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Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
RESEARCH ARTICLE

A calcineurin B-like protein participates in low oxygen signalling in rice

Viet The Ho A B , Anh Nguyet Tran A , Francesco Cardarelli C , Pierdomenico Perata A and Chiara Pucciariello A D
+ Author Affiliations
- Author Affiliations

A PlantLab, Institute of Life Sciences, Scuola Superiore Sant’Anna, Piazza Martiri della Libertà 33, 56127 Pisa, Italy.

B Present address: Faculty of Biotechnology and Environmental Technology, University of Food Industry, Ho Chi Minh City, Vietnam.

C NEST, Istituto Nanoscienze – CNR and Scuola Normale Superiore, Piazza San Silvestro 12, 56127 Pisa, Italy.

D Corresponding author. Email: c.pucciariello@sssup.it

Functional Plant Biology 44(9) 917-928 https://doi.org/10.1071/FP16376
Submitted: 29 October 2016  Accepted: 17 May 2017   Published: 23 June 2017

Abstract

Following the identification of the calcineurin B-like interacting protein kinase 15 (CIPK15), which is a regulator of starch degradation, the low O2 signal elicited during rice germination under submergence has been linked to the sugar sensing cascade and calcium (Ca2+) signalling. CIPK proteins are downstream effectors of calcineurin B-like proteins (CBLs), which act as Ca2+ sensors, whose role under low O2 has yet to be established. In the present study we describe CBL4 as a putative CIPK15 partner, transcriptionally activated under low O2 in rice coleoptiles. The transactivation of the rice embryo CBL4 transcript and CBL4 promoter was influenced by the Ca2+ blocker ruthenium red (RR). The bimolecular fluorescence complementation (BiFC) assay associated to fluorescence recovery after photobleaching (FRAP) analysis confirmed that CBL4 interacts with CIPK15. The CBL4-CIPK15 complex is localised in the cytoplasm and the plasma-membrane. Experiments in protoplasts showed a dampening of α-amylase 3 (RAMY3D) expression after CBL4 silencing by artificial miRNA. Our results suggest that under low O2, the Ca2+ sensor CBL4 interacts with CIPK15 to regulate RAMY3D expression in a Ca2+-dependent manner.

Additional keywords: α-amylase, calcium, CBL4, CIPK15, low oxygen, Oryza sativa.


References

Alpi A, Beevers H (1983) Effects of O2 concentration on rice seedling. Plant Physiology 71, 30–34.
Effects of O2 concentration on rice seedling.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL3sXmvV2gug%3D%3D&md5=182b1072148be3e9e8fff156e48adacaCAS |

Banti V, Mafessioni F, Loreti E, Alpi A, Perata P (2010) The heat-inducible transcription factor HsfA2 enhances anoxia tolerance in Arabidopsis. Plant Physiology 152, 1471–1483.
The heat-inducible transcription factor HsfA2 enhances anoxia tolerance in Arabidopsis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXmsF2lsbk%3D&md5=92df2cf796ae1e5f415464061c2d9ec1CAS |

Batistič O, Sorek N, Schultke S, Yalovsky S, Kudla J (2008) Dual fatty acyl modification determines the localization and plasma membrane targeting of CBL/CIPK Ca2+ signaling complexes in Arabidopsis. The Plant Cell 20, 1346–1362.
Dual fatty acyl modification determines the localization and plasma membrane targeting of CBL/CIPK Ca2+ signaling complexes in Arabidopsis.Crossref | GoogleScholarGoogle Scholar |

Batistič O, Waadt R, Steinhorst L, Held K, Kudla J (2010) CBL-mediated targeting of CIPKs facilitates the decoding of calcium signals emanating from distinct cellular stores. The Plant Journal 61, 211–222.
CBL-mediated targeting of CIPKs facilitates the decoding of calcium signals emanating from distinct cellular stores.Crossref | GoogleScholarGoogle Scholar |

Chaves-Sanjuan A, Sanchez-Barrena MJ, Gonzalez-Rubio JM, Moreno M, Ragel P, Jimenez M, Pardo JM, Martinez-Ripoll M, Quintero FJ, Albert A (2014) Structural basis of the regulatory mechanism of the plant CIPK family of protein kinases controlling ion homeostasis and abiotic stress. Proceedings of the National Academy of Sciences of the United States of America 111, E4532–E4541.
Structural basis of the regulatory mechanism of the plant CIPK family of protein kinases controlling ion homeostasis and abiotic stress.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXhs1yhtb%2FJ&md5=834f8fbb9bf48cb0e872e1c5c5390b0dCAS |

DeFalco TA, Bender KW, Snedden WA (2010) Breaking the code: Ca2+ sensors in plant signaling. The Biochemical Journal 425, 27–40.
Breaking the code: Ca2+ sensors in plant signaling.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhs1Sru7vM&md5=ddf61a29ac3e79995cb397a89f37c951CAS |

Di Rienzo C, Piazza V, Gratton E, Beltram F, Cardarelli F (2014) Probing short-range protein Brownian motion in the cytoplasm of living cells. Nature Communications 5, 5891
Probing short-range protein Brownian motion in the cytoplasm of living cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2MXksVersr0%3D&md5=a687678f33af1dcfd58ef8b5b40d6d22CAS |

Gonzali S, Loreti E, Cardarelli F, Novi G, Parlanti S, Pucciariello C, Bassolino L, Banti V, Licausi F, Perata P (2015) Universal stress protein HRU1 mediates ROS homeostasis under anoxia. Nature Plants 1, 15151
Universal stress protein HRU1 mediates ROS homeostasis under anoxia.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2MXhvFertLzM&md5=5f5e055cbbf63acdadba8087203c93f8CAS |

Gu Z, Ma B, Jiang Y, Chen Z, Su X, Zhang H (2008) Expression analysis of the calcineurin B-line gene family in rice (Oryza sativa L.) under environmental stresses. Gene 415, 1–12.
Expression analysis of the calcineurin B-line gene family in rice (Oryza sativa L.) under environmental stresses.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXlsFCqtLk%3D&md5=1fce0c31c6941e81d9f77a99efe2e818CAS |

Hashimoto K, Eckert C, Anschütz U, Scholz M, Held K, Waadt R, Reyer A, Hippler M, Becker D, Kudla J (2012) Phosphorylation of calcineurin B-like (CBL) calcium sensor proteins by their CBL-interacting protein kinases (CIPKs) is required for full activity of CBL-CIPK complexes toward their target proteins. Journal of Biological Chemistry 287, 7956–7968.
Phosphorylation of calcineurin B-like (CBL) calcium sensor proteins by their CBL-interacting protein kinases (CIPKs) is required for full activity of CBL-CIPK complexes toward their target proteins.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XjsFCkur0%3D&md5=3cbd1b8466586efeda0e485bcb0e6072CAS |

Horie T, Costa A, Kim TH, Han MJ, Horie R, Leung HY, Miyao A, Hirochika H, An G, Schroeder JI (2007) Rice OsHKT2;1 transporter mediates large Na+ influx component into K+-starved roots for growth. EMBO Journal 26, 3003–3014.
Rice OsHKT2;1 transporter mediates large Na+ influx component into K+-starved roots for growth.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXms1WntLg%3D&md5=4c762c650474ab3469a11f9220307e62CAS |

Hruz T, Laule O, Szabo G, Wessendorp F, Bleuler S, Oertle L, Widmayer P, Gruissem W, Zimmermann P (2008) Genevestigator V3: a reference expression database for the meta-analysis of transcriptomes. Advances in Bioinformatics 2008, 420747
Genevestigator V3: a reference expression database for the meta-analysis of transcriptomes.Crossref | GoogleScholarGoogle Scholar |

Kanwar P, Sanyal SK, Tokas I, Yadav AK, Pandey A, Kapoor S, Pandey GK (2014) Comprehensive structural, interaction and expression analysis of CBL and CIPK complement during abiotic stresses and development in rice. Cell Calcium 56, 81–95.
Comprehensive structural, interaction and expression analysis of CBL and CIPK complement during abiotic stresses and development in rice.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXhtF2js7%2FJ&md5=f3f44f45a04347ddb128ecd19c8b52bdCAS |

Kaplan B, Davydov O, Knight H, Galon Y, Knight MR, Fluhr R, Fromn H (2006) Rapid transcriptome changes induced by cytosolic Ca2+ transients reveal ABRE-related sequences as Ca2+- responsive cis elements in Arabidopsis. The Plant Cell 18, 2733–2748.
Rapid transcriptome changes induced by cytosolic Ca2+ transients reveal ABRE-related sequences as Ca2+- responsive cis elements in Arabidopsis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28Xht1ejurjP&md5=e9be0acda3dd349f3c427b8aaed8ea6bCAS |

Karimi M, Inze’ D, Depicker A (2002) GATEWAY vectors for Agrobacterium-mediated plant transformation. Trends in Plant Science 7, 193–195.
GATEWAY vectors for Agrobacterium-mediated plant transformation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38Xjtl2ktrk%3D&md5=419c33527b983f1d52328f2f6004d4aeCAS |

Kolukisaoglu U, Weinl S, Blazervic D, Batistic O, Kudla J (2004) Calcium sensors and their interacting protein kinases: genomics of the Arabidopsis and rice CBL-CIPK signaling networks. Plant Physiology 134, 43–58.
Calcium sensors and their interacting protein kinases: genomics of the Arabidopsis and rice CBL-CIPK signaling networks.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXhtVagtbo%3D&md5=f96bee0ed04f21a12843833f2514eef3CAS |

Kretzschmar T, Pelayo MA, Trijatmiko KR, Gabunada LF, Alam R, Jimenez R, Mendioro MS, Slamet-Loedin IH, Sreenivasulu N, Bailey-Serres J, Ismail AM, Mackill DJ, Septiningsih EM (2015) A trehalose-6-phosphate phosphatase enhances anaerobic germination tolerance in rice. Nature Plants 1, 15124
A trehalose-6-phosphate phosphatase enhances anaerobic germination tolerance in rice.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2MXhvFClsLbL&md5=27c9993cc3681bc5e9f1ca6b01485ca2CAS |

Kudahettige NP, Pucciariello C, Alpi A, Perata P (2011) Regulatory interplay of the Sub1A and CIPK15 pathways in the regulation of α-amylase production in flooded rice plants. Plant Biology 13, 611–619.
Regulatory interplay of the Sub1A and CIPK15 pathways in the regulation of α-amylase production in flooded rice plants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXot1Ogu7w%3D&md5=3cad957d9e5a4722a461d48388cd9abfCAS |

Kurusu T, Hamada J, Nokajima H, Kitagawa Y, Kiyoduka M, Takahashi A, Hanamata S, Ohno R, Hayashi T, Okada K, Koga J, Hirochika H, Yamane H, Kuchitsu K (2010) Regulation of microbe-associated molecular pattern-induced hypersensitive cell death, phytoalexin production, and defense gene expression by calcineurin B-like protein-interacting protein kinases, OsCIPK14/15, in rice cultured cells. Plant Physiology 153, 678–692.
Regulation of microbe-associated molecular pattern-induced hypersensitive cell death, phytoalexin production, and defense gene expression by calcineurin B-like protein-interacting protein kinases, OsCIPK14/15, in rice cultured cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXnvVamt70%3D&md5=e8ae61c9b53aaab15a1856fbd3f75a5bCAS |

Lee K-W, Chen P-W, Lu C-A, Chen S, Ho T-HD, Yu S-M (2009) Coordinated responses to oxygen and sugar deficiency allow rice seedlings to tolerate flooding. Science Signaling 2, ra61
Coordinated responses to oxygen and sugar deficiency allow rice seedlings to tolerate flooding.Crossref | GoogleScholarGoogle Scholar |

Licausi F, Weits DA, Pant BD, Scheible WR, Geigenberger P, van Dongen JT (2011) Hypoxia responsive gene expression is mediated by various subsets of transcription factors and miRNAs that are determined by the actual oxygen availability. New Phytologist 190, 442–456.
Hypoxia responsive gene expression is mediated by various subsets of transcription factors and miRNAs that are determined by the actual oxygen availability.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXmt1Gktb0%3D&md5=c7486ebf0d0233b60f1406dfa73c8f85CAS |

Lin CR, Lee KW, Chen CY, Hong YF, Chen JL, Lu CA, Chen KT, Ho TH, Yu SM (2014) SnRK1A-interacting negative regulators modulate the nutrient starvation signaling sensor SnRK1 in source–sink communication in cereal seedlings under abiotic stress. The Plant Cell 26, 808–827.
SnRK1A-interacting negative regulators modulate the nutrient starvation signaling sensor SnRK1 in source–sink communication in cereal seedlings under abiotic stress.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXmtVOisbo%3D&md5=d0a51e8a3d6bacf67d304d89b0e521c4CAS |

Lokdarshi A, Conner WC, McClintock C, Li T, Roberts DM (2016) Arabidopsis CML38, a calcium sensor that localizes to ribonucleoprotein complexes under hypoxia stress. Plant Physiology 170, 1046–1059.
Arabidopsis CML38, a calcium sensor that localizes to ribonucleoprotein complexes under hypoxia stress.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC28Xht1WjsbbL&md5=415b1ba6f35fcb204d13dec8ebae2cf4CAS |

Loreti E, Yamaguchi J, Alpi A, Perata P (2003) Sugar modulation of α-amylase genes under anoxia. Annals of Botany 91, 143–148.
Sugar modulation of α-amylase genes under anoxia.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXitVCksLs%3D&md5=0668d1fa70e77b8003010da8d19fbc99CAS |

Luan S (2009) The CBL-CIPK network in plant calcium signaling. Trends in Plant Science 14, 37–42.
The CBL-CIPK network in plant calcium signaling.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXntVyhuw%3D%3D&md5=f13003d38d3974b0156319bfd172b504CAS |

Martínez-Atienza J, Jiang X, Garciadeblas B, Mendoza I, Zhu JK, Pardo JM, Quitero FJ (2007) Conservation of the salt overly sensitive pathway in rice. Plant Physiology 143, 1001–1012.
Conservation of the salt overly sensitive pathway in rice.Crossref | GoogleScholarGoogle Scholar |

Nie X, Durnin DC, Igamberdiev AU, Hill RD (2006) Cytosolic calcium is involved in the regulation of barley hemoglobin gene expression. Planta 223, 542–549.
Cytosolic calcium is involved in the regulation of barley hemoglobin gene expression.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XivFWjsbs%3D&md5=775a4a9b10a76238d2bb58dd5d2e40cbCAS |

Ossowski S, Schwab R, Weigel D (2008) Gene silencing in plants using artificial microRNAs and other small RNAs. The Plant Journal 53, 674–690.
Gene silencing in plants using artificial microRNAs and other small RNAs.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXivFaqsbc%3D&md5=c141c1fceed1bc6c0af3d96f16fa6723CAS |

Perata P, Pozueta-Romero J, Akazawa T, Yamaguchi J (1992) Effect of anoxia on starch breakdown in rice and wheat seeds. Planta 188, 611–618.
Effect of anoxia on starch breakdown in rice and wheat seeds.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK3sXhvFensw%3D%3D&md5=2716e37846d44e6e0bfcdc14b381a11aCAS |

Perata P, Guglielminetti L, Alpi A (1997) Mobilization of endosperm reserves in cereal seeds under anoxia. Annals of Botany 79, 49–56.
Mobilization of endosperm reserves in cereal seeds under anoxia.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2sXhtlWrs7s%3D&md5=a6baa80317d6e1ae1b230ccee5bea50aCAS |

Piao H-L, Xuan Y-H, Park SH, Je BI, Park SJ, Park SH, Kim CM, Huang J, Wang GK, Kim MJ, Kang SM, Lee I-J, Kwon T-R, Kim YH, Yeo U-S, Yi G, Son DY, Han CD (2010) OsCIPK31, a CBL-interacting protein kinase is involved in germination and seedling growth under abiotic stress condition in rice plants. Molecules and Cells 30, 19–27.
OsCIPK31, a CBL-interacting protein kinase is involved in germination and seedling growth under abiotic stress condition in rice plants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXptFGmtbs%3D&md5=fc07b5aab92afbaa67d8ed51c1a38a3fCAS |

Pineros M, Tester M (1997) Calcium channels in higher plant cells: selectivity, regulation and pharmacology. Journal of Experimental Botany 48, 551–577.
Calcium channels in higher plant cells: selectivity, regulation and pharmacology.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2sXjs1eqsro%3D&md5=b143657798bbe30bd2ec6133b4a0179eCAS |

Rombauts S, Déhais P, Montagu M-V, Rouzé P (1999) PlantCARE, a plant cis-acting regulatory element data. Nucleic Acids Research 27, 295–296.
PlantCARE, a plant cis-acting regulatory element data.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXpsVKgtQ%3D%3D&md5=2900db486a4aab9b5687267a04296832CAS |

Saeed AI, Sharov V, White J, Li J, Liang W, Bhagabati N, Braisted J, Klapa M, Currier T, Thiagarajan M, Sturn A, Snuffin M, Rezantsev A, Popov D, Ryltsov A, Kostukovich E, Borisovsky I, Liu Z, Vinsavich A, Trush V, Quackenbush J (2003) TM4: A free, open-source system for microarray data management and analysis. BioTechniques 34, 374–378.

Sedbrook JC, Kronebusch PJ, Borisy GG, Trewavas AJ, Masson PH (1996) Transgenic AEQUORIN reveals organ-specific cytosolic Ca2+ responses to anoxia in Arabidopsis thaliana seedlings. Plant Physiology 111, 243–257.
Transgenic AEQUORIN reveals organ-specific cytosolic Ca2+ responses to anoxia in Arabidopsis thaliana seedlings.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK28XivFekuro%3D&md5=e15f6b7bf949479b1da8fbdfefcc76dbCAS |

Sorenson R, Bailey-Serres J (2014) Selective mRNA sequestration by OLIGOURIDYLATE-BINDING PROTEIN 1 contributes to translational control during hypoxia in Arabidopsis. Proceedings of the National Academy of Sciences of the United States of America 111, 2373–2378.
Selective mRNA sequestration by OLIGOURIDYLATE-BINDING PROTEIN 1 contributes to translational control during hypoxia in Arabidopsis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXisFyisL0%3D&md5=25e67a5c9391384feb422e6edfd786d8CAS |

Subbaiah CC, Bush DS, Sachs MM (1994a) Elevation of cytosolic calcium precedes anoxic gene expression in maize suspension-cultured cells. The Plant Cell 6, 1747–1762.
Elevation of cytosolic calcium precedes anoxic gene expression in maize suspension-cultured cells.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXivVWkt7k%3D&md5=71e0537c0ba8875fb1bd9f6346d397dfCAS |

Subbaiah CC, Zhang J, Sachs MM (1994b) Involvement of intracellular calcium in anaerobic gene expression and survival of maize seedlings. Plant Physiology 105, 369–376.
Involvement of intracellular calcium in anaerobic gene expression and survival of maize seedlings.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2cXktVGhtb4%3D&md5=246a6df5336f87589d5eebce74b6c261CAS |

Tsuji H, Nakazono M, Saisho D, Tsutsumi N, Hirai A (2000) Transcript levels of the nuclear-encoded respiratory genes in rice decrease by oxygen deprivation: evidence for involvement of calcium in expression of the alternative oxidase 1a gene. FEBS Letters 471, 201–204.
Transcript levels of the nuclear-encoded respiratory genes in rice decrease by oxygen deprivation: evidence for involvement of calcium in expression of the alternative oxidase 1a gene.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3cXitlOlurw%3D&md5=beae1371a686fd734a4a015051e82941CAS |

Wang F, Chen ZH, Liu X, Colmer TD, Zhou M, Shabala S (2016) Tissue-specific root ion profiling reveals essential roles of the CAX and ACA calcium transport systems in response to hypoxia in Arabidopsis. Journal of Experimental Botany 67, 3747–3762.
Tissue-specific root ion profiling reveals essential roles of the CAX and ACA calcium transport systems in response to hypoxia in Arabidopsis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC28XhsF2jsr7N&md5=654af58297ff985b130bb4a606ddb26cCAS |

Warthmann N, Ossowski S, Schwab R, Weigel D (2013) Artificial microRNAs for specific gene silencing in rice. Methods in Molecular Biology 956, 131–149.
Artificial microRNAs for specific gene silencing in rice.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2MXitFalsb0%3D&md5=47e902fef9fe2647262b76c47099e84bCAS |

Xiang Y, Huang Y, Xiong L (2007) Characterization of stress-responsive CIPK genes in rice for stress tolerance improvement. Plant Physiology 144, 1416–1428.
Characterization of stress-responsive CIPK genes in rice for stress tolerance improvement.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXot1OlsLw%3D&md5=2dd3bd7a9676c09a3bdca6d7f958602bCAS |

Yang W, Kong Z, Omo-Ikerodah E, Xu W, Li Q, Xue Y (2008) Calcineurin B-like interacting protein kinase OsCIPK23 functions in pollination and drought stress responses in rice (Oryza sativa L.). Journal of Genetics and Genomics 35, 531–543.
Calcineurin B-like interacting protein kinase OsCIPK23 functions in pollination and drought stress responses in rice (Oryza sativa L.).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXht1aitbrJ&md5=3eec5da66c2332b7cd1cf72842e3dcaeCAS |

Yemelyanov VV, Shishova MS, Chirkova TV, Lindberg SM (2011) Anoxia-induced elevation of cytosolic Ca2+ concentration depends of different Ca2+ sources in rice and wheat protoplast. Planta 234, 271–280.
Anoxia-induced elevation of cytosolic Ca2+ concentration depends of different Ca2+ sources in rice and wheat protoplast.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXptlyjsbk%3D&md5=60a8c18cb760b136b8e618608d7784eaCAS |

Yoo S-D, Cho Y-H, Sheen J (2007) Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nature Protocols 2, 1565–1572.
Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXhtFahur3I&md5=5672eaf09fd03dfa6773f1dfe54e2946CAS |

Yu S-M, Lo S-F, Ho T-HD (2015) Source–sink communication: regulated by hormone, nutrient, and stress cross-signaling. Trends in Plant Science 20, 844–857.
Source–sink communication: regulated by hormone, nutrient, and stress cross-signaling.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2MXhslKktL7F&md5=77a0b74a692b06cae6d2089403c3de99CAS |

Zhang Y, Su J, Duan S, Ao Y, Dai J, Liu J, Wang P, Li Y, Liu B, Feng D (2011) A highly efficient rice green tissue protoplast system for transient gene expression and studying light/chloroplast-related processes. Plant Methods 7, 30–44.
A highly efficient rice green tissue protoplast system for transient gene expression and studying light/chloroplast-related processes.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXhs1yjsrrO&md5=c37273074957520fcfed7234a377bba8CAS |