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Loss of copy number and expression of transgene during meiosis in Pyropia tenera

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Abstract

Pyropia is a genus of red algae that is cultivated as a commercial food and is a model organism in marine biotechnology. Its habitat is threatened by global climate change, resulting in considerable interest in developing productive strains that are tolerant to the changing climate. Such strains will be valuable only if they are genetically stable, but introduced genetic traits often are either weakened or completely lost in progeny. To determine and understand the likely fate of a newly acquired trait in Pyropia, we generated PyGUS transgenic P. tenera and analyzed the expression of the transgene and its copy number through its sexual life cycle. The vegetative and diploid cells of parent lines showed stable expression of the PyGUS gene, but approximately 30% of progeny had no histochemical staining for GUS and reduced levels of PyGUS mRNA, even though the transgene was present. Genomic qPCR analysis revealed that the copy number of the PyGUS gene was reduced in gametophyte thalli, which were generated after meiosis. We conclude that the PyGUS gene was markedly unstable during meiosis, and that loss of gene copies and the altered expression of the transgene occurred mostly during meiotic recombination. We anticipate that these results will benefit understanding of molecular mechanisms that confer stability to genetic traits in P. tenera and related species.

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References

  • Blouin NA, Brodie JA, Grossman AC, Xu P, Brawley SH (2011) Porphyra: a marine crop shaped by stress. Trends Plant Sci 16(1):29–37

    Article  CAS  Google Scholar 

  • Burzycki GM, Waaland JR (1987) On the position of meiosis in the life history of Porphyra torta (Rhodophyta). Bot Mar 30:5–10

  • Cai X, Xu SS (2007) Meiosis-driven genome variation in plants. Curr Genom 8(3):151–161

    Article  CAS  Google Scholar 

  • Cerutti H, Johnson AM, Gillham NW, Boynton JE (1997) Epigenetic silencing of a foreign gene in nuclear transformants of Chlamydomonas. Plant Cell 9(6):925–945

    Article  CAS  Google Scholar 

  • Choi S, Hwang MS, Im SO, Kim NJ, Jeong WJ, Park EJ, Gong YG, Choi DW (2013) Transcriptome sequencing and comparative analysis of the gametophyte thalli of Pyropia tenera under normal and high temperature conditions. J Appl Phycol 25(4):1237–1246

    Article  CAS  Google Scholar 

  • Cnudde F, Gerats T (2005) Meiosis : Inducing variation by reduction. Plant Biol 7:321–341

    Article  CAS  Google Scholar 

  • Ding H, Zhang B, Yan X (2016) Isolation and characterization of a heat-resistant strain with high yield of Pyropia yezoensis Ueda (Bangiales, Rhodophyta). Aquacult Fish 1:24–33

    Article  Google Scholar 

  • Fukuda S, Mikami K, Uji T, Park EJ, Ohba T, Asada K, Kitade Y, Endo H, Kato I, Saga N (2008) Factors influencing efficiency of transient gene expression in the red macrophyte Porphyra yezoensis. Plant Sci 174(3):329–339

    Article  CAS  Google Scholar 

  • Galagan JE, Calvo SE, Borkovich KA, Selker EU, Read ND, Jaffe D, FitzHugh W, Ma LJ et al (2003) The genome sequence of the filamentous fungus Neurospora crassa. Nature 422:859–868

    Article  CAS  Google Scholar 

  • Garcia N, Li Y, Dooner HK, Messing J (2017) Maize defective kernel mutant generated by insertion of a Ds element in a gene encoding a highly conserved TTI2 cochaperone. Proc Natl Acad Sci USA 114:5165–5170

    Article  CAS  Google Scholar 

  • Höck J, Weinmann L, Ender C, Rüdel S, Kremmer E, Raabe M, Urlaub H, Meister G (2007) Proteomic and functional analysis of Argonaute-containing mRNA-protein complexes in human cells. EMBO Rep 8(11):1052–1060

    Article  Google Scholar 

  • Huang LB, Yan XH (2019) Construction of a genetic linkage map in Pyropia yezoensis (Bangiales, Rhodophyta) and QTL analysis of several economic traits of blades. PLoS ONE 14(3):e0209128

    Article  CAS  Google Scholar 

  • Im S, Choi S, Hwang MS, Park EJ, Jeong WJ, Choi DW (2015) De novo assembly of transcriptome from the gametophyte of the marine red algae Pyropia seriata and identification of abiotic stress response genes. J Appl Phycol 27(3):1343–1353

    Article  CAS  Google Scholar 

  • Im S, Lee HN, Jung HS, Yang S, Park EJ, Hwang MS, Jeong WJ, Choi DW (2017) Transcriptome-based identification of the desiccation response genes in marine red algae Pyropia tenera (Rhodophyta) and enhancement of abiotic stress tolerance by PtDRG2 in Chlamydomonas. Mar Biotechnol 19(3):232–245

    Article  CAS  Google Scholar 

  • Kim NG (1999) Culture studies of Porphyra dentata and P. pseudolinearis (Bangiales, Rhodophyta), two dioecious species from Korea. Hydrobiologia 398:127–135

    Google Scholar 

  • Kim EJ, Ma XR, Cerutti H (2015) Gene silencing in microalgae: mechanisms and biological roles. Bioresour Technol 184:23–32

    Article  CAS  Google Scholar 

  • Korogodina VL, Korogodin VI, Simonyan NV, Maiorova ES (1995) Characteristics of spontaneous revertants in haploid yeast. Yeast 11:701–711

    Article  CAS  Google Scholar 

  • Lee DW, Pratt RJ, McLaughlin M, Aramayo R (2003) An argonaute-like protein is required for meiotic silencing. Genetics 164(2):821–828

    CAS  PubMed  PubMed Central  Google Scholar 

  • Li Z, Bock R (2018) Replication of bacterial plasmids in the nucleus of the red alga Porphyridium purpureum. Nat Commun 9:3451

    Article  Google Scholar 

  • Lim JM, Ahn JW, Hwangbo K, Choi DW, Park EJ, Hwang MS, Liu JR, Jeong WJ (2013) Development of cyan fluorescent protein (CFP) reporter system in green alga Chlamydomonas reinhardtii and macroalgae Pyropia sp. Plant Biotechnol Rep 7(3):407–414

    Article  Google Scholar 

  • Ma JH, Miura A (1984) Observations of the nuclear division in the conchospores and their germlings in Porphyra yezoensis Ueda. Jpn J Phycol 32:373–378 (in Japanese with English abstract)

    Google Scholar 

  • Matzke MA, Primig M, Trnovsky J, Matzke AJ (1989) Reversible methylation and inactivation of marker genes in sequentially transformed tobacco plants. EMBO J 8:643–649

    Article  CAS  Google Scholar 

  • Meza TJ, Kamfjord D, Håkelien AM, Evans I, Godager LH, Mandal A, Jakobsen KS, Aalen RB (2001) The frequency of silencing in Arabidopsis thaliana varies highly between progeny of siblings and can be influenced by environmental factors. Transgenic Res 10(1):53–67

    Article  CAS  Google Scholar 

  • Mikami K, Li L, Takahashi M (2012) Monospore-based asexual life cycle in Porphyra yezoensis. In: Mikami K (ed) Porphyra yezoensis: frontiers in physiological and molecular biological research. Nova Science, New York, pp 15–37

  • Mochizuki K, Gorovsky MA (2004) Small RNAs in genome rearrangement in Tetrahymena. Curr Opin Genet Dev 14:1–7

    Article  Google Scholar 

  • Ohme M, Miura A (1988) Tetrad analysis in conchospore germlings of Porphyra yezoensis (Rhodophyta, Bangiales). Plant Sci 57:135–140

    Article  Google Scholar 

  • Rajeev KS, Anunanthini P, Ramalingam S (2015) Epigenetic silencing in transgenic plants. Front Plant Sci 6:693

    Google Scholar 

  • Sahoo D, Tang X, Yarish C (2002) Porphyra—the economic seaweed as a new experimental system. Curr Sci 83:1313–1316

    Google Scholar 

  • Shi J, Chen Y, Xu Y, Ji D, Chen C, Xie C (2017) Differential proteomic analysis by iTRAQ reveals the mechanism of Pyropia haitanensis responding to high temperature stress. Sci Rep 7:44734

    Article  CAS  Google Scholar 

  • Shimizu A, Morishima K, Kobayashi M, Kunimoto M, Nakayama I (2008) Identification of Porphyra yezoensis (Rhodophyta) meiosis by DNA quantification using confocal laser scanning microscopy. J Appl Phycol 20:83–88

    Article  CAS  Google Scholar 

  • Shin JA, Miura A (1990) Estimation of the degree of self-fertilization in Porphyra yezoensis (Bangiales, Rhodophyta). Hydrobiologia 204(1):397–400

    Article  Google Scholar 

  • Shin YJ, Lim JM, Park JH, Choi DW, Hwang MS, Park EJ, Min SR, Kim SW, Jeong WJ (2016) Characterization of PyGUS gene silencing in the red macroalga Pyropia yezoensis. Plant Biotechnol Rep 10(6):359–367

    Article  Google Scholar 

  • Shin YJ, Min SR, Kang DY, Lim JM, Park EJ, Hwang MS, Choi DW, Ahn JW, Park YI, Jeong WJ (2018) Characterization of high temperature-tolerant strains of Pyropia yezoensis. Plant Biotechnol Rep 12:365–373

    Article  Google Scholar 

  • Son SH, Ahn JW, Uji T, Choi DW, Park EJ, Hwang MS, Liu JR, Choi DS, Mikami K, Jeong WJ (2012) Development of an expression system using the heat shock protein 70 promoter in the red macroalga Porphyra tenera. J Appl Phycol 24(1):79–87

    Article  CAS  Google Scholar 

  • Song MS, Rossi JJ (2017) Molecular mechanisms of Dicer: endonuclease and enzymatic activity. Biochem J 474(10):1603–1618

    Article  CAS  Google Scholar 

  • Turner DJ, Miretti M, Rajan D, Fiegler H, Carter NP, Blayney ML, Beck S, Hurles ME (2008) The rates of de novo meiotic deletions and duplications causing several genomic disorders in the male germline. Nat Genet 40(1):90–95

    Article  CAS  Google Scholar 

  • Uji T, Hirata R, Fukuda S, Mizuta H, Saga N (2014) A codon-optimized bacterial antibiotic gene used as selection marker for stable nuclear transformation in the marine red alga Pyropia yezoensis. Mar Biotechnol 16(3):251–255

    Article  CAS  Google Scholar 

  • Wang YX, Copnhaver GP (2018) Meiotic recombination: mixing it up in plants. Annu Rev Plant Biol 69:577–609

    Article  CAS  Google Scholar 

  • Yan XH, He LH, Aruga Y (2007) Karyological observations on the occurrence of meiosis in the life cycle of Porphyra haitanensis Chang et Zeng (Bangiales, Rhodophyta). Bot Mar 50:257–263

  • Yao MC, Fuller P, Xi X (2003) Programmed DNA deletion as an RNA-guided system of genome defense. Science 300:1581–1584

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the Golden Seed Project, Ministry of Agriculture, Food, and Rural Affairs (MAFRA) and Rural Development Administration (RDA) (213008-05-3-SB820), and a grant from the KRIBB Research Initiative program.

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Authors and Affiliations

  1. Plant System Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 34141, Republic of Korea

    Jong-Min Lim, Yoon Ju Shin, Sokyong Jung, Seung A. Choi, Sung Ran Min & Won-Joong Jeong

  2. Department of Biology Education, Chonnam National University, Gwangju, 61186, Republic of Korea

    Dong-Woog Choi

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  1. Jong-Min Lim
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  2. Yoon Ju Shin
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  3. Sokyong Jung
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  4. Seung A. Choi
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  5. Dong-Woog Choi
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  6. Sung Ran Min
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  7. Won-Joong Jeong
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Correspondence to Won-Joong Jeong.

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Lim, JM., Shin, Y.J., Jung, S. et al. Loss of copy number and expression of transgene during meiosis in Pyropia tenera. Plant Biotechnol Rep 13, 653–661 (2019). https://doi.org/10.1007/s11816-019-00563-y

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  • DOI: https://doi.org/10.1007/s11816-019-00563-y

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