( 1r^ =fgdc`ku pn ;h · 2020. 3. 11. · y. ishii & s. maruyama 1 ( 1r^ =fgdc`ku pn ;h ¼ 9 ^...

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Y. Ishii & S. Maruyama1 (1R^=fgDC`kUPN;H ƼĹɞėƸŤ ūÌþćþćɂƫãDžćƽNjDž 980-8578 ČðƷÚĜɍǻÍǸěąɍǻ 6-3 Yuu Ishii, Shinichiro Maruyama Methods for genetic transformation of coral symbiont algae: Current status and future directions Keywords: algae, cnidarian animals, endosymbiosis, Symbiodinium, transformation Graduate School of Life Sciences, Tohoku University 6-3, Aramaki Aza-Aoba, Aoba-ku, Sendai, Miyagi, 980-8578, Japan DOI: 10.24480/bsj-review.8c 5 .0012 4 poa_^t3M1R^l ppoa_^ ÐǛǭǽɖ*)+ƎƬƘǰŶÉƝ&µƫɀ§):LjĆìɞ%5ÂǭÉƝɜJ{MPO BXI[wJ('ɝ)Ǜǭ¶µƫ:ƒɏƄǽɖɜ Symbiodinium spp. ɝ+ȅǾǽ&â,;ɜêɝɞȝŰɘ *ƛĝƛĝƎñ*ƫĽǗ=Ő:ſƫƬǫ&$*þ(ĨÄ=ň"$:POƿ)$ɞ µƫǽ%:ȅǾǽ*²ÜĿ%¢8;De}I* 90%+Ď%:PO)×Ƒ;$:&5 Ȋ<;$:ɜ 2007 ɝ(ġǩ)+ɞȅǾǽɜzooxanthella ɝ&+ƎƬƘǰŶÉƝ)µƫ:ɛǵ( Ƿȅǵ*ÐǛǭǽɖ*Ǥdž%:ɞŧlj%+ÂǭÉƝ)µƫ: Symbiodinium spp. =Ƣǩ*ȅǾ ǽ&$Ń ê ÂǭÉƝ&µƫ:ȅǾǽ ɜAɝƛĝƎñ*POƿɜB, Cɝ ȅǾǽ=µƫ:ȩƿPO ɜÂǭÉƝȾǶǾǣBRPO ƴgbxPBPODžɝɜDɝ ȅ Ǿǽ=µƫVBYFBX I[wJɜExaiptasia pallidaɝ ɜE ɝƲ˹Ʃ)79Ɏµƫ ƠĽ)VBYFBXI[ wJɜF ɝȅǾǽŻģêɜɣƨħ ǛǭɣȬƋǛǭɝɜ GɝȅǾǽ ɜSymbiodinium sp. clade BɝƨħǛ ǭɜ HɝȅǾǽɜ Symbiodinium sp. clade BɝȬƋǛǭ

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Page 1: ( 1R^ =fgDC`kU PN ;H · 2020. 3. 11. · Y. Ishii & S. Maruyama 1 ( 1R^ =fgDC`kU PN ;H ¼ 9 ^ ¸ d kÌþ þ B «ã Å ½ Ë Å 980-8578 ð · Ú M ûÍ ø M û6-3 Yuu Ishii, Shinichiro

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Ƽ�Ĺɞ�ėƸ�Ť�

ūÌþćþćɂƫãDžćƽNjDž �980-8578ČðƷ�ÚĜɍǻÍǸěąɍǻ6-3

Yuu Ishii, Shinichiro Maruyama

Methods for genetic transformation of coral symbiont algae: Current status and future directions

Keywords: algae, cnidarian animals, endosymbiosis, Symbiodinium, transformation Graduate School of Life Sciences, Tohoku University

6-3, Aramaki Aza-Aoba, Aoba-ku, Sendai, Miyagi, 980-8578, Japan DOI: 10.24480/bsj-review.8c5.00124

poa_^t3M�1R��^l p�poa_^���

ÐǛǭǽɖ*�)+ƎƬƘǰŶÉƝ&µƫɀ§):Lj�Ćì�ɞ�%5ÂǭÉƝɜJ{M�P�O�

BXI�[wJ('ɝ)Ǜǭ¶µƫ�:ƒɏƄǽɖɜSymbiodinium spp.ɝ+ȅǾǽ&â,;ɜêɝɞȝŰɘ

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�Ƿȅǵ*ÐǛǭǽɖ*Ǥdž%:�ɞŧlj%+ÂǭÉƝ)µƫ�:Symbiodinium spp. =Ƣǩ*ȅǾ

ǽ&�$Ń��

ê� ÂǭÉƝ&µƫ�:ȅǾǽ

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ɜEɝ �Ʋ˹Ʃ�)79Ɏµƫ

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wJɜFɝȅǾǽŻģêɜ�ɣƨħ

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ǭɜHɝȅǾǽɜSymbiodinium sp.

clade BɝȬƋǛǭ

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Apt, K.E., Kroth, P.G., & Grossman A.R. 1996. Stable nuclear transformation of the diatom Phaeodactylum

tricornutum. Mol. Gen. Genet. 252:572–579.

Baek, K., Kim, D.H., Jeong, J., Sim, S.J., Melis, A., et al. 2016. DNA-free two-gene knockout in Chlamydomonas

reinhardtii via CRISPR-Cas9 ribonucleoproteins. Sci. Rep. 6:30620. Baumgarten, S., Simakov, O., Esherick, L.Y., Liew, Y.J., Lehnert, E.M., et al. 2015. The genome of Aiptasia, a sea

anemone model for coral symbiosis. Proc. Natl. Acad. Sci. U.S.A. 112:11893–11898.

Brown, B.E. 1997. Coral bleaching: causes and consequences. Coral Reefs. 16:S129–138. Chen, H.L., Li, S.S, Huang, R., & Tsai, H. 2008. Conditional production of a functional fish growth hormone in the

transgenic line of Nannochloropsis oculata (Eustigmatophyceae). J. Phycol. 44:768–776.

Chen, Y., Wang, Y., Sun, Y., Zhang, L., & Li, W. 2001. Highly efficient expression of rabbit neutrophil peptide-1 gene in Chlorella ellipsoidea cells. Curr. Genet. 39:365–370.

Chow, K.C., & Tung, W.L. 1999. Electrotransformation of Chlorella vulgaris. Plant Cell Rep. 18:778–780.

Clough, S.J, & Bent, A.F. 1998. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J. 16:735–743.

Dawson, H.N., Burlingame, R., & Cannons, A.C. 1997. Stable transformation of Chlorella: rescue of nitrate reductase-

deficient mutants with the nitrate reductase gene. Curr. Microbiol. 35:356–362. Debuchy, R., Purton, S., & Rochaix, J.D. 1989. The argininosuccinate lyase gene of Chlamydomonas reinhardtii: an

important tool for nuclear transformation and for correlating the genetic and molecular maps of the ARG7 locus.

EMBO J. 8:2803–2809. Doetsch, N.A., Favreau, M.R., Kuscuoglu, N., & Thompson, M.D. 2001. Chloroplast transformation in Euglena

gracilis: splicing of a group III twintron transcribed from a transgenic psbK operon. Curr. Genet. 39:49–60.

Dunahay, T.G., Jarvis, E.E., & Roessler, P.G. 1995. Genetic transformation of the diatoms Cyclotella cryptica and Navicula saprophila. J. Phycol. 31:1004–1012.

Fernández-Robledo, J.A, Lin, Z., & Vasta, G.R. 2008. Transfection of the protozoan parasite Perkinsus marinus. Mol.

Biochem. Parasitol. 157:44–53. Hambleton, E.A., Guse, A., & Pringle, J.R. 2014. Similar specificities of symbiont uptake by adults and larvae in an

anemone model system for coral biology. J. Exp. Biol. 217:1613–1619.

Hiei, Y., Ohta, S., Komari, T., & Kumashiro, T. 1994. Efficient transformation of rice (Oryza sativa L.) mediated by agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J. 6:271–282.

Hirakawa, Y., & Ishida, K. 2010. Internal plastid-targeting signal found in a RubisCO small subunit protein of a

chlorarachniophyte alga. Plant J. 64:402–410. Hirakawa, Y., Kofuji, R., & Ishida, K. 2008. Transient transformation of a chlorarachniophyte alga, Lotharella

amoebiformis (Chlorarachniophyceae), with uida and egfp reporter genes. J Phycol. 44:814–820

Page 7: ( 1R^ =fgDC`kU PN ;H · 2020. 3. 11. · Y. Ishii & S. Maruyama 1 ( 1R^ =fgDC`kU PN ;H ¼ 9 ^ ¸ d kÌþ þ B «ã Å ½ Ë Å 980-8578 ð · Ú M ûÍ ø M û6-3 Yuu Ishii, Shinichiro

������� �� ��� �� ��

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�� Æ 2007ɟǽɖ30®ğ*DzƙÛ –ǽɖ�8ȇ:ƫƝȫË�îƨ�ƪõ–ɟūƎþćºƜ�ɟDŽĂęɟ Kindle, K.L. 1990. High-frequency nuclear transformation of Chlamydomonas reinhardtii. Proc. Natl. Acad. Sci. U.S.A.

87:1228–1232. Lapidot, M., Raveh, D., Sivan, A., Arad, S.M., & Shapira, M. 2002. Stable chloroplast transformation of the unicellular

red alga Porphyridium species. Plant Physiol. 129:7–12.

Li, J.F., Norville, J.E., Aach, J., McCormack, M., Zhang, D., et al. 2013. Multiplex and homologous recombination-mediated genome editing in Arabidopsis and Nicotiana benthamiana using guide RNA and Cas9. Nat. Biotechnol.

31:688–691.

Lin, S., Cheng, S., Song, B., Zhong, X., Lin, X., et al. 2015. The Symbiodinium kawagutii genome illuminates dinoflagellate gene expression and coral symbiosis. Science. 350:691–694.

Maruyama, S., Shoguchi, E., Satoh, N., & Minagawa, J. 2015. Diversification of the light-harvesting complex gene

family via intra- and intergenic duplications in the coral symbiotic alga Symbiodinium. PLoS ONE. 10:e0119406. Minoda, A., Sakagami, R., Yagisawa, F., Kuroiwa, T., & Tanaka, K. 2004. Improvement of culture conditions and

evidence for nuclear transformation by homologous recombination in a red alga, Cyanidioschyzon merolae 10D.

Plant Cell Physiol. 45:667–671. Miyahara, M., Aoi, M., Inoue-Kashino, N., Kashino, Y., & Ifuku, K. 2013. Highly efficient transformation of the diatom

Phaeodactylum tricornutum by multi-pulse electroporation. Biosci. Biotechnol. Biochem. 77:874–876.

Mohamed, A.R., Cumbo, V., Harii, S., Shinzato, C., Chan, C.X., et al. 2016. The transcriptomic response of the coral Acropora digitifera to a competent Symbiodinium strain: the symbiosome as an arrested early phagosome. Mol. Ecol.

25:3127–3141.

Muto, M., Fukuda, Y., Nemoto, M., Yoshino, T., Matsunaga, T., & Tanaka, T. 2013. Establishment of a genetic transformation system for the marine pennate diatom Fistulifera sp. strain JPCC DA0580--a high triglyceride

producer. Mar. Biotechnol. 15:48–55.

Nekrasov, V., Staskawicz, B., Weigel, D., Jones, J.D.G., & Kamoun, S. 2013. Targeted mutagenesis in the model plant Nicotiana benthamiana using Cas9 RNA-guided endonuclease. Nat Biotechnol. 31:691–693.

Oakley, C.A., Ameismeier, M.F., Peng, L., Weis, V.M., Grossman, A.R., & Davy, S.K. 2016. Symbiosis induces

widespread changes in the proteome of the model cnidarian Aiptasia. Cell Microbiol. 18:1009–1023. Ohnuma, M., Yokoyama, T., Inouye, T., Sekine, Y., & Tanaka, K. 2008. Polyethylene glycol (PEG)-mediated transient

gene expression in a red alga, Cyanidioschyzon merolae 10D. Plant Cell Physiol. 49:117–120.

Ortiz-Matamoros, M.F., Villanueva, M.A., & Islas-Flores, T. 2015a. Transient transformation of cultured photosynthetic dinoflagellates (Symbiodinium spp.) with plant-targeted vectors. Cien. Mar. 41:21-32.

Ortiz-Matamoros, M.F., Islas-Flores, T., Voigt, B., Menzel, D., Baluška, F., & Villanueva, M.A. 2015b. Heterologous

DNA uptake in cultured Symbiodinium spp. aided by Agrobacterium tumefaciens. PLoS ONE. 10:e0132693 Poulsen, N., Chesley, P.M., & Kröger, N. 2006. Molecular genetic manipulation of the diatom Thalassiosira

pseudonana (bacillariophyceae). J. Phycol. 42:1059–1065

Poulsen, N., & Kröger, N. 2005. A new molecular tool for transgenic diatoms: control of mRNA and protein biosynthesis by an inducible promoter-terminator cassette. FEBS J. 272:3413–3423

Reid, W., & O'Brochta, D.A. 2016. Applications of genome editing in insects. Curr. Opin. Insect Sci. 13:43–54.

Page 8: ( 1R^ =fgDC`kU PN ;H · 2020. 3. 11. · Y. Ishii & S. Maruyama 1 ( 1R^ =fgDC`kU PN ;H ¼ 9 ^ ¸ d kÌþ þ B «ã Å ½ Ë Å 980-8578 ð · Ú M ûÍ ø M û6-3 Yuu Ishii, Shinichiro

������� �� ��� �� ��

������������� ����� ��

Y. Ishii & S. Maruyama�8

Rowan, R., & Powers, D.A. 1991. A molecular genetic classification of zooxanthellae and the evolution of animal-algal

symbioses. Science. 251:1348–1351.

Sakamoto, H., Kita, K., & Matsuzaki, M. 2016. Drug selection using bleomycin for transfection of the oyster-infecting parasite Perkinsus marinus. Parasitol. Int. 65:563–566

Sawahel, W., Onde, S., Knight, C., & Cove, D. 1992. Transfer of foreign DNA into Physcomitrella patens protonemal

tissue by using the gene gun. Plant Mol. Biol. Rep. 10:314–315. Schaefer, D., Zryd, J.P., Knight, C.D., Cove, D.J. 1991. Stable transformation of the moss Physcomitrella patens. Mol.

Gen. Genet. 226:418–424.

Schaefer, D.G., & Zryd, J.P. 1997. Efficient gene targeting in the moss Physcomitrella patens. Plant J. 11:1195–1206 Schiedlmeier, B., Schmitt, R., Müller, W., Kirk, M.M., Gruber, H., et al. 1994. Nuclear transformation of Volvox carteri.

Proc. Natl. Acad. Sci. U.S.A. 91:5080–5084.

Shan, Q., Wang, Y., Li, J., Zhang, Y., Chen, K., et al. 2013. Targeted genome modification of crop plants using a CRISPR-Cas system. Nat. Biotechnol. 31:686–688.

Shin, S.E., Lim, J.M., Koh, H.G., Kim, E.K., Kang, N.K., et al. 2016. CRISPR/Cas9-induced knockout and knock-in

mutations in Chlamydomonas reinhardtii. Sci. Rep. 6:27810 Shinzato, C., Shoguchi, E., Kawashima, T., Hamada, M., Hisata, K., et al. 2011. Using the Acropora digitifera genome

to understand coral responses to environmental change. Nature. 476:320–323.

ĒØ Ű� 2014. ƒɏƄǽɖ*ơƞ(ŲMft. ÒƫÉƝć�ɈȒ 47, 5-12. Shoguchi, E., Shinzato, C., Kawashima, T., Gyoja, F., Mungpakdee, S., et al. 2013. Draft assembly of the Symbiodinium

minutum nuclear genome reveals dinoflagellate gene structure. Curr. Biol. 23:1399–1408.

Sun, Y., Gao, X., Li, Q., Zhang, Q., & Xu, Z. 2006. Functional complementation of a nitrate reductase defective mutant of a green alga Dunaliella viridis by introducing the nitrate reductase gene. Gene. 377:140–149.

Sun, Y., Yang, Z., Gao, X., Li, Q., Zhang, Q., & Xu, Z. 2005. Expression of foreign genes in Dunaliella by

electroporation. Mol. Biotechnol. 30:185–192.

ȗȌ ­ÿɞ�Ø � 2009. ȩƿP�O)µƫ�:ȅǾǽ*»ăǗǡćƳƽNj)ɀ�:~ky�. śŧP

�Oƿć�Ȓ� ǐ10ěɞ13-23. Takahashi, S., Whitney, S., Itoh, S., Maruyama, T., & Badger, M. 2008. Heat stress causes inhibition of the de novo

synthesis of antenna proteins and photobleaching in cultured Symbiodinium. Proc. Natl. Acad. Sci. U.S.A. 105:4203–

4208

Tan, C., Qin, S., Zhang, Q., & Jiang, P. 2005. Establishment of a micro-particle bombardment transformation system for Dunaliella salina. J. Microbiol. 43:361–365

ten Lohuis, M.R., & Miller, D.J. 1998. Genetic transformation of dinoflagellates (Amphidinium and Symbiodinium):

expression of GUS in microalgae using heterologous promoter constructs. Plant J. 13:427–435. Teng, C., Qin, S., Liu, J., Yu, D., Liang, C., & Tseng, C. 2002. Transient expression of lacZ in bombarded unicellular

green alga Haematococcus pluvialis. J. Appl. Phycol. 14:497–500 �

Page 9: ( 1R^ =fgDC`kU PN ;H · 2020. 3. 11. · Y. Ishii & S. Maruyama 1 ( 1R^ =fgDC`kU PN ;H ¼ 9 ^ ¸ d kÌþ þ B «ã Å ½ Ë Å 980-8578 ð · Ú M ûÍ ø M û6-3 Yuu Ishii, Shinichiro

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