• Title/Summary/Keyword: GBSSI

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Molecular Characterization of Granule-Bound Starch Synthase (GBSSI) gene of Waxy Locus Mutants in Japonica Rice (Oryza sativa L.)

  • Sohn, Seong-Han;Rhee, Yong;Hwang, Duk-Ju;Lee, Sok-Young;Lee, Jung-Ro;Lee, Yeon-Hee;Shin, Young-Seop;Jeung, Ji-Ung;Kim, Myung-Ki
    • Korean Journal of Breeding Science
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    • v.42 no.1
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    • pp.1-10
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    • 2010
  • Five mutants were investigated at the molecular level to determine the factors responsible for mutated endosperm types. They were classified as high (HA) or low amylose (LA) phenotypes based on the amylose content in endosperm. The five were previously produced from Ilpum and Shindongjin cultivar treated with N-methyl-N-nitrosourea and gamma-ray irradiation, respectively. Analysis of the genomic structure and expression of Granule-bounded Starch Synthase I (GBSSI) genes revealed that mutants generally showed a higher incidence of nucleotide transition than transversion, and the $A:T{\rightarrow}G:C$ transition was particularly prevalent. The rates of nucleotide substitution in HA mutants were generally higher than those in the LA mutants, leading to higher substitutions of amino acid in the HA mutants. Neither nucleotide substitutions interfering with intron splicing or causing early termination of protein translation were found, nor any large-sized deletions or additions were found in all the mutants. In principle, amylose content can be regulated by three factors: internal alterations of GBSSI protein, the strength of gene expression, and other unknown external factors. Our results indicate that the endosperm mutants from Shindongjin arose from internal alterations of GBSSI proteins, which may be the result of amino acid substitutions. On the other hand, the Ilpum mutants might be principally caused by the alteration of gene expression level. Analysis of another three glutinous cultivars revealed that the major factor leading to glutinous phenotypes is the 23-bp duplicative motif (5'-ACGGGTTCCAGGGCCTCAAGCCC-3') commonly found in exon 2, which results in the premature termination of protein translation leading to the production of a non-functional GBSSI enzyme.

Granule-Bound Starch Synthase I (GBSSI): An Evolutionary Perspective and Haplotype Diversification in Rice Cultivars

  • Sang-Ho Chu;Gi Whan Baek;Yong-Jin Park
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.219-219
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    • 2022
  • Granule-bound starch synthase I (GBSSI), encoded by the waxy gene, is responsible for the accumulation of amylose during the development of starch granules in rice endosperm. Despite many findings on waxy alleles, the genetic diversity and evolutionary studies are still not fully explored regarding their functional effects. Comprehensive evolutionary analyses were performed to investigate the genetic variations and relatedness of the GBSSI gene in 374 rice accessions composed of 54 wild accessions and 320 bred cultivars (temperate japonica, tropical japonica, indica, aus, aromatic, and admixture). GBSS1 coding regions were analyzed from a VCF file retrieved from whole-genome resequencing data, and eight haplotypes were identified in the GBSSI coding region of 320 bred cultivars. The genetic diversity indices revealed the most negative Tajima's D value in the tropical-japonica, followed by the aus and temperate-japonica, while Tajima's D values in indica were positive, indicating balancing selection. Diversity reduction was noticed in temperate japonica (0.0003) compared to the highest one (wild, 0.0044), illustrating their higher genetic differentiation by FST-value (0.604). The most positive Tajima's D value was observed in indica (0.5224), indicating the GBSSI gene domestication signature under balancing selection. In contrast, the lowest and negative Tajima's D value was found in tropical japonica (-0.5291), which might have experienced a positive selection and purified due to the excess of rare alleles. Overall, our study offers insights into haplotype diversity and evolutionary fingerprints of GBSSI. It ako provides genomic information to increase the starch content of cooked rice.

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Modification of amylose content of sweetpotato starch by RNAi technique

  • Shimada, Takiko;Otani, Motoyasu;Hamada, Tatsurou;Kim, Sun-Hyung
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.351-355
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    • 2005
  • In the storage roots of sweetpotato (Ipomoea batatas (L.) Lam. cv. Kokei 14), 10 to 20% of starch is essentially unbranched linear amylose and the other major component is branched amylopectin. Amylose is produced by the enzyme GBSSI (granule bound starch synthase I), whereas amylopectin is produced by a concerted action of soluble starch synthase and starch branching enzymes (SBEI and SBEII). We constructed double-stranded RNA (dsRNA) interference vectors of GBSSI and IbSBEII and introduced them into sweetpotato genome via Agrobacterium-mediated gene transformation. The endogenous GBSSI expression was inhibited by dsRNA of GBSSI in 73 % of transgenic plants giving rise to the storage tubers containing amylopectin but not amylose. On the other hand, all sweetpotato plants transformed with dsRNA of IbSBEII contained a larger amount of amylose than the non-transgenic control (up to 25% compared to 10% in the controls). The RNA interference (RNAi) is effectively inhibited the gene expression in thestarch metabolic pathway and modified the characteristics of starch in sweetpotato.

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Variation of Amylose Content Using dsRNAi Vector by Targeting 3'-UTR Region of GBSSI Gene in Rice (GBSSI 유전자 3'UTR 영역의 발현 억제 dsRNAi 벡터를 이용한 아밀로스함량 조절 벼 개발)

  • Park, Hyang-Mi;Choi, Man-Soo;Chun, Areum;Lee, Jeung-Heui;Kim, Myeong-Ki;Kim, Yeon-Gyu;Shin, Dong-Bum;Lee, Jang-Yong;Kim, Yul-Ho
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.515-524
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    • 2010
  • The amylose content of starch is a major factor in the texture of cooked cereal grains. Therefore, down-regulation of amylose synthesis is one of the alternative method to improve eating quality of rice. We developed transgenic rice plants designed to suppress granule-bound starch synthase I(GBSSI) gene using RNA interference(RNAi) technology. Transgenic plants with RNAi vector containing the 3'-UTR region of GBSSI showed a lower amylose content in rice endosperm than that of wild-type. The range of amylose content was 5.9~9.0% in the transgenic plants, whereas that of wild-type was 17.7~18.0%. Transgenic rices showed the decrease of short chain and the increase of long chain by analyzing chain length distribution of amylopectin in the endosperm. In the SEM micrographs, we found that compound starch granules in whole grains of the wild-type rice were readily split during fracturing, while the starch granules in RNAi-transgenic lines showed small voluminous, non-angular rounded bodies.

Use of Single-leaf Cutting in the Study of the Expression of Starch Synthesis and Modification Genes in Sweetpotato

  • Kim Sun-Hyung;Hamada Tatsuro;Otani Matoyasu;Koga Hironori;Shimada Takiko
    • Journal of Plant Biotechnology
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    • v.7 no.2
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    • pp.123-127
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    • 2005
  • The evaluation of source potential and sink strength is the generally large and laborious sample size required to adequately assess anyone of the parameters in field-grown sweetpotato. For this purpose we used the rooted single-leaf cuttings with petioles, because the source and sink organs are restricted in this system. The rooted single-leaf cutting of sweetpotato provides a unique source-sink model system, and is established within about 50 days after planting. In this study, the sink potential of sweetpotato tubers was examined based on the expression of genes for starch synthesis (AGPase) and modification (SBEII and GBSSI) in single rooted leaf plant. The gene expression patterns of GBSSI, SBEII and AGPase at various developmental stages and in different types of root tissues presented. These results suggest that the rooted single-rooted method can be used an ideal model system to study physiological and biochemical mechanisms in sweetpotato.

Allelic Variation of Glutenin, Granule-Bound Starch Synthase l and Puroindoline in Korean Wheat Cultivar

  • Park, Chul-Soo;Pena, Roberto J.;Baik, Byung-Kee;Kang, Chon-Sik;Heo, Hwa-Young;Cheong, Young-Keun;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.54 no.2
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    • pp.181-191
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    • 2009
  • To investigate the genetic variation of high-and low-molecular-weight glutenin subunits (BMW-GS and LMW-GS), granule-bound starch synthase I (GBSSI) and puroindoline in 24 Korean wheat cultivars. At the BMW-GS compositions, three Glu-A1 alleles, five Glu-B1 alleles and three Glu-D1 alleles were identified. The high frequency of alleles at each locus was Glu-A1c allele (15 cultivars), Glu-B1b allele (16 cultivars) and Glu-D1f allele (16 cultivars). Four alleles were identified at the Glu-A3 and Glu-B3 loci and three at Glu-D3 locus and Glu-A3d, Glu-B3d and Glu-D3a were mainly found at each Glu-3 locus. Glu-A3d, Glu-B3d, Glu-D3b or c (4 cultivars, respectively) and Glu-A3d, Glu-B3d, Glu-D3a and Glu-A3c, Glu-B3d or h, Glu-D3a (3 cultivar, respectively) were predominantly found in Korean wheats. At the GBSS compositions, 2 waxy wheat cultivars, Shinmichal and Shinmichal1, showed null alleles on the Wx loci and other cultivars were wild type in GBSS compositions. At the puroindoline gene compositions, Korean wheat cultivars carried 3 genotypes, which 10 cultivars (41.7%) were Pina-D1a and Pinb-D1a, 11 cultivars (45.8%) had Pina-D1a and Pinb-D1b and 3 cultivars (12.5%) carried Pina-D1b and Pinb-D1a. These genetic variations could present the information to improve flour and end-use quality in Korean wheat breeding programs.

Disentangling Evolutionary Pattern and Haplotype Distribution of Starch Synthase III-1 (SSIIIb) in Korean Rice Collection

  • Bhagwat Nawade ;Yong-Jin Park
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.214-214
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    • 2022
  • Soluble starch synthases (SSs) elongate α-glucans from ADP-Glc to the glucan nonreducing ends and play a critical role in synthesizing resistant starch in the rice. A total of 10 SSs isoforms were reported in rice, including granules-bound starch synthase I (GBSSI), GBSSII, starch synthase I (SSI), SSIIa (SSII-3), SSIIb (SSII-2), SSIIc (SSII-1), SSIIIa (SSIII-2), SSIIIb (SSIII-1), SSIVa (SSIV-1), and SSIVb (SSIV-2). SSIII proteins are involved in forming the B chain and elongating cluster filling chains in amylopectin metabolism. The functions of SSIIIb (SSIII-1) are less clear as compared to SSs. Here, we sought to shed light on the genetic diversity profiling of the SSIII-1 gene in 374 rice accessions composed of 54 wild-type accessions and 320 bred cultivars (temperate japonica, indica, tropical japonica, aus, aromatic, and admixture). In total, 17 haplotypes were identified in the SSIII-1 coding region of 320 bred cultivars, while 44 haplotypes were detected from 54 wild-type accessions. The genetic diversity indices revealed the most negative Tajima's D value in the temperate-japonica, followed by the wild type, while Tajima's D values in other ecotypes were positive, indicating balancing selection. Nucleotide diversity in the SSIII-1 region was highest in the wild group (0.0047) while lowest in temperate-japonica. Lower nucleotide diversity in the temperate-japonica is evidenced by the negative Tajima's D and suggested purifying selection. The fixation index (FST) revealed a very high level of gene flow (low FST) between the tropical-japonica and admixture groups (FST=-0.21) followed by admixture and wild groups (-0.04), indica and admixture groups (0.02), while low gene flow with higher FST estimates between the temperate-japonica and aus groups (0.72), tropical-japonica and aromatic groups (0.71), and temperate-japonica and admixture groups (0.52). Taken together, our study offers insights into haplotype diversity and evolutionary fingerprints of SSIII-1. It provides genomic information to increase the resistant starch content of cooked rice.

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The change of grain quality and starch assimilation of rice under future climate conditions according to RCP 8.5 scenario (RCP 8.5 시나리오에 따른 미래 기후조건에서 벼의 품질 및 전분 동화 특성 변화)

  • Sang, Wan-Gyu;Cho, Hyeoun-Suk;Kim, Jun-Hwan;Shin, Pyong;Baek, Jae-Kyeong;Lee, Yun-Ho;Cho, Jeong-Il;Seo, Myung-Chul
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.20 no.4
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    • pp.296-304
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    • 2018
  • The objective of this study was to analyze the impact of climate change on rice yield and quality. Experiments were conducted using SPAR(Soil-Plant-Atmosphere-Research) chambers, which was designed to create virtual future climate conditions, in the National Institute of Crop Science, Jeonju, Korea, in 2016. In the future climate conditions($+2.8^{\circ}C$ temp, 580 ppm $CO_2$) of year 2051~2060 according to RCP 8.5 scenario, elevated temperature and $CO_2$ accelerated the heading date by about five days than the present climate conditions, resulted in a high temperature environment during grain filling stage. Rice yield decreased sharply in the future climate conditions due to the high temperature induced poor ripening. And the spikelet numbers, ripening ratio, and 1000-grain weight of brown rice were significantly decreased compared to control. The rice grain quality was also decreased sharply, especially due to the increased immature grains. In the future climate conditions, expression of starch biosynthesis-related genes such as granule-bound starch synthase(GBSSI, GBSSII, SSIIa, SSIIb, SSIIIa), starch branching enzyme(BEIIb) and ADP-glucose pyrophosphorylase(AGPS1, AGPS2, AGPL2) were repressed in developing seeds, whereas starch degradation related genes such as ${\alpha}-amylase$(Amy1C, Amy3D, Amy3E) were induced. These results suggest that the reduction in yield and quality of rice in the future climate conditions is likely caused mainly by the poor grain filling by high temperature. Therefore, it is suggested to develop tolerant cultivars to high temperature during grain filling period and a new cropping system in order to ensure a high quality of rice in the future climate conditions.