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Re-programming of gene expression in the CS8 rice line over-expressing ADPglucose pyrophosphorylase induces a suppressor of starch biosynthesis.
Plant J. 2019 03; 97(6):1073-1088.PJ

Abstract

The CS8 transgenic rice (Oryza sativa L.) lines expressing an up-regulated glgC gene produced higher levels of ADPglucose (ADPglc), the substrate for starch synthases. However, the increase in grain weight was much less than the increase in ADPglc levels suggesting one or more downstream rate-limiting steps. Endosperm starch levels were not further enhanced in double transgenic plants expressing both glgC and the maize brittle-1 gene, the latter responsible for transport of ADPglc into the amyloplast. These studies demonstrate that critical processes within the amyloplast stroma restrict maximum carbon flow into starch. RNA-seq analysis showed extensive re-programming of gene expression in the CS8 with 2073 genes up-regulated and 140 down-regulated. One conspicuous gene, up-regulated ~15-fold, coded for a biochemically uncharacterized starch binding domain-containing protein (SBDCP1) possessing a plastid transit peptide. Confocal microscopy and transmission electron microscopy analysis confirmed that SBDCP1 was located in the amyloplasts. Reciprocal immunoprecipitation and pull-down assays indicated an interaction between SBDCP1 and starch synthase IIIa (SSIIIa), which was down-regulated at the protein level in the CS8 line. Furthermore, binding by SBDCP1 inhibited SSIIIa starch polymerization activity in a non-competitive manner. Surprisingly, artificial microRNA gene suppression of SBDCP1 restored protein expression levels of SSIIIa in the CS8 line resulting in starch with lower amylose content and increased amylopectin chains with a higher degree of polymerization. Collectively, our results support the involvement of additional non-enzymatic factors such as SBDCP in starch biosynthesis.

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Authors+Show Affiliations

Institute of Biological Chemistry, Washington State University, Pullman, WA, 99164, USA.

Institute of Biological Chemistry, Washington State University, Pullman, WA, 99164, USA.

Faculty of Bioresource Science, Akita Prefectural University, Shimoshinjo-Nakano, Akita-City, 010-0195, Japan.

Institute of Biological Chemistry, Washington State University, Pullman, WA, 99164, USA.

Institute of Biological Chemistry, Washington State University, Pullman, WA, 99164, USA.

Institute of Biological Chemistry, Washington State University, Pullman, WA, 99164, USA.

Institute of Biological Chemistry, Washington State University, Pullman, WA, 99164, USA.

Institute of Biological Chemistry, Washington State University, Pullman, WA, 99164, USA.

Institute of Biological Chemistry, Washington State University, Pullman, WA, 99164, USA.

Faculty of Bioresource Science, Akita Prefectural University, Shimoshinjo-Nakano, Akita-City, 010-0195, Japan.

Institute of Biological Chemistry, Washington State University, Pullman, WA, 99164, USA.

MeSH

Carbohydrate MetabolismDown-RegulationEndospermGene ExpressionGene Expression ProfilingGlucose-1-Phosphate AdenylyltransferaseOryzaPlant ProteinsPlants, Genetically ModifiedPlastidsStarchStarch SynthaseUp-RegulationZea mays

Pub Type(s)

Journal Article

Research Support, Non-U.S. Gov't

Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

30523657

Citation

Cakir, Bilal, et al. "Re-programming of Gene Expression in the CS8 Rice Line Over-expressing ADPglucose Pyrophosphorylase Induces a Suppressor of Starch Biosynthesis." The Plant Journal : for Cell and Molecular Biology, vol. 97, no. 6, 2019, pp. 1073-1088.

Cakir B, Tian L, Crofts N, et al. Re-programming of gene expression in the CS8 rice line over-expressing ADPglucose pyrophosphorylase induces a suppressor of starch biosynthesis. Plant J. 2019;97(6):1073-1088.

Cakir, B., Tian, L., Crofts, N., Chou, H. L., Koper, K., Ng, C. Y., Tuncel, A., Gargouri, M., Hwang, S. K., Fujita, N., & Okita, T. W. (2019). Re-programming of gene expression in the CS8 rice line over-expressing ADPglucose pyrophosphorylase induces a suppressor of starch biosynthesis. The Plant Journal : for Cell and Molecular Biology, 97(6), 1073-1088. https://doi.org/10.1111/tpj.14180

Cakir B, et al. Re-programming of Gene Expression in the CS8 Rice Line Over-expressing ADPglucose Pyrophosphorylase Induces a Suppressor of Starch Biosynthesis. Plant J. 2019;97(6):1073-1088. PubMed PMID: 30523657.

* Article titles in AMA citation format should be in sentence-case

TY - JOUR T1 - Re-programming of gene expression in the CS8 rice line over-expressing ADPglucose pyrophosphorylase induces a suppressor of starch biosynthesis. AU - Cakir,Bilal, AU - Tian,Li, AU - Crofts,Naoko, AU - Chou,Hong-Li, AU - Koper,Kaan, AU - Ng,Chun-Yeung, AU - Tuncel,Aytug, AU - Gargouri,Mahmoud, AU - Hwang,Seon-Kap, AU - Fujita,Naoko, AU - Okita,Thomas W, Y1 - 2019/01/14/ PY - 2018/09/27/received PY - 2018/11/23/revised PY - 2018/11/27/accepted PY - 2018/12/14/pubmed PY - 2020/4/9/medline PY - 2018/12/8/entrez KW - ADPglucose KW - CBM20 KW - Oryza sativa L. KW - SSIIIa KW - starch synthesis SP - 1073 EP - 1088 JF - The Plant journal : for cell and molecular biology JO - Plant J VL - 97 IS - 6 N2 - The CS8 transgenic rice (Oryza sativa L.) lines expressing an up-regulated glgC gene produced higher levels of ADPglucose (ADPglc), the substrate for starch synthases. However, the increase in grain weight was much less than the increase in ADPglc levels suggesting one or more downstream rate-limiting steps. Endosperm starch levels were not further enhanced in double transgenic plants expressing both glgC and the maize brittle-1 gene, the latter responsible for transport of ADPglc into the amyloplast. These studies demonstrate that critical processes within the amyloplast stroma restrict maximum carbon flow into starch. RNA-seq analysis showed extensive re-programming of gene expression in the CS8 with 2073 genes up-regulated and 140 down-regulated. One conspicuous gene, up-regulated ~15-fold, coded for a biochemically uncharacterized starch binding domain-containing protein (SBDCP1) possessing a plastid transit peptide. Confocal microscopy and transmission electron microscopy analysis confirmed that SBDCP1 was located in the amyloplasts. Reciprocal immunoprecipitation and pull-down assays indicated an interaction between SBDCP1 and starch synthase IIIa (SSIIIa), which was down-regulated at the protein level in the CS8 line. Furthermore, binding by SBDCP1 inhibited SSIIIa starch polymerization activity in a non-competitive manner. Surprisingly, artificial microRNA gene suppression of SBDCP1 restored protein expression levels of SSIIIa in the CS8 line resulting in starch with lower amylose content and increased amylopectin chains with a higher degree of polymerization. Collectively, our results support the involvement of additional non-enzymatic factors such as SBDCP in starch biosynthesis. SN - 1365-313X UR - https://www.unboundmedicine.com/medline/citation/30523657/Re_programming_of_gene_expression_in_the_CS8_rice_line_over_expressing_ADPglucose_pyrophosphorylase_induces_a_suppressor_of_starch_biosynthesis_ DB - PRIME DP - Unbound Medicine ER -