A mutant of rice lacking the leaf large subunit of ADP-glucose pyrophosphorylase has drastically reduced leaf starch content but grows normally

2007 ◽  
Vol 34 (6) ◽  
pp. 480 ◽  
Author(s):  
Sandrine Rösti ◽  
Brendan Fahy ◽  
Kay Denyer
Keyword(s):  
Environmental Conditions ◽  
No Reduction ◽  
Starch Content ◽  
Large Subunit ◽  
Starch Synthesis ◽  
Small Subunit ◽  
Wild Type ◽  
Gene Encoding ◽  
Subunit Protein ◽  
Adp Glucose Pyrophosphorylase

A mutant of rice was identified with a Tos17 insertion in OsAPL1, a gene encoding a large subunit (LSU) of ADP-glucose pyrophosphorylase (AGPase). The insertion prevents production of a normal transcript from OsAPL1. Characterisation of the mutant (apl1) showed that the LSU encoded by OsAPL1 is required for AGPase activity in rice leaf blades. In mutant leaf blades, the AGPase small subunit protein is not detectable and the AGPase activity and starch content are reduced to <1 and <5% of that in wild type blades, respectively. The mutation also leads to a reduction in starch content in the leaf sheaths but does not significantly affect AGPase activity or starch synthesis in other parts of the plant. The sucrose, glucose and fructose contents of the leaves are not affected by the mutation. Despite the near absence of starch in the leaf blades, apl1 mutant rice plants grow and develop normally under controlled environmental conditions and show no reduction in productivity.

A rice mutant lacking a large subunit of ADP-glucose pyrophosphorylase has drastically reduced starch content in the culm but normal plant morphology and yield

2012 ◽  
Vol 39 (12) ◽  
pp. 1068 ◽  
Author(s):  
Frederick R. Cook ◽  
Brendan Fahy ◽  
Kay Trafford
Keyword(s):  
No Reduction ◽  
Oryza Sativa L ◽  
Starch Content ◽  
Large Subunit ◽  
Plant Morphology ◽  
Gene Encoding ◽  
Rice Mutant ◽  
Carbohydrate Storage ◽  
Adp Glucose Pyrophosphorylase ◽  
Storage Pools

A mutant of rice (Oryza sativa L.) was identified with a Tos17 insertion in Os05g50380, a gene encoding a plastidial large subunit (LSU) of ADP-glucose pyrophosphorylase (AGPase) that was previously called OsAPL3 or OsAGPL1. The insertion prevents the production of a normal transcript. Characterisation of the mutant showed that this LSU is required for 97% of the starch synthesised in the flowering stem (culm), approximately half of the AGPase activity in developing embryos and that it contributes to AGPase activity in the endosperm. Despite the near absence of starch in the culms and reduced starch content in the embryos, the mutant rice plants grow and develop normally, and show no reduction in productivity. The starch content of leaves is increased in the mutant, revealing plasticity in the distribution of photosynthates among different temporary carbohydrate storage pools within the plant.

Cloning and expression analysis of cDNAs encoding ADP-glucose pyrophosphorylase large and small subunits from hulless barley (Hordeum vulgare L. var. nudum)

2018 ◽  
Vol 73 (5-6) ◽  
pp. 191-197 ◽  
Author(s):  
Dongmei Li ◽  
Zhimin Yang ◽  
Xinchun Liu ◽  
Zhen Song ◽  
Zongyun Feng ◽  
...  
Keyword(s):  
Accumulation Rate ◽  
Large Subunit ◽  
Starch Synthesis ◽  
Small Subunit ◽  
Cloning And Expression ◽  
Starch Accumulation ◽  
Hordeum Vulgare L ◽  
Hulless Barley ◽  
Expression Levels ◽  
Adp Glucose Pyrophosphorylase

Abstract As an important plateau cereal crop, hulless barley is the principal food for the Tibetan people in China. ADP-glucose pyrophosphorylase (AGPase) is considered as the key enzyme for starch biosynthesis in plants. In this study, cDNAs encoding the small subunit (SSU I) and large subunit (LSU I) of AGPase were isolated from hulless barley. The results showed that SSU I and LSU I were 1438 and 1786 bp in length with a complete open reading frame (ORF) of 1419 and 1572 bp. The ORF-encoded polypeptides of 472 and 523 amino acids were having calculated molecular masses of 52.01 and 58.23 kDa, and the pI values were 5.59 and 6.30. In addition, phylogenetic analysis showed that SSU I and LSU I had the same phylogenetic trends with some species. Furthermore, expression levels in different growth periods and tissues of two hulless barley varieties were analyzed by quantitative reverse transcription-polymerase chain reaction. Gene expression levels of SSU I and LSU I were consistent with the total starch accumulation rate in endosperm. In conclusion, our data confirmed that SSU I and LSU I played an important role in hulless barley starch synthesis.

Molecular Cloning and Spatial Expression of an ApL1 cDNA for the Large Subunit of ADP-Glucose Pyrophosphorylase from Arabidopsis thaliana

1999 ◽  
Vol 54 (5-6) ◽  
pp. 353-358 ◽  
Author(s):  
Leszek A. Kleszkowski ◽  
Lubomir N. Sokolov ◽  
Cheng Luo ◽  
Per Villand
Keyword(s):  
Arabidopsis Thaliana ◽  
Large Subunit ◽  
Critical Role ◽  
Starch Synthesis ◽  
Transit Peptide ◽  
Homologous Proteins ◽  
Peptide Cleavage ◽  
Reading Frame ◽  
Calculated Molecular Weight ◽  
Adp Glucose Pyrophosphorylase

Abstract A cDNA, A p L 1a , corresponding to a homologue of the large subunit of ADP-glucose pyrophosphorylase (AG Pase), has been isolated/characterised by screening a cDNA library prepared from leaves of Arabidopsis thaliana, followed by rapid amplification of cDNA 3′-ends (3′-RACE). Within the 1685 nucleotide-long sequence (excluding polyA tail), an open reading frame encodes a protein of 522 amino acids (aa), with a calculated molecular weight of 57.7 kDa. The derived aa sequence does not contain any discernible transit peptide cleavage site motif, similarly to two other recently sequenced full-length Arabidopsis homo-logues for AGPase, and shows ca. 58–78 % identity to homologous proteins from other plants/tissues. The corresponding gene was found (rosette and stem leaves, stems, flowers and fruits), consistent with its critical role in starch synthesis in

Modulation of procaspase-7 self-activation by PEST amino acid residues of the N-terminal prodomain and intersubunit linker

2017 ◽  
Vol 95 (6) ◽  
pp. 634-643
Author(s):  
Juliano Alves ◽  
Miguel Garay-Malpartida ◽  
João M. Occhiucci ◽  
José E. Belizário
Keyword(s):  
Amino Acid ◽  
Large Subunit ◽  
Small Subunit ◽  
Cleavage Sites ◽  
Amino Acid Residues ◽  
Wild Type ◽  
Caspase 7 ◽  
Caspase Family ◽  
The Impact

Procaspase-7 zymogen polypeptide is composed of a short prodomain, a large subunit (p20), and a small subunit (p10) connected to an intersubunit linker. Caspase-7 is activated by an initiator caspase-8 and -9, or by autocatalysis after specific cleavage at IQAD198↓S located at the intersubunit linker. Previously, we identified that PEST regions made of amino acid residues Pro (P), Glu (E), Asp (D), Ser (S), Thr (T), Asn (N), and Gln (Q) are conserved flanking amino acid residues in the cleavage sites within a prodomain and intersubunit linker of all caspase family members. Here we tested the impact of alanine substitution of PEST amino acid residues on procaspase-7 proteolytic self-activation directly in Escherichia coli. The p20 and p10 subunit cleavage were significantly delayed in double caspase-7 mutants in the prodomain (N18A/P26A) and intersubunit linker (S199A/P201A), compared with the wild-type caspase-7. The S199A/P201A mutants effectively inhibited the p10 small subunit cleavage. However, the mutations did not change the kinetic parameters (kcat/KM) and optimal tetrapeptide specificity (DEVD) of the purified mutant enzymes. The results suggest a role of PEST-amino acid residues in the molecular mechanism for prodomain and intersubunit cleavage and caspase-7 self-activation.

scholarly journals NUCLEAR MUTATION INCREASES STREPTOMYCIN AND SPECTINOMYCIN SENSITIVITY IN CHLAMYDOMONAS

Genetics ◽  
1978 ◽  
Vol 88 (4) ◽  
pp. 643-650
Author(s):  
Robert W Lee ◽  
Jan A Sapp
Keyword(s):  
Large Subunit ◽  
Small Subunit ◽  
Wild Type ◽  
Resistance Mutations ◽  
Erythromycin Resistance ◽  
Spectinomycin Resistance ◽  
Nuclear Mutation ◽  
Resistant Strains ◽  
Nuclear Locus ◽  
Type Strains

ABSTRACT A spontaneously arising nuclear mutation, ss-1, has been identified in Chlamydomonas reinhardtii that decreases both streptomycin and spectinomycin resistance levels about 10-fold after its introduction into all wild-type, streptomycin-resistant and spectinomycin-resistant strains examined. The mutations for resistance map to nuclear and uniparentally inherited (chloroplast) loci. In contrast, no modification of erythromycin resistance was detected after introducing ss-1 into wild-type strains or into strains carrying nuclear or uniparentally inherited erythromycin-resistance mutations. We suggest that ss-1 affects the small subunit of the chloroplast ribosome because others have shown that streptomycin and spectinomycin resistance in C. reinhardtii are associated with this subunit, whereas erythromycin resistance is associated with the large subunit. ss-1 shows no linkage with the nuclear locus for streptomycin resistance.

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 A Novel Spirooxindole Derivative Inhibits the Growth of Leishmania donovani Parasites bothIn VitroandIn Vivoby Targeting Type IB Topoisomerase

2016 ◽  
Vol 60 (10) ◽  
pp. 6281-6293 ◽  
Author(s):  
Sourav Saha ◽  
Chiranjit Acharya ◽  
Uttam Pal ◽  
Somenath Roy Chowdhury ◽  
Kahini Sarkar ◽  
...  
Keyword(s):  
Leishmania Donovani ◽  
Nuclear Dna ◽  
Large Subunit ◽  
Peritoneal Macrophages ◽  
Small Subunit ◽  
Drug Resistant ◽  
Wild Type ◽  
Content Type ◽  
Fluorescence Studies ◽  
Topoisomerase Ib

ABSTRACTVisceral leishmaniasis is a fatal parasitic disease, and there is an emergent need for development of effective drugs against this neglected tropical disease. We report here the development of a novel spirooxindole derivative,N-benzyl-2,2′α-3,3′,5′,6′,7′,7α,α′-octahydro-2methoxycarbonyl-spiro[indole-3,3′-pyrrolizidine]-2-one (compound 4c), which inhibitsLeishmania donovanitopoisomerase IB (LdTopIB) and kills the wild type as well as drug-resistant parasite strains. This compound inhibits catalytic activity of LdTopIB in a competitive manner. Unlike camptothecin (CPT), the compound does not stabilize the DNA-topoisomerase IB cleavage complex; rather, it hinders drug-DNA-enzyme covalent complex formation. Fluorescence studies show that the stoichiometry of this compound binding to LdTopIB is 2:1 (mole/mole), with a dissociation constant of 6.65 μM. Molecular docking with LdTopIB using the stereoisomers of compound 4c produced two probable hits for the binding site, one in the small subunit and the other in the hinge region of the large subunit of LdTopIB. This spirooxindole is highly cytotoxic to promastigotes ofL. donovaniand also induces apoptosis-like cell death in the parasite. Treatment with compound 4c causes depolarization of mitochondrial membrane potential, formation of reactive oxygen species inside parasites, and ultimately fragmentation of nuclear DNA. Compound 4c also effectively clears amastigote forms of wild-type and drug-resistant parasites from infected mouse peritoneal macrophages but has less of an effect on host macrophages. Moreover, compound 4c showed strong antileishmanial efficacies in the BALB/c mouse model of leishmaniasis. This compound potentially can be used as a lead for developing excellent antileishmanial agents against emerging drug-resistant strains of the parasite.

scholarly journals A single gene encodes two different transcripts for the ADP-glucose pyrophosphorylase small subunit from barley (Hordeum vulgare)

1996 ◽  
Vol 313 (1) ◽  
pp. 149-154 ◽  
Author(s):  
Tine THORBJØRNSEN ◽  
Per VILLAND ◽  
Leszek A. KLECZKOWSKI ◽  
Odd-Arne OLSEN
Keyword(s):  
Hordeum Vulgare ◽  
Genomic Library ◽  
Single Gene ◽  
Starch Synthesis ◽  
Transit Peptide ◽  
Small Subunit ◽  
Peptide Sequence ◽  
Subunit Gene ◽  
Starchy Endosperm ◽  
Adp Glucose Pyrophosphorylase

ADP-glucose pyrophosphorylase (AGPase), a heterotetrameric enzyme composed of two small and two large subunits, catalyses the first committed step of starch synthesis in plant tissues. In an attempt to learn more about the organization and expression of the small-subunit gene of AGPase, we have studied the small-subunit transcripts as well as the structure of the gene encoding these transcripts in barley (Hordeum vulgare L. cv. Bomi). Two different transcripts (bepsF1 and blps14) were identified: bepsF1 was abundantly expressed in the starchy endosperm but not in leaves, whereas blps14 was isolated from leaves but was also found to be present at a moderate level in the starchy endosperm. The sequences for the two transcripts are identical over approx. 90% of the length, with differences being confined solely to their 5ʹ ends. In blps14, the unique 5ʹ end is 259 nt long and encodes a putative plastid transit peptide sequence. For the 178-nt 5ʹ end of bepsF1, on the other hand, no transit peptide sequence could be recognized. A lambda clone that hybridized to the AGPase transcripts was isolated from a barley genomic library and characterized. The restriction map has suggested a complex organization of the gene, with alternative exons encoding the different 5ʹ ends of the two transcripts followed by nine exons coding for the common part of the transcripts. The sequence of a portion of the genomic clone, covering the alternative 5ʹ-end exons as well as upstream regions, has verified that both transcripts are encoded by the gene. The results suggest that the small-subunit gene of barley AGPase transcribes two different mRNAs by a mechanism classified as alternative splicing.

Cloning and characterization of Agp1, the gene encoding the small subunit of ADP-glucose pyrophosphorylase from wheat and its relatives

Biologia ◽  
2017 ◽  
Vol 72 (12) ◽  
Author(s):  
Xiao-Wei Zhang ◽  
Qing Wang ◽  
Ling-Ling Zhang ◽  
Xiao-Juan Zhong ◽  
Qian-Tao Jiang ◽  
...  
Keyword(s):  
Cdna Sequence ◽  
Small Subunit ◽  
Full Length ◽  
Gene Encoding ◽  
Full Length Cdna ◽  
S Gene ◽  
Limiting Step ◽  
Adp Glucose Pyrophosphorylase

AbstractADP-glucose pyrophosphorylase (AGP) consists of two large (AGP-L) and two small (AGP-S) subunits; it regulates the limiting step in the biosynthesis of starch. Here, we isolated the full-length cDNA sequence of the AGP-S gene (designed as

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