scholarly journals NTRC and Thioredoxin f Overexpression Differentially Induces Starch Accumulation in Tobacco Leaves

Plants ◽  
2019 ◽  
Vol 8 (12) ◽  
pp. 543 ◽  
Author(s):  
María Ancín ◽  
Luis Larraya ◽  
Alicia Fernández-San Millán ◽  
Jon Veramendi ◽  
Tessa Burch-Smith ◽  
...  
Keyword(s):  
Starch Content ◽  
Starch Synthesis ◽  
Genetically Engineered ◽  
Starch Accumulation ◽  
Starch Metabolism ◽  
Reductive Activation ◽  
Amylolytic Enzyme ◽  
Tobacco Plants ◽  
Tobacco Leaves ◽  
The Impact

Thioredoxin (Trx) f and NADPH-dependent Trx reductase C (NTRC) have both been proposed as major redox regulators of starch metabolism in chloroplasts. However, little is known regarding the specific role of each protein in this complex mechanism. To shed light on this point, tobacco plants that were genetically engineered to overexpress the NTRC protein from the chloroplast genome were obtained and compared to previously generated Trx f-overexpressing transplastomic plants. Likewise, we investigated the impact of NTRC and Trx f deficiency on starch metabolism by generating Nicotiana benthamiana plants that were silenced for each gene. Our results demonstrated that NTRC overexpression induced enhanced starch accumulation in tobacco leaves, as occurred with Trx f. However, only Trx f silencing leads to a significant decrease in the leaf starch content. Quantitative analysis of enzyme activities related to starch synthesis and degradation were determined in all of the genotypes. Zymographic analyses were additionally performed to compare the amylolytic enzyme profiles of both transplastomic tobacco plants. Our findings indicated that NTRC overexpression promotes the accumulation of transitory leaf starch as a consequence of a diminished starch turnover during the dark period, which seems to be related to a significant reductive activation of ADP-glucose pyrophosphorylase and/or a deactivation of a putative debranching enzyme. On the other hand, increased starch content in Trx f-overexpressing plants was connected to an increase in the capacity of soluble starch synthases during the light period. Taken together, these results suggest that NTRC and the ferredoxin/Trx system play distinct roles in starch turnover.

scholarly journals Transcriptome Analysis and Identification of Genes Associated with Starch Metabolism in Castanea henryi Seed (Fagaceae)

2020 ◽  
Vol 21 (4) ◽  
pp. 1431 ◽  
Author(s):  
Bin Liu ◽  
Ruqiang Lin ◽  
Yuting Jiang ◽  
Shuzhen Jiang ◽  
Yuanfang Xiong ◽  
...  
Keyword(s):  
Seed Germination ◽  
Candidate Genes ◽  
Starch Content ◽  
Sugar Content ◽  
Soluble Sugar ◽  
Starch Accumulation ◽  
Transcriptional Level ◽  
Potential Candidate ◽  
Starch Metabolism ◽  
Carbohydrate Storage

Starch is the most important form of carbohydrate storage and is the major energy reserve in some seeds, especially Castanea henryi. Seed germination is the beginning of the plant’s life cycle, and starch metabolism is important for seed germination. As a complex metabolic pathway, the regulation of starch metabolism in C. henryi is still poorly understood. To explore the mechanism of starch metabolism during the germination of C. henryi, we conducted a comparative gene expression analysis at the transcriptional level using RNA-seq across four different germination stages, and analyzed the changes in the starch and soluble sugar contents. The results showed that the starch content increased in 0–10 days and decreased in 10–35 days, while the soluble sugar content continuously decreased in 0–30 days and increased in 30–35 days. We identified 49 candidate genes that may be associated with starch and sucrose metabolism. Three ADP-glucose pyrophosphorylase (AGPase) genes, two nucleotide pyrophosphatase/phosphodiesterases (NPPS) genes and three starch synthases (SS) genes may be related to starch accumulation. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to validate the expression levels of these genes. Our study combined transcriptome data with physiological and biochemical data, revealing potential candidate genes that affect starch metabolism during seed germination, and provides important data about starch metabolism and seed germination in seed plants.

scholarly journals Starch synthesis and gelatinization properties of potato tubers

2022 ◽  
Vol 52 (4) ◽  
Author(s):  
Wang Su ◽  
Guangji Ye ◽  
Yun Zhou ◽  
Jian Wang
Keyword(s):  
Gene Expression ◽  
Enzyme Activity ◽  
Starch Granule ◽  
Starch Content ◽  
Starch Synthesis ◽  
Starch Accumulation ◽  
Enzyme Gene ◽  
Middle Stage ◽  
Pasting Temperature ◽  
Starch Synthesis Enzyme

ABSTRACT: Biosynthesis is the only source of potato starch which is an important raw material for food processing, modified starch and biomass energy. However, it is not clear about the evolution of starch synthesis with tuber development in potato. The present study evaluated the differences of starch synthesis and gelatinization properties of potato tubers with different starch content. Relative to cultivars of medium and low starch content, cultivars of high starch content showed significantly higher SBEII gene expression, AGPase and SSS enzyme activity, and total starch content after middle stage of starch accumulation, and had smaller average starch granule size during whole process of tuber development, and had higher pasting temperature before late stages of tuber growth, and had lower pasting temperature after middle stage of starch accumulation. Path analysis showed that, after middle stage of starch accumulation, effects on starch gelatinization of cultivars with high, medium and low starch content represented starch synthesis enzyme activity > starch accumulation > starch granule distribution > starch synthesis enzyme gene expression, starch synthesis enzyme gene expression > starch synthesis enzyme activity > starch accumulation > starch granule distribution, starch synthesis enzyme gene expression > starch granule distribution > starch synthesis enzyme activity > starch accumulation, respectively. In the study, phases existed in the starch biosynthesis of potato tuber, and the starch quality and its formation process were different among varieties with different starch content. The findings might contribute to starch application and potato industries.

scholarly journals Expression of Maize MADS Transcription Factor ZmES22 Negatively Modulates Starch Accumulation in Rice Endosperm

2019 ◽  
Vol 20 (3) ◽  
pp. 483 ◽  
Author(s):  
Kangyong Zha ◽  
Haoxun Xie ◽  
Min Ge ◽  
Zimeng Wang ◽  
Yu Wang ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Mrna Expression ◽  
Starch Content ◽  
Soluble Sugar ◽  
Starch Synthesis ◽  
Synthesis Process ◽  
Starch Accumulation ◽  
Rice Grain ◽  
Altered Expression ◽  
Rice Endosperm

As major component in cereals grains, starch has been one of the most important carbohydrate consumed by a majority of world’s population. However, the molecular mechanism for regulation of biosynthesis of starch remains elusive. In the present study, ZmES22, encoding a MADS-type transcription factor, was modestly characterized from maize inbred line B73. ZmES22 exhibited high expression level in endosperm at 10 days after pollination (DAP) and peaked in endosperm at 20 DAP, indicating that ZmES22 was preferentially expressed in maize endosperm during active starch synthesis. Transient expression of ZmES22 in tobacco leaf revealed that ZmES22 protein located in nucleus. No transactivation activity could be detected for ZmES22 protein via yeast one-hybrid assay. Transformation of overexpressing plasmid 35S::ZmES22 into rice remarkedly reduced 1000-grain weight as well as the total starch content, while the soluble sugar was significantly higher in transgenic rice lines. Moreover, overexpressing ZmES22 reduced fractions of long branched starch. Scanning electron microscopy images of transverse sections of rice grains revealed that altered expression of ZmES22 also changed the morphology of starch granule from densely packed, polyhedral starch granules into loosely packed, spherical granules with larger spaces. Furthermore, RNA-seq results indicated that overexpressing ZmES22 could significantly influence mRNA expression levels of numerous key regulatory genes in starch synthesis pathway. Y1H assay illustrated that ZmES22 protein could bind to the promoter region of OsGIF1 and downregulate its mRNA expression during rice grain filling stages. These findings suggest that ZmES22 was a novel regulator during starch synthesis process in rice endosperm.

Enhanced Carbohydrate Metabolism and Salinity Tolerance of Tobacco Plants Overexpressing a Vacuolar H+-ATPase Subunit B2 (DoVHAb2) Gene from Yam (Dioscorea opposita Thunb.)

2021 ◽  
Vol 15 (4) ◽  
pp. 504-513
Author(s):  
Ying Shao ◽  
Yanfang Zhang ◽  
Shuchun Guo ◽  
Lingmin Zhao ◽  
Xiaohua Sun ◽  
...  
Keyword(s):  
Salinity Tolerance ◽  
Starch Content ◽  
Nacl Stress ◽  
Vital Role ◽  
Full Length ◽  
Starch Metabolism ◽  
Wild Type ◽  
Tobacco Plants ◽  
Atpase Subunit ◽  
Functional Studies

ATP synthase plays a vital role in plant growth and stress tolerance, functional studies of DoVHAb2 in yam tuber starch metabolism and salinity tolerance have so far not been reported. Full-length cloning and analysis of DoVHAb2 were conducted to ascertain its function. The gDNAs were cloned and analyzed. Quantitative real-time polymerase chain reaction (qRT-PCR) was probed in yam tuber developmental stage and different organs of DoVHAb2. Transient expression vector was constructed and injected into tobacco leaves to observe the subcellular localization of genes. Overexpressed fusion vector of gene was constructed and transformed into tobacco by Agrobacterium-mediated method to identify the function of DoVHAb2 gene. In the results, the full-length of DoVHAb2 was 1926 bp, encoding 488 amino acids, and it was divided into 14 exons and 15 introns, its highest expression was found in tubers than in stems and leaves in yam, the yam DoVHAb2 protein was localized to cytoplasm. The starch content, ADP glucose pyrophosphorylase, starch synthase and ATPase activity were significantly higher in transgenic tobacco plants than in the wild-type. The transgenic plants also had higher leaf differentiation rate, soluble protein content, superoxide dismutase and peroxidase activities, and lower malondialdehyde content than the wild type under NaCl stress. In conclusion, the results indicated that overexpression of DoVHAb2 enhanced starch metabolism and conferred salinity tolerance.

Suppression of starch synthesis in rice stems splays tiller angle due to gravitropic insensitivity but does not affect yield

2015 ◽  
Vol 42 (1) ◽  
pp. 31 ◽  
Author(s):  
Masaki Okamura ◽  
Tatsuro Hirose ◽  
Yoichi Hashida ◽  
Ryu Ohsugi ◽  
Naohiro Aoki
Keyword(s):  
Double Mutant ◽  
Oryza Sativa L ◽  
Starch Content ◽  
Large Subunit ◽  
Starch Synthesis ◽  
Starch Accumulation ◽  
Gravitropic Response ◽  
Rice Mutant ◽  
Matter Production ◽  
Tiller Angle

In rice (Oryza sativa L.), tiller angle – defined as the angle between the main culm and its side tillers – is one of the important factors involved in light use efficiency. To clarify the relationship between tiller angle, gravitropism and stem-starch accumulation, we investigated the shoot gravitropic response of a low stem-starch rice mutant which lacks a large subunit of ADP-glucose pyrophosphorylase (AGP), called OsAGPL1 and exhibits relatively spread tiller angle. The insensitive gravitropic response exhibited by the mutant led us to the conclusion that insensitivity of gravitropism caused by stem-starch reduction splayed the tiller angle. Furthermore, since another AGP gene called OsAGPL3 was expressed at considerable levels in graviresponding sites, we generated a double mutant lacking both OsAGPL1 and OsAGPL3. The double mutant exhibited still lower stem-starch content, less sensitive gravitropic response and greater tiller angle spread than the single mutants. This indicated that the expansion of the tiller angle caused by the reduction in starch level was intense according to the extent of the reduction. We found there were no significant differences between the double mutant and wild-type plants in terms of dry matter production. These results provided new insight into the importance of stem-starch accumulation and ideal plant architecture.

scholarly journals NnABI4-Mediated ABA Regulation of Starch Biosynthesis in Lotus (Nelumbo nucifera Gaertn)

2021 ◽  
Vol 22 (24) ◽  
pp. 13506
Author(s):  
Peng Wu ◽  
Ailian Liu ◽  
Yongyan Zhang ◽  
Kai Feng ◽  
Shuping Zhao ◽  
...  
Keyword(s):  
Theoretical Basis ◽  
Plant Hormone ◽  
Nuclear Protein ◽  
Starch Content ◽  
Starch Synthesis ◽  
Starch Biosynthesis ◽  
Luciferase Assay ◽  
Starch Accumulation ◽  
Exogenous Aba ◽  
Total Starch

Starch is an important component in lotus. ABA is an important plant hormone, which plays a very crucial role in regulating plant starch synthesis. Using ‘MRH’ as experimental materials, the leaves were sprayed with exogenous ABA before the rhizome expansion. The results showed that stomatal conductance and transpiration rate decreased while net photosynthetic rate increased. The total starch content of the underground rhizome of lotus increased significantly. Meanwhile, qPCR results showed that the relative expression levels of NnSS1, NnSBE1 and NnABI4 were all upregulated after ABA treatment. Then, yeast one-hybrid and dual luciferase assay suggested that NnABI4 protein can promote the expression of NnSS1 by directly binding to its promoter. In addition, subcellular localization results showed that NnABI4 encodes a nuclear protein, and NnSS1 protein was located in the chloroplast. Finally, these results indicate that ABA induced the upregulated expression of NnABI4, and NnABI4 promoted the expression of NnSS1 and thus enhanced starch accumulation in lotus rhizomes. This will provide a theoretical basis for studying the molecular mechanism of ABA regulating starch synthesis in plant.

scholarly journals Over-Expressing TaSPA-B Reduces Prolamin and Starch Accumulation in Wheat (Triticum aestivum L.) Grains

2020 ◽  
Vol 21 (9) ◽  
pp. 3257 ◽  
Author(s):  
Dandan Guo ◽  
Qiling Hou ◽  
Runqi Zhang ◽  
Hongyao Lou ◽  
Yinghui Li ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
Leucine Zipper ◽  
Protein Biosynthesis ◽  
Starch Content ◽  
Starch Synthesis ◽  
Triticum Aestivum L ◽  
Luciferase Reporter ◽  
Starch Accumulation ◽  
Basic Leucine Zipper ◽  
Wheat Lines

Starch and prolamin composition and content are important indexes for determining the processing and nutritional quality of wheat (Triticum aestivum L.) grains. Several transcription factors (TFs) regulate gene expression during starch and protein biosynthesis in wheat. Storage protein activator (TaSPA), a member of the basic leucine zipper (bZIP) family, has been reported to activate glutenin genes and is correlated to starch synthesis related genes. In this study, we generated TaSPA-B overexpressing (OE) transgenic wheat lines. Compared with wild-type (WT) plants, the starch content was slightly reduced and starch granules exhibited a more polarized distribution in the TaSPA-B OE lines. Moreover, glutenin and ω- gliadin contents were significantly reduced, with lower expression levels of related genes (e.g., By15, Dx2, and ω-1,2 gliadin gene). RNA-seq analysis identified 2023 differentially expressed genes (DEGs). The low expression of some DEGs (e.g., SUSase, ADPase, Pho1, Waxy, SBE, SSI, and SS II a) might explain the reduction of starch contents. Some TFs involved in glutenin and starch synthesis might be regulated by TaSPA-B, for example, TaPBF was reduced in TaSPA-B OE-3 lines. In addition, dual-luciferase reporter assay indicated that both TaSPA-B and TaPBF could transactivate the promoter of ω-1,2 gliadin gene. These results suggest that TaSPA-B regulates a complex gene network and plays an important role in starch and protein biosynthesis in wheat.

scholarly journals Microbial Volatile-Induced Accumulation of Exceptionally High Levels of Starch in Arabidopsis Leaves Is a Process Involving NTRC and Starch Synthase Classes III and IV

2011 ◽  
Vol 24 (10) ◽  
pp. 1165-1178 ◽  
Author(s):  
Jun Li ◽  
Ignacio Ezquer ◽  
Abdellatif Bahaji ◽  
Manuel Montero ◽  
Miroslav Ovecka ◽  
...  
Keyword(s):  
Time Course ◽  
Starch Content ◽  
Thioredoxin Reductase ◽  
Redox Status ◽  
Starch Synthase ◽  
Reduced Form ◽  
Starch Accumulation ◽  
Knockout Mutants ◽  
Microbial Volatiles ◽  
The Impact

Microbial volatiles promote the accumulation of exceptionally high levels of starch in leaves. Time-course analyses of starch accumulation in Arabidopsis leaves exposed to fungal volatiles (FV) emitted by Alternaria alternata revealed that a microbial volatile-induced starch accumulation process (MIVOISAP) is due to stimulation of starch biosynthesis during illumination. The increase of starch content in illuminated leaves of FV-treated hy1/cry1, hy1/cry2, and hy1/cry1/cry2 Arabidopsis mutants was many-fold lower than that of wild-type (WT) leaves, indicating that MIVOISAP is subjected to photoreceptor-mediated control. This phenomenon was inhibited by cordycepin and accompanied by drastic changes in the Arabidopsis transcriptome. MIVOISAP was also accompanied by enhancement of the total 3-phosphoglycerate/Pi ratio, and a two- to threefold increase of the levels of the reduced form of ADP-glucose pyrophosphorylase. Using different Arabidopsis knockout mutants, we investigated the impact in MIVOISAP of downregulation of genes directly or indirectly related to starch metabolism. These analyses revealed that the magnitude of the FV-induced starch accumulation was low in mutants impaired in starch synthase (SS) classes III and IV and plastidial NADP-thioredoxin reductase C (NTRC). Thus, the overall data showed that Arabidopsis MIVOISAP involves a photocontrolled, transcriptionally and post-translationally regulated network wherein photoreceptor-, SSIII-, SSIV-, and NTRC-mediated changes in redox status of plastidial enzymes play important roles.

Starch synthesis and localization in post-germination Pinusedulis seedlings

1994 ◽  
Vol 24 (7) ◽  
pp. 1457-1463 ◽  
Author(s):  
J. Brad Murphy ◽  
Mark F. Hammer
Keyword(s):  
Seed Germination ◽  
Starch Content ◽  
Starch Synthesis ◽  
Minor Component ◽  
Dry Weight ◽  
Starch Synthase ◽  
Starch Accumulation ◽  
Low Levels ◽  
Starch Conversion ◽  
A Minor

Following pine seed germination, lipids in the megagametophyte are converted to sucrose, which is transported to the emerging seedling to support its growth. In several conifer species, an increase in the seedling starch content following germination has been reported. To further characterize this phenomenon, starch accumulation and localization, starch synthase (EC 2.4.1.21) activity (both soluble and granule-bound), and partitioning of exogenous 14C-sucrose were determined following germination of pinyon (Pinusedulis Engelm.) seeds. Starch was a minor component in dry embryos, accounting for only 3% of the dry weight. Starch levels increased 22-fold and 15-fold in the cotyledons and hypocotyl, respectively, by 8 days after germination. Starch accumulated to 65% of the dry weight in the cotyledons and 46% in the hypocotyl. The root and epicotyl accumulated relatively low levels of starch, only about 7%. Starch was localized primarily in the cortex and pith of the hypocotyl, the cortex of the cotyledons, and the root cap. Only granule-bound starch synthase showed a significant increase in activity during germination, and its changes more closely followed the pattern of starch accumulation. Exogenous 14C-sucrose was partitioned primarily into starch. After a 24-h labeling period, starch in both the cotyledons and hypocotyl accounted for 38% of total label (61% of the incorporated label) in these organs. In the roots, starch accounted for only 2.5 and 14%, respectively, of the total and incorporated label. The spatial and temporal pattern of starch accumulation closely paralleled previously reported patterns for the activity of sucrose synthase, which is apparently associated with the sucrose–starch conversion. Starch accumulation in the seedling accounts for approximately 50% of the sucrose transported from the megagametophyte following pinyon seed germination. Thus, starch appears to serve as an important transitory carbon pool for the growing seedling and may serve additional functions during seedling development.

scholarly journals Increasing Starch Accumulation via Genetic Modification of the ADP-glucose Pyrophosphorylase

2009 ◽  
Author(s):  
Arthur Schaffer ◽  
Jack Preiss ◽  
Marina Petreikov ◽  
Ilan Levin
Keyword(s):  
Starch Content ◽  
Large Subunit ◽  
Tomato Fruit ◽  
Sugar Content ◽  
Soluble Sugar ◽  
Starch Synthesis ◽  
Regulatory Subunit ◽  
Starch Accumulation ◽  
Research Project ◽  
Immature Fruit

The overall objective of the research project was to utilize biochemical insights together with both classical and molecular genetic strategies to improve tomato starch accumulation. The proposal was based on the observation that the transient starch accumulation in the immature fruit serves as a reservoir for carbohydrate and soluble sugar content in the mature fruit, thereby impacting on fruit quality. The general objectives were to optimize AGPase function and activity in developing fruit in order to increase its transient starch levels. The specific research objectives were to: a) perform directed molecular evolution of the limiting enzyme of starch synthesis, AGPase, focussing on the interaction of its regulatory and catalytic subunits; b) determine the mode of action of the recently identified allelic variant for the regulatory subunit in tomato fruit that leads to increased AGPase activity and hence starch content. During the course of the research project major advances were made in understanding the interaction of the small and large subunits of AGPase, in particular the regulatory roles of the different large subunits, in determining starch synthesis. The research was performed using various experimental systems, including bacteria and Arabidopsis, potato and tomato, allowing for broad and meaningful conclusions to be drawn. A novel discovery was that one of the large subunits of tomato AGPase is functional as a monomer. A dozen publications describing the research were published in leading biochemical and horticultural journals. The research results clearly indicated that increasing AGPase activity temporally in the developing fruit increase the starch reservoir and, subsequently, the fruit sugar content. This was shown by a comparison of the carbohydrate balance in near-isogenic tomato lines differing in a gene encoding for the fruit-specific large subunit (LS1). The research also revealed that the increase in AGPase activity is due to a temporal extension of LS1 gene expression in the developing fruit which in turn stabilizes the limiting heterotetrameric enzyme, leading to sustained starch synthesis. This genetic variation can successfully be utilized in the breeding of high quality tomatoes.

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