scholarly journals CRISPR/Cas9-Mediated Knockout of Galactinol Synthase-Encoding Genes Reduces Raffinose Family Oligosaccharide Levels in Soybean Seeds

2020 ◽  
Vol 11 ◽  
Author(s):  
Huy Le ◽  
Nhung Hong Nguyen ◽  
Dong Thị Ta ◽  
Thao Nhu Thi Le ◽  
Thao Phuong Bui ◽  
...  
Keyword(s):  
Double Mutant ◽  
Crop Improvement ◽  
Soybean Seed ◽  
Dry Weight ◽  
Soluble Carbohydrates ◽  
Soluble Carbohydrate ◽  
Soybean Seeds ◽  
Galactinol Synthase ◽  
Target Sites ◽  
Mutant Lines

Raffinose family oligosaccharides (RFOs) are major soluble carbohydrates in soybean seeds that cannot be digested by human and other monogastric animals. Hence, a major goal is to reduce RFO levels to improve the nutritional quality of soybean. In this study, we utilized a dual gRNAs CRISPR/Cas9 system to induce knockouts in two soybean galactinol synthase (GOLS) genes, GmGOLS1A and its homeolog GmGOLS1B. Genotyping of T0 plants showed that the construct design was efficient in inducing various deletions in the target sites or sequences spanning the two target sites of both GmGOLS1A and GmGOLS1B genes. A subset of induced alleles was successfully transferred to progeny and, at the T2 generation, we identified null segregants of single and double mutant genotypes without off-target induced mutations. The seed carbohydrate analysis of double mutant lines showed a reduction in the total RFO content of soybean seed from 64.7 mg/g dry weight to 41.95 mg/g dry weight, a 35.2% decrease. On average, the stachyose content, the most predominant RFO in soybean seeds, decreased by 35.4% in double mutant soybean, while the raffinose content increased by 41.7%. A slight decrease in verbascose content was also observed in mutant lines. Aside from changes in soluble carbohydrate content, some mutant lines also exhibited increased protein and fat contents. Otherwise, no difference in seed weight, seed germination, plant development and morphology was observed in the mutants. Our findings indicate that GmGOLS1A and GmGOLS1B contribute to the soybean oligosaccharide profile through RFO biosynthesis pathways, and are promising targets for future investigation, as well as crop improvement efforts. Our results also demonstrate the potential in using elite soybean cultivars for transformation and targeted genome editing.

scholarly journals Arsenic Application Changed Growth, Photo-Synthetic Pigments and Antioxidant Enzymes Activity in Sorghum (Sorghum bicolor L.) Under Salinity Stress

2019 ◽  
Vol 50 (3) ◽  
pp. 155-163 ◽  
Author(s):  
B. Talebi ◽  
M. Heidari ◽  
H. Ghorbani
Keyword(s):  
Plant Biomass ◽  
Dry Weight ◽  
Carotenoid Content ◽  
Soluble Carbohydrates ◽  
Soluble Carbohydrate ◽  
Guaiacol Peroxidase ◽  
Anthocyanin Content ◽  
Synthetic Pigments ◽  
Root Tissues ◽  
Gpx Activity

Abstract The elevation of arsenic (As) content in soils is of considerable concern with respect to its uptake by plant and subsequent entry into wildlife and human food chains. The treatment of sorghum seedlings with As as NaH2As4O. 7H2O at various concentrations (A1 = 0, A2 = 20, A3 = 40 and A4 = 60 mg As kg−1 soil) and salinity at four different levels (S1 = 0, S2 = 3, S3 = 6 and S3 = 9 dS m−1) reduced fresh and dry weights of sorghum plants. The co-application of As and salinity increased the guaiacol peroxidase (GPX) activity in shoot and root tissues. The highest GPX activity in shoot and root tissues was obtained at S2A4 and S3A3 treatments, respectively. The activity of catalase (CAT) in shoot was not changed, but unlike the GPX activity, salinity and As decreased the CAT activity in root tissues. Concerning the photosynthesis pigments, salinity had no effect on the chlorophyll ‘a’, chlorophyll ‘b’ and carotenoid content in leaves, but the As treatment significantly decreased the content of both chlorophyll types. Salinity increased the anthocyanin content in leaves. There were negative correlation between soluble carbohydrates (r2 = −0.78**) and stomata conductance (r2 = −0.45**) and dry weight of the plant biomass in this study. By increasing the salinity and As concentration in root medium, soluble carbohydrate in leaves increased but salinity decreased the leaf stomata conductance.

Genotypic variation in water-soluble carbohydrate accumulation in wheat

2006 ◽  
Vol 33 (9) ◽  
pp. 799 ◽  
Author(s):  
Sari A. Ruuska ◽  
Greg J. Rebetzke ◽  
Anthony F. van Herwaarden ◽  
Richard A. Richards ◽  
Neil A. Fettell ◽  
...  
Keyword(s):  
Genotypic Variation ◽  
Dry Weight ◽  
Grain Filling ◽  
Water Soluble ◽  
Soluble Carbohydrates ◽  
Soluble Carbohydrate ◽  
Broad Sense Heritability ◽  
Carbohydrate Accumulation ◽  
Sequential Mechanism ◽  
Water Soluble Carbohydrate

The water-soluble carbohydrate (WSC) that accumulates in the stems of wheat during growth can be an important contributor to grain filling, particularly under conditions when assimilation is limited, such as during end-of-season drought. WSC concentration was measured at anthesis across a diverse set of wheat genotypes over multiple environments. Environmental differences in WSC concentration were large (means for the set ranging between 108 and 203 mg g–1 dry weight), and there were significant and repeatable differences in WSC accumulation among genotypes (means ranging from 112 to 213 mg g–1 dry weight averaged across environments), associated with large broad-sense heritability (H = 0.90 ± 0.12). These results suggest that breeding for high WSC should be possible in wheat. The composition of the WSC, examined in selected genotypes, indicated that the variation in total WSC was attributed mainly to variation in the fructan component, with the other major soluble carbohydrates, sucrose and hexose, varying less. The degree of polymerisation (DP) of fructo-oligosaccharides was up to ~13 in samples where higher levels of WSC were accumulated, owing either to genotype or environment, but the higher DP components (DP > 6) were decreased in samples of lower total WSC. The results are consistent with fructan biosynthesis occurring via a sequential mechanism that is dependent on the availability of sucrose, and differences in WSC contents of genotypes are unlikely to be due to major mechanistic differences.

scholarly journals (295) Relationship Between Macromolecule and Metal Ion Accumulation in Two Salt-stressed Cultivars of Sweetpotato under In Vitro Conditions

HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1079A-1079
Author(s):  
Devi Prasad V. Potluri
Keyword(s):  
Salt Stress ◽  
Metal Ion ◽  
Ipomoea Batatas ◽  
Dry Weight ◽  
Soluble Carbohydrates ◽  
Soluble Carbohydrate ◽  
Shoot Height ◽  
Sodium Accumulation ◽  
Carbohydrate Levels

Two cultivars of sweetpotato [Ipomoea batatas (L.) Lam.], Commensal and Salyboro, were subjected to salt stress using axillary bud cultures. The salt levels ranged from 0–150 mM. After 10 weeks of growth, plantlet shoot height, dry weight, number of nodes, levels of proline, soluble carbohydrate, and protein; and metal ions sodium, potassium, magnesium, and calcium, were measured. In both cultivars, proline accumulation was higher in the shoot. There was a positive correlation between the increase in soluble carbohydrates and proteins in `Commnesal', but not in `Salyboro'. More sodium accumulated in the shoots of `Salyboro' compared to `Commensal'. The accumulation of sodium reduced the calcium and potassium, but not magnesium levels. Increase in sodium levels correlated with the increase in soluble carbohydrate levels is `Salyboro', but not in `Commensal'. A similar trend was evident with praline and sodium accumulation. Based on these and previous results, the cultivar `Salyboro' appears to be more susceptible to salt stress.

scholarly journals Soybean seed galactinol synthase activity as determined by a novel colorimetric assay

2000 ◽  
Vol 12 (3) ◽  
pp. 203-212 ◽  
Author(s):  
MARLUCI RIBEIRO ◽  
CARLOS R. FELIX ◽  
SILENE DE PAULINO LOZZI
Keyword(s):  
Colorimetric Assay ◽  
Colorimetric Method ◽  
Soybean Seed ◽  
Maximum Activity ◽  
Soybean Seeds ◽  
Colorimetric Determination ◽  
Raffinose Oligosaccharides ◽  
Galactinol Synthase ◽  
Isotopic Method ◽  
Key Enzyme

Galactinol synthase (GS) is a key enzyme for the biosynthesis of raffinose oligosaccharides (RO) which are the flatulence factors present in soybean seeds and several other legumes. Understanding of soybean seed GS properties is, therefore, of biotechnological interest. The GS enzyme catalyses formation of galactinol and UDP from UDP-gal and myo-inositol. This enzyme is currently assayed by an isotopic method. We have then idealized a more convenient method for GS assay based on the indirect colorimetric determination of the UDP formed which is then hydrolyzed by exogenous apyrase and the resulting Pi quantified by a modification of the colorimetric method of Fiske & SubbaRow. The color developed is stable, and the method is suitable for detection of very low GS activity. The GS activity profiles of developing soybean seeds determined by the isotopic and the colorimetric methods are closely related. The GS enzyme was partially purified (46-fold) by treatment of seed extract with MnCl2, sequential chromatographies on DEAE-Sepharose, Phenyl-Sepharose CL-4B and Q-Sepharose columns. The crude and the partially purified enzyme showed maximum activity at pH 7.0 and 50 ºC. Dithiothreitol and MnCl2 enhanced considerably the activity of the partially purified enzyme. While UDP-glc could be hydrolyzed by the enzyme at a reative activity corresponding to 49% of that calculated for UDP-gal, UDP-man and sucrose were completely ineffective as alternative substrates.

scholarly journals Pengaruh Priming pada Vigor Benih Kedelai (Glycine max. [L] Merrill) yang Dikecambahkan pada Tanah Masam

2021 ◽  
Vol 20 (2) ◽  
pp. 120
Author(s):  
Nabila Lutfiah ◽  
Agustiansyah . ◽  
Paul B. Timotiwu
Keyword(s):  
Acid Soil ◽  
Seedling Emergence ◽  
Soybean Seed ◽  
Acid Soils ◽  
Dry Weight ◽  
Soybean Seeds ◽  
Best Response ◽  
Normal Seedling ◽  
Level Of Significance ◽  
The University

The need of soybeans continuously increases due to the increasing of human population. The germination of soybeans seeds in the acid soil such as ultisol soil often experienced barrier. One of the ways to improve seed germination in the acid soils is to give priming treatment on the seeds. The research was conducted in the green house of the Integrated Field Laboratory, Faculty of Agriculture, the University of Lampung. It was conducted from December 2020 to March 2021. The research used factorial treatment that was applied in the Completely Random Design with three replicates. The Tukey’s Honestly Significance Difference with 5% level of significance was used to compare the average of treaments. Data analyzing was done by using R studio statistics program. The first factor  was kinds of priming which consisted of without priming as a controle, water priming within 12 hours, KNO3 1% within 3 hours, KNO3 2% within 6 hours, GA3 50 ppm within 14 hours, GA3 100 ppm within 14 hours, and PEG 7,5% within 24 hours. The second factor was three soybean seed varieties, i.e. Anjasmoro, Burangrang, and Grobogan. Results showed that Giberelin priming of 50 ppm for 14 hours was the best priming in seedling emergence in the ultisol soils. The Burangrang soybean seeds showed the best T50% and the number of radicles appeared in the first day. The Anjasmoro soybean seeds showed the best response in the dry weight of normal seedling and hypocotyls lenght of normal seedling in the ultisol soils.Key Words: Priming, soybean, ultisol, variety

scholarly journals Genetic Analyses of Soluble Carbohydrate Concentrations in Onion Bulbs

2009 ◽  
Vol 134 (6) ◽  
pp. 618-623 ◽  
Author(s):  
Steven Raines ◽  
Cynthia Henson ◽  
Michael J. Havey
Keyword(s):  
Dry Weight ◽  
Interval Mapping ◽  
Field Trials ◽  
Chromosome 8 ◽  
Soluble Solids ◽  
Soluble Carbohydrates ◽  
Soluble Carbohydrate ◽  
Onion Bulbs ◽  
Bulb Dormancy ◽  
Chromosome Regions

Fructans are the main soluble carbohydrate in onion (Allium cepa) bulbs and their concentrations show significant correlations with dry weights and pungency. In previous research, we identified regions on chromosomes 5 and 8 associated with higher amounts of soluble carbohydrates in onion bulbs. In this research, we estimated the genetic effects and interactions between these two chromosome regions using larger inbred families grown in field trials over 3 years. Bulbs were evaluated 30 and 90 days after harvest (DAH) for dry weights and soluble carbohydrates. Fructan concentrations decreased significantly between 30 and 90 DAH, consistent with loss of bulb dormancy over this period. Dry weights were negatively correlated with fructose and positively correlated with sucrose and fructans. Analyses of variance and interval mapping revealed that the region on chromosome 5 affected bulb dry weight, whereas the region on chromosome 8 significantly affected both dry weight and fructan concentrations. Regions on chromosomes 5 and 8 showed dominance for increased dry weights and/or soluble solids. Interactions between regions on chromosomes 5 and 8 were significant only for sucrose and the fructan neokestose, indicating that these regions independently contribute to higher amounts of soluble carbohydrates. These results demonstrate that onions with low concentrations of soluble carbohydrates were developed by selecting for relatively few recessively inherited chromosome regions.

QTL affecting soluble carbohydrate concentrations in stored onion bulbs and their association with flavor and health-enhancing attributes

Genome ◽  
2004 ◽  
Vol 47 (3) ◽  
pp. 463-468 ◽  
Author(s):  
Michael J Havey ◽  
Claudio R Galmarini ◽  
Ali Fuat Gökçe ◽  
Cynthia Henson
Keyword(s):  
Dry Weight ◽  
Interval Mapping ◽  
Field Trials ◽  
Soluble Carbohydrates ◽  
Soluble Carbohydrate ◽  
Onion Bulbs ◽  
Invertase Gene ◽  
Trait Locus ◽  
Genomic Regions ◽  
Group D

Onion bulbs accumulate fructans, a type of soluble carbohydrate associated with lower rates of colorectal cancers. Higher fructan concentrations in bulbs are correlated with higher pungency, longer dormancy, and greater onion-induced antiplatelet activity (OIAA). We analyzed replicated field trials of a segregating family for types and concentrations of soluble carbohydrates in onion bulbs 90 days after harvest. Means were adjusted using dry weight as the covariant to reveal highly significant (P < 0.001) differences among parents and families for glucose, fructose, sucrose, and the fructans 1-kestose, neokestose, and (6G,1)-nystose. Fructan concentrations showed significant (P < 0.05) phenotypic correlations with each other and with sucrose, pungency, and OIAA. These observations are consistent with the hypothesis that onion bulbs accumulating fructans take up or retain less water, concentrating both soluble carbohydrates and thiosulfinates responsible for pungency and OIAA. Interval mapping of family means from the covariant analyses revealed regions on linkage groups A and D significantly (LOD > 2.68) affecting soluble carbohydrate concentrations. The enzyme catalyzing the first step of fructan polymerization, 1-sucrose-sucrose fructosyltransferase (1-SST), mapped independently of these genomic regions. One region on linkage group D near an acid-invertase gene was significantly (LOD = 3.45) associated with sucrose concentrations. This study reveals that the accumulation of sucrose in stored onion bulbs may allow for the combination of sweeter flavor with significant OIAA.Key words: quantitative trait locus, fructans, thiosulfinates, reducing sugars.

Cyclitol galactosides in embryos of buckwheat stem–leaf–seed explants fed D-chiro-inositol, myo-inositol or D-pinitol

2005 ◽  
Vol 15 (4) ◽  
pp. 329-338 ◽  
Author(s):  
Janet M. Ma ◽  
Marcin Horbowicz ◽  
Ralph L. Obendorf
Keyword(s):  
Effective Means ◽  
Dry Weight ◽  
Insulin Dependent Diabetes Mellitus ◽  
Control Treatment ◽  
Soluble Carbohydrates ◽  
Dependent Diabetes Mellitus ◽  
Soluble Carbohydrate ◽  
Mature Embryos ◽  
Legume Seeds ◽  
Stem Leaf

Crop seeds accumulate soluble carbohydrates as part of their maturation process. In legume seeds, the major soluble carbohydrates are sucrose and its galactosides raffinose, stachyose and verbascose. In buckwheat (Fagopyrum esculentum Moench) seeds, the major soluble carbohydrates are sucrose and galactosides of d-chiro-inositol, named fagopyritols. This study was conducted to determine changes in soluble carbohydrate accumulation in embryos of buckwheat seeds after feeding solutions containing the free cyclitols D-chiro-inositol, myo-inositol and d-pinitol to stem–leaf–seed explants. Feeding D-chiro-inositol to explants resulted in a fourfold to fivefold increase in the accumulation of free d-chiro-inositol, fagopyritol A1 and fagopyritol B1 in embryos of mature seeds, but resulted in 30% less embryo dry weight compared to the control treatment without cyclitols. Feeding myo-inositol to buckwheat explants increased d-chiro-inositol in leaves and increased accumulation of fagopyritol A1 and fagopyritol B1 fivefold in embryos, fagopyritol A2 and fagopyritol B2 fourfold; fagopyritol A3 and fagopyritol B3 were also detected, with no reduction in accumulated embryo dry weight. Feeding d-pinitol to buckwheat explants resulted in accumulation of free d-pinitol in mature embryos, but not galactopinitols. D-Pinitol, galactopinitol A and galactopinitol B were not detected in embryos from explants fed solutions without d-pinitol. Feeding d-pinitol also resulted in reduced D-chiro-inositol accumulation by buckwheat seeds. The results indicate that myo-inositol may be the precursor to d-chiro-inositol synthesis, and fagopyritols accumulated in response to d-chiro-inositol availability in the embryo. We suggest that increasing myo-inositol in buckwheat maternal tissues may be an effective means to enhance the accumulation of D-chiro-inositol and fagopyritols in seeds, compounds that may be beneficial for the treatment of non-insulin-dependent diabetes mellitus.

scholarly journals Reserve mobilization in soybean seeds under water restriction after storage

2020 ◽  
Vol 42 ◽  
Author(s):  
Thaísa Fernanda Oliveira ◽  
Heloisa Oliveira dos Santos ◽  
Rafaela Aparecida de Carvalho ◽  
Hellismar Wakson da Silva ◽  
Raquel Maria de Oliveira Pires ◽  
...  
Keyword(s):  
Dry Matter ◽  
Reduction Rate ◽  
Soybean Seed ◽  
Dry Weight ◽  
Soybean Seeds ◽  
Water Restriction ◽  
Reserve Mobilization ◽  
Germination Process ◽  
One Year ◽  
Total Seed

Abstract: Germination does not guarantee plant establishment if there is no efficient and enough reserve mobilization from seeds to the initial seedling growth, mainly in adverse situations. Therefore, this study aimed to evaluate the mobilization of reserves in soybean seeds under water restriction. Four soybean seed lots previously stored for one year were used in the study. To simulate water stress, we used polyethylene glycol (PEG) solutions at concentrations of 0.0, -0.1, and -0.2 MPa. The number of normal, infected and abnormal seedlings were evaluated, along with the number of dead and hard seeds. The radicle, hypocotyl, and cotyledons were separated to determine the dry weight based on the total seed reserve available for mobilization. From this value, the seed reserve reduction, relative dry matter yield, seed reserve reduction rate, and reserve conversion efficiency into dry seedling weight were determined. The increase in water restriction elevates the reserve consumption during the germination process. Vigorous seeds are more efficient in mobilizing and converting grains reserve in dry seedling mass. In less vigorous seeds, mainly those on water restriction, the increase in reserve consumption does not result in conversion into dry seedling mass.

Water status influences common events of soluble carbohydrate accumulation during soybean seed development and germination

2002 ◽  
Vol 80 (3) ◽  
pp. 262-270 ◽  
Author(s):  
Albert T Modi ◽  
Miller B McDonald ◽  
John G Streeter
Keyword(s):  
Seed Germination ◽  
Water Content ◽  
Seed Development ◽  
Water Status ◽  
Soybean Seed ◽  
Soluble Carbohydrates ◽  
Soluble Carbohydrate ◽  
Radicle Protrusion ◽  
Glycine Max L ◽  
Three Phases

Seed development and germination are two distinct physiological stages that are normally separated by a metabolically quiescent period in orthodox seeds. Comparison of seed water status during these two processes and how it influences the biochemical activities remains unclear. The objective of this study was to compare soybean (Glycine max (L.) Merrill cv. Ohio FG1) seed development and germination, including the first 6 h after radicle protrusion, with respect to soluble carbohydrate occurrence at different stages characterized by water content and osmolality. Cyclitols and sugars were monitored at nine stages of development and during the first 30 h of germination. Three phases of water loss and osmolality increase found during seed development were correlated with three phases of water absorption and osmolality decrease during seed germination. This study provided evidence that soybean seed cotyledons and axes have similar patterns of water content and osmolality during seed development and germination and that three major events of soluble carbohydrate occurrence are shared by soybean seed parts during development and germination.Key words: seed development, seed germination, soluble carbohydrates, soybean, water status.

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