? A callus transformation system for gene functional studies in soybean
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    2017, Vol. 16 Issue (09): 1913-1922     DOI: 10.1016/S2095-3119(16)61621-0
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A callus transformation system for gene functional studies in soybean
XU Kun1*, ZHANG Xiao-mei1*, FAN Cheng-ming1, CHEN Fu-lu1, ZHU Jin-long1, ZHANG Shi-long2, CHEN Qing-shan3, FU Yong-fu1
1 Key Lab of Soybean Biology (Beijing), Ministry of Agriculture/National Key Facility of Crop Gene Resource and Genetic Improvement/Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China
2 Bijie Agricultural Science Research Institute of Guizhou, Bijie 551700, P.R.China
3 College of Agriculture, Northeast Agricultural University, Harbin 150030, P.R.China
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Abstract     Obtaining transgenic plants is a common method for analyzing gene function. Unfortunately, stable genetic transformation is difficult to achieve, especially for plants (e.g., soybean), which are recalcitrant to genetic transformation. Transient expression systems, such as Arabidopsis protoplast, Nicotiana leaves, and onion bulb leaves are widely used for gene functional studies. A simple method for obtaining transgenic soybean callus tissues was reported recently. We extend this system with simplified culture conditions to gene functional studies, including promoter analysis, expression and subcellular localization of the target protein, and protein-protein interaction. We also evaluate the plasticity of this system with soybean varieties, different vector constructs, and various Agrobacterium strains. The results indicated that the callus transformation system is efficient and adaptable for gene functional investigation in soybean genotype-, vector-, and Agrobacterium strain-independent modes. We demonstrated an easy set-up and practical homologous strategy for soybean gene functional studies.
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Key wordssoybean callus     gene function studies     transformation     
Received: 2016-09-14; Published: 2017-01-24

This work was supported by the Transgenic Programs, China (2014ZX0800930B and 2016ZX08009-001) and the National Natural Science Found of China (31371703).

Corresponding Authors: Correspondence FU Yong-fu, Tel/Fax: +86-10-82105864, E-mail: fufu19cn@163.com, fuyongfu@caas.cn   
About author: XU Kun, E-mail: xkxq8996@163.com; ZHANG Xiao-mei, E-mail: zxmzh@163.com; * These authors contributed equally to this study.
Cite this article:   
XU Kun, ZHANG Xiao-mei, FAN Cheng-ming, CHEN Fu-lu, ZHU Jin-long, ZHANG Shi-long, CHEN Qing-shan, FU Yong-fu. A callus transformation system for gene functional studies in soybean[J]. Journal of Integrative Agriculture, 2017, 16(09): 1913-1922.
http://www.chinaagrisci.com/Jwk_zgnykxen/EN/ 10.1016/S2095-3119(16)61621-0      or     http://www.chinaagrisci.com/Jwk_zgnykxen/EN/Y2017/V16/I09/1913
[1] Abel S, Theologis A. 1994. Transient transformation of Arabidopsis leaf protoplasts: A versatile experimental system to study gene expression. The Plant Journal, 5, 421-427.
[2] Altpeter F, Baisakh N, Beachy R, Bock R, Capell T, Christou P, Daniell H, Datta K, Datta S, Dix P J. 2005. Particle bombardment and the genetic enhancement of crops: Myths and realities. Molecular Breeding, 15, 305-327.
[3] Asai T, Tena G, Plotnikova J, Willmann M R, Chiu W L, Gomez-Gomez L, Boller T, Ausubel F M, Sheen J. 2002. MAP kinase signalling cascade in Arabidopsis innate immunity. Nature, 415, 977-983.
[4] Assmann S M, Simoncini L, Schroeder J I. 1985. Blue light activates electrogenic ion pumping in guard cell protoplasts of Vicia faba L. Nature, 318, 285-287.
[5] Birch R G. 1997. Plant transformation: Problems and strategies for practical application. Annual Review of Plant Biology, 48, 297-326.
[6] Chen Q J, Zhou H M, Chen J, Wang X C. 2006. Using a modified TA cloning method to create entry clones. Analytical Biochemistry, 358, 120-125.
[7] Chiu W l, Niwa Y, Zeng W, Hirano T, Kobayashi H, Sheen J. 1996. Engineered GFP as a vital reporter in plants. Current Biology, 6, 325-330.
[8] Cho Y H, Yoo S D, Sheen J. 2006. Regulatory functions of nuclear hexokinase1 complex in glucose signaling. Cell, 127, 579-589.
[9] Christou P. 1995. Strategies for variety-independent genetic transformation of important cereals, legumes and woody species utilizing particle bombardment. Euphytica, 85, 13-27.
[10] Christou P, McCabe D E, Swain W F. 1988. Stable transformation of soybean callus by DNA-coated gold particles. Plant Physiology, 87, 671-674.
[11] Cocking E C. 1961. A method for the isolation of plant protoplasts and vacuoles. Nature, 187, 962-963.
[12] Damm B, Schmidt R, Willmitzer L. 1989. Efficient transformation of Arabidopsis thaliana using direct gene transfer to protoplasts. Molecular and General Genetics, 217, 6-12.
[13] Di R, Purcell V, Collins G B, Ghabrial S A. 1996. Production of transgenic soybean lines expressing the bean pod mottle virus coat protein precursor gene. Plant Cell Reports, 15, 746-750.
[14] Duangporn P, Siripong P. 2009. Effect of auxin and cytokinin on phyllanthusol a production by callus cultures of phyllanthus acidus skeels. American-Eurasian Journal of Agricultural and Environmental Science, 5, 258-263.
[15] Earley K W, Haag J R, Pontes O, Opper K, Juehne T, Song K, Pikaard C S. 2006. Gateway-compatible vectors for plant functional genomics and proteomics. The Plant Journal, 45, 616-629.
[16] Gaudin V, Lütticke S, Jouanin L. 1994. A reporter gene under the control of tms or aux promoters is differentially expressed in tobacco and barley protoplasts. Plant Cell Reports, 13, 155-158.
[17] Geiss-Friedlander R, Melchior F. 2007. Concepts in sumoylation: A decade on. Nature Reviews Molecular Cell Biology, 8, 947-956.
[18] Godwin I, Todd G, Ford-Lloyd B, Newbury H J. 1991. The effects of acetosyringone and pH on Agrobacterium-mediated transformation vary according to plant species. Plant Cell Reports, 9, 671-675.
[19] Hecker C M, Rabiller M, Haglund K, Bayer P, Dikic I. 2006. Specification of SUMO1- and SUMO2-interacting motifs. Journal of Biological Chemistry, 281, 16117-16127.
[20] 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. The Plant Journal, 6, 271-282.
[21] Hinchee M A, Connor-Ward D V, Newell C A, McDonnell R E, Sato S J, Gasser C S, Fischhoff D A, Re D B, Fraley R T, Horsch R B. 1988. Production of transgenic soybean plants using Agrobacterium-mediated DNA transfer. Nature Biotechnology, 6, 915-922.
[22] Hwang I, Sheen J. 2001. Two-component circuitry in Arabidopsis cytokinin signal transduction. Nature, 413, 383-389.
[23] Jiang N, Jeon E H, Pak J H, Ha T J, Baek I Y, Jung W S, Lee J H, Kim D H, Choi H K, Cui Z. 2010. Increase of isoflavones in soybean callus by Agrobacterium-mediated transformation. Plant Biotechnology Reports, 4, 253-260.
[24] Kerscher O. 2007. SUMO junction - What’s your function? EMBO Reports, 8, 550-555.
[25] Liu D, Liu S, Chang D, Wang L, Wang D, Wang N N. 2013. A simple and efficient method for obtaining transgenic soybean callus tissues. Acta Physiologiae Plantarum, 35, 2113-2125.
[26] Lukatkin A. 2010. Use of maize callus cultures for assessing chilling stress resistance. Russian Agricultural Sciences, 36, 331-333.
[27] Luo G, Hepburn A, Widholm J. 1994. A simple procedure for the expression of genes in transgenic soybean callus tissue. Plant Cell Reports, 13, 632-636.
[28] Marion J, Bach L, Bellec Y, Meyer C, Gissot L, Faure J D. 2008. Systematic analysis of protein subcellular localization and interaction using high-throughput transient transformation of Arabidopsis seedlings. The Plant Journal, 56, 169-179.
[29] Masson J, Paszkowski J. 1992. The culture response of Arabidopsis thaliana protoplasts is determined by the growth conditions of donor plants. The Plant Journal, 2, 829-833.
[30] Mccullen C A, Binns A N. 2006. Agrobacterium tumefaciens and plant cell interactions and activities required for interkingdom macromolecular transfer. Annual Review of Cell & Developmental Biology, 22, 101-127.
[31] Meurer C, Dinkins R, Collins G. 1998. Factors affecting soybean cotyledonary node transformation. Plant Cell Reports, 18, 180-186.
[32] Mizoi J, Ohori T, Moriwaki T, Kidokoro S, Todaka D, Maruyama K, Kusakabe K, Osakabe Y, Shinozaki K, Yamaguchi-Shinozaki K. 2013. GmDREB2A; 2, a canonical DEHYDRATION-RESPONSIVE ELEMENT-BINDING PROTEIN2-yype transcription factor in soybean, is posttranslationally regulated and mediates dehydration-responsive element-dependent gene expression. Plant Physiology, 161, 346-361.
[33] Nakagawa T, Kurose T, Hino T, Tanaka K, Kawamukai M, Niwa Y, Toyooka K, Matsuoka K, Jinbo T, Kimura T. 2007. Development of series of gateway binary vectors, pGWBs, for realizing efficient construction of fusion genes for plant transformation. Journal of Bioscience & Bioengineering, 104, 34-41.
[34] Nelson B K, Cai X, Nebenführ A. 2007. A multicolored set of in vivo organelle markers for co-localization studies in Arabidopsis and other plants. The Plant Journal, 51, 1126-1136.
[35] Olhoft P M, Donovan C M, Somers D A. 2006. Soybean (Glycine max) transformation using mature cotyledonary node explants. Agrobacterium Protocols, 343, 385-396.
[36] Olhoft P M, Flagel L E, Donovan C M, Somers D A. 2003. Efficient soybean transformation using hygromycin B selection in the cotyledonary-node method. Planta, 216, 723-735.
[37] Owens L D, Cress D E. 1985. Genotypic variability of soybean response to Agrobacterium strains harboring the Ti or Ri plasmids. Plant Physiology, 77, 87-94.
[38] Paz M M, Shou H, Guo Z, Zhang Z, Banerjee A K, Wang K. 2004. Assessment of conditions affecting Agrobacterium-mediated soybean transformation using the cotyledonary node explant. Euphytica, 136, 167-179.
[39] Ramesh A, Sharma S K, Joshi O, Khan I. 2011. Phytase, phosphatase activity and p-nutrition of soybean as influenced by inoculation of bacillus. Indian Journal of Microbiology, 51, 94-99.
[40] Roslan H A, Salter M G, Wood C D, White M R, Croft K P, Robson F, Coupland G, Doonan J, Laufs P, Tomsett A B. 2001. Characterization of the ethanol-inducible alc gene-expression system in Arabidopsis thaliana. The Plant Journal, 28, 225-235.
[41] Salter M G, Paine J A, Riddell K V, Jepson I, Greenland A J, Caddick M X, Tomsett A B. 1998. Characterisation of the ethanol-induciblealcgene expression system for transgenic plants. The Plant Journal, 16, 127-132.
[42] Schmutz J, Cannon S B, Schlueter J, Ma J, Mitros T, Nelson W, Hyten D L, Song Q, Thelen J J, Cheng J. 2010. Genome sequence of the palaeopolyploid soybean. Nature, 463, 178-183.
[43] Sheikholeslam S N, Weeks D P. 1987. Acetosyringone promotes high efficiency transformation of Arabidopsis thaliana explants by Agrobacterium tumefaciens. Plant Molecular Biology, 8, 291-298.
[44] Sheludko Y V. 2008. Agrobacterium-mediated transient expression as an approach to production of recombinant proteins in plants. Recent Patents on Biotechnology, 2, 198-208.
[45] Shih M D, Hsieh J S, Hsing Y I. 2001. Regulation of the soybean GmPM9 promoter in callus tissue. Journal of Genetics and Molecular Biology, 12, 125-137.
[46] Sparkes I A, Runions J, Kearns A, Hawes C. 2006. Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants. Nature Protocols, 1, 2019-2025.
[47] Trick H, Finer J. 1998. Sonication-assisted Agrobacterium-mediated transformation of soybean [Glycine max (L.) Merrill] embryogenic suspension culture tissue. Plant Cell Reports, 17, 482-488.
[48] Trick H N, Finer J J. 1997. SAAT: Sonication-assisted Agrobacterium-mediated transformation. Transgenic Research, 6, 329-336.
[49] Tzfira T, Li J, Lacroix B, Citovsky V. 2004. Agrobacterium T-DNA integration: Molecules and models. Trends in Genetics, 20, 375-383.
[50] Ueki S, Lacroix B, Krichevsky A, Lazarowitz S G, Citovsky V. 2008. Functional transient genetic transformation of Arabidopsis leaves by biolistic bombardment. Nature Protocols, 4, 71-77.
[51] Wang S, Tiwari S B, Hagen G and Guilfoyle T J. 2005. AUXIN RESPONSE FACTOR7 restores the expression of auxin-responsive genes in mutant Arabidopsis leaf mesophyll protoplasts. The Plant Cell, 17, 1979-1993.
[52] Wang X, Fan C, Zhang X, Zhu J, Fu Y F. 2013. BioVector, a flexible system for gene specific-expression in plants. BMC Plant Biology, 13, 198.
[53] Wang Y C, Klein T M, Fromm M, Cao J, Sanford J C, Wu R. 1988. Transient expression of foreign genes in rice, wheat and soybean cells following particle bombardment. Plant Molecular Biology, 11, 433-439.
[54] Wu H, Sparks C, Amoah B, Jones H. 2003. Factors influencing successful Agrobacterium-mediated genetic transformation of wheat. Plant Cell Reports, 21, 659-668.
[55] Xiao C, Chen F, Yu X, Lin C, Fu Y F. 2009. Over-expression of an AT-hook gene, AHL22, delays flowering and inhibits the elongation of the hypocotyl in Arabidopsis thaliana. Plant Molecular Biology, 71, 39-50.
[56] Xu X Y, Shi G X, Wang J, Zhang L L, Kang Y N. 2011. Copper-induced oxidative stress in Alternanthera philoxeroides callus. Plant Cell Tissue & Organ Culture, 106, 243-251.
[57] Yamada T, Takagi K, Ishimoto M. 2012. Recent advances in soybean transformation and their application to molecular breeding and genomic analysis. Breeding Science, 61, 480.
[58] Yang Y, Li R, Qi M. 2000. In vivo analysis of plant promoters and transcription factors by agroinfiltration of tobacco leaves. The Plant Journal, 22, 543-551.
[59] 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.
[60] Zheng Y, Schumaker K S, Guo Y. 2012. Sumoylation of transcription factor MYB30 by the small ubiquitin-like modifier E3 ligase SIZ1 mediates abscisic acid response in Arabidopsis thaliana. Proceedings of the National Academy of Sciences of the United States of America, 109, 12822-12827.
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