基金supported by the National High Technology Research and Development Program of China(Grant No.2006AA100101)National Science and Technology Support Program of China(Grant No. 2006BAD01A01-5)+1 种基金Special Program for Rice Scientific Research,Ministry of Agriculture,China(Grant No. nyhyzx 07-001-006)Super Rice Breeding and Demonstration Program,Ministry of Agriculture,China and Jiangsu Agricultural Scientific Self-innovation Fund,China(Grant No.CX[07]603)
文摘The translucent endosperm trait in a japonica rice variety ＇Kantou 194＇ is controlled by a Wx-mq gene which is allelic to Wx locus by genetic analysis and allelic test. The amylose content analysis showed that an intermediate amylose content between those of glutinous and non-glutinous rice existed in endosperm of homozygous Wx-mq genotype. The slight changes of amylose content in different varieties and F1 grains with an identical Wx-mq genotype might be influenced by dissimilar genetic background. To identify the Wx-mq genotype simply and rapidly, a cleaved amplified polymorphic sequence （CAPS） marker was designed. The result from the molecular detection indicated that it could be used for marker-assisted selection for low amylose content varieties in rice breeding.
基金supported by the National Natural Science Foundation of China(30270814)the Teaching and Research Award Program for Outstanding Young Teachers in Higher Education Institutions of MOE,R R.Chinathe Group Project of Natural Science Foundation of Guangdong Province(200023).
文摘By using the laser scanning confocal microscope and plastic （Leica 7022 historesin embedding kit） semi-thin sectioning technique, comparative studies on the development of endosperm were carried out between autotetraploid and diploid rices. About one third of the ovaries in the autotetraploid showed normal endosperm development as those in the diploid. In these ovaries, one of the polar nuclei would fuse with the sperm nucleus, and the primary endosperm nucleus formed and underwent the first division in 4 hours after pollination; the anticlinal wall began to grow centripetally between the free nuclei starting from the wall ingrowths of the embryo sac near the micropylar end, and some of the phragmoplasts formed transformed into periclinal walls. In addition, some of the cell wall situated in the middle of the endosperm appeared to originate from phragmoplasts, whereas others seemed to develop randomly without the obvious formation of phragmoplasts. Cellulose began to accumulate in the wall of aleurone cell layer at 6 days after pollination. The cellulose wall of the cells of the aleurone cell layer appeared to have completely formed within 7 to 8 days after pollination. On the other hand, about two thirds of the ovaries in the autotetraploid showed abnormality in endosperm development with various types, such as non-fertilization, abnormal fertilization, endosperm development-delay and non-synchronization in the development of cellulose wall of cells of the aleurone layer. These abnormalities usually resulted in decreased seed setting in autotetraploid rice.
基金supported by the Fundamental Research Funds for the Central Universities (XDJK2013C023)the Chongqing Postdoctoral Science Foundation (Xm201344)+2 种基金the China Postdoctoral Science Foundation (2014M552303)the Research Fund for the Doctoral Program of Southwest University (SWU112037)the Research Fund for the Doctoral Program of Higher Education (2011182120011)
文摘ZAG2 has been identified as a maternally expressed imprinted gene in maize endosperm.Our study revealed that paternally inherited ZAG2 alleles were imprinted in maize endosperm and embryo at 14 days after pollination(DAP), and consistently imprinted in endosperm at 10, 12, 16, 18, 20, 22, 24, 26, and 28 DAP in reciprocal crosses between B73 and Mo17. ZAG2 alleles were also imprinted in reciprocal crosses between Zheng 58 and Chang7-2 and between Huang C and 178. ZAG2 alleles exhibited differential imprinting in hybrids of 178 × Huang C and B73 × Mo17, while in other hybrids ZAG2 alleles exhibited binary imprinting. The tissue-specific expression pattern of ZAG2 showed that ZAG2 was expressed at a high level in immature ears, suggesting that ZAG2 plays important roles in not only kernel but ear development.
基金Supported by National Natural Science Foundation of China(30900874)Natural Science Foundation of Hunan Province(11JJ3026)+1 种基金Excellent Youth Fund Project of Hunan Department of Education(13B052)Crop Science Open Fund Project(ZWKF201504)
文摘To ascertain the effect mechanism of high temperature after anthesis on rice quality, the experiment was conducted with two rice lines, the heat-tolerant line 996 and heat-sensitive line 4628, with high temperature and optimal temperature in the growth chamber to investigate the effect of high temperature stress after anthesis on rice appearance quality, milling quality, cooking and eating quality and starch granule structure of endo- sperm. The result showed that milled rice rate, head rice rate, amylose content and gel consistency of both lines decreased under high temperature stress after anthesis, while the ratio of grain length to width, chalky rate, chalkiness, protein content increased. Under high temperature treatments, gelatinization temperature, final viscosity, set back and peak time increased, breakdown decreased, Mg content and K content increased, Mg/K ra- tio decreased. Under same treatment, the extent of rice quality of heat tolerant line 996 affected by high temperature was lower than that of heat sensitive line 4628. Under high temperature stress after anthesis, starch granule arranged untightly, most of starch granules existed in the form of a single starch endosperm, refractive index decreased, transparency reduced, and led to the formation of chalk. Under high temperature stress af- ter anthesis, the increase of protein content, the decrease of Mg/K, the changes of rice RVA profile characteristics and starch granule structure of endosperm could be the main reason for the decrease of rice cooking and eating quality and appearance quality.
基金supported by the Key Program of the Development of Variety of Genetically Modified Organisms(Grant Nos.2009ZX08001-019B and 2008ZX08001-006)the Special Program for Rice Scientific Research of Ministry of Agriculture(Grant No.nyhyzx 07-001-006)+1 种基金the Key Support Program of Science and Technology of Jiangsu Province(Grant No.BE2008354)the Self-directed Innovation Fund of Agricultural Science and Technology in Jiangsu Province,China(Grant No.CX[09]634)
文摘A high-yielding japonica rice variety, Wuyunjing 7, bred in Jiangsu Province, China as a female parent was crossed with a Japanese rice variety Kantou 194, which carries a rice stripe disease resistance gene Stv-b＇ and a translucent endosperm mutant gene Wx-mq. From F2 generations, a sequence characterized amplified region （SCAR） marker tightly linked with Stv-b＇ and a cleaved amplified polymorphic sequence （CAPS） marker for Wx-mq were used for marker-assisted selection. Finally, a new japonica rice line, Ning 9108, with excellent agronomic traits was obtained by multi-generational selection on stripe disease resistance and endosperm appearance. The utilization of the markers from genes related to rice quality and disease resistance was helpful not only for establishing a marker-assisted selection system of high-quality and disease resistance for rice but also for providing important intermediate materials and rapid selection method for good quality, disease resistance and high yield in rice breeding.
基金This work was supported by grants from the National Natural Science Foundation of China(31901427)the National Transgenic Science and Technology Program(2019ZX08010-003)+1 种基金the Agricultural Science and Technology Innovation Program of CAAS(CAAS-ZDXT2018001)the Young Talent of CAAS to Yulong Ren.
文摘Endosperm as the storage organ of starch and protein in cereal crops largely determines grain yield and quality.Despite the fact that several pentatricopeptide repeat(PPR)proteins required for endosperm development have been identified in rice,the molecular mechanisms of many P-type PPR proteins in endosperm development remains unclear.Here,we isolated a rice floury endosperm mutant ppr5 that developed small starch grains and an abnormal aleurone layer,accompanied by decreased starch,protein,and amylose contents.Map-based cloning combined with a complementation test demonstrated that PPR5 encodes a P-type PPR protein that is localized to the mitochondria.The mutation in PPR5 caused reduced splicing efficiency of mitochondrial NADH dehydrogenase 4(nad4)gene intron 3 and reduced complex I assembly and activity.Loss of PPR5 function greatly upregulated expression of alternative oxidases(AOXs),reduced ATP production,and affected mitochondrial morphology.We demonstrate that PPR5,as a P-type PPR protein,is required for mitochondrial function and endosperm development by controlling the cis-splicing of mitochondrial nad4 intron 3.
基金supported by China National Natural Science Foundation (Grant Nos. 31471472 and 31301303)Natural Science Foundation of Zhejiang Province,China (Grant Nos. LY13C130003 and LY14C130012)
文摘The major storage substance in rice endosperm is starch, which accounts for 80% of dry matter weight. In this study, rice mutant flo7, selected from the progeny of Nipponbare＇s tissue culture, displayed floury and opaque endosperm. Compared with its corresponding wild type （WT） Nipponbare, the mutant rio7 produced longer, narrower, thinner and lighter grains. The levels of glucose, fructose and sucrose in the mutant flo7 endosperm were higher than those in the WT endosperm, whereas the protein content was not affected. With respect to both amylose content and gel consistency, the mutant flo7 was lower than WT, but its alkali value was higher. Scanning electron microscopic examinations showed that the endosperm of the mutant flo7 contained irregular, loosely packed and compound starch granules. Genetic analysis indicated that the mutant phenotype was determined by a single recessive nuclear gene The flo7 locus was mapped to a region on the long arm of chromosome 12, within a 95.1 kb interval defined by the markers C2-11 and C5-15. There are 13 open reading frames in the mapping interval. Transcription profiling of the developing grains showed that a number of genes involved in starch synthesis were affected differently in the mutant flo7.
基金Supported by Key Project of Science and Technology in Henan Province(152102110100,152102110036)National Natural Science Foundation of China(U1604110,U1404319,31270727,31600992)+3 种基金Nanhu Scholars Program for Young Scholars of XYNU(2016056)Major Science and Technology Project in Henan Province(121100110200)Students Scientific Research Fund of Xinyang Normal University(2015-DXS-158)Fund of Institute for Conservation and Utilization of Agro-bioresources in Dabie Mountains(2016020)
文摘Chalkiness characters affect not only the grain appearance,milling,eating and cooking qualities but also the grain nutritional quality in rice,thus it is one of the most important traits in rice. It is very important for us to investigate the relation of the chalkiness formation and the development of endosperm structure and starch granule of different rice varieties. Here,we have investigated the chalkiness characters such as chalkiness rate,chalkiness degree and chalkiness area in 15 japonica rice varieties from southern Henan. Furthermore,the endosperm structure and starch granules of rice grain were also observed with scanning electron microscope. The results showed that the 15 japonica varieties have a significantly linear relationship between the chalkiness rate and chalkiness degree. Among the varieties,the biggest difference is the chalkiness rate,the second is the chalkiness area,and the last is the chalkiness degree. Moreover,there is a certain correlation between the distribution of starch granules,the arrangement of endosperm cells and the occurrence of grain chalkiness in the different rice varieties. For the same variety,the starch granules of chalky and non-chalky grains have obvious difference,while the starch granules from the transparent part of chalky rice and non-chalky rice do not have significant difference. The results would provide useful references for the improvement of grain quality in rice.
文摘To reveal the suitability of using mature embryos as an explant source in wheat tissue culture, mature embryos from eight common wheat cultivars （Triticum aestivum L. cv.） were cultured with or without endosperm to test their efficiency of callus induction and plant regeneration. When embryos were cultured together with endosperm （endosperm-supported culture, ES）, the percentage of callus induction was significantly lower than that when embryos were cultured in the absence of endosperm （non-endosperm-supported culture, NES）. This pattern was evident in most genotypes, regardless of whether 2 or 8 mg L^-1 2,4-D was added in the NES culture. However, in ES culture, more induced calli were differentiated into distinct green spots and they further developed into plantlets. Thus, more plants were regenerated in ES culture than in the NES treatment. Most of the eight tested genotypes showed a significant difference in callus induction rate and plantlet regeneration in both ES and NES cultures. In addition, the enzymatic activity of oxalate oxidase in the callus of ES culture condition was obviously higher than that in the callus of NES culture condition, suggesting that the activity of oxalate oxidase may be a parameter for selection of calli with potential for plantlet regeneration. These results indicate that wheat mature embryos are valuable explants for highly efficient callus induction and plant regeneration, if proper treatment and medium are used.
基金supported by the National Natural Science Foundation of China(30070363)the Foundation for Doctorate Research of Ministry of Education,China(200005041).
文摘Morphological variations of the nucleus in starchy endosperm cell were observed by theelectron-transmisson microscope during endosperm development in rice. Along with thedevelopment of the starchy endosperm, the nuclei of the cells showed chromatin condensation,the typical feature of programmed cell death (PCD). The nuclei also showed nucleusdeformation, disruption of nuclear envelope, nucleoplasm leaking into the cytoplasm andnucleus disintegration resulting in nuclear residue formation. From the nucleus deformationto the nucleus disintegration, the morphological changes of the nucleus were orderlyprogressive. This indicated that the cell death of starchy endosperm in rice wasprogrammed cell death. Evans Blue staining observation showed that the cell death wasinitially detected in the central part of starchy endosperm in rice, then expandedoutward. The activities of superoxide dismutase (SOD) and catalase (CAT) in rice starchyendosperm both descended continuously as development progressed. The analysis of DNA ofrice starchy endosperm did not show the presence of DNA laddering. The above resultsshowed that the cell death of starchy endosperm in rice was a special form of PCD.
基金This work was financially supported in part by grants from National Science and Technology Support Project of China(2016YFD0101205)the Natural Science Foundation of Jiangsu Province,China(BK20160586)+1 种基金National Transgenic Major Project of China(2019ZX08010-004)as well as Six Talent Peaks Project of Jiangsu Province,China(NY-020).
文摘Endosperm mutants are critical to the studies on both starch synthesis and metabolism and genetic improvement of starch quality in maize.In the present study,a novel maize endosperm mutant A0178 of natural variation was used as the experimental material and identified and then characterized.Through phenotypic identification,genetic analysis,main ingredients measurement and embryo rescue,development of genetic mapping population from A0178,the endosperm mutant gene was located.The results showed that the mutant exhibited extremely low germination ability as attributed to the inhibited embryo development,and amounts of sugars were accumulated in the mutant seeds and more sugars content was detected at 23 days after pollination(DAP)in A0178 than B73.Employing genetic linkage analysis,the mutant trait was mapped in the bin 5.04 on chromosome 5.Sequence analysis showed that two sites of base transversion and insertion presented in the protein coding region and non-coding region of the mutant brittle-1(bt1),the adenylate translocator encoding gene involved in the starch synthesis.The single base insertion in the coding region cause frameshift mutation,early termination and lose of function of Brittle-1(BT1).All results suggested that bt1 is a novel allelic gene and the causal gene of this endosperm mutant,providing insights on the mechanism of endosperm formation in maize.
