scholarly journals Simultaneous antisense inhibition of two starch-synthase isoforms in potato tubers leads to accumulation of grossly modified amylopectin

1999 ◽  
Vol 338 (2) ◽  
pp. 515-521 ◽  
Author(s):  
James R. LLOYD ◽  
Volker LANDSCHÜTZE ◽  
Jens KOSSMANN
Keyword(s):  
Transgenic Plants ◽  
Amylose Content ◽  
Starch Synthase ◽  
Antisense Inhibition ◽  
Starch Granules ◽  
Potato Tubers ◽  
Chain Lengths ◽  
Antisense Construct ◽  
Total Starch ◽  
Long Chains

A chimaeric antisense construct was used to reduce the activities of the two major starch-synthase isoforms in potato tubers simultaneously. A range of reductions in total starch-synthase activities were found in the resulting transgenic plants, up to a maximum of 90% inhibition. The reduction in starch-synthase activity had a profound effect on the starch granules, which became extremely distorted in appearance compared with the control lines. Analysis of the starch indicated that the amounts produced in the tubers, and the amylose content of the starch, were not affected by the reduction in activity. In order to understand why the starch granules were distorted, amylopectin was isolated and the constituent chain lengths analysed. This indicated that the amylopectin was very different to that of the control. It contained more chains of fewer than 15 glucose units in length, and fewer of between 15 and 80 glucose units. In addition, the amylopectin contained more very long chains. Amylopectin from plants repressed in just one of the activities of the two starch-synthase isoforms, which we have reported upon previously, were also analysed. Using a technique different to that used previously we show that both isoforms also affect the amylopectin, but in a way that is different to when both isoforms are repressed together.

scholarly journals Identification and molecular characterization of mutant line deficiency in three waxy proteins of common wheat (Triticum aestivum L.)

2019 ◽  
Author(s):  
Qian Liu ◽  
Yaping Hu ◽  
Mengyun Hu ◽  
Lijing Sun ◽  
Xiyong Chen ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
Amylose Content ◽  
Premature Stop Codon ◽  
Triticum Aestivum L ◽  
Starch Granules ◽  
Null Line ◽  
Wild Type ◽  
Expression Levels ◽  
Waxy Proteins ◽  
Total Starch

Abstract Background: Starch is the main component of wheat (Triticum aestivum L.) grain and a key factor in determining wheat processing quality. The Wx gene encodes the granule bound starch synthase I (GBSS I) and is the sole gene responsible for amylose synthesis. Results: A waxy mutant (Wx-null) was isolated by screening M3 seeds derived from 1.0% EMS mutagenized materials with I2-KI staining of endosperm starch. SDS-PAGE electrophoresis confirmed that the Wx-null line lacked all three waxy proteins. DNA sequencing revealed three SNPs and a 3-bp InDel in the first exon, and a 16-bp InDel at the junction region of the first Wx-A1 intron from the Wx-null line. Six SNPs were identified in Wx-B1 gene of Wx-null line compared to the wild-type Gao 8901, including four missense mutations. One nonsense mutation was found at position 857 in the fourth exon, which resulted in a premature stop codon. Expression levels of Wx genes were dramatically reduced in the Wx-null line. Nonsense-mediated mRNA decay (NMD) may be triggered to degrade the non-functional Wx mRNA. There were no detectable differences in granule size and morphology between Wx-null and wild-type, but the Wx-null line contained a larger proportion of B-type starch granules. The amylose content of the Wx-null line (0.22%) was remarkably lower compared to the wild-type Gao 8901 (20.82%). Total starch is also lower in the Wx-null line. Conclusions: All three waxy proteins were non-functional in the Wx-null line. NMD may be the cause for reduced expression levels of Wx genes in the Wx-null line. The Wx-null line exhibited more B-type starch granules, dramatically lower amylose content, and decreased total starch. The Wx-null line may provide a potential waxy material with high agronomic performance in wheat breeding programs.

scholarly journals Characterization, Functional Properties, and Resistant Starch of Freshwater Macrophytes

2021 ◽  
Vol 2021 ◽  
pp. 1-12
Author(s):  
Farahin N. N. Syed ◽  
Muta H. Zakaria ◽  
Japar S. Bujang ◽  
Annie Christianus
Keyword(s):  
Thermal Properties ◽  
Functional Properties ◽  
Resistant Starch ◽  
Amylose Content ◽  
Starch Content ◽  
Rheological Behaviour ◽  
Nelumbo Nucifera ◽  
Starch Granules ◽  
Starch Properties ◽  
Total Starch

Several aquatic macrophytes such as Colocasia esculenta, Eleocharis dulcis, Nelumbo nucifera, Sagittaria sagittifolia, Trapa bispinosa, and Typha angustifolia possessed carbohydrate mainly in their storage and reproductive parts. Starch morphology, total starch, and amylose content of these six freshwater plant species were determined. Their functional properties, i.e., starch crystallinity, thermal properties, and rheological behaviour were assessed. Large starch granules were in N. nucifera rhizome (>15 μm), medium-sized was N. nucifera seed (8-18 μm), while the rest of the starches were small starch granules (<8 μm). Shapes of the starch granules varied from oval and irregular with centric hilum to elongated granules with the eccentric hilum. Eleocharis dulcis corm starch had significantly higher total starch content (90.87%), followed by corms of C. esculenta (82.35%) and S. sagittifolia (71.71%). Nelumbo nucifera seed starch had significantly higher amylose content (71.45%), followed by T. angustifolia pollen (36.47%). In comparison, the waxy starch was in N. nucifera rhizome (7.63%), T. bispinosa seed (8.83%), C. esculenta corm (10.61%), and T. angustifolia rhizome (13.51%). Higher resistant starch was observed mostly in rhizomes of N. nucifera (39.34%)>T. angustifolia (37.19%) and corm parts of E. dulcis (37.41%)>S. sagittifolia (35.09%) compared to seed and pollen starches. The XRD profiles of macrophytes starches displayed in all the corms and N. nucifera seed had A-type crystallinity. The T. bispinosa seed had CA-type, whereas the rest of the starches exhibited CB-type crystallinity. Waxy starches of C. esculenta corm had higher relative crystallinity (36.91%) and viscosity (46.2 mPa s) than regular starches. Based on thermal properties, high-amylose of N. nucifera seed and T. angustifolia pollen resulted in higher gelatinization enthalpy (19.93 and 18.66 J g-1, respectively). Starch properties showed equally good potential as commercial starches in starch-based food production based on their starch properties and functionality.

Antisense inhibition of a tomato meiotic proteinase suggests functional redundancy of proteinases during microsporogenesis

Genome ◽  
2001 ◽  
Vol 44 (4) ◽  
pp. 644-650 ◽  
Author(s):  
C Daniel Riggs ◽  
Katherine Zeman ◽  
Rhoda DeGuzman ◽  
Annette Rzepczyk ◽  
Andrew A Taylor
Keyword(s):  
Cell Death ◽  
Transgenic Plants ◽  
Serine Proteinase ◽  
Anther Development ◽  
Antisense Inhibition ◽  
Gene Encoding ◽  
Tomato Gene ◽  
Antisense Construct ◽  
Development Proceeds

Anther development in angiosperms culminates in the programmed cell death of specific tissues to facilitate the release of pollen. Despite a wealth of morphological descriptions of this process, there have been few reports on the regulation of dehiscence or the coordination of events between tissues. We have cloned an anther-specific tomato gene encoding a serine proteinase that is expressed during meiosis and late microsporogenesis. The conceptualized tomato meiotic proteinase (TMP) is a member of a family of genes that exhibit characteristics of mammalian proprotein convertases. To examine the role of TMP in microsporogenesis, we generated transgenic plants harboring an antisense construct of the gene. Some of these plants produced little or no detectable TMP, yet no phenotypic abnormalities were observed. Zymogram analyses revealed that multiple proteinases are present in mature anthers and that proteinase activity increases as development proceeds. Taken together, these data indicate that the role of TMP during microsporogenesis, if any, may be compensated for by other proteinases.Key words: anther, development, pollen, proteinase, subtilase, tapetum.

scholarly journals Genome-Wide Identification and Genetic Variations of the Starch Synthase Gene Family in Rice

Plants ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1154
Author(s):  
Hongjia Zhang ◽  
Seong-Gyu Jang ◽  
San Mar Lar ◽  
Ah-Rim Lee ◽  
Fang-Yuan Cao ◽  
...  
Keyword(s):  
Gene Family ◽  
Catalytic Domain ◽  
Amylose Content ◽  
Expression Patterns ◽  
Starch Synthase ◽  
Eating Quality ◽  
Segmental Duplications ◽  
Genome Wide ◽  
The Relationship

Starch is a major ingredient in rice, and the amylose content of starch significantly impacts rice quality. OsSS (starch synthase) is a gene family related to the synthesis of amylose and amylopectin, and 10 members have been reported. In the present study, a synteny analysis of a novel family member belonging to the OsSSIV subfamily that contained a starch synthase catalytic domain showed that three segmental duplications and multiple duplications were identified in rice and other species. Expression data showed that the OsSS gene family is involved in diverse expression patterns. The prediction of miRNA targets suggested that OsSS are possibly widely regulated by miRNA functions, with miR156s targeted to OsSSII-3, especially. Haplotype analysis exhibited the relationship between amylose content and diverse genotypes. These results give new insight and a theoretical basis for the improved amylose content and eating quality of rice.

scholarly journals Production of very-high-amylose cassava by post-transcriptional silencing of branching enzyme genes

2018 ◽  
Author(s):  
Wenzhi Zhou ◽  
Shanshan Zhao ◽  
Shutao He ◽  
Qiuxiang Ma ◽  
Xinlu Lu ◽  
...  
Keyword(s):  
Amylose Content ◽  
Length Distribution ◽  
Constitutive Expression ◽  
Raw Material ◽  
Distribution Analysis ◽  
Starch Granules ◽  
Storage Roots ◽  
Chain Length Distribution ◽  
Transgenic Cassava ◽  
High Amylose

AbstractHigh amylose starch, a desired raw material in the starch industry, can be produced by plants deficient in the function of branching enzymes (BEs). Here we report the production of transgenic cassava plants with starches containing up to 50% amylose due to the constitutive expression of hair-pin dsRNAs targeting the BE1 or BE2 genes. A significant decrease in BE transcripts was confirmed in these transgenic plants by quantitative real-time RT-PCR. The absence of BE1 protein in the BE1-RNAi plant lines (BE1i) and a dramatically lower level of BE2 protein in the BE2-RNAi plant lines (BE2i) were further confirmed by Western blot assays. All transgenic plant lines were grown up in the field, but with reduced biomass production of the above-ground parts and storage roots compared to wild type (WT). Considerably high amylose content in the storage roots of BE2i plant lines was achieved, though not in BE1i plant lines. Storage starch granules of BE1i and BE2i plants had similar morphology as WT, however, the size of BE1i starch granules were bigger than that of WT. Comparisons of amylograms and thermograms of all three sources of storage starches revealed dramatic changes to the pasting properties and a higher melting temperature for BE2i starches. Glucan chain length distribution analysis showed a slight increase in chains of DP>36 in BE1i lines and a dramatic increase in glucan chains between DP 10-20 and DP>40 in BE2i lines, compared to that of WT starch. Furthermore, BE2i starches displayed a B-type X-ray diffraction pattern instead of the A-type pattern found in BE1i and WT starches. Therefore, cassava BE1 and BE2 function differently in storage root starch biosynthesis; silencing of cassava BE1 or BE2 caused various changes to starch physico-chemical properties and amylopectin structure. We also report that remarkably high amylose content in cassava starch has been first obtained in transgenic cassava by silencing of BE2 expression, thus showing a high potential for future industrial utilization.

scholarly journals A null allele of granule bound starch synthase (Wx-B1) may be one of the major genes controlling chapatti softness

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0246095
Author(s):  
Venkatesh Chunduri ◽  
Natasha Sharma ◽  
Monika Garg
Keyword(s):  
Null Allele ◽  
Amylose Content ◽  
Association Studies ◽  
Starch Synthase ◽  
Near Isogenic Lines ◽  
Processing Quality ◽  
Major Genes ◽  
Northern India ◽  
Granule Bound Starch Synthase ◽  
Tearing Strength

Chapatti (unleavened flatbread) is a staple food in northern India and neighboring countries but the genetics behind its processing quality are poorly understood. To understand the genes determining chapatti quality, differentially expressed genes were selected from microarray data of contrasting chapatti cultivars. From the gene and trait association studies, a null allele of granule bound starch synthase (GBSS; Wx-B1) was found to be associated with low amylose content and good chapatti quality. For validation, near-isogenic lines (NILs) of this allele were created by marker assisted backcross (MAB) breeding. Background screening indicated 88.2 to 96.7% background recovery in 16 selected BC3F5 NILs. Processing quality and sensory evaluation of selected NILs indicated improvement in chapatti making quality. Traits that showed improvement were mouthfeel, tearing strength and softness indicating that the Wx-B1 may be one of the major genes controlling chapatti softness.

scholarly journals Triple null mutations in starch synthase SSIIa gene homoeologs lead to high amylose and resistant starch in hexaploid wheat

2020 ◽  
Author(s):  
Adam Schoen ◽  
Anupama Joshi ◽  
Vijay K Tiwari ◽  
Bikram S. Gill ◽  
Nidhi Rawat
Keyword(s):  
Resistant Starch ◽  
Amylose Content ◽  
Starch Content ◽  
Wheat Variety ◽  
Starch Synthesis ◽  
Starch Synthase ◽  
Single Individual ◽  
Individual Genome ◽  
Mutant Genotype ◽  
Knock Out

Abstract Background: Lack of nutritionally appropriate foods is one of the leading causes of obesity in the US and worldwide. Wheat (Triticum aestivum) provides 20% of the calories consumed daily across the globe. The nutrients in the wheat grain come primarily from the starch composed of amylose and amylopectin. Resistant starch content, which is known to have significant human health benefits, can be increased by modifying starch synthesis pathways. Starch synthase enzyme SSIIa, also known as starch granule protein isoform-1 (SGP-1), is integral to the biosynthesis of the branched and readily digestible glucose polymer amylopectin. The goal of this work was to develop a triple null mutant genotype for SSIIa locus in the elite hard red winter wheat variety ‘Jagger’ and evaluate the effect of the knock-out mutations on resistant starch content in grains with respect to wild type. Results: Knock-out mutations in SSIIa in the three genomes of wheat variety ‘Jagger’ were identified using TILLING. Subsequently, these loss-of function mutations on A, B, and D genomes were combined by crossing to generate a triple knockout mutant genotype Jag-ssiia-∆ABD. The Jag-ssiia-∆ABD had an amylose content of 35.70% compared to 31.15% in Jagger, leading to ~118% increase in resistant starch in the Jag-ssiia-∆ABD genotype of Jagger wheat. The single individual genome mutations also had various effects on starch composition. Conclusions: Our full null Jag-ssiia-∆ABD mutant showed a significant increase in RS without the shriveled grain phenotype seen in other ssiia knockouts in elite wheat cultivars. Moreover, this study shows the potential for developing nutritionally improved foods in a non-GM approach. Since all the mutants have been developed in an elite wheat cultivar, their adoption in production and supply will be feasible in future.

Multiple effects of the starch synthase II mutation in developing wheat endosperm

2007 ◽  
Vol 34 (5) ◽  
pp. 431 ◽  
Author(s):  
Behjat Kosar-Hashemi ◽  
Zhongyi Li ◽  
Oscar Larroque ◽  
Ahmed Regina ◽  
Makoto Yamamori ◽  
...  
Keyword(s):  
Triticum Aestivum L ◽  
Endosperm Development ◽  
Starch Synthase ◽  
Starch Granules ◽  
Branching Enzyme ◽  
Wild Type ◽  
Drastic Reduction ◽  
Branching Patterns ◽  
Soluble Phase ◽  
Mutant Lines

A line of wheat (Triticum aestivum L.), sgp-1, that does not express starch synthase II (SSII, also known as SGP-1) has previously been reported. In this study, F1 derived doubled haploid lines with homozygous wild type or mutant alleles for SGP-1 genes were identified from a cross between the original mutant and a wild type Australian cultivar. Analysis of the starch granules showed that in the mutant lines they are markedly distorted from 15 days postanthesis during grain development. Starch branching patterns showed an increase in the proportion of short chains (DP 6–10) at an earlier stage, but this increase became much more pronounced at 15 days postanthesis and persisted until maturity. There was also a consistent and drastic reduction throughout seed development in the relative amounts of starch branching enzyme II (SBEII, comprising SBEIIa and SBEIIb) and starch synthase I (SSI) bound to the starch granules. In the soluble phase, however, there was relatively little change in the amount of SBEIIb, SBEIIa or SSI protein. Therefore loss of SSII specifically leads to the loss of SBEIIb, SBEIIa and SSI protein in the granule-bound phase and the effect of this mutation is clearly manifest from the mid-stage of endosperm development in wheat.

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