Improvement of recombinant miraculin production in transgenic tomato by crossbreeding-based genetic background modification

Transgenic Research - 2022
Kyoko Hiwasa-Tanase1,2, Suzuno Ohmura3, Natsumi Kitazawa3, Azusa Ono3, Takeshi Suzuki3, Hiroshi Ezura2,1
1Faculty of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Japan
2Tsukuba-Plant Innovation Research Center, University of Tsukuba, Tsukuba, Japan
3Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Japan

Tóm tắt

An important optimization step in plant-based recombinant protein production systems is the selection of an appropriate cultivar after a potential host has been determined. Previously, we have shown that transgenic tomatoes of the variety ‘Micro-Tom’ accumulate incredibly high levels of miraculin (MIR) due to the introduction of MIR gene controlled by a CaMV35S promoter and a heat-shock protein terminator. However, ‘Micro-Tom’ is unsuitable for commercial production of MIR as it is a dwarf cultivar characterized by small-sized fruit and poor yield. Here, we used the crossbreeding approach to transfer the high MIR accumulation trait of transgenic ‘Micro-Tom’ tomatoes to ‘Natsunokoma’ and ‘Aichi First’, two commercial cultivars producing medium and large fruit sizes, respectively. Fruits of the resultant crossbred lines were larger (~ 95 times), but their miraculin accumulation levels (~ 1,062 μg/g fresh mass) were comparable to the donor cultivar, indicating that the high miraculin accumulation trait was preserved regardless of fruit size or cultivar. Further, the transferred trait resulted in a 3–4 fold increase in overall miraculin production than that of the previously reported line 5B. These findings demonstrate the effectiveness of crossbreeding in improving MIR production in tomatoes and could pave the way for a more efficient production of recombinant proteins in other plants.

Từ khóa


Tài liệu tham khảo

Cheniclet C, Rong WY, Causse M, Frangne N, Bolling L, Bordeaux VS, Ornon V (2005) Cell expansion and endoreduplication show a large genetic variability in pericarp and contribute strongly to tomato fruit growth. Plant Physiol 139:1984–1994. https://doi.org/10.1104/pp.105.068767

Cunha NB, Murad AM, Cipriano TM, Araujo ACG, Aragao FJL, Leite A, Vianna GR, McPhee TR, Souza GHMF, Waters MJ, Rech EL (2011) Expression of functional recombinant human growth hormone in transgenic soybean seeds. Transgenic Res 20:811–826. https://doi.org/10.1007/s11248-010-9460-z

Daniell H, Streatfield SJ, Wycoff K (2001) Medical molecular farming: production of antibodies, biopharmaceuticals and edible vaccines in plants. Trends Plant Sci 6:219–226. https://doi.org/10.1016/S1360-1385(01)01922-7

Dong OX, Ronald P (2021) Targeted DNA insertion in plants. Proc Natl Acad Sci USA 118:e2004834117. https://doi.org/10.1073/pnas.2004834117

Ezura H, Hiwasa-Tanase K (2018) Mass production of the taste-modifying protein miraculin in transgenic plants. In: Merillon JM, Ramawat K (eds) Sweeteners, reference series in phytochemistry. Springer, Cham, pp 1–18

Conley AJ, Zhu H, Le LC, Jevnikar AM, Lee BH, Brandle JE, Menassa R (2011) Recombinant protein production in a variety of Nicotiana hosts: a comparative analysis. Plant Biotechnol J 9:434–444. https://doi.org/10.1111/j.1467-7652.2010.00563.x

Hanson WD (1959) Early generation analysis of lengths of heterozygous chromosome segments around a locus held heterozygous with backcrossing or selfing. Genetics 44:833–837

Hirai T, Kurokawa N, Duhita N, Hiwasa-Tanase K, Kato K, Kato K, Ezura H (2011) The HSP terminator of Arabidopsis thaliana induces extremely high-level accumulation of miraculin protein in transgenic tomato. J Agric Food Chem 59:9942–9949. https://doi.org/10.1021/jf202501e

Hirai T, Sato M, Toyooka K, Sun HJ, Yano M, Ezura H (2010) Miraculin, a taste-modifying protein is secreted into intercellular spaces in plant cells. J Plant Physiol 167:209–215. https://doi.org/10.1016/j.jplph.2009.08.001

Hiwasa-Tanase K, Hirai T, Kato K, Duhita N, Ezura H (2012) From miracle fruit to transgenic tomato: mass production of the taste-modifying protein miraculin in transgenic plants. Plant Cell Rep 31:513–525. https://doi.org/10.1007/s00299-011-1197-5

Hiwasa-Tanase K, Yano T, Kon T, Terakawa T, Ezura H (2021) Environmental risk assessment of transgenic miraculin-accumulating tomato in a confined field trial in Japan. Plant Biotechnol 38:421–431. https://doi.org/10.5511/plantbiotechnology.21.1021a

Horn ME, Woodard SL, Howard JA (2004) Plant molecular farming: systems and products. Plant Cell Rep 22:711–720. https://doi.org/10.1007/s00299-004-0767-1

Hospital F (2001) Size of donor chromosome segments around introgressed loci and reduction of linkage drag in marker-assisted backcross programs. Genetics 158:1363–1379. https://doi.org/10.1093/genetics/158.3.1363

Ito K, Asakura T, Morita Y, Nakajima K, Koizumi A, Shimizu-Ibuka A, Masuda K, Ishiguro M, Terada T, Maruyama J, Kitamoto K, Misaka T, Abe K (2007) Microbial production of sensory-active miraculin. Biochem Biophys Res Commun 360:407–411. https://doi.org/10.1016/j.bbrc.2007.06.064

Ito K, Sugawara T, Koizumi A, Nakajima K, Shimizu-Ibuka A, Shiroishi M, Asada H, Yurugi-Kobayashi T, Shimamura T, Asakura T, Masuda K, Ishiguro M, Misaka T, Iwata S, Kobayashi T, Abe K (2010) Bulky high-mannose-type N-glycan blocks the taste-modifying activity of miraculin. Biochim Biophys Acta 1800:986–992. https://doi.org/10.1016/j.bbagen.2010.06.003

Joseph JA, Akkermans S, Nimmegeers P, Van Impe JF (2019) Bioproduction of the recombinant sweet protein thaumatin: current state of the art and perspectives. Front Microbiol 10:695. https://doi.org/10.3389/fmicb.2019.00695

Kajiura H, Hiwasa-Tanase K, Ezura H, Fujiyama K (2018) Comparison of the N-glycosylation on recombinant miraculin expressed in tomato plants with native miraculin. Plant Biotechnol 35:375–379. https://doi.org/10.5511/plantbiotechnology.18.1023a

Kajiura H, Hiwasa-Tanase K, Ezura H, Fujiyama K (2022) Effect of fruit maturation on N-glycosylation of plant-derived native and recombinant miraculin. Plant Physiol Biochem 178:70–79. https://doi.org/10.1016/j.plaphy.2022.02.026

Kant R (2005) Sweet proteins – Potential replacement for artificial low calorie sweeteners. Nutr J 4:5. https://doi.org/10.1186/1475-2891-4-5

Kato K, Maruyama S, Hirai T, Hiwasa-Tanase K, Mizoguchi T, Goto E, Ezura H (2011) A trial of production of the plant-derived high-value protein in a plant factory. Plant Signal Behav 6:1172–1179. https://doi.org/10.4161/psb.6.8.16373

Kim YW, Hirai T, Kato K, Hiwasa-Tanase K, Ezura H (2010a) Gene dosage and genetic background affect miraculin accumulation in transgenic tomato fruits. Plant Biotechnol 27:333–338. https://doi.org/10.5511/plantbiotechnology.27.333

Kim YW, Kato K, Hirai T, Hiwasa-Tanase K, Ezura H (2010b) Spatial and developmental profiling of miraculin accumulation in transgenic tomato fruits expressing the miraculin gene constitutively. J Agric Food Chem 58:282–286. https://doi.org/10.1021/jf9030663

Kolotilin I, Kaldis A, Pereira EO, Laberge S, Menassa R (2013) Optimization of transplastomic production of hemicellulases in tobacco: effects of expression cassette configuration and tobacco cultivar used as production platform on recombinant protein yields. Biotechnol Biofuels 6:65. https://doi.org/10.1186/1754-6834-6-65

Kurihara K, Beidler LM (1968) Taste-modifying protein from miracle fruit. Science 161:1241–1243. Doi: https://doi.org/10.1126/science.161.3847.1241

Kurihara K, Beidler LM (1969) Mechanism of the action of taste-modifying protein. Nature 222:1176–1179. https://doi.org/10.1038/2221176a0

Lu Y, Tian Y, Shen R, Yao Q, Wang M, Chen M, Dong J, Zhang T, Li F, Lei M, Zhu J-K (2020) Targeted, efficient sequence insertion and replacement in rice. Nat Biotechnol 38:1402–1407. https://doi.org/10.1038/s41587-020-0581-5

Ma X, Zhu Q, Chen Y, Liu YG (2016) CRISPR/Cas9 platforms for genome editing in plants: Developments and Applications. Mol Plant 9:961–974. https://doi.org/10.1016/j.molp.2016.04.009

Maliga P (2003) Progress towards commercialization of plastid transformation technology. Trends Biotechnol 121:20–28. https://doi.org/10.1016/S0167-7799(02)00007-0

Matsuyama T, Satoh M, Nakata R, Aoyama T, Inoue H (2009) Functional expression of miraculin, a taste-modifying protein in Escherichia coli. J Biochem 145:445–450. Doi: https://doi.org/10.1093/jb/mvn184

Meyers B, Zaltsman A, Lacroix B, Kozolovsky SV, Krichevsky A (2010) Nuclear and plastid genetic engineering of plants: comparison of opportunities and challenges. Biotechnol Adv 28:747–756. https://doi.org/10.1016/j.biotechadv.2010.05.022

Mooradian AD, Smith M, Tokuda M (2017) The role of artificial and natural sweeteners in reducing the consumption of table sugar: a narrative review. Clin Nutr ESPEN 18:1–8. https://doi.org/10.1016/j.clnesp.2017.01.004

Nagaya S, Kawamura K, Shinmyo A, Kato K (2010) The HSP terminator of Arabidopsis thaliana increases gene expression in plant cells. Plant Cell Physiol 51:328–332. https://doi.org/10.1093/pcp/pcp188

Naveira H, Barbadilla A (1992) The theoretical distribution of lengths of intact chromosome segments around a locus held heterozygous with backcrossing in a diploid species. Genetics 130:205–209. https://doi.org/10.1093/genetics/130.1.205

Obembe OO, Popoola JO, Leelavathi S, Reddy SV (2011) Advances in plant molecular farming. Biotechnol Adv 29:210–222. https://doi.org/10.1016/j.biotechadv.2010.11.004

Ono A, Hiwasa-Tanase K, Nonaka S, Ezura H (2021) The accumulation of recombinant miraculin is independent of fruit size in tomato. Plant Biotech 38:161–165. https://doi.org/10.5511/plantbiotechnology.20.0904a

Park YJ, Han JE, Jung LH, YJ, Murthy HN, Park SY, (2020) Large-scale production of recombinant miraculin protein in transgenic carrot callus suspension cultures using air-lift bioreactors. AMB Expr 10:140. https://doi.org/10.1186/s13568-020-01079-3

Park YJ, Han JE, Lee LH, JY, Ho TT, Park SY, (2021) Production of recombinant miraculin protein in carrot callus via Agrobacterium-mediated transformation. Plant Cell Tiss Organ Cult 145:615–623. https://doi.org/10.1007/s11240-021-02032-3

Peng RH, Yao QH, Xiong AS, Cheng ZM, Li Y (2006) Codon modifications and an endoplasmic reticulum-targeting sequence additively enhance expression of an Aspergillus phytase gene in transgenic canola. Plant Cell Rep 25:124–132. https://doi.org/10.1007/s00299-005-0036-y

Russell C, Grimes C, Baker P, Sievert K, Lawrence MA (2021) The drivers, trends and dietary impacts of non-nutritive sweeteners in the food supply: a narrative review. Nutr Res Rev 34:185–208. https://doi.org/10.1017/s0954422420000268

Saito T, Ariizumi T, Okabe Y, Asamizu E, Hiwasa-Tanase K, Fukuda N, Mizoguchi T, Yamazaki Y, Aoki K, Ezura H (2011) TOMATOMA: A novel tomato mutant database distributing Micro-Tom mutant collections. Plant Cell Physiol 52:283–296. https://doi.org/10.1093/pcp/pcr004

Schillberg S, Twyman RM, Fischer R (2005) Opportunities for recombinant antigen and antibody expression in transgenic plants—technology assessment. Vaccine 23:1764–1769. https://doi.org/10.1016/j.vaccine.2004.11.002

Stam P, Zeven AC (1981) The theoretical proportion of the donor genome in near-isogenic lines of self-fertilizers bred by backcrossing. Euphytica 30:227–238

Sugaya T, Yano M, Sun HJ, Hirai T, Ezura H (2008) Transgenic strawberry expressing a taste-modifying protein, miraculin. Plant Biotechnol 25:329–333. https://doi.org/10.5511/plantbiotechnology.25.329

Sun HJ, Cui ML, Ma B, Ezura H (2006) Functional expression of the taste-modifying protein, miraculin, in transgenic lettuce. FEBS Lett 580:620–626. https://doi.org/10.1016/j.febslet.2005.12.080

Sun HJ, Kataoka H, Yano M, Ezura H (2007) Genetically stable expression of functional miraculin, a new type of alternative sweetener, in transgenic tomato plants. Plant Biotechnol J 5:768–777. https://doi.org/10.1111/j.1467-7652.2007.00283.x

Toews I, Lohner S, Küllenberg DG, Sommer H, Meerpohl JJ (2019) Association between intake of non-sugar sweeteners and health outcomes: systematic review and meta-analyses of randomized and non-randomized controlled trials and observational studies. BMJ 364:k4718. https://doi.org/10.1136/bmj.k4718

Tourrette E, Falque M, Martin OC (2021) Enhancing backcross programs through increased recombination. Genet Sel Evol 53:25. https://doi.org/10.1186/s12711-021-00619-0

Yano M, Hirai T, Kato K, Hiwasa-Tanase K, Fukuda N, Ezura H (2010) Tomato is a suitable material for producing recombinant miraculin protein in genetically stable manner. Plant Sci 178:469–473. https://doi.org/10.1016/j.plantsci.2010.02.016

Yarra R (2020) Plastome engineering in vegetable crops: current status and future prospects. Mol Biol Rep 47:8061–8074. https://doi.org/10.1007/s11033-020-05770-3

Young ND, Tanksley SD (1989) RFLP analysis of the size of chromosomal segments retained around the Tm-2 locus of tomato during backcross breeding. Theor Appl Genet 77:353–359

Thông tin
Thông tin xuất bản

Transgenic Research

Thông tin tác giả