scholarly journals Shifting carbon flux from non-transient starch to lipid allows oil accumulation in transgenic tobacco leaves

2020 ◽  
Author(s):  
Kevin L. Chu ◽  
Lauren M. Jenkins ◽  
Sally R. Bailey ◽  
Shrikaar Kambhampati ◽  
Somnath Koley ◽  
...  
Keyword(s):  
Transgenic Tobacco ◽  
Carbon Flux ◽  
Soluble Sugars ◽  
Dry Weight ◽  
Starch Accumulation ◽  
Leaf Biomass ◽  
Wild Type ◽  
Oil Accumulation ◽  
Tobacco Leaves ◽  
Source Sink

AbstractPlant leaf biomass is composed predominantly of carbohydrate and protein with less than 5% dry weight allocated to lipid and less than 1% of total lipid in the form of triacylglycerols (TAGs). The combined overexpression of multiple genes involved in different aspects of TAG synthesis and stabilization can result in TAG accumulation to over 30% dry weight in tobacco leaves, presumably requiring many metabolic adjustments within plant cells. The metabolic consequences to the combined source and sink capacities of high oil accumulating transgenic tobacco leaves compared to wild-type were inspected across development and photoperiod by utilizing foliar biomass components and 13CO2 flux through central carbon intermediates. Lipid biosynthesis was investigated through assessment of acyl-acyl carrier protein (ACP) pools using a recently derived quantification method that was extended to accommodate isotopic labeling. Lipids accumulated stepwise over plant development in the high-oil leaves, with 13CO2-labeling studies confirming increased carbon flux to lipids. The large increase in lipid content was concurrent with a decrease in foliar starch, with limited contribution from non-sucrose soluble sugars, indicating a redirection of carbon from starch to lipids. Starch accumulated non-transiently with plant age in wild-type leaves, suggesting an inherent capacity for a developmentally-regulated carbon sink in tobacco leaves that may have enabled the programmed altered carbon partitioning to lipids in transgenics. These studies provide insight into the metabolic plasticity of dual source-sink leaves over development and may in part explain recent successful leaf lipid engineering efforts in tobacco.One sentence summaryEngineering high oil accumulation in tobacco leaves is enabled by inherent source-sink plasticity associated with non-transient foliar starch accumulation over development.

scholarly journals Modification of Petunia Seedling Carbohydrate Partitioning by Irradiance

1992 ◽  
Vol 117 (3) ◽  
pp. 477-480
Author(s):  
David F. Graper ◽  
Will Healy
Keyword(s):  
Petunia Hybrida ◽  
Soluble Sugars ◽  
Dry Weight ◽  
Invertase Activity ◽  
Acid Invertase ◽  
Starch Accumulation ◽  
Photon Flux ◽  
Photosynthetic Photon Flux ◽  
Total Carbohydrate ◽  
Relative Growth

Petunia × hybrida Villm. `Red Flash' plants received either 10 or 20 mol·day-1 photosynthetic photon flux (PPF) in growth chambers at: 175 μmol·m-2·s-1 for 16 hours, 350 μmol·m-2·s-1 for 8 or 16 hours, or 350 μmol·m-2 s-1 for 8 hours plus 8 hours of incandescent photoperiod extension (5 μmol·m-2·s-1 PPF). The irradiation components of peak, total, and duration were examined. Doubling total PPF increased total carbohydrate (CHO) production by 60%, seedling dry weight (DW) by 30%, rate of seedling growth by 25%, and acid invertase activity by 50% compared to the other treatments, once the seedlings had reached the two-leaf stage. Seedlings receiving 20 mol·day-1 PPF partitioned 14% more CHO into ethanol soluble sugars rather than starch, which may explain the increase in relative growth rate observed with supplemental irradiance treatments. Extending the photoperiod for 8 hours with 5 μmol·m-2·s-1 PPF reduced total CHO production by 50% compared to the same treatment without photoperiodic lighting. Treatment with 350 μmol·m-2·s-1 for 8 hours resulted in the highest O2 evolution (8.8 μmol O2/min per dm2). Increasing the photoperiod from 8 to 16 hours gave the lowest rate of O2 evolution (4.5 μmol O2/min per dm2). Previous reports of the importance of photosynthetic period in controlling partitioning between starch and sugars may have simply observed a decreasing rate of starch accumulation due to increased total PPF.

scholarly journals Effect of CdCl2 on the Growth and Antioxidases Activities in Transgenic Metallothionein and Wild Type Tobacco Leaves

HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 822A-822
Author(s):  
Jiping Sheng* ◽  
Lin Shen ◽  
Binggen Ru
Keyword(s):  
Heavy Metals ◽  
Transgenic Plant ◽  
Transgenic Tobacco ◽  
Wild Type ◽  
Plant Weight ◽  
Tobacco Leaves ◽  
Environmental Cleaning ◽  
Cd Tolerance

Metallothioneins (MTs) has selective capability to bind heavy metals such as Cd and Pb. Former study in our lab showed that MT gene from mouse was transferred into tobacco to absorb more heavy metals from soil. This study was conducted to plant transgenic tobacco and wild type tobacco on MS media with 20 μmol·L-1 CdCl2. Transgenic tobacco grew strong, whereas the growth of wild type tobacco was severely prohibited. At 21st day, an average single transgenic plant weight was 1.5 times higher than that of wild type, and its height was also 1.33 higher. The activities of antioxidases, such as POD, CAT, PPO in transgenic tobacco leaves showed significant lower than that of wild type, which was 32.3%, 43.3%, 187.5% lower respectively. The results indicated that the transgenic MT tobacco had higher Cd tolerance, and a promising future in the application of environmental cleaning.

scholarly journals The T-DNA Oncogene A4-orf8 from Agrobacterium rhizogenes A4 Induces Abnormal Growth in Tobacco

2005 ◽  
Vol 18 (3) ◽  
pp. 205-211 ◽  
Author(s):  
Marie Umber ◽  
Bernadette Clément ◽  
Léon Otten
Keyword(s):  
Agrobacterium Rhizogenes ◽  
Soluble Sugars ◽  
Starch Accumulation ◽  
Wild Type ◽  
Abnormal Growth ◽  
Regenerated Plants ◽  
Physical Linkage ◽  
Dark Green ◽  
Characteristic Growth ◽  
Fad Binding

The related orf8 and iaaM T-DNA genes from Agrobacterium are each composed of two distinct parts. The 5′ parts (called Norf8 or NiaaM) encode a 200-amino-acid (aa) sequence with homology to various T-DNA oncoproteins such as RolB, RolC, and 6b. The 3′ parts (Corf8 or CiaaM) encode a 550-aa sequence with homology to IaaM proteins from Pseudomonas and Pantoea spp. Whereas iaaM genes encode flavin adenine dinucleotide (FAD)-dependent tryptophan 2-monooxygenases that catalyze the synthesis of indole-3-acetamide (IAM), A4-orf8 from Agrobacterium rhizogenes A4 does not. Plants expressing a 2x35S-A4-Norf8 construct accumulate soluble sugars and starch. We now have regenerated plants that express the full-size 2x35S-A4-orf8 and the truncated 2x35S-A4-Corf8 gene. 2x35S-A4-Corf8 plants accumulate starch and show reduced growth like 2x35SA4-Norf8 plants but, in addition, display a novel set of characteristic growth modifications. These consist of leaf hypertrophy and hyperplasia (blisters); thick, dark-green leaves; thick stems; and swollen midveins. Mutations in the putative FAD-binding site of A4-Orf8 did not affect the blister syndrome. Plants expressing 2x35S-A4-Corf8 had a normal phenotype but contained less starch and soluble sugars than did wild-type plants. When 2x35S-A4-Corf8 plants were crossed to starch-accumulating 2x35S-A4-Norf8 plants with reduced growth, A4-Corf8 partially restored growth and reduced starch accumulation. A4-Corf8xA4-Norf8 crosses did not lead to the blister syndrome, suggesting that this requires physical linkage of the A4-NOrf8 and A4-COrf8 sequences.

ELONGATED HYPOCOTYL 5 mediates blue light-induced starch degradation in tomato

2020 ◽  
Author(s):  
Han Dong ◽  
Chaoyi Hu ◽  
Chaochao Liu ◽  
Jiachun Wang ◽  
Yanhong Zhou ◽  
...  
Keyword(s):  
Blue Light ◽  
Soluble Sugars ◽  
Light Signal ◽  
Starch Accumulation ◽  
Starch Degradation ◽  
Sugar Accumulation ◽  
Wild Type ◽  
Mobility Shift ◽  
Storage Carbohydrate ◽  
In The Wild

Abstract Starch is the major storage carbohydrate in plants, and its metabolism in chloroplasts depends mainly on light. However, the mechanism through which photoreceptors regulate starch metabolism in chloroplasts is unclear. In this study, we found that the cryptochrome 1a (CRY1a)-mediated blue light signal is critical for regulating starch accumulation by inducing starch degradation through the HY5 transcription factor in the chloroplasts in tomato. cry1a mutants and HY5-RNAi plants accumulated more starch and presented lower transcript levels of starch degradation-related genes in their leaves than did the wild-type (WT) plants. Blue light significantly induced the transcription of starch degradation-related genes in the wild-type and CRY1a- or HY5-overexpressing plants but had little effect in the cry1a and HY5-RNAi plants. Dual-luciferase assays, electrophoretic mobility shift assays (EMSA) and chromatin immunoprecipitation (ChIP)-qPCR revealed that HY5 could activate the starch degradation-related genes PWD, BAM1, BAM3, BAM8, MEX1 and DPE1 by directly binding to their promoters. Silencing of HY5 and these starch degradation-related genes in CRY1a-overexpressing plants led to increased accumulation of starch and decreased accumulation of soluble sugars. These findings presented here not only deepen our understanding of how light control starch degradation and sugar accumulation but also allow us to explore potential targets for improving crop quality.

scholarly journals `Hass' Avocado Carbohydrate Fluctuations. II. Fruit Growth and Ripening

1999 ◽  
Vol 124 (6) ◽  
pp. 676-681 ◽  
Author(s):  
Xuan Liu ◽  
Paul W. Robinson ◽  
Monica A. Madore ◽  
Guy W. Witney ◽  
Mary Lu Arpaia
Keyword(s):  
Fruit Ripening ◽  
Sugar Content ◽  
Soluble Sugar ◽  
Soluble Sugars ◽  
Fruit Size ◽  
Storage Period ◽  
Dry Weight ◽  
Persea Americana ◽  
Oil Accumulation ◽  
Low Temperature Storage

Changes in soluble sugar and starch reserves in avocado (Persea americana Mill. on `Duke 7' rootstock) fruit were followed during growth and development and during low temperature storage and ripening. During the period of rapid fruit size expansion, soluble sugars accounted for most of the increase in fruit tissue biomass (peel: 17% to 22%, flesh: 40% to 44%, seed: 32% to 41% of the dry weight). More than half of the fruit total soluble sugars (TSS) was comprised of the seven carbon (C7) heptose sugar, D-mannoheptulose, and its polyol form, perseitol, with the balance being accounted for by the more common hexose sugars, glucose and fructose. Sugar content in the flesh tissues declined sharply as oil accumulation commenced. TSS declines in the seed were accompanied by a large accumulation of starch (≈30% of the dry weight). During postharvest storage at 1 or 5 °C, TSS in peel and flesh tissues declined slowly over the storage period. Substantial decreases in TSS, and especially in the C7 sugars, was observed in peel and flesh tissues during fruit ripening. These results suggest that the C7 sugars play an important role, not only in metabolic processes associated with fruit development, but also in respiratory processes associated with postharvest physiology and fruit ripening.

Starch reduction in rice stems due to a lack of OsAGPL1 or OsAPL3 decreases grain yield under low irradiance during ripening and modifies plant architecture

2013 ◽  
Vol 40 (11) ◽  
pp. 1137 ◽  
Author(s):  
Masaki Okamura ◽  
Tatsuro Hirose ◽  
Yoichi Hashida ◽  
Tohru Yamagishi ◽  
Ryu Ohsugi ◽  
...  
Keyword(s):  
Grain Yield ◽  
Plant Architecture ◽  
Oryza Sativa L ◽  
Large Subunit ◽  
Dry Weight ◽  
Leaf Sheath ◽  
Starch Accumulation ◽  
Wild Type ◽  
Gene Encoding ◽  
Rice Mutant

Starch accumulated in rice (Oryza sativa L.) stems before heading as nonstructural carbohydrates (NSCs) is reported to be important for improving and stabilising grain yield. To evaluate the importance of stem starch, we investigated a retrotransposon (Tos17) insertion rice mutant lacking a gene encoding a large subunit of ADP-glucose pyrophosphorylase (AGP) called OsAGPL1 or OsAPL3. The AGP activity and starch contents of the mutant were drastically reduced in the stem (i.e. leaf sheath and culm) but not in the leaf blade or endosperm. This starch reduction in the leaf sheaths of the mutant was complemented by the introduction of wild-type OsAGPL1. These results strongly suggest that OsAGPL1 plays a principal role in stem starch accumulation. Field experimentations spanning 2 years revealed that the mutant plants were shorter than the wild-type plants. Moreover, the tiller number and angle were larger in the mutant plants than the wild-type plants, but the dry weight at heading stage was not different. The grain yield was slightly lower in control plots without shading treatment. However, this difference increased substantially with shading. Therefore, stem starch is indispensable for normal ripening under low irradiance conditions and probably contributes to the maintenance of appropriate plant architecture.

Co-expression of AGPS and AGPL genes encoded for AGPase from cassava to enhance starch accumulation in transgenic tobacco

2016 ◽  
Vol 14 (2) ◽  
pp. 287-293
Author(s):  
Nguyễn Văn Đoài ◽  
Nguyễn Minh Hồng ◽  
Lê Thu Ngọc ◽  
Nguyễn Thị Thơm ◽  
Nguyễn Đình Trọng ◽  
...  
Keyword(s):  
Transgenic Tobacco ◽  
Higher Plants ◽  
Starch Content ◽  
Genetically Modified Crops ◽  
Starch Accumulation ◽  
35S Promoter ◽  
Effective Strategies ◽  
Plant Expression Vector ◽  
Transgenic Lines ◽  
Cassava Varieties

The AGPase (ADP-Glucose pyrophosphorylase) is one of the ubiquitous enzymes catalyzing the first step in starch biosynthesis. It plays an important role in regulation and adjusts the speed of the entire cycle of glycogen biosynthesis in bacteria and starch in plants. In higher plants, it is a heterotetramer and tetrameric enzyme consisting two large subunits (AGPL) and two small subunits (AGPS) and encoded by two genes. In this paper, both AGPS and AGPL genes were sucessfully isolated from cassava varieties KM140 and deposited in Genbank with accession numbers KU243124 (AGPS) and KU243122 (AGPL), these two genes were fused with P2a and inserted into plant expression vector pBI121 under the control of 35S promoter. The efficient of this construct was tested in transgenic N. tabacum. The presence and expression of AGPS and AGPL in transgenic plants were confirmed by PCR and Western hybridization. The starch content was quantified by the Anthrone method. Transgenic plant analysis indicated that that two targeted genes were expressed simultaneously in several transgenic tobacco lines under the control of CaMV 35S promoter.  The starch contents in 4 analyzed tobacco transgenic lines displays the increase 13-116%  compared to WT plants. These results indicated that the co-expression of AGPS and AGPL is one of effective strategies for enhanced starch production in plant. These results can provide a foundation for developing other genetically modified crops to increase starch accumulation capacity.

Effects of an Autoregulatory Senescence-inhibitor Gene Construct on Nicotiana alata Link and Otto.

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 519d-519 ◽  
Author(s):  
Kenneth R. Schroeder ◽  
Dennis P. Stimart
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Plant Height ◽  
Dry Weight ◽  
Nicotiana Alata ◽  
Wild Type ◽  
Gene Encoding ◽  
Delayed Senescence ◽  
Gene Construct ◽  
Shoot Dry Weight ◽  
Inhibitor System

Nicotiana alata Link and Otto. was transformed via Agrobacterium tumefaciens encoding a senescence-specific promoter SAG12 cloned from Arabidopsis thaliana fused to a Agrobacterium tumefaciens gene encoding isopentenyl transferase (IPT) that catalyzes cytokinin synthesis. This was considered an autoregulatory senescence-inhibitor system. In 1996, we reported delayed senescence of intact flowers by 2 to 6 d and delayed leaf senescence of transgenic vs. wild-type N. alata. Further evaluations in 1997 revealed several other interesting effects of the SAG12-IPT gene construct. Measurement of chlorophyll content of mature leaves showed higher levels of both chlorophyll a and b in transgenic material under normal fertilization and truncated fertilization regimes. At 4 to 5 months of age transgenic plants expressed differences in plant height, branching, and dry weight. Plant height was reduced by 3 to 13 cm; branch counts increased 2 to 3 fold; and shoot dry weight increased up to 11 g over wild-type N. alata. These observations indicate the system is not tightly autoregulated and may prove useful to the floriculture industry for producing compact and more floriferous plants.

scholarly journals Rootstocks Overexpressing StNPR1 and StDREB1 Improve Osmotic Stress Tolerance of Wild-Type Scion in Transgrafted Tobacco Plants

2021 ◽  
Vol 22 (16) ◽  
pp. 8398
Author(s):  
Yasmine S. Hezema ◽  
Mukund R. Shukla ◽  
Alok Goel ◽  
Murali M. Ayyanath ◽  
Sherif M. Sherif ◽  
...  
Keyword(s):  
Osmotic Stress ◽  
Transgenic Tobacco ◽  
Physiological Status ◽  
Wild Type ◽  
Long Distance ◽  
Graft Union ◽  
Tobacco Plants ◽  
Osmotic Stress Tolerance ◽  
And Function ◽  
Gene Expression Levels

In grafted plants, the movement of long-distance signals from rootstocks can modulate the development and function of the scion. To understand the mechanisms by which tolerant rootstocks improve scion responses to osmotic stress (OS) conditions, mRNA transport of osmotic responsive genes (ORGs) was evaluated in a tomato/potato heterograft system. In this system, Solanum tuberosum was used as a rootstock and Solanum lycopersicum as a scion. We detected changes in the gene expression levels of 13 out of the 21 ORGs tested in the osmotically stressed plants; of these, only NPR1 transcripts were transported across the graft union under both normal and OS conditions. Importantly, OS increased the abundance of StNPR1 transcripts in the tomato scion. To examine mRNA mobility in transgrafted plants, StNPR1 and StDREB1 genes representing the mobile and non-mobile transcripts, respectively, were overexpressed in tobacco (Nicotiana tabacum). The evaluation of transgenic tobacco plants indicated that overexpression of these genes enhanced the growth and improved the physiological status of transgenic plants growing under OS conditions induced by NaCl, mannitol and polyethylene glycol (PEG). We also found that transgenic tobacco rootstocks increased the OS tolerance of the WT-scion. Indeed, WT scions on transgenic rootstocks had higher ORGs transcript levels than their counterparts on non-transgenic rootstocks. However, neither StNPR1 nor StDREB1 transcripts were transported from the transgenic rootstock to the wild-type (WT) tobacco scion, suggesting that other long-distance signals downstream these transgenes could have moved across the graft union leading to OS tolerance. Overall, our results signify the importance of StNPR1 and StDREB1 as two anticipated candidates for the development of stress-resilient crops through transgrafting technology.

scholarly journals Overexpression of vesicle-associated membrane protein PttVAP27-17 as a tool to improve biomass production and the overall saccharification yields in Populus trees

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Madhavi Latha Gandla ◽  
Niklas Mähler ◽  
Sacha Escamez ◽  
Tomas Skotare ◽  
Ogonna Obudulu ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Biomass Production ◽  
Enzymatic Saccharification ◽  
Dry Weight ◽  
Populus Tremula ◽  
Transgenic Trees ◽  
Wild Type ◽  
Glucose Yield ◽  
Transgenic Lines ◽  
Wild Type Control

Abstract Background Bioconversion of wood into bioproducts and biofuels is hindered by the recalcitrance of woody raw material to bioprocesses such as enzymatic saccharification. Targeted modification of the chemical composition of the feedstock can improve saccharification but this gain is often abrogated by concomitant reduction in tree growth. Results In this study, we report on transgenic hybrid aspen (Populus tremula × tremuloides) lines that showed potential to increase biomass production both in the greenhouse and after 5 years of growth in the field. The transgenic lines carried an overexpression construct for Populus tremula × tremuloides vesicle-associated membrane protein (VAMP)-associated protein PttVAP27-17 that was selected from a gene-mining program for novel regulators of wood formation. Analytical-scale enzymatic saccharification without any pretreatment revealed for all greenhouse-grown transgenic lines, compared to the wild type, a 20–44% increase in the glucose yield per dry weight after enzymatic saccharification, even though it was statistically significant only for one line. The glucose yield after enzymatic saccharification with a prior hydrothermal pretreatment step with sulfuric acid was not increased in the greenhouse-grown transgenic trees on a dry-weight basis, but increased by 26–50% when calculated on a whole biomass basis in comparison to the wild-type control. Tendencies to increased glucose yields by up to 24% were present on a whole tree biomass basis after acidic pretreatment and enzymatic saccharification also in the transgenic trees grown for 5 years on the field when compared to the wild-type control. Conclusions The results demonstrate the usefulness of gene-mining programs to identify novel genes with the potential to improve biofuel production in tree biotechnology programs. Furthermore, multi-omic analyses, including transcriptomic, proteomic and metabolomic analyses, performed here provide a toolbox for future studies on the function of VAP27 proteins in plants.

Sign in / Sign up

Export Citation Format

Share Document