Granule-bound Starch Synthase I Research Articles

Exploring the synergy of enzymes, nutrients, and gene networks in rice starch granule biogenesis.

Rice is a primary food source almost for more than 50% of the total world's population. Glycemic index (GI) is high in most of the rice varieties, limiting their consumption by diabetic and obese people. As a result, developing new rice varieties with low GI necessitates a thorough understanding of starch biogenesis gene expression and its interrelationship. A total 200 rice genotypes were analyzed for total starch content (TSC), amylopectin content (APC), and amylose content (AC). The clustering of these rice genotypes was done based on their AC. Further, these genotypes were categorized into three groups up to 10% amylose-low, 10-26% amylose-medium, and more than 26% amylose-high. Among them, six genotypes 1 from low AC (NJ-72), 2 from medium AC (UPRI-2003-18, PRR-126), and 3 from high AC (RNRM-7, Urvashi and Ananga) were selected. The genotypes selected from the medium and high AC groups were having 2% amylose variation among themselves respectively and they were further used to study the level of RS, protein content (PC), fatty acid (FA) profiles, and granule morphology along with low group sample. Resistant starch (RS) content ranged from 0.33-2.75%, and fatty acid profiling revealed high levels of palmitic, linoleic, and oleic acids. The degree of crystallinity and APC% were found to be positively correlated. Ananga, the genotype with the highest RS, displayed compact starch granules. Further, NJ-72 showing low RS and Ananga with high RS were selected for investigation of enzymatic activities of starch biosynthesis, metabolites accumulation, and expressions of 20 starch biogenesis genes in developing endosperm. Starch branching enzymes (SBE) and starch synthase (SS) activities peaked at 13 days after anthesis (DAA), while starch debranching enzymes (DBE) were most active at 18 DAA. In Ananga, TSC, AC, APC, and RS levels progressively increased from 3 to 23 DAA. Ananga showed 1.25-fold upregulation of granule-bound starch synthase I (GBSSI) at 18DAA. Higher expressions of SSI and SBEIIb were observed in NJ-72 at 13DAA. PUL2 was predominantly expressed followed by ISA1. GBSSI was positively correlated with both AC and RS while SS, SBE, and DBE were positively related to APC. This research could lead to the development of rice varieties with improved nutritional qualities, such as higher RS content, which is beneficial for human health due to its role in lowering glycemic response and promoting gut health. Additionally, the study provides insights into how the modulation of key genes and enzymes can affect starch composition, offering strategies to breed rice varieties tailored for specific dietary needs or industrial applications.

Deciphering the Role of Waxy Gene Mutations in Enhancing Rice Grain Quality.

Amylose content (AC) stands as a pivotal determinant of rice grain quality, primarily governed by the Waxy gene (Wx). The allelic variation within this gene, particularly the presence of the Wxmp allele derived from the ancestral Wxmq allele, significantly influences AC and is prevalent among soft japonica rice varieties in southern China. Although both alleles are associated with lower AC, there remains a paucity of detailed understanding regarding the interplay between specific functional single nucleotide polymorphisms (SNPs) within these alleles and the overarching rice grain quality. To investigate this, we engineered three distinct transgenic rice lines, each harboring the Wxmp, Wxmq, or Wxb-5c alleles in the background of the glutinous rice cultivar Nip(wx). This suite of transgenic rice lines showcased varying degrees of grain transparency inversely correlated to AC, which in turn influenced other physicochemical properties of the rice grains, such as taste value of cooked rice, gel consistency, and starch pasting properties. Additionally, analyses of gene expression and enzyme activity revealed that the functional SNPs, Ex4-53G to A and Ex5-53T to C, lead to a decline in the activity of granule-bound starch synthase I (GBSSI) without altering expression levels.

Creating a zero amylose barley with high soluble sugar content by genome editing.

Amylose biosynthesis is strictly associated with granule-bound starch synthase I (GBSSI) encoded by the Waxy gene. Mutagenesis of single bases in the Waxy gene, which induced by CRISPR/Cas9 genome editing, caused absence of intact GBSSI protein in grain of the edited line. The amylose and amylopectin contents of waxy mutants were zero and 31.73%, while those in the wild type were 33.50% and 39.00%, respectively. The absence of GBSSI protein led to increase in soluble sugar content to 37.30% compared with only 10.0% in the wild type. Sucrose and β-glucan, were 39.16% and 35.40% higher in waxy mutants than in the wild type, respectively. Transcriptome analysis identified differences between the wild type and waxy mutants that could partly explain the reduction in amylose and amylopectin contents and the increase in soluble sugar, sucrose and β-glucan contents. This waxy flour, which showed lower final viscosity and setback, and higher breakdown, could provide more option for food processing.

OsLESV and OsESV1 promote transitory and storage starch biosynthesis to determine rice grain quality and yield

Transitory starch is an important carbon source in leaves, and its biosynthesis and metabolism are closely related to grain quality and yield. The molecular mechanisms controlling leaf transitory starch biosynthesis and degradation and their effects on rice (Oryza sativa) quality and yield remain unclear. Here, we show that OsLESV and OsESV1, the rice orthologs of AtLESV and AtESV1, are associated with transitory starch biosynthesis in rice. The total starch and amylose contents in leaves and endosperms are significantly reduced, and the final grain quality and yield are compromised in oslesv and osesv1 single and oslesv esv1 double mutants. Furthermore, we found that OsLESV and OsESV1 bind to starch, and this binding depends on a highly conserved C-terminal tryptophan-rich region that acts as a starch-binding domain. Importantly, OsLESV and OsESV1 also interact with the key enzymes of starch biosynthesis, granule-bound starch synthase I (GBSSI), GBSSII, and pyruvate orthophosphote dikiase (PPDKB), to maintain their protein stability and activity. OsLESV and OsESV1 also facilitate the targeting of GBSSI and GBSSII from plastid stroma to starch granules. Overexpression of GBSSI, GBSSII, and PPDKB can partly rescue the phenotypic defects of the oslesv and osesv1 mutants. Thus, we demonstrate that OsLESV and OsESV1 play a key role in regulating the biosynthesis of both leaf transitory starch and endosperm storage starch in rice. These findings deepen our understanding of the molecular mechanisms underlying transitory starch biosynthesis in rice leaves and reveal how the transitory starch metabolism affects rice grain quality and yield, providing useful information for the genetic improvement of rice grain quality and yield.

The elite eating quality alleles Wxb and ALKb are regulated by OsDOF18 and coordinately improve head rice yield.

Head rice yield (HRY) measures rice milling quality and determines final grain yield and commercial value. Here, we report that two major quantitative trait loci for milling quality in rice, qMq-1 and qMq-2, represent allelic variants of Waxylv/Waxyb (hereafter Wx) encoding Granule-Bound Starch Synthase I (GBSSI) and Alkali Spreading Value ALKc/ALKb encoding Soluble Starch Synthase IIa (SSIIa), respectively. Complementation and overexpression transgenic lines in indica and japonica backgrounds confirmed that Wx and ALK coordinately regulate HRY by affecting amylose content, the number of amylopectin branches, amyloplast size, and thus grain filling and hardness. The transcription factor OsDOF18 acts upstream of Wx and ALK by activating their transcription. Furthermore, rice accessions with Wxb and ALKb alleles showed improved HRY over those with Wxlv and ALKc. Our study not only reveals the novel molecular mechanism underlying the formation of HRY but also provides a strategy for breeding rice cultivars with improved HRY.

Development of a model for granule-bound starch synthase I activity using free-energy calculations

Starch is a branched polymer of glucose with two components, both of which have (1 → 4)-α linear links and (1 → 6)-α branch points: amylopectin, of high molecular weight with many short branches, and amylose, of lower molecular weight and only a few long-chain branches. Granule-bound starch synthase I (GBSSI) is one of the main enzymes controlling amylose synthesis and chain-length distribution. As production of different GBSSI mutants is time-consuming and laborious, molecular dynamics (MD) simulations are used here to predict the binding of different GBSSI mutants to a representative amylose fragment. The simulations were atomistic, with explicit solvent and docking, a method successfully used to understand the binding of wild-type GBSSI to amylose fragments. The binding of GBSSI to G5 (a pentasaccharide amylose fragment) is combined with free-energy calculations employing a thermodynamic integration method to predict the effects of mutations on enzyme activity. Ten GBSSI mutants with different enzyme activities were analyzed to find the structural and energy changes among different single amino-acid mutants and their possible relationship to starch characteristics. Comparing the structural changes and the relative binding free energy of G5 to the wild type GBSSI and GBSSI mutants, it was found that mutants with negative binding energy (lower than −2.0 kcal/mol) are more likely to have higher enzyme activity and amylose content compared to the wild type. This theoretical paper used simulations and robust free energy calculations to interpret in planta data with potential predictions as to what mutants might be generated to give desired properties. This study can be used to help develop grains with improved functional properties.

Molecular and genetic bases of rice cooking and eating quality: An updated review

AbstractBackground and ObjectivesRice grain quality is a primary determinant of its market price and consumer acceptance. Although milling quality, appearance, cooking and eating quality (CEQ), and nutritional quality represent the main features of grain quality, rice CEQ is the most important feature in terms of price. Starch physicochemical properties and sensory evaluation have been widely applied to predict and assess rice CEQ. Understanding the genetic and molecular bases of CEQ formation will facilitate improvements in rice quality through a molecular breeding strategy.FindingsThe major genes responsible for rice CEQ formation have already been characterized in previous studies. Waxy (Wx) encoding granule‐bound starch synthase I (GBSSI) controls the apparent amylose content (AAC), starch synthase IIa (SSIIa) controls the gelatinization temperature, and fragrant gene (fgr) controls the aroma of cooked rice. Many natural variations (allelic variants) have been identified in these genes among rice germplasm. The protein content in rice grains is not only responsible for the nutritional quality but also CEQ. Two major genes that control the protein content have been identified and cloned. Pyramiding of different alleles by marker‐assisted selection and creation of new alleles by genome editing technology have facilitated improvements in new rice varieties with desirable CEQ.ConclusionsIn addition to presenting updates on the advances made in the important CEQ genes, we have identified some future challenges. These include the need to exploit new alleles in these genes, especially in Wx, to confer low AAC with a transparent appearance; identification of alleles in Wx that can improve the texture of cooked rice with high AAC and high‐resistant starch; formulation of methods to manipulate genes and modify agronomic practices to reduce the protein content to improve CEQ; and development of methods to breed climate‐smart rice with good, stable CEQ in changing environments.Significance and NoveltyThis article identifies some priorities for future research, which can enhance our understanding of the molecular basis of CEQ to improve this important rice quality attribute.

Genome-Wide Association Study of Starch Properties in Local Thai Rice.

Rice (Oryza sativa L.) is the main source of energy for humans and a staple food of high cultural significance for much of the world's population. Rice with highly resistant starch (RS) is beneficial for health and can reduce the risk of disease, especially type II diabetes. The identification of loci affecting starch properties will facilitate breeding of high-quality and health-supportive rice. A genome-wide association study (GWAS) of 230 rice cultivars was used to identify candidate loci affecting starch properties. The apparent amylose content (AAC) among rice cultivars ranged from 7.04 to 33.06%, and the AAC was positively correlated with RS (R2 = 0.94) and negatively correlated with rapidly available glucose (RAG) (R2 = -0.73). Three loci responsible for starch properties were detected on chromosomes 1, 6, and 11. On chromosome 6, the most significant SNP corresponded to LOC_Os06g04200 which encodes granule-bound starch synthase I (GBSSI) or starch synthase. Two novel loci associated with starch traits were LOC_Os01g65810 and LOC_Os11g01580, which encode an unknown protein and a sodium/calcium exchanger, respectively. The markers associated with GBSSI and LOC_Os11g01580 were tested in two independent sets of rice populations to confirm their effect on starch properties. The identification of genes associated with starch traits will further the understanding of the molecular mechanisms affecting starch in rice and may be useful in the selection of rice varieties with improved starch.

Three Starch Synthase IIa (SSIIa) Alleles Reveal the Effect of SSIIa on the Thermal and Rheological Properties, Viscoelasticity, and Eating Quality of Glutinous Rice

Glutinous rice accumulates amylose-free starch and is utilized for rice cakes and crackers, owing to the loss of the Waxy gene which encodes granule-bound starch synthase I (GBSSI). Starch synthase IIa (SSIIa) elongates amylopectin chains with a degree of polymerization (DP) of 6-12 to 13-24 and greatly influences starch properties. To elucidate the relationship between the branch length of amylopectin and the thermal and rheological properties, viscoelasticity, and eating quality of glutinous rice, three allelic near isogenic lines with high, low, or no SSIIa activity were generated (designated as SS2a wx, ss2aL wx, and ss2a wx, respectively). Chain length distribution analyses revealed that ss2a wx exhibited the highest short chain (DP < 12) number and lowest gelatinization temperature, whereas SS2a wx showed the opposite results. Gel filtration chromatography showed that the three lines contained essentially no amylose. Viscoelasticity analyses of rice cakes stored at low temperature for different durations revealed that ss2a wx maintained softness and elasticity for up to 6 days, while SS2a wx hardened within 6 h. Sensory evaluation was consistent with mechanical evaluation. The relationship of amylopectin structure with the thermal and rheological properties, viscoelasticity, and eating quality of glutinous rice is discussed.

Genetic and molecular analysis of starch physicochemical properties and its relationship with endosperm modification in quality protein maize

Quality protein maize (QPM) was created converting the soft opaque-2 endosperm into a vitreous phenotype, but the mechanisms involved in this modification are not completely understood. Recombinant inbred lines (RILs) derived from K0326Y QPM and W64Ao2 were used to identify quantitative trait loci (QTL) associated with starch physicochemical properties. RILs contrasting in vitreousness were also used to evaluate the expression of starch biosynthesis genes. Mapping identified 5-6 QTLs for each trait that explained 66 % of the phenotypic variation. Three QTLs on bins 4.05, 5.04, and 9.03 were found close to the starch biosynthesis genes Brittle-2 (Bt2), Amylose extender-1 (Ae1), and Waxy-1 (Wx1), respectively. The expression of Wx1 was three-fold greater in K0326Y QPM than W64Ao2 and eight-fold higher in vitreous than opaque RILs, which corresponded with the greater accumulation of granule bound starch synthase I (GBSSI) and the higher amylose content observed in the vitreous lines. The increased synthesis of amylose may result in starch granules with more amorphous regions that favor their compaction. These results suggest that endosperm modification in QPM is associated with the synthesis of starch containing more amylose during kernel development, which may facilitate the packing of the starch granules resulting in the vitreous phenotype.

Regulation of Amylose Content by Single Mutations at an Active Site in the Wx-B1 Gene in a Tetraploid Wheat Mutant.

The granule-bound starch synthase I (GBSSI) encoded by the waxy gene is responsible for amylose synthesis in the endosperm of wheat grains. In the present study, a novel Wx-B1 null mutant line, M3-415, was identified from an ethyl methanesulfonate-mutagenized population of Chinese tetraploid wheat landrace Jianyangailanmai (LM47). The gene sequence indicated that the mutated Wx-B1 encoded a complete protein; this protein was incompatible with the protein profile obtained using sodium dodecyl sulfate–polyacrylamide gel electrophoresis, which showed the lack of Wx-B1 protein in the mutant line. The prediction of the protein structure showed an amino acid substitution (G470D) at the edge of the ADPG binding pocket, which might affect the binding of Wx-B1 to starch granules. Site-directed mutagenesis was further performed to artificially change the amino acid at the sequence position 469 from alanine (A) to threonine (T) (A469T) downstream of the mutated site in M3-415. Our results indicated that a single amino acid mutation in Wx-B1 reduces its activity by impairing its starch-binding capacity. The present study is the first to report the novel mechanism underlying Wx-1 deletion in wheat; moreover, it provided new insights into the inactivation of the waxy gene and revealed that fine regulation of wheat amylose content is possible by modifying the GBSSI activity.

Diurnal changes in starch molecular structures and expression profiles of starch biosynthesis enzymes in rice developing seeds

The diurnal changes in the expression profiles of starch synthesis related enzymes (SSREs) has been previously studied in transitory starches, while its influences on storage starch molecular structures in the rice endosperm during seed development have not been elucidated. In this study, the changes in the transcript levels of starch synthesis related genes (SSRGs), the protein abundances and enzyme activities of SSREs as well as starch molecular structures in rice endosperm at 10 days after flowering (DAF) over the diurnal cycle were analyzed. It was found that the expression profiles of SSRG and the protein contents of SSREs displayed different diurnal patterns between two indica rice varieties with medium- and high-amylose content (AC), respectively. The expression levels of SSRGs were higher in the light time, and most SSREs also accumulated during this period except debranching enzymes. Amylose synthesis displayed distinct diurnal patterns in two rice varieties, which is attributed to the diurnal changes in the protein content of granule-bound starch synthase I (GBSSI), but amylopectin chain-length distributions (CLDs) remained unaltered due to its vast numbers of branches. The results provide the first step to understand the roles of each enzyme isoform involved in starch synthesis in response to diurnal regulation in rice endosperm.

Protein targeting to starch 1, a functional protein of starch biosynthesis in wheat (Triticum aestivum L.)

TaPTST1, a wheat homolog of AtPTST1 containing CBM can interact with GBSSI and regulate starch metabolism in wheat endosperm. In cereal endosperm, native starch comprising amylose and amylopectin is synthesized by the coordinated activities of several pathway enzymes. Amylose in starch influences its physio-chemical properties resulting in several human health benefits. The Granule-Bound Starch Synthase I (GBSSI) is the most abundant starch-associated protein. GBSSI lacks dedicated Carbohydrate-binding module (CBM). Previously, Protein Targeting ToStarch 1 (PTST1) was identified as a crucial protein for the localization of GBSSI to the starch granules in Arabidopsis. The function of its homologous protein in the wheat endosperm is not known. In this study, TaPTST1, an AtPTST1 homolog, containing a CBM and a coiled-coil domain was identified in wheat. Protein-coding nucleotide sequence of TaPTST1 from Indian wheat variety 'C 306' was cloned and characterized. Homology modelling and molecular docking suggested the potential interaction of TaPTST1 with glucans and GBSSI. The TaPTST1 expression was higher in wheat grain than the other tissues, suggesting a grain-specific function. In vitro binding assays demonstrated different binding affinities of TaPTST1 for native starch, amylose, and amylopectin. Furthermore, the immunoaffinity pull-down assay revealed that TaPTST1 directly interacts with GBSSI, and the interaction is mediated by a coiled-coil domain. The direct protein-protein interaction was further confirmed by bimolecular fluorescence complementation assay (BiFC) in planta. Based on our findings we postulate a functional role for TaPTST1 in starch metabolism by targeting GBSSI to starch granules in wheat endosperm.

The Effect and Regulation Mechanism of Powdery Mildew on Wheat Grain Carbon Metabolism

AbstractThe effects of powdery mildew on gene expressions and activities of carbon (C) metabolism enzymes in Zhengmai 103 (resistant) and Xi'nong 979 (susceptible) are studied. Under severe powdery mildew, expressions and activities of sucrose synthase, uridine‐diphosphate glucose pyrophosphorylase, glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH), and cytosolic α‐1,4‐glucan phosphorylase decrease at 20 or 25–30 days after anthesis, and total soluble sugars content also decreases. It indicates that the first part of C metabolism (sucrose degradation) changes and total C flux reduces. Meanwhile, decreases in expressions and activities of adenosine‐diphosphate glucose pyrophosphorylase (AGPase), soluble starch synthase (SSS), starch‐branching enzyme I and II (SBEI and SBEII), and plastidic α‐1,4‐glucan phosphorylase coincide with decreases in amylopectin content, whereas increases in granule‐bound starch synthase I (GBSSI) expression and activity coincide with increases in amylose content. It shows that the second part of C metabolism (starch synthesis) changes. Therefore, powdery mildew alters C metabolism. In addition, it also induces defense responses in grains, such as the up‐regulation of a defense‐related protein endogenous α‐amylase/subtilisin inhibitor. Since GAPDH, AGPase, SSS, SBEII, and GBSSI are regulated by redox, changes of C metabolism are mainly regulated by cellular redox status.

Impact of Drought Stress on Grains Filling in Rice and its Management: A Review

Rice (Oryza sativa L.) is the world’s single most important staple food source for over half of the world’s population, whose demand is expected to increase as human population increases almost exponentially every year. There are different kinds of abiotic and biotic stresses that a rice plant endures. Among the different forms of environmental stresses, drought hampers the production of rice grains notably. The reality of climate change is so stark that more and more droughts are expected to be evident in the distant future. Water, being a requisite for rice cultivation, is gradually becoming a highly limited resource across the earth. In comparison to other cereal crops, rice is more sensitive to the soil water content. Decrease of soil water content or the unavailability of water triggers drought stress in rice. Therefore, the management of drought stress for the growth of rice plants is essential as even moderate stress during reproductive growth may result in a drastic reduction of grain yield. The distinct changes brought about by drought stress not only range from morphological to physiological changes but also from biochemical to molecular changes as well. A number of starch synthesis related enzymes like granule bound starch synthase I (GBSSI), soluble starch synthase IIa (SSIIa) play important role in grain filling, which are also affected by limited water under drought environment. Due to water scarcity, the rate of photosynthesis decreases, as a result of which dry matter accumulation decreases, which ultimately affects grain filling in rice. Drought causes delayed flowering as well as spikelet sterility, followed by low grain yield during harvesting. The prevalence of drought over longer periods causes reactive oxygen species to form within the rice plants which subsequently denatures proteins and nucleic acids. In many places where drought continues for longer periods, rice plants die resulting in huge loss of grain yields. The effective strategies that have been adopted involves-the development of stable drought tolerant rice varieties through plant breeding programs, the development of short duration rice cultivars which have short growing cycle and harvesting periods that may help the plant to escape drought, the development of rice plants having deep root system such that it can penetrate the soil to find the available moisture and water required for growth, as well as the development of transgenic drought tolerant rice plants which over express specific genes to withstand drought, are few of the effective management strategies that need special attention. Thus, it can be said that drought management for rice production is a crucial step towards food security and food production in the world. The present discussions will emphasize on the negative impact of drought in rice plants and also discuss possible key management practices that can address the existing and imminent drought related problems on rice plants.

Functional Interactions between Enzymes Involved in Amylose and Amylopectin Biosynthesis in Rice Based on Mathematical Models.

Starch biosynthesis is controlled by multiple enzymes, including granule-bound starch synthase I (GBSSI), soluble starch synthases (SSs), branching enzymes (BEs), and debranching enzymes (DBEs). Although the role of individual isoforms has been primarily elucidated, the precise information about how they work together in the synthesis of specific amylose and amylopectin chains is still unclear. In this study, starch molecular chain-length distributions (CLDs) of five rice varieties with different amylose contents were measured by fluorophore-assisted carbohydrate electrophoresis and size-exclusion chromatography and fitted with two mathematical models, and the protein abundance of 11 starch synthesis-related enzymes was measured by western blotting. The correlation between model fitting parameters of amylose and amylopectin CLDs demonstrated that amylose and amylopectin syntheses are closely dependent. GBSSI could interact with BEI, BEIIb, SSIIa, SSIVb, ISA1, PUL, and PHO1 to synthesize the amylopectin intermediate and long chains as well as amylose chains. In addition, the interaction among SSIVb and SSI, SSIIa, BEI, BEIIb, ISA1, and PUL possibly suggests that SSIVb assists them to synthesize the amylopectin chains. The results can help understand the mechanisms about the functional interaction of different enzyme isoforms in starch biosynthesis.

Haplotype Variations and Evolutionary Analysis of the Granule-Bound Starch Synthase I Gene in the Korean World Rice Collection

Granule-bound starch synthase I (GBSSI) is responsible for Waxy gene encoding the, which is involved in the amylose synthesis step of starch biosynthesis. We investigated the genotypic and haplotypic variations of GBSSI (Os06g0133000) gene, including its evolutionary relatedness in the nucleotide sequence level using single-nucleotide polymorphisms (SNPs), indels, and structural variations (SVs) from 475 Korean World Rice Collection (KRICE_CORE), which comprised 54 wild rice and 421 cultivated represented by 6 ecotypes (temperate japonica, indica, tropical japonica, aus, aromatic, and admixture) or in another way by 3 varietal types (landrace, weedy, and bred). The results revealed that 27 of 59 haplotypes indicated a total of 12 functional SNPs (fSNPs), identifying 9 novel fSNPs. According to the identified novel fSNPs, we classified the entire rice collection into three groups: cultivated, wild, and mixed (cultivated and wild) rice. Five novel fSNPs were localized in wild rice: four G/A fSNPs in exons 2, 9, and 12 and one T/C fSNP in exon 13. We also identified the three previously reported fSNPs, namely, a G/A fSNP (exon 4), an A/C fSNP (exon 6), and a C/T fSNP (exon 10), which were observed only in cultivated rice, whereas an A/G fSNP (exon 4) was observed exclusively in wild rice. All-against-all comparison of four varietal types or six ecotypes of cultivated rice with wild rice showed that the GBSSI diversity was higher only in wild rice (π = 0.0056). The diversity reduction in cultivated rice can be useful to encompass the origin of this gene GBSSI during its evolution. Significant deviations of positive (wild and indica under balancing selection) and negative (temperate and tropical japonica under purifying selection) Tajima's D values from a neutral model can be informative about the selective sweeps of GBSSI genome insights. Despite the estimation of the differences in population structure and principal component analysis (PCA) between wild and subdivided cultivated subgroups, an inbreeding effect was quantified by FST statistic, signifying the genetic relatedness of GBSSI. Our findings of a novel wild fSNPS can be applicable for future breeding of waxy rice varieties. Furthermore, the signatures of selective sweep can also be of informative into further deeper insights during domestication.

Genotype-dependent and heat-induced grain chalkiness in rice correlates with the expression patterns of starch biosynthesis genes.

Starch biosynthesis is a complex process underlying grain chalkiness in rice in a genotype-dependent manner. Coordinated expression of starch biosynthesis genes is important for producing translucent rice grains, while disruption in this process leads to opaque or chalky grains. To better understand the dynamics of starch biosynthesis genes in grain chalkiness, six rice genotypes showing variable chalk levels were subjected to gene expression analysis during reproductive stages. In the chalky genotypes, peak expression of the large subunit genes of ADP-glucose pyrophosphorylase (AGPase), encoding the first key step in starch biosynthesis, occurred in the stages before grain filling commenced, creating a gap with the upregulation of starch synthase genes, granule bound starch synthase I (GBSSI) and starch synthase IIA (SSIIA). Whereas, in low-chalk genotypes, AGPase large subunit genes expressed at later stages, generally following the expression patterns of GBSSI and SSIIA. However, heat treatment altered the expression in a genotype-dependent manner that was accompanied by transformed grain morphology and increased chalkiness. The suppression of AGPase subunit genes during early grain filling stages was observed in the chalky genotypes or upon heat treatment, which could result in a limited pool of ADP-Glucose for synthesizing amylose and amylopectin, the major components of the starch. This suboptimal starch biosynthesis process could subsequently lead to inefficient grain filling and air pockets that contribute to chalkiness. In summary, this study suggests a mechanism of grain chalkiness based on the expression patterns of the starch biosynthesis genes in rice.

Molecular characterization of Hedera (Araliaceae) from Atlantic Iberian Peninsula

The Atlantic territories of western Europe and their surrounding areas are cohabited by two different species of ivies (Hedera), which are morphologically very similar, although they present different ploidy levels: Hedera helix (2x) and Hedera hibernica (4x). Concerning the northwest Atlantic Iberian territories and their surrounding areas, there are discrepancies regarding the identity of Hedera individuals at the specific level, since they have been identified as exclusively H. hibernica, but also as belonging to both species. In this context, we have aimed to determine whether the Hedera found in Atlantic Iberian Peninsula (Cantabrian Mountains territories and their surrounding areas) belong to H. helix, H. hibernica or to both. In order to achieve this, high-copy nuclear marker Internal Transcribed Spacer (ITS) and low-copy nuclear marker Granule-bound starch synthase I (GBSSI) were analyzed and compared to the results of Hedera samples from Central Europe and the Spanish Mediterranean Basin. Combined analyses of ITS-GBSSI datasets discriminate the species Hedera helix and Hedera hibernica, and our data suggest that H. hibernica is the only representative of Hedera in the Atlantic Iberian territories.

Analysis of Genetic Potential of Banyuwangi Local Rice (Oryza sativa L.) Based on Relative Expression of Homogentisate Geranylgeranyl Transferase (HGGT) and Granule-Bound Starch Synthase I (GBSSI) Gene

Rice nutrition including vitamin and amylose contents become important aspect for many people around the world. Rice with high amylose content (low glycemic index) is good for those with Diabetes mellitus. Tocotrienol, one precursor of Vitamin-E biosynthesis is catalyzed by enzymes encoded HGGT, while amylose biosynthesis is catalyzed by enzymes encoded GBSSI. The objective of this study was to find rice varieties with high tocotrienol and/or amylose content based on the expression of HGGT and GBSSI among eight Banyuwangi local rice varieties. Relative expression of HGGT and GBSSI was measured by qRT-PCR and analyzed using 2ΔCt method. Statistical analysis resulted in the significantly different of HGGT and GBSSI relative expression among samples. Relative expression of HGGT from the highest to the lowest were demonstrated by Hitam Melik, Hitam Pekat, Blambangan A3, Merah Bali, Blambangan A2, Berlian, Janur Kuning, and SOJ A3, respectively; while relative expression of GBSSI from the highest to the lowest were demonstrated by Hitam Melik, Hitam Pekat, SOJ A3, Janur Kuning, Berlian, Merah Bali, Blambangan A3, and Blambangan A2, respectively. Based on this research we conclude that Hitam Melik potentially produces higher tocotrienol and lower glycemic index than other studied varieties.