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Transgenic Lilium longiflorum plants containing the bar-uidA gene controlled by the rice RPC1, Agrobacterium rolD, mas2, and CaMV 35S promoters

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Abstract

Lilies are grown in the field for bulb production where they are susceptible to infection by Pratylenchus penetrans, the root lesion nematode. These migratory nematodes feed on roots and often physically disrupt the root tissues when many nematodes enter and move throughout the root to feed. The goal of this study was to find a root-specific or root-preferred promoter because of our interest in genetic engineering of lilies for nematode resistance. Several promoters, the Agrobacterium rolD and mas2, CaMV 35S, and rice RPC1 were examined in transgenic Lilium longiflorum, Easter lily, plants that contained the bar-uidA fusion gene under each promoter. Histochemical staining showed that the rolD, mas2, and CaMV 35S promoters directed gus gene expression in the cortex and stele of transformed roots. Gus expression was limited to the stele of roots with the rice RPC1 promoter making it unsuitable for engineering nematode resistance. The mas2 promoter was found to be expressed primarily in roots rather than shoots whereas the CaMV 35S promoter expressed well in both roots and shoots. Levels of gus specific activity were relatively high for both the mas2 and CaMV 35S promoter in roots but low with the rolD promoter. Because the mas2 promoter had relatively high levels of gus specific activity in roots and not shoots, and expression was throughout cortex and stele tissues of the roots, mas2 appears to be a promising promoter for engineering resistance to root lesion nematodes in Lilium longiflorum.

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References

  • Azadi P, Bagheri H, Nalousi AM, Nazari F, Chandler SF (2016) Current status and biotechnological advances in genetic engineering of ornamental plants. Biotech Adv 34:1073–1090

    Article  Google Scholar 

  • Bakhshaie M, Khosravi S, Azadi P, Bagheri H, van Tuyl JM (2016) Biotechnological advances in Lilium. Plant Cell Rep 35:1799–1826

    Article  CAS  PubMed  Google Scholar 

  • Bouché N, Fait A, Zik M, Fromm H (2004) The root-specific glutamate decarboxylase (GAD1) is essential for sustaining GABA levels in Arabidopsis. Plant Mol Biol 55:315–325

    Article  PubMed  Google Scholar 

  • Cohen A (2011) Biotechnology in lilies-dreams vs. reality. Acta Hort 900:149–160

    Article  Google Scholar 

  • Conjin C (2014) Developments in the control of lily diseases. Acta Hort 1027:213–229

    Article  Google Scholar 

  • Davies EL, MacGuidwin AE (2005) Lesion nematode disease. https://www.apsnet.org/edcenter/intropp/lessons/Nematodes/Pages/LesionNematode.aspx

  • Dellaporta S, Wood J, Hicks J (1983) A plant DNA minipreparation. Version II. Plant Mol Biol Rpt 1:19–21

    Article  CAS  Google Scholar 

  • Elmayan T, Tepfer M (1995) Evaluation in tobacco of the organ specificity and strength of the rolD promoter, domain A of the 35S promoter and the 35S2 promoter. Trans Res 4:388–396

    Article  CAS  Google Scholar 

  • Escobar C, De Meutter J, Aristizábal FA, Sanz-Alférez S, del Campo FF et al (1999) Isolation of the LEMM19 gene and promoter analysis during a compatible plant-nematode interaction. Mol Plant Microbe Int 12:440–449

    Article  CAS  Google Scholar 

  • FloraHolland in facts and Figs (2015) Koninklijke Cooperatieve Bloemenvelling FloraHolland U.A. https://www.royalfloralholland.com/en

  • Furtado A, Henry RJ, Takaiwa F (2008) Comparison of promoters in transgenic rice. Plant Biotech J 6:679–693

    Article  CAS  Google Scholar 

  • Goddijn OJM, Lindsey K, van der Lee FM, Klap JC, Sigmons PC (1993) Differential gene expression in nematode-induced feeding structures of transgenic plants harbouring promoter-gusA fusion constructs. Plant J 4:863–973

    Article  CAS  PubMed  Google Scholar 

  • Green J, Vain P, Fearnehough MT, Worland B, Snape JW, Atkinson HJ (2002) Analysis of the expression patterns of the Arabidopsis thaliana tubulin-1 and Zea mays ubiquitin- promoters in rice plants in association with nematode infection. Physiol Mol Plant Path 60:197–205

    Article  CAS  Google Scholar 

  • Guevara-García A, Mosqueda-Cano G, Argüello-Astorga G, Simpson J, Herrera-Estrella L (1993) Tissue-specific and wound-inducible pattern of expression of the mannopine synthase promoter is determined by the interaction between positive and negative cis–regulatory elements. Plant J 4:49594–49505

    Article  Google Scholar 

  • Guevara-García A, López-Bucio J, Herrera-Estrella L (1999) The mannopine synthase promoter contains vectorial cis-regulatory elements that act as enhancers and silencers. Mol Gen Genet 262:608–617

    Article  PubMed  Google Scholar 

  • Hensgens LAM, Fornerod MWJ, Rueb S, Winkler AA, van der Veen S, Schilperoort RA (1992) Translation controls the expression level of a chimaeric reporter gene. Plant Mol Biol 20:921–938

    Article  CAS  PubMed  Google Scholar 

  • Huang L-y, Zhang F, Qin Q, Wang W-s, Zhang T, Fu B-y (2015) Identification and validation of root-specific promoters in rice. J Int Agric 14:1–10

    Article  CAS  Google Scholar 

  • Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS-fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jeong H-J, Jung K-H (2015) Rice tissue-specific promoters and condition-dependent promoters for effective translational application. J Int Plant Biol 57:913–924

    Article  CAS  Google Scholar 

  • Kakrana A, Kumar A, Satheesh V, Abdin MZ, Subramaniam K, Bhattacharya RC, Srinivasan R, Sirohi A, Jain PK (2017) Identification, validation and utilization of novel nematode- responsive root-specific promoters in Arabidopsis for inducing host-delivered RNAi mediated root-knot nematode resistance. Front Plant Sci 8:article 2049

    Article  Google Scholar 

  • Kamo K, Blowers A (1999) Tissue specificity and expression level of gusA under rolD, mannopine synthase and translation elongation factor 1 subunit α promoters in transgenic Gladiolus plants. Plant Cell Rep 18:809–815

    Article  CAS  Google Scholar 

  • Koyama T, Ono T, Shimizu M, Jinbo T, Mizuno R, Tomita K, Mitsukawa N, Kawazu T, Kimura T, Ohmiya K, Sakka K (2005) Promoter of Arabidopsis thaliana phosphate transporter gene drives root-specific expression of transgene in rice. J Biosci Bioeng 99:38–42

    Article  CAS  PubMed  Google Scholar 

  • Langridge WHR, Fitzgerald KJ, Koncz C, Schell J, Szalay AA (1989) Dual promoter of Agrobacterium tumefaciens mannopine synthase genes is regulated by plant growth hormones. Proc Natl Acad Sci USA 86:3219–3223

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Leach F, Aoyagi K (1991) Promoter analysis of the highly expressed rolC and rolD root- inducing genes of Agrobacterium rhizogenes: enhancer and tissue-specific DNA determinants are dissociated. Plant Sci 79:69–76

    Article  CAS  Google Scholar 

  • Li Y, Liu S, Yu Z, Liu Y, Wu P (2013) Isolation and characterization of two novel root-specific promoters in rice (Oryza sativa L.). Plant Sci 207:37–44

    Article  CAS  PubMed  Google Scholar 

  • Lilley CG, Urwin PE, Johnston KA, Atkinson HJ (2004) Preferential expression of a plant cystatin at nematode feeding sites confers resistance to Meloidogyne incognita and Globodera pallida. Plant Biotech J 2:3–12

    Article  CAS  Google Scholar 

  • Lilley CJ, Wang D, Atkinson HJ, Urwin PE (2011) Effective delivery of a nematode-repellent peptide using a root-cap-specific promoter. Plant Biotech J 9:151–161

    Article  CAS  Google Scholar 

  • Liu J, Zhang J, Xu B, Jia C, Zhang J, Tan G, Jin Z (2011) Regeneration and production of transgenic Lilium longiflorum via Agrobacterium tumefaciens. Vitro Cell Dev Biol-Plant 47:348–356

    Article  Google Scholar 

  • Luo HL, Luo LP, Guan BC, Li EX, Xiong J, Sun BT, Peng K, Yang BY (2014) Evaluation of candidate references genes for RT-qPCR in lily (Lilium brownii). J Hort Sci Biotech 89:345–351

    Article  CAS  Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

    Google Scholar 

  • Matsuyama T, Yasumura N, Funakoshi M, Yamada Y, Hashimoto T (1999) Maize genes specifically expressed in the outermost cells of the root cap. Plant Cell Physiol 40:469–476

    Article  CAS  PubMed  Google Scholar 

  • Mauro ML, Trovato M, De Paolis A, Gallelli A, Costantino P, Altamura MM (1996) The plant oncogene rolD stimulates flowering in transgenic tobacco plants. Dev Biol 180:693–700

    Article  CAS  PubMed  Google Scholar 

  • Mauro ML, Constantino P, Bettini PP (2017) The never ending story of rol genes: a century after. Plant Cell Tiss Org Cult 131:201–212

    Article  CAS  Google Scholar 

  • McElroy D, Zhang W, Cao J, Wu R (1990) Isolation of an efficient actin promoter for use in rice transformation. Plant Cell 2:163–171

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mizutani M, Watanabe S, Nakagawa T, Maeshima M (2006) Aquaporin NIP2; 1 is mainly localized to the ER membrane and shows root-specific accumulation in Arabidopsis thaliana. Plant Cell Physiol 47:1420–1426

    Article  CAS  PubMed  Google Scholar 

  • Murakami T, Anzai H, Imai S, Satoh A, Nagaoka K, Thompson CJ (1986) The bialaphos biosynthetic genes of Streptomyces hygroscopicus: molecular cloning and characterization of the gene cluster. Mol Gen Genet 205:42–50

    Article  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid assays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Onyango SO, Roderick H, Tripathi JN, Collins R, Atkinson HJ, Oduor RO, Tripathi L (2016) The ZmRCP-1 promoter of maize provides root tip specific expression of transgenes in plantain. J Biol Res-Thessaloniki 23:4 https://doi.org/10.1186/s40709-016-0041-z

    Article  CAS  Google Scholar 

  • Opperman CH, Taylor CG, Conkling MA (1994) Root-knot nematode directed expression of a plant root-specific gene. Science 263:221–223

    Article  CAS  PubMed  Google Scholar 

  • Roderick H, Tripathi L, Babirye A, Wang D, Tripathi J, Urwin PE, Atkinson HJ (2012) Generation of transgenic plantain (Musa spp.) with resistance to plant pathogenic nematodes. Mol Plant Path 13:842–851

    Article  CAS  Google Scholar 

  • Trovato M, Maras B, Linhares F, Costantino P (2001) The plant oncogene rolD encodes a functional ornithine cyclodeaminase. Proc Natl Acad Sci USA 98:13449–13453

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Urwin PE, Lilley CJ, McPherson MJ, Atkinson HJ (1997) Resistance to both cyst and root knot nematodes conferred by transgenic Arabidopsis expressing a modified plant cystatin. Plant J 12:455–461

    Article  CAS  PubMed  Google Scholar 

  • USDA National Agricultural Statistics Service. (2016) Floriculture Crops. 2015 Summary. https://www.agmrc.org/commodities-products/specialty-crops/floriculture

  • Vieira P, Wantoch S, Lilley CJ, Chitwood DJ, Atkinson HJ, Kamo K (2015a) Expression of a cystatin transgene can confer resistance to the root lesion nematodes in Lilium longiflorum cv. ‘Nellie White’ Trans Res 24:423–432

    Google Scholar 

  • Vieira P, Eves-van den Akker S, Verma R, Wantoch S, Eisenback JD, Kamo K (2015b) The Pratylenchus penetrans transcriptome as a source for the development of alternative control strategies: mining for putative genes involved in parasitism and evaluation of in planta RNAi. PLoS ONE 10:e0144674

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vieira P, Mowery J, Kilcrease J, Eisenback JD, Kamo K (2017) Characterization of Lilium longiflorum cv. ‘Nellie White’ infection with root-lesion nematode Pratylenchus penetrans by bright-field and transmission electron microscopy. J Nemat 49:2–11

    Article  CAS  Google Scholar 

  • Vissenberg K, Oyama M, Osato Y, Yokoyama R, Verbelen J-P, Nishitani K (2005) Differential expression of AtXTH, AtXTH18, AtXTY19, AtXTH20 genes in Arabidopsis roots. Physiological roles in specification in cell wall construction. Plant Cell Physiol 46:192–200

    Article  CAS  PubMed  Google Scholar 

  • Walawage SL, Britton MT, Leslie CA, Uratsu SL, Li Y, Dandekar AM (2013) Stacking resistance to crown gall and nematodes in walnut rootstocks. BMC Genom 14:668–680

    Article  CAS  Google Scholar 

  • Wally O, Jayaraj J, Punga ZK (2008) Comparative expression of β-glucuronidase with five different promoters in transgenic carrot (Daucus carota L.) root and leaf tissues. Plant Cell Rep 27:279–287

    Article  CAS  PubMed  Google Scholar 

  • Westerdahl BB, Giraud D, Etter S, Riddle LJ, Anderson CA (1998) Problems associated with crop rotation for management of Pratylenchus penetrans on Easter Lily. J Nemat 30(4S):581–589

    CAS  Google Scholar 

  • Won S-K, Lee Y-J, Lee H-Y, Heo Y-K, Cho M, Cho H-T (2009) Cis-element-and transcription-based screening of root hair-specific genes and their functional characterization in Arabidopsis. Plant Physiol 150:1459–1473

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xue G-P, Rae AL, White RG, Drenth J, Richardson T, McIntrye CL (2016) A strong root-specific expression system for stable transgene expression in bread wheat. Plant Cell Rep 35:469–481

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

Anne O’Connor and Sarah Shih are thanked for technical assistance. Mention of a trademark, proprietary product or vendor does not imply its approval to the exclusion of other products or vendors that may also be suitable.

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Authors and Affiliations

  1. Floral and Nursery Plants Research Unit, USDA, National Arboretum, Beltsville, MD, USA

    Kathryn Kamo

  2. Crop Improvement and Genetics Research, USDA-ARS, Western Regional Research Center, Albany, CA, USA

    Roger Thilmony

  3. Electron and Confocal Microscopy Unit, USDA, Beltsville, MD, USA

    Gary Bauchan

Authors
  1. Kathryn Kamo
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  2. Roger Thilmony
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  3. Gary Bauchan
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Contributions

KK planned the study, made the transformed lilies, did histochemical gus staining, and wrote the manuscript. RT provided input and the rice RPC1 construct and sequence and read and reviewed the manuscript. GB took photos of histochemically stained lilies and made the root cross sections.

Corresponding author

Correspondence to Kathryn Kamo.

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Conflict of interest

None of the authors have any conflict of interest with this research.

Ethical approval

This research does not involve animal or human subjects. All authors have reviewed the manuscript and given their consent to submit. The manuscript has been approved for submission to a journal by the required USDA officials.

Additional information

Communicated by T. Winkelmann.

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Kamo, K., Thilmony, R. & Bauchan, G. Transgenic Lilium longiflorum plants containing the bar-uidA gene controlled by the rice RPC1, Agrobacterium rolD, mas2, and CaMV 35S promoters. Plant Cell Tiss Organ Cult 136, 303–312 (2019). https://doi.org/10.1007/s11240-018-1515-5

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  • DOI: https://doi.org/10.1007/s11240-018-1515-5

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