Effects of Proteome Rebalancing and Sulfur Nutrition on the Accumulation of Methionine Rich δ-zein in Transgenic Soybeans

Overview

Journal Front Plant Sci

Date 2014 Nov 27

PMID 25426134

Citations 13

Authors

Won-Seok Kim

Joseph M Jez

Hari B Krishnan

Affiliations

Soon will be listed here.

Abstract

Expression of heterologous methionine-rich proteins to increase the overall sulfur amino acid content of soybean seeds has been only marginally successful, presumably due to low accumulation of transgenes in soybeans or due to gene silencing. Proteome rebalancing of seed proteins has been shown to promote the accumulation of foreign proteins. In this study, we have utilized RNAi technology to suppress the expression of the β-conglycinin, the abundant 7S seed storage proteins of soybean. Western blot and 2D-gel analysis revealed that β-conglycinin knockdown line (SAM) failed to accumulate the α', α, and β-subunits of β-conglycinin. The proteome rebalanced SAM retained the overall protein and oil content similar to that of wild-type soybean. We also generated transgenic soybean lines expressing methionine-rich 11 kDa δ-zein under the control of either the glycinin or β-conglycinin promoter. The introgression of the 11 kDa δ-zein into β-conglycinin knockdown line did not enhance the accumulation of the 11 kDa δ-zein. However, when the same plants were grown in sulfur-rich medium, we observed 3- to 16-fold increased accumulation of the 11 kDa δ-zein. Transmission electron microscopy observation revealed that seeds grown in sulfur-rich medium contained numerous endoplasmic reticulum derived protein bodies. Our findings suggest that sulfur availability, not proteome rebalancing, is needed for high-level accumulation of heterologous methionine-rich proteins in soybean seeds.

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References

Hagan N, Upadhyaya N, Tabe L, Higgins T . The redistribution of protein sulfur in transgenic rice expressing a gene for a foreign, sulfur-rich protein. Plant J. 2003; 34(1):1-11. DOI: 10.1046/j.1365-313x.2003.01699.x. View

Friedman A, Long S, Brown S, Buikema W, Ausubel F . Construction of a broad host range cosmid cloning vector and its use in the genetic analysis of Rhizobium mutants. Gene. 1982; 18(3):289-96. DOI: 10.1016/0378-1119(82)90167-6. View

Galili G, Amir R . Fortifying plants with the essential amino acids lysine and methionine to improve nutritional quality. Plant Biotechnol J. 2013; 11(2):211-22. DOI: 10.1111/pbi.12025. View

Hacham Y, Avraham T, Amir R . The N-terminal region of Arabidopsis cystathionine gamma-synthase plays an important regulatory role in methionine metabolism. Plant Physiol. 2002; 128(2):454-62. PMC: 148908. DOI: 10.1104/pp.010819. View

Tada Y, Utsumi S, Takaiwa F . Foreign gene products can be enhanced by introduction into low storage protein mutants. Plant Biotechnol J. 2006; 1(6):411-22. DOI: 10.1046/j.1467-7652.2003.00038.x. View

Cunha N, Araujo A, Leite A, Murad A, Vianna G, Rech E . Correct targeting of proinsulin in protein storage vacuoles of transgenic soybean seeds. Genet Mol Res. 2010; 9(2):1163-70. DOI: 10.4238/vol9-2gmr849. View

Bilyeu K, Zeng P, Coello P, Zhang Z, Krishnan H, Bailey A . Quantitative conversion of phytate to inorganic phosphorus in soybean seeds expressing a bacterial phytase. Plant Physiol. 2007; 146(2):468-77. PMC: 2245832. DOI: 10.1104/pp.107.113480. View

Ding S, Huang L, Wang Y, Sun H, Xiang Z . High-level expression of basic fibroblast growth factor in transgenic soybean seeds and characterization of its biological activity. Biotechnol Lett. 2006; 28(12):869-75. DOI: 10.1007/s10529-006-9018-6. View

Harada J, Barker S, Goldberg R . Soybean beta-conglycinin genes are clustered in several DNA regions and are regulated by transcriptional and posttranscriptional processes. Plant Cell. 1989; 1(4):415-25. PMC: 159773. DOI: 10.1105/tpc.1.4.415. View

10.

Nielsen N, Dickinson C, Cho T, Thanh V, Scallon B, Fischer R . Characterization of the glycinin gene family in soybean. Plant Cell. 1989; 1(3):313-28. PMC: 159764. DOI: 10.1105/tpc.1.3.313. View

11.

Shigemitsu T, Ozaki S, Saito Y, Kuroda M, Morita S, Satoh S . Production of human growth hormone in transgenic rice seeds: co-introduction of RNA interference cassette for suppressing the gene expression of endogenous storage proteins. Plant Cell Rep. 2011; 31(3):539-49. DOI: 10.1007/s00299-011-1191-y. View

12.

Frizzi A, Caldo R, Morrell J, Wang M, Lutfiyya L, Brown W . Compositional and transcriptional analyses of reduced zein kernels derived from the opaque2 mutation and RNAi suppression. Plant Mol Biol. 2010; 73(4-5):569-85. DOI: 10.1007/s11103-010-9644-1. View

13.

Gayler K, Sykes G . Effects of nutritional stress on the storage proteins of soybeans. Plant Physiol. 1985; 78(3):582-5. PMC: 1064779. DOI: 10.1104/pp.78.3.582. View

14.

Tabe L, Wirtz M, Molvig L, Droux M, Hell R . Overexpression of serine acetlytransferase produced large increases in O-acetylserine and free cysteine in developing seeds of a grain legume. J Exp Bot. 2009; 61(3):721-33. PMC: 2814105. DOI: 10.1093/jxb/erp338. View

15.

Wesley S, Helliwell C, Smith N, Wang M, Rouse D, Liu Q . Construct design for efficient, effective and high-throughput gene silencing in plants. Plant J. 2001; 27(6):581-90. DOI: 10.1046/j.1365-313x.2001.01105.x. View

16.

Kim W, Chronis D, Juergens M, Schroeder A, Hyun S, Jez J . Transgenic soybean plants overexpressing O-acetylserine sulfhydrylase accumulate enhanced levels of cysteine and Bowman-Birk protease inhibitor in seeds. Planta. 2011; 235(1):13-23. DOI: 10.1007/s00425-011-1487-8. View

17.

Taylor M, Chapman R, Beyaert R, Hernandez-Sebastia C, Marsolais F . Seed storage protein deficiency improves sulfur amino acid content in common bean (Phaseolus vulgaris L.): redirection of sulfur from gamma-glutamyl-S-methyl-cysteine. J Agric Food Chem. 2008; 56(14):5647-54. DOI: 10.1021/jf800787y. View

18.

Tabe L, Droux M . Sulfur assimilation in developing lupin cotyledons could contribute significantly to the accumulation of organic sulfur reserves in the seed. Plant Physiol. 2001; 126(1):176-87. PMC: 102292. DOI: 10.1104/pp.126.1.176. View

19.

Takahashi M, Uematsu Y, Kashiwaba K, Yagasaki K, Hajika M, Matsunaga R . Accumulation of high levels of free amino acids in soybean seeds through integration of mutations conferring seed protein deficiency. Planta. 2003; 217(4):577-86. DOI: 10.1007/s00425-003-1026-3. View

20.

Thanh V, Shibasaki K . Major proteins of soybean seeds. A straightforward fractionation and their characterization. J Agric Food Chem. 1976; 24(6):1117-21. DOI: 10.1021/jf60208a030. View