Repression of barley cathepsins, HvPap-19 and HvPap-1, differentially alters grain composition and delays germination

2021 ◽  
Author(s):  
Andrea Gomez-Sanchez ◽  
M Estrella Santamaria ◽  
Pablo Gonzalez-Melendi ◽  
Aleksandra Muszynska ◽  
Christiane Matthess ◽  
...  
Keyword(s):  
Sucrose Concentration ◽  
Starch Content ◽  
Cysteine Proteases ◽  
Artificial Microrna ◽  
Grain Composition ◽  
Knock Down ◽  
Germination Process ◽  
Lower Protein ◽  
Transgenic Lines ◽  
Livestock Feeding

Abstract During barley germination, cysteine-proteases are essential in the mobilization of storage compounds providing peptides and amino acids to sustain embryo growth until photosynthesis is completely established. Knock-down barley plants, generated by artificial microRNA, for the cathepsins B- and F-like, HvPap-19 and HvPap-1 genes, respectively, showed less cysteine protease activities and consequently lower protein degradation. The functional redundancy between proteases triggered an enzymatic compensation associated with an increase in serine-protease activities in both knock-down grains, which was not sufficient to maintain germination rates and behaviour. Concomitantly, these transgenic lines showed alterations in the accumulation of protein and carbohydrates in the grain. While the total amount of protein increased in both transgenic lines, the starch content decreased in HvPap-1 knock-down lines and the sucrose concentration was reduced in silenced HvPap-19 grains. Consequently, phenotypes of HvPap-1 and HvPap-19 artificial microRNA lines showed a delay in the grain germination process. These data expand the potential of exploring the properties of barley proteases to be selectively modified and used for brewing or livestock feeding industry.

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.

Osmotic Adjustment: Effect of Water Stress on Carbohydrates in Leaves, Stems and Roots of Apple

1995 ◽  
Vol 22 (5) ◽  
pp. 747 ◽  
Author(s):  
Z Wang ◽  
B Quebedeaux ◽  
GW Stutte
Keyword(s):  
Water Stress ◽  
Osmotic Adjustment ◽  
Sucrose Concentration ◽  
Starch Content ◽  
Soluble Carbohydrates ◽  
Soluble Carbohydrate ◽  
Malus Domestica Borkh ◽  
Mature Leaves ◽  
Young Leaves ◽  
Response To Water

Potted apple (Malus domestica Borkh. cv. Jonathan) trees were subjected to water stress in a greenhouse. Midday leaf water potential (ΨW), osmotic potential (ΨS), soluble carbohydrates, and starch content of expanding and mature leaves, stems, and roots were measured to determine whether active osmotic adjustment occurred and if water stress affected carbohydrate metabolism. Mature leaves had the highest total soluble carbohydrate level (357 mM) and lowest Ψ (-1.85 MPa), followed by young leaves (278 mM, -1.58 MPa), stems (115 mM, -1.02 MPa), and roots (114 mM, -0.87 MPa). Sorbitol was the major component in all organs ranging from 53% of total soluble carbohydrate in young leaves to 73% in mature leaves. When ΨW decreased from -1.0 to -3.2 MPa, active osmotic adjustments of 0.3-0.4 MPa were observed in mature leaves, stems, and roots while a significantly higher adjustment of 1.0 MPa was detected in young leaves 5 days after the initiation of water stress. Sorbitol levels in leaves and stems gradually increased as ΨW decreased from -1.0 to -2.5 MPa, and then remained relatively stable or decreased slightly as ΨW decreased from -2.5 to -3.2 MPa. However, the percentage of soluble carbohydrate as sorbitol in roots decreased in response to water stress. Sucrose concentration decreased in mature leaves and stems, but increased in young leaves and roots as ΨW decreased. Starch concentrations in stems and roots also decreased as water stress developed. The sorbitol to sucrose ratios increased in mature leaves, but decreased in roots in response to water stress.

An analysis of the role of the ShSUT1 sucrose transporter in sugarcane using RNAi suppression

2017 ◽  
Vol 44 (8) ◽  
pp. 795 ◽  
Author(s):  
Donna Glassop ◽  
Jiri Stiller ◽  
Graham D. Bonnett ◽  
Christopher P. L. Grof ◽  
Anne L. Rae
Keyword(s):  
Sucrose Concentration ◽  
Amplicon Sequencing ◽  
Phloem Loading ◽  
Gene Transcript ◽  
Direct Role ◽  
Rnai Technology ◽  
Transgenic Lines ◽  
Sucrose Mobilisation ◽  
And Storage

The role of ShSUT1 in sucrose mobilisation and storage in sugarcane was investigated by employing RNAi technology to reduce the expression of this gene. Transcript profiling in non-transformed plants showed an alignment between expression and sucrose concentration, with strongest expression in source leaves and increasing expression through the daylight period of a diurnal cycle. Five transgenic plant lines were produced with reduced ShSUT1 expression ranging from 52 to 92% lower than control plants. Differential suppression of ShSUT1 sequence variants in the highly polyploid sugarcane genome were also investigated. Amplicon sequencing of the ShSUT1 variants within the transgenic lines and controls showed no preferential suppression with only minor differences in the proportional expression of the variants. A range of altered sugar, fibre and moisture contents were measured in mature leaf and internode samples, but no phenotype was consistently exhibited by all five transgenic lines. Phenotypes observed indicate that ShSUT1 does not play a direct role in phloem loading. ShSUT1 is likely involved with retrieving sucrose from intercellular spaces for transport and storage.

scholarly journals Aplicação de farinha de banana verde na substituição parcial da farinha de trigo em pão de forma

2018 ◽  
Vol 39 (6) ◽  
pp. 2399 ◽  
Author(s):  
Eliseth De Souza Viana ◽  
Andrea Dos Santos Souza ◽  
Ronielli Cardoso Reis ◽  
Vânia Jesus dos Santos de Oliveira
Keyword(s):  
Resistant Starch ◽  
Purchase Intention ◽  
Starch Content ◽  
Sensory Acceptance ◽  
Light Yellow ◽  
Significant Difference ◽  
Lower Protein ◽  
Banana Flour ◽  
Light Yellow Color ◽  
Green Banana

The objective of this work was to develop a sliced bread product with green banana flour in order to obtain high resistant starch content and good acceptance. Four bread formulations were studied, at concentrations of 0% (control), 15%, 20% and 25% green banana flour (Terra Maranhão variety). The green banana flour (GBF) was characterized as to color (L * coordinate and C * and h * parameters), and flour and bread were characterized with respect to chemical composition and total and resistant starch contents. Sensory acceptance of breads was investigated for the attributes color, flavor, aroma, texture, overall acceptance and purchase intention. The GBF presented light yellow color, with values of L * = 81.32, C * = 18.37 and h * = 89.51. The values (d. b.) for the other characteristics were: yield, 25.17%; moisture, 6.69%; ash, 1.60%; protein, 2.66%; lipids, 0.61%; total starch, 67.49%; and resistant starch, 56.29%. There was no significant difference (p > 0.05) between the breads containing GBF, and the sliced breads showed lower protein value (8.64%) than the control (10.60%) and higher resistant starch content (2.91%) compared to the control (0.65%). Addition of GBF to bread did not interfere with the moisture, ash and lipid contents of the products. Breads with concentrations of 15 and 20% GBF received acceptance higher than 90% for all sensory attributes. The use of GBF in sliced bread at the 15% and 20% levels resulted in a product with high sensory acceptance (greater than 90%) and with resistant starch content 4.2 times higher than conventional sliced bread.

The expression pattern of the Pho1a genes encoding plastidic starch phosphorylase correlates with the degradation of starch during fruit ripening in green-fruited and red-fruited tomato species

2019 ◽  
Vol 46 (12) ◽  
pp. 1146 ◽  
Author(s):  
Maria A. Slugina ◽  
Anna V. Shchennikova ◽  
Elena Z. Kochieva
Keyword(s):  
Developmental Stages ◽  
Sucrose Concentration ◽  
Starch Content ◽  
Nucleotide Polymorphisms ◽  
Starch Phosphorylase ◽  
Ripe Fruit ◽  
Single Nucleotide ◽  
Tomato Species ◽  
Genes Encoding ◽  
Polymorphism Level

Genes encoding plastidic starch phosphorylase Pho1a were identified in 10 tomato species (Solanum section Lycopersicon). Pho1a genes showed higher variability in green-fruited than in red-fruited tomato species, but had an extremely low polymorphism level compared with other carbohydrate metabolism genes and an unusually low ratio of intron to exon single nucleotide polymorphisms (SNPs). In red-fruited species, Pho1a was expressed in all analysed tissues, including fruit at different developmental stages, with the highest level in mature green fruit, which is strong sink organ importing sucrose and accumulating starch. In green-fruited species Solanum peruvianum and Solanum arcanum, the Pho1a expression level was similar in mature green and ripe fruit, whereas in Solanum chmielewskii, it was higher in ripe fruit, and in Solanum habrochaites, the dynamics of fruit-specific Pho1a expression was similar to that in red-fruited tomatoes. During fruit development, in red-fruited Solanum lycopersicum, sucrose level was low, the monosaccharide content increased; in green-fruited S. peruvianum, the sucrose concentration increased and those of monosaccharides decreased. In both species, the starch content and Pho1a expression were downregulated. The evolutionary topology based on Pho1a sequences was consistent with the current division of tomatoes into red-fruited and green-fruited species, except for S. habrochaites.

scholarly journals Starch metabolism in tubers of transgenic potato (Solanum tuberosum) with increased ADPglucose pyrophosphorylase*

1996 ◽  
Vol 320 (2) ◽  
pp. 493-498 ◽  
Author(s):  
Lee J. SWEETLOVE ◽  
Michael M. BURRELL ◽  
Tom ap REES
Keyword(s):  
Solanum Tuberosum ◽  
Starch Content ◽  
Starch Synthesis ◽  
Fold Increase ◽  
Transgenic Potato ◽  
Detectable Effect ◽  
Adpglucose Pyrophosphorylase ◽  
Response Coefficient ◽  
Transgenic Lines

The aim of this work was to use tubers from transgenic lines of potato (Solanum tuberosum) containing increased amounts of ADPglucose pyrophosphorylase to study the role of this enzyme in the control of starch synthesis. A 4–5-fold increase in activity of the enzyme, achieved by transformation with the Escherichia coli ADPglucose pyrophosphorylase gene glgC-16, had no detectable effect on the starch content of developing or mature tubers. No significant effects were found on the contents of ADPglucose, UDPglucose, glucose 1-phosphate, glucose 6-phosphate, PPi, ATP and ADP. Flux from [U-14C]sucrose, supplied to tubers still attached to the plant, to starch increased roughly in proportion to the increase in ADPglucose pyrophosphorylase activity. These measurements of flux gave a response coefficient close to 1 for the activity of the pyrophosphorylase in respect of starch synthesis. Pulse–chase experiments with [U-14C]sucrose showed that the increased flux into starch in the transformed tubers was accompanied by an increased rate of starch turnover. Further experiments suggested that the increased turnover was associated with an increase in the capacity of the tubers to degrade starch.

scholarly journals Synthesis and transfer of RNAi construct for potential knock-down of gene expression of root-knot nematode (Meloidogyne graminicola) to rice (Oryza sativa L.)

2020 ◽  
Vol 19 (04) ◽  
pp. 36-44
Author(s):  
Phong V. Nguyen
Keyword(s):  
Expression Vector ◽  
Parasitic Nematode ◽  
Oryza Sativa L ◽  
Artificial Microrna ◽  
Root Knot Nematode ◽  
Rnai Construct ◽  
Meloidogyne Graminicola ◽  
Knock Down ◽  
Plant Parasitic

Effectors play key roles in the parasitism of the plant-parasitic nematode. Silencing the effector-coding genes was applied to study the function and role of nematode effectors. In this study, the Mgra16281 gene (ID: MK322955.1) encoding an effector with the unknown function was cloned from the rice root-knot nematode Meloidogyne graminicola isolated in Long An province. To knock-down the expression of this gene, an artificial microRNA was synthesized based on the Osa-MIR528 precursor and inserted into an expression vector. This microRNA can be expressed in rice to investigate the function of MGRA16281 of root-knot nematode via host-induced gene silencing approach (HIGS).

scholarly journals Papain-like cysteine proteases are required for the regulation of photosynthetic gene expression and acclimation to high light stress

2021 ◽  
Vol 72 (9) ◽  
pp. 3441-3454
Author(s):  
Sarah Alomrani ◽  
Karl J Kunert ◽  
Christine H Foyer
Keyword(s):  
Gene Expression ◽  
Large Subunit ◽  
Light Stress ◽  
D1 Protein ◽  
Carbon Assimilation ◽  
Cysteine Proteases ◽  
High Light ◽  
High Light Stress ◽  
Photosynthetic Gene ◽  
Transgenic Lines

AbstractChloroplasts are considered to be devoid of cysteine proteases. Using transgenic Arabidopsis lines expressing the rice cystatin, oryzacystatin I (OC-I), in the chloroplasts (PC lines) or cytosol (CYS lines), we explored the hypothesis that cysteine proteases regulate photosynthesis. The CYS and PC lines flowered later than the wild type (WT) and accumulated more biomass after flowering. In contrast to the PC rosettes, which accumulated more leaf chlorophyll and carotenoid pigments than the WT, the CYS lines had lower amounts of leaf pigments. High-light-dependent decreases in photosynthetic carbon assimilation and the abundance of the Rubisco large subunit protein, the D1 protein, and the phosphorylated form of D1 proteins were attenuated in the CYS lines and reversed in the PC lines relative to the WT. However, the transgenic lines had higher amounts of LHC, rbcs, pasbA, and pasbD transcripts than the WT, and also showed modified chloroplast to nucleus signalling. We conclude that cysteine proteases accelerate the reconfiguration of the chloroplast proteome after flowering and in response to high-light stress. Inhibition of cysteine proteases, such as AtCEP1, slows chloroplast protein degradation and stimulates photosynthetic gene expression and chloroplast to nucleus signalling, enhancing stress tolerance traits.

scholarly journals Effect of Germination on Nutritional Composition of Common Buckwheat (Fagopyrum esculentum Moench)

2021 ◽  
pp. 1-7
Author(s):  
K. Shreeja ◽  
S. Suchiritha Devi ◽  
W. Jessie Suneetha ◽  
B. Neeraja Prabhakar
Keyword(s):  
Nutritional Value ◽  
Reducing Sugars ◽  
Starch Content ◽  
Nutritional Composition ◽  
Growth Trend ◽  
Germination Process ◽  
The World ◽  
Fagopyrum Esculentum Moench ◽  
Sodium And Potassium ◽  
Buckwheat Flour

There is a growth trend in buckwheat consumption, as it has high nutritional value. Buckwheat is the main form consumed, although the consumption of buckwheat sprouts is increasingly popular in the world. Buckwheat grains were germinated at 30°C for 48 hours with 12 hours of soaking in BOD incubator followed by drying at 60°C. Germination process has increased moisture, protein and crude fiber (11.03% - 12.77%, 10.22% - 12.14% and 0.92% - 1.44%). Minerals such as sodium and potassium were also increased. Total and reducing sugars were increased and starch content has decreased with the germination process. Germinated buckwheat flour can be used in the formulations of nutraceutical foods.

Sign in / Sign up

Export Citation Format

Share Document