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1.
Planta ; 260(1): 19, 2024 Jun 05.
Article in English | MEDLINE | ID: mdl-38839605

ABSTRACT

MAIN CONCLUSION: A mutation was first found to cause the great generation of glutelin precursors (proglutelins) in rice (Oryza sativa L.) endosperm, and thus referred to as GPGG1. The GPGG1 was involved in synthesis and compartmentation of storage proteins. The PPR-like gene in GPGG1-mapped region was determined as its candidate gene. In the wild type rice, glutelins and prolamins are synthesized on respective subdomains of rough endoplasmic reticulum (ER) and intracellularly compartmentalized into different storage protein bodies. In this study, a storage protein mutant was obtained and characterized by the great generation of proglutelins combining with the lacking of 13 kD prolamins. A dominant genic-mutation, referred to as GPGG1, was clarified to result in the proteinous alteration. Novel saccular composite-ER was shown to act in the synthesis of proglutelins and 14 kD prolamins in the mutant. Additionally, a series of organelles including newly occurring several compartments were shown to function in the transfer, trans-plasmalemmal transport, delivery, deposition and degradation of storage proteins in the mutant. The GPGG1 gene was mapped to a 67.256 kb region of chromosome 12, the pentatricopeptide repeat (PPR)-like gene in this region was detected to contain mutational sites.


Subject(s)
Endosperm , Glutens , Mutation , Oryza , Oryza/genetics , Oryza/metabolism , Endosperm/genetics , Endosperm/metabolism , Glutens/genetics , Glutens/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Prolamins/genetics , Prolamins/metabolism , Seed Storage Proteins/genetics , Seed Storage Proteins/metabolism , Endoplasmic Reticulum/metabolism , Chromosome Mapping , Genome, Plant/genetics
2.
New Phytol ; 243(1): 213-228, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38715414

ABSTRACT

Arabidopsis lamin analogs CROWDED NUCLEIs (CRWNs) are necessary to maintain nuclear structure, genome function, and proper plant growth. However, whether and how CRWNs impact reproduction and genome-wide epigenetic modifications is unknown. Here, we investigate the role of CRWNs during the development of gametophytes, seeds, and endosperm, using genomic and epigenomic profiling methods. We observed defects in crwn mutant seeds including seed abortion and reduced germination rate. Quadruple crwn null genotypes were rarely transmitted through gametophytes. Because defects in seeds often stem from abnormal endosperm development, we focused on crwn1 crwn2 (crwn1/2) endosperm. These mutant seeds exhibited enlarged chalazal endosperm cysts and increased expression of stress-related genes and the MADS-box transcription factor PHERES1 and its targets. Previously, it was shown that PHERES1 expression is regulated by H3K27me3 and that CRWN1 interacts with the PRC2 interactor PWO1. Thus, we tested whether crwn1/2 alters H3K27me3 patterns. We observed a mild loss of H3K27me3 at several hundred loci, which differed between endosperm and leaves. These data indicate that CRWNs are necessary to maintain the H3K27me3 landscape, with tissue-specific chromatin and transcriptional consequences.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Endosperm , Gene Expression Regulation, Plant , Histones , Mutation , Reproduction , Arabidopsis/genetics , Arabidopsis/physiology , Arabidopsis Proteins/metabolism , Arabidopsis Proteins/genetics , Histones/metabolism , Endosperm/genetics , Endosperm/metabolism , Mutation/genetics , Seeds/genetics , Seeds/growth & development , Cell Nucleus/metabolism , Methylation
3.
BMC Plant Biol ; 24(1): 458, 2024 May 27.
Article in English | MEDLINE | ID: mdl-38797860

ABSTRACT

BACKGROUND: The endosperm serves as the primary source of nutrients for maize (Zea mays L.) kernel embryo development and germination. Positioned at the base of the endosperm, the transfer cells (TCs) of the basal endosperm transfer layer (BETL) generate cell wall ingrowths, which enhance the connectivity between the maternal plant and the developing kernels. These TCs play a crucial role in nutrient transport and defense against pathogens. The molecular mechanism underlying BETL development in maize remains unraveled. RESULTS: This study demonstrated that the MYB-related transcription factor ZmMYBR29, exhibited specific expression in the basal cellularized endosperm, as evidenced by in situ hybridization analysis. Utilizing the CRISPR/Cas9 system, we successfully generated a loss-of-function homozygous zmmybr29 mutant, which presented with smaller kernel size. Observation of histological sections revealed abnormal development and disrupted morphology of the cell wall ingrowths in the BETL. The average grain filling rate decreased significantly by 26.7% in zmmybr29 mutant in comparison to the wild type, which impacted the dry matter accumulation within the kernels and ultimately led to a decrease in grain weight. Analysis of RNA-seq data revealed downregulated expression of genes associated with starch synthesis and carbohydrate metabolism in the mutant. Furthermore, transcriptomic profiling identified 23 genes that expressed specifically in BETL, and the majority of these genes exhibited altered expression patterns in zmmybr29 mutant. CONCLUSIONS: In summary, ZmMYBR29 encodes a MYB-related transcription factor that is expressed specifically in BETL, resulting in the downregulation of genes associated with kernel development. Furthermore, ZmMYBR29 influences kernels weight by affecting the grain filling rate, providing a new perspective for the complementation of the molecular regulatory network in maize endosperm development.


Subject(s)
Edible Grain , Endosperm , Gene Expression Regulation, Plant , Plant Proteins , Transcription Factors , Zea mays , Zea mays/genetics , Zea mays/growth & development , Zea mays/metabolism , Transcription Factors/genetics , Transcription Factors/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Edible Grain/genetics , Edible Grain/growth & development , Edible Grain/metabolism , Endosperm/genetics , Endosperm/growth & development , Endosperm/metabolism , Cell Wall/metabolism , Cell Wall/genetics , Seeds/genetics , Seeds/growth & development , Seeds/metabolism , CRISPR-Cas Systems
4.
Mol Plant ; 17(5): 788-806, 2024 May 06.
Article in English | MEDLINE | ID: mdl-38615195

ABSTRACT

During maize endosperm filling, sucrose not only serves as a source of carbon skeletons for storage-reserve synthesis but also acts as a stimulus to promote this process. However, the molecular mechanisms underlying sucrose and endosperm filling are poorly understood. In this study, we found that sucrose promotes the expression of endosperm-filling hub gene Opaque2 (O2), coordinating with storage-reserve accumulation. We showed that the protein kinase SnRK1a1 can attenuate O2-mediated transactivation, but sucrose can release this suppression. Biochemical assays revealed that SnRK1a1 phosphorylates O2 at serine 41 (S41), negatively affecting its protein stability and transactivation ability. We observed that mutation of SnRK1a1 results in larger seeds with increased kernel weight and storage reserves, while overexpression of SnRK1a1 causes the opposite effect. Overexpression of the native O2 (O2-OE), phospho-dead (O2-SA), and phospho-mimetic (O2-SD) variants all increased 100-kernel weight. Although O2-SA seeds exhibit smaller kernel size, they have higher accumulation of starch and proteins, resulting in larger vitreous endosperm and increased test weight. O2-SD seeds display larger kernel size but unchanged levels of storage reserves and test weight. O2-OE seeds show elevated kernel dimensions and nutrient storage, like a mixture of O2-SA and O2-SD seeds. Collectively, our study discovers a novel regulatory mechanism of maize endosperm filling. Identification of S41 as a SnRK1-mediated phosphorylation site in O2 offers a potential engineering target for enhancing storage-reserve accumulation and yield in maize.


Subject(s)
Endosperm , Plant Proteins , Sucrose , Zea mays , Zea mays/metabolism , Zea mays/genetics , Endosperm/metabolism , Phosphorylation , Plant Proteins/metabolism , Plant Proteins/genetics , Sucrose/metabolism , Gene Expression Regulation, Plant , Protein Serine-Threonine Kinases/metabolism , Protein Serine-Threonine Kinases/genetics , Transcription Factors/metabolism , Transcription Factors/genetics , DNA-Binding Proteins/metabolism , DNA-Binding Proteins/genetics , Seeds/metabolism , Seeds/genetics , Seeds/growth & development
5.
J Biosci Bioeng ; 138(1): 44-53, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38614830

ABSTRACT

Kopyor is a coconut with unique characteristics from Indonesia, one of the largest coconut producers in the world. Kopyor is an edible mature coconut with soft endosperm. Although this fruit is one of the most popular coconuts in the world, there are limited studies on its properties, including its sensory attributes and metabolite profiles. This study investigates the characteristics of kopyor using sensory evaluation, a widely targeted metabolomics approach, and multivariate analysis. The liquid (water) and solid (flesh) endosperms were collected as the samples. The results showed that kopyor has characteristics that distinguish it from normal mature and young coconuts. Kopyor water has a milky, creamy, nutty, bitter, and astringent taste with an oily aftertaste and mouthfeel. Kopyor flesh is soft and moist and gives a sandy mouth feel. This study analyzed the sensory attributes of the kopyor endosperm for the first time and compared it with those of normal mature and young coconuts. A gas chromatography mass spectrometry analysis showed that kopyor contained wider variety of metabolites than normal coconuts of the same age. Based on the differential analysis and orthogonal projections to latent structures-regression, kopyor water was characterized by the accumulation of flavor-related metabolites, such as amino acids and organic acids, which contributed to its sensory complexity. This study solidified the effects of maturation and endosperm type on metabolite accumulation in kopyor endosperm. This pioneering information will lead to the future use of kopyor and other unique coconuts worldwide for food, contributing to the sustainability of the coconut industry.


Subject(s)
Cocos , Gas Chromatography-Mass Spectrometry , Metabolomics , Taste , Cocos/chemistry , Metabolomics/methods , Indonesia , Endosperm/metabolism , Endosperm/chemistry , Humans
6.
New Phytol ; 242(6): 2635-2651, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38634187

ABSTRACT

Endosperm is the main storage organ in cereal grain and determines grain yield and quality. The molecular mechanisms of heat shock proteins in regulating starch biosynthesis and endosperm development remain obscure. Here, we report a rice floury endosperm mutant flo24 that develops abnormal starch grains in the central starchy endosperm cells. Map-based cloning and complementation test showed that FLO24 encodes a heat shock protein HSP101, which is localized in plastids. The mutated protein FLO24T296I dramatically lost its ability to hydrolyze ATP and to rescue the thermotolerance defects of the yeast hsp104 mutant. The flo24 mutant develops more severe floury endosperm when grown under high-temperature conditions than normal conditions. And the FLO24 protein was dramatically induced at high temperature. FLO24 physically interacts with several key enzymes required for starch biosynthesis, including AGPL1, AGPL3 and PHO1. Combined biochemical and genetic evidence suggests that FLO24 acts cooperatively with HSP70cp-2 to regulate starch biosynthesis and endosperm development in rice. Our results reveal that FLO24 acts as an important regulator of endosperm development, which might function in maintaining the activities of enzymes involved in starch biosynthesis in rice.


Subject(s)
Endosperm , Gene Expression Regulation, Plant , Mutation , Oryza , Plant Proteins , Starch , Oryza/genetics , Oryza/metabolism , Oryza/growth & development , Endosperm/metabolism , Endosperm/growth & development , Starch/metabolism , Starch/biosynthesis , Plant Proteins/metabolism , Plant Proteins/genetics , Mutation/genetics , Protein Binding , Plastids/metabolism , Genetic Complementation Test , Adenosine Triphosphate/metabolism , Adenosine Triphosphate/biosynthesis , Thermotolerance , Transcription Factors
7.
Curr Biol ; 34(8): R308-R312, 2024 04 22.
Article in English | MEDLINE | ID: mdl-38653196

ABSTRACT

Flowering plants, also known as angiosperms, emerged approximately 150 to 200 million years ago. Since then, they have undergone rapid and extensive expansion, now encompassing around 90% of all land plant species. The remarkable diversification of this group has been a subject of in-depth investigations, and several evolutionary innovations have been proposed to account for their success. In this primer, we will specifically focus on one such innovation: the advent of seeds containing endosperm.


Subject(s)
Biological Evolution , Magnoliopsida , Reproduction , Magnoliopsida/physiology , Magnoliopsida/genetics , Reproduction/physiology , Endosperm/physiology , Seeds/physiology
8.
Int J Biol Macromol ; 267(Pt 2): 131663, 2024 May.
Article in English | MEDLINE | ID: mdl-38636760

ABSTRACT

Palm seedlings are visually selected from mature fruits in a slow process that leads to nonuniform germination and high embryo mortality. In this study, we determined the levels of monosaccharides, their crystallinity, and their role in the formation of Euterpe edulis endosperm during seed maturation. Seeds harvested from 108 to 262 days after anthesis (DAA) were analyzed morphologically, physiologically, and chemically to measure soluble and insoluble lignins, ashes, structural carbohydrates, degree of crystallinity, and endo-ß-mannanase. The seeds achieved maximum germination and vigor at 164 DAA. During the early stages, only compounds with a low structural order were formed. The contents of soluble and insoluble lignins, ashes, glucans, and galactans decreased during maturation. Those of mannans, the main structural carbohydrate in the endosperm, increased along with the degree of crystallinity, as suggested by a mannan-I-type X-ray diffraction pattern. Similarly, endo-ß-mannanase activity peaked at 262 DAA. The superior physiological outcome of seeds and seedlings at 164 DAA implies a 98-day shorter harvesting time. The state of mannans during seed maturation could be used as a marker to improve seedling production by E. edulis.


Subject(s)
Arecaceae , Germination , Mannans , Seeds , Seeds/growth & development , Seeds/chemistry , Mannans/chemistry , Arecaceae/chemistry , Arecaceae/growth & development , Trees , Lignin/chemistry , Lignin/metabolism , Endosperm/chemistry , Endosperm/metabolism , Seedlings/growth & development
9.
BMC Plant Biol ; 24(1): 184, 2024 Mar 12.
Article in English | MEDLINE | ID: mdl-38475704

ABSTRACT

Using the blueberry cultivar "Powderblue" after pollination, fruits at different developmental stages were collected for study. The transverse and longitudinal diameters, individual fruit weight, and fruit water content were measured during their development. Employing tissue sectioning and microscopy techniques, we systematically studied the morphological features and anatomical structures of the fruits and seeds at various developmental stages, aiming to elucidate the cytological patterns during blueberry fruit development. The results of our study revealed that the "Powderblue" blueberry fruit growth and development followed a double "S" curve. Mature "Powderblue" blueberries were blue-black in color, elliptical in shape, with five locules, an inferior ovary, and an average fruit weight of 1.73 ± 0.17 g, and a moisture content of 78.865 ± 0.9%. Blueberry fruit flesh cells were densely arranged with no apparent intercellular spaces, and mesocarp cells accounted for 52.06 ± 7.4% of fruit cells. In the early fruit development stages, the fruit flesh cells were rapidly dividing, significantly increasing in number but without greatly affecting the fruit's morphological characteristics. During the later stages of fruit development, the expansion of the fruit flesh cells became prominent, resulting in a noticeable increase in the fruit's dimensions. Except for the epidermal cells, cells in all fruit tissues showed varying degrees of rupture as fruit development progressed, with the extent of cell rupture increasing, becoming increasingly apparent as the fruit gradually softened. Additionally, numerous brachysclereids (stone cells) appeared in the fruit flesh cells. Stone cells are mostly present individually in the fruit flesh tissue, while in the placental tissue, they often group together. The "Powderblue" blueberry seeds were light brown, 4.13 ± 0.42 mm long, 2.2 ± 0.14 mm wide, with each fruit containing 50-60 seeds. The "Powderblue" seeds mainly consisted of the seed coat, endosperm, and embryo. The embryo was located at the chalazal end in the center of the endosperm and was spatially separated. The endosperm, occupying the vast majority of the seed volume, comprised both the chalazal and outer endosperm, and the endosperm developed and matured before the embryo. As the seed developed, the seed coat was gradually lignified and consisted of palisade-like stone cells externally and epidermal layer cells internally.


Subject(s)
Blueberry Plants , Fruit , Pregnancy , Female , Humans , Blueberry Plants/chemistry , Placenta , Seeds , Endosperm
10.
BMC Plant Biol ; 24(1): 196, 2024 Mar 18.
Article in English | MEDLINE | ID: mdl-38494545

ABSTRACT

BACKGROUND: Chalkiness is a common phenotype induced by various reasons, such as abiotic stress or the imbalance of starch synthesis and metabolism during the development period. However, the reason mainly for one gene losing its function such as NAC (TFs has a large family in rice) which may cause premature is rarely known to us. RESULTS: The Ko-Osnac02 mutant demonstrated an obviously early maturation stage compared to the wild type (WT) with 15 days earlier. The result showed that the mature endosperm of Ko-Osnac02 mutant exhibited chalkiness, characterized by white-core and white-belly in mature endosperm. As grain filling rate is a crucial factor in determining the yield and quality of rice (Oryza sativa, ssp. japonica), it's significant that mutant has a lower amylose content (AC) and higher soluble sugar content in the mature endosperm. Interestingly among the top DEGs in the RNA sequencing of N2 (3DAP) and WT seeds revealed that the OsBAM2 (LOC_Os10g32810) expressed significantly high in N2 mutant, which involved in Maltose up-regulated by the starch degradation. As Prediction of Protein interaction showed in the chalky endosperm formation in N2 seeds (3 DAP), seven genes were expressed at a lower-level which should be verified by a heatmap diagrams based on DEGs of N2 versus WT. The Tubulin genes controlling cell cycle are downregulated together with the MCM family genes MCM4 ( ↓), MCM7 ( ↑), which may cause white-core in the early endosperm development. In conclusion, the developing period drastically decreased in the Ko-Osnac02 mutants, which might cause the chalkiness in seeds during the early endosperm development. CONCLUSIONS: The gene OsNAC02 which controls a great genetic co-network for cell cycle regulation in early development, and KO-Osnac02 mutant shows prematurity and white-core in endosperm.


Subject(s)
Endosperm , Oryza , Endosperm/metabolism , Starch/metabolism , Seeds/genetics , Edible Grain/genetics , Homeostasis , Oryza/metabolism , Gene Expression Regulation, Plant
11.
J Mol Graph Model ; 129: 108761, 2024 06.
Article in English | MEDLINE | ID: mdl-38552302

ABSTRACT

ADP-glucose pyrophosphorylase plays a pivotal role as an allosteric enzyme, essential for starch biosynthesis in plants. The higher plant AGPase comparises of a pair of large and a pair of small subunits to form a heterotetrameric complex. Growing evidence indicates that each subunit plays a distinct role in regulating the underlying mechanism of starch biosynthesis. In the rice genome, there are four large subunit genes (OsL1-L4) and three small subunit genes (OsS1, OsS2a, and OsS2b). While the structural assembly of cytosolic rice AGPase subunits (OsL2:OsS2b) has been elucidated, there is currently no such documented research available for plastidial rice AGPases (OsL1:OsS1). In this study, we employed protein modeling and MD simulation approaches to gain insights into the structural association of plastidial rice AGPase subunits. Our results demonstrate that the heterotetrameric association of OsL1:OsS1 is very similar to that of cytosolic OsL2:OsS2b and potato AGPase heterotetramer (StLS:StSS). Moreover, the yeast-two-hybrid results on OsL1:OsS1, which resemble StLS:StSS, suggest a differential protein assembly for OsL2:OsS2b. Thus, the regulatory and catalytic mechanisms for plastidial AGPases (OsL1:OsS1) could be different in rice culm and developing endosperm compared to those of OsL2:OsS2b, which are predominantly found in rice endosperm.


Subject(s)
Oryza , Glucose-1-Phosphate Adenylyltransferase/genetics , Glucose-1-Phosphate Adenylyltransferase/chemistry , Glucose-1-Phosphate Adenylyltransferase/metabolism , Oryza/genetics , Endosperm/genetics , Endosperm/metabolism , Computer Simulation , Starch/metabolism , Protein Subunits/metabolism
12.
Plant Physiol ; 195(2): 1365-1381, 2024 May 31.
Article in English | MEDLINE | ID: mdl-38471799

ABSTRACT

Several starch synthesis regulators have been identified, but these regulators are situated in the terminus of the regulatory network. Their upstream regulators and the complex regulatory network formed between these regulators remain largely unknown. A previous study demonstrated that NAM, ATAF, and CUC (NAC) transcription factors, OsNAC20 and OsNAC26 (OsNAC20/26), redundantly and positively regulate the accumulation of storage material in rice (Oryza sativa) endosperm. In this study, we detected OsNAC25 as an upstream regulator and interacting protein of OsNAC20/26. Both OsNAC25 mutation and OE resulted in a chalky seed phenotype, decreased starch content, and reduced expression of starch synthesis-related genes, but the mechanisms were different. In the osnac25 mutant, decreased expression of OsNAC20/26 resulted in reduced starch synthesis; however, in OsNAC25-overexpressing plants, the OsNAC25-OsNAC20/26 complex inhibited OsNAC20/26 binding to the promoter of starch synthesis-related genes. In addition, OsNAC20/26 positively regulated OsNAC25. Therefore, the mutual regulation between OsNAC25 and OsNAC20/26 forms a positive regulatory loop to stimulate the expression of starch synthesis-related genes and meet the great demand for starch accumulation in the grain filling stage. Simultaneously, a negative regulatory loop forms among the 3 proteins to avoid the excessive expression of starch synthesis-related genes. Collectively, our findings demonstrate that both promotion and inhibition mechanisms between OsNAC25 and OsNAC20/26 are essential for maintaining stable expression of starch synthesis-related genes and normal starch accumulation.


Subject(s)
Gene Expression Regulation, Plant , Oryza , Plant Proteins , Starch , Transcription Factors , Oryza/genetics , Oryza/metabolism , Starch/metabolism , Starch/biosynthesis , Plant Proteins/genetics , Plant Proteins/metabolism , Transcription Factors/metabolism , Transcription Factors/genetics , Endosperm/metabolism , Endosperm/genetics
13.
Plant J ; 118(6): 2124-2140, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38551088

ABSTRACT

The basal region of maize (Zea mays) kernels, which includes the pedicel, placenta-chalazal, and basal endosperm transfer layers, serves as the maternal/filial interface for nutrient transfer from the mother plant to the developing seed. However, transcriptome dynamics of this maternal/filial interface remain largely unexplored. To address this gap, we conducted high-temporal-resolution RNA sequencing of the basal and upper kernel regions between 4 and 32 days after pollination and deeply analyzed transcriptome dynamics of the maternal/filial interface. Utilizing 790 specifically and highly expressed genes in the basal region, we performed the gene ontology (GO) term and weighted gene co-expression network analyses. In the early-stage basal region, we identified five MADS-box transcription factors (TFs) as hubs. Their homologs have been demonstrated as pivotal regulators at the maternal/filial interface of rice or Arabidopsis, suggesting their potential roles in maize kernel development. In the filling-stage basal region, numerous GO terms associated with transcriptional regulation and transporters are significantly enriched. Furthermore, we investigated the molecular function of three hub TFs. Through genome-wide DNA affinity purification sequencing combined with promoter transactivation assays, we suggested that these three TFs act as regulators of 10 basal-specific transporter genes involved in the transfer of sugars, amino acids, and ions. This study provides insights into transcriptomic dynamic and regulatory modules of the maternal/filial interface. In the future, genetic investigation of these hub regulators must advance our understanding of maternal/filial interface development and function.


Subject(s)
Gene Expression Regulation, Plant , Plant Proteins , Seeds , Transcriptome , Zea mays , Zea mays/genetics , Zea mays/growth & development , Zea mays/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Seeds/genetics , Seeds/growth & development , Seeds/metabolism , Transcription Factors/genetics , Transcription Factors/metabolism , Endosperm/genetics , Endosperm/growth & development , Endosperm/metabolism , Gene Regulatory Networks , Gene Expression Profiling
14.
Peptides ; 177: 171184, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38432550

ABSTRACT

It is ideal to ingest bioactive substances from daily foods to stay healthy. Rice is the staple food for almost half of the human population. We found that an orally administered enzymatic digest of rice endosperm protein exhibits antidepressant-like effects in the tail suspension test (TST) using mice. A comprehensive peptide analysis of the digest using liquid chromatography-tandem mass spectrometry was performed, and a tridecapeptide QQFLPEGQSQSQK, detected in the digest, was chemosynthesized. Oral administration of the tridecapeptide exhibited antidepressant-like effects at a low dose comparable to classical antidepressant in the TST. This also exhibited anti-depressant-like effect in the forced swim test. We named it rice endosperm-derived antidepressant-like peptide (REAP). Intriguingly, intraperitoneal administration had no effect. Orally administered REAP(8-13) but not REAP(1-7) exhibited antidepressant-like activity, suggesting that the C-terminal structure is important for the antidepressant-like effect. We confirmed the presence of REAP, corresponding to rice glutelin type B4(130-142) and B5(130-142), in the digest. The effects of REAP were blocked by both dopamine D1 and D2 antagonists. These results suggest that it exerts its antidepressant-like activity through activation of the dopamine system. Taken together, oral administration of a novel tridecapeptide exhibited antidepressant-like effects via the dopamine system. This is the first report of a rice-derived peptide that exhibits antidepressant-like effects.


Subject(s)
Antidepressive Agents , Endosperm , Oryza , Oryza/chemistry , Animals , Antidepressive Agents/pharmacology , Antidepressive Agents/chemistry , Antidepressive Agents/administration & dosage , Mice , Endosperm/chemistry , Administration, Oral , Male , Plant Proteins/chemistry , Plant Proteins/pharmacology , Depression/drug therapy , Peptides/chemistry , Peptides/pharmacology , Peptides/administration & dosage
16.
Transgenic Res ; 33(1-2): 47-57, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38451380

ABSTRACT

Cellobiohydrolase II (CBH II) is an exo-glucanase that is part of a fungal mixture of enzymes from a wood-rot fungus, Trichoderma reesei. It is therefore difficult to purify and to establish a specific activity assay. The gene for this enzyme, driven by the rice Os glutelin promoter, was transformed into High II tissue culture competent corn, and the enzyme accumulated in the endosperm of the seed. The transgenic line recovered from tissue culture was bred into male and female elite Stine inbred corn lines, stiff stalk 16083-025 (female) and Lancaster MSO411 (male), for future production in their hybrid. The enzyme increases its accumulation throughout its 6 generations of back crosses, 27-266-fold between T1 and T2, and 2-10-fold between T2 and T3 generations with lesser increases in T4-T6. The germplasm of the inbred lines replaces the tissue culture corn variety germplasm with each generation, with the ultimate goal of producing a high-yielding hybrid with the transgene. The CBH II enzyme was purified from T5 inbred male grain 10-fold to homogeneity with 47.5% recovery. The specific activity was determined to be 1.544 units per µg protein. The corn-derived CBH II works in biopolishing of cotton by removing surface fibers to improve dyeability and increasing glucose from corn flour for increasing ethanol yield from starch-based first-generation processes.


Subject(s)
Cellulase , Trichoderma , Cellulose 1,4-beta-Cellobiosidase/genetics , Cellulose 1,4-beta-Cellobiosidase/metabolism , Zea mays/genetics , Zea mays/metabolism , Endosperm/genetics , Endosperm/metabolism , Trichoderma/genetics , Trichoderma/metabolism , Plant Breeding , Cellulase/genetics
17.
Proc Natl Acad Sci U S A ; 121(14): e2321612121, 2024 Apr 02.
Article in English | MEDLINE | ID: mdl-38530890

ABSTRACT

To preserve germination ability, plant seeds must be protected from environmental stresses during the storage period. Here, we demonstrate that autophagy, an intracellular degradation system, maintains seed germination ability in Arabidopsis thaliana. The germination ability of long-term (>5 years) stored dry seeds of autophagy-defective (atg) mutant and wild-type (WT) plants was compared. Long-term stored (old) seeds of atg mutants showed lower germination ability than WT seeds, although short-term stored (new) seeds of atg mutants did not show such a phenotype. After removal of the seed coat and endosperm from old atg mutant seeds, the embryos developed into seedlings. Autophagic flux was maintained in endosperm cells during the storage period, and autophagy defect resulted in the accumulation of oxidized proteins and accelerated endosperm cell death. Consistent with these findings, the transcripts of genes, ENDO-ß-MANNANASE 7 and EXPANSIN 2, which are responsible for degradation/remodeling of the endosperm cell wall during germination, were reduced in old atg mutant seeds. We conclude that autophagy maintains endosperm quality during seed storage by suppressing aging-dependent oxidative damage and cell death, which allows the endosperm to perform optimal functions during germination, i.e., cell wall degradation/remodeling, even after long-term storage.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Arabidopsis/genetics , Endosperm/genetics , Germination/physiology , Seeds/genetics , Arabidopsis Proteins/metabolism , Autophagy , Gene Expression Regulation, Plant
18.
Int J Biol Macromol ; 261(Pt 2): 129948, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38311140

ABSTRACT

In present study, we characterized the formation, interfacial rheology, and storage stability of emulsions stabilized by microendosperm maize-derived zein (M-Zein)/whey protein isolate fiber (WPIF) nanoparticles. Microendosperm maize is a newly developed, oleic acid-rich oilseed resource. Recent research has shown that M-Zein possesses unique hydrophobic properties. Combining it with WPIF may enhance its performance as a stabilizer. Optimization of weight ratios for M-Zein/WPIF composites, guided by particle size analysis, fluorescence spectroscopy, three-phase contact angle (θ), and interfacial rheological analysis, revealed that a 4: 6 mass ratio at pH 7 yielded favorable wettability (θ = 91.2°). Interfacial rheology analysis showed that the combination of WPIF reduced M-Zein's interfacial tension to 7.2 mN/m and 36.7 mN/m at oil-water and air-water interfaces, respectively. The M-Zein/WPIF complex exhibited an elastic protein layer at the oil-water interface. Further investigations into nanoparticle concentration, oil phase volume, and pH revealed that emulsions containing 3 % nanoparticles (w/w), 50 % oil phase volume, and pH 7 showed the best storage stability. This research highlights the development of M-Zein/WPIF composited nanoparticles with superior storage stability and interfacial rheology. Additionally, it introduces a novel application for M-Zein, which elevates the value proposition of microendosperm maize.


Subject(s)
Nanoparticles , Zein , Emulsions/chemistry , Zein/chemistry , Zea mays , Whey Proteins , Endosperm , Particle Size , Rheology , Water/chemistry , Nanoparticles/chemistry
19.
Food Chem ; 444: 138597, 2024 Jun 30.
Article in English | MEDLINE | ID: mdl-38310783

ABSTRACT

Polar lipids have biosynthetic pathways which intersect and overlap with triacylglycerol biosynthesis; however, polar lipids have not been well characterized in the developing endosperms of oat with high oil accumulation. The polar lipids in endosperms of oat and wheat varieties having different oil contents were analyzed and compared at different developmental stages. Our study shows that the relative contents of polar lipid by mass were decreased more slowly in wheat than in oat. Phosphatidylcholine and phosphatidylethanolamine were the major phospholipids, which showed similar abundance and gradual decreases during endosperm development in oat and wheat, while lysophospholipids were noticeably higher in oat. Monogalactosyldiacylglycerol showed a gradual increase in wheat and a decrease in oat during endosperm development. The relative contents of some polar lipid species and their unsaturation index were significantly different in their endosperms. These characteristics of polar lipids might indicate an adaption of oat to accommodate oil accumulation.


Subject(s)
Avena , Endosperm , Endosperm/metabolism , Avena/metabolism , Triticum , Lipidomics , Phosphatidylcholines/metabolism
20.
Science ; 383(6683): 646-653, 2024 Feb 09.
Article in English | MEDLINE | ID: mdl-38330116

ABSTRACT

In multicellular organisms, sexual reproduction relies on the formation of highly differentiated cells, the gametes, which await fertilization in a quiescent state. Upon fertilization, the cell cycle resumes. Successful development requires that male and female gametes are in the same phase of the cell cycle. The molecular mechanisms that reinstate cell division in a fertilization-dependent manner are poorly understood in both animals and plants. Using Arabidopsis, we show that a sperm-derived signal induces the proliferation of a female gamete, the central cell, precisely upon fertilization. The central cell is arrested in S phase by the activity of the RETINOBLASTOMA RELATED1 (RBR1) protein. Upon fertilization, delivery of the core cell cycle component CYCD7;1 causes RBR1 degradation and thus S phase progression, ensuring the formation of functional endosperm and, consequently, viable seeds.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Endosperm , Gametogenesis, Plant , Paternal Inheritance , Arabidopsis/cytology , Arabidopsis/growth & development , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Cell Division , Endosperm/cytology , Endosperm/physiology
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