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Journal of Integrative Agriculture  2015, Vol. 14 Issue (10): 1980-1991    DOI: 10.1016/S2095-3119(15)61059-0
Physiology·Biochemistry·Cultivation·Tillage Advanced Online Publication | Current Issue | Archive | Adv Search |
Over-expression of GhDWF4 gene improved tomato fruit quality and accelerated fruit ripening
 YE  Shu-e, LI  Fang, LI  Xian-bi, HONG  Qi-bin, ZHAI  Yun-lan, HU  Ming-yu, WEI  Ting, DENG  Sha-sha, PEI  Yan, LUO  Ming
1、Key Laboratory of Biotechnology and Crop Quality Improvement, Ministry of Agriculture/Biotechnology Research Center,
Southwest University, Chongqing 400716, P.R.China
2、Citrus Research Institute, Chinese Academy of Agricultural Sciences/National Citrus Engineering Research Center, Chongqing
400712, P.R.China
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摘要  Brassinosteroids (BRs), a class of steroidal phytohormones are essential for many biological processes in plant. However, little is known about their roles in fruit development. Tomato is a highly valuable vegetable and has been adopted as the model species for studying fruit growth, development, and ripening. To understand the role of endogenous BRs in the development of tomato fruit, the expression patterns of three homologues of DWF4 gene were investigated and the transgenic tomato plants were generated in which the GhDWF4 gene from upland cotton (Gossypium hirsutum L.) was ectopically expressed. The contents of main quality components were analyzed in fruits of transgenic tomato line and non-transgenic line (control plant, CP) when the fruit was mature. SlCYP90B3 that possesses high homology with GhDWF4 preferentially expressed in mature fruit. Significantly higher contents of soluble sugar, soluble proteins, and vitamin C were obtained in fruit of transgenic tomato lines compared with those in the CP. Furthermore, overexpressing GhDWF4 promoted fruit growth and ripening. The weight per fruit was increased by about 23% in transgenic lines. In addition, overexpressing GhDWF4 promoted the germination of transgenic tomato seeds and hypocotyl elongation of seedlings. These results indicated that overexpressing GhDWF4 gene in tomato could increase the contents of many nutrients in fruit and accelerate fruit ripening. It is suggested that increased endogenous BRs in fruit affect the growth and development of tomato fruit and therefore improved the nutrient quality of tomato.

Abstract  Brassinosteroids (BRs), a class of steroidal phytohormones are essential for many biological processes in plant. However, little is known about their roles in fruit development. Tomato is a highly valuable vegetable and has been adopted as the model species for studying fruit growth, development, and ripening. To understand the role of endogenous BRs in the development of tomato fruit, the expression patterns of three homologues of DWF4 gene were investigated and the transgenic tomato plants were generated in which the GhDWF4 gene from upland cotton (Gossypium hirsutum L.) was ectopically expressed. The contents of main quality components were analyzed in fruits of transgenic tomato line and non-transgenic line (control plant, CP) when the fruit was mature. SlCYP90B3 that possesses high homology with GhDWF4 preferentially expressed in mature fruit. Significantly higher contents of soluble sugar, soluble proteins, and vitamin C were obtained in fruit of transgenic tomato lines compared with those in the CP. Furthermore, overexpressing GhDWF4 promoted fruit growth and ripening. The weight per fruit was increased by about 23% in transgenic lines. In addition, overexpressing GhDWF4 promoted the germination of transgenic tomato seeds and hypocotyl elongation of seedlings. These results indicated that overexpressing GhDWF4 gene in tomato could increase the contents of many nutrients in fruit and accelerate fruit ripening. It is suggested that increased endogenous BRs in fruit affect the growth and development of tomato fruit and therefore improved the nutrient quality of tomato.
Keywords:  brassinosteroids       GhDWF4       tomato       fruit ripening       fruit quality       transgenic plant  
Received: 27 October 2014   Accepted:
Fund: 

This work was supported by the Natural Science Foundation of Chongqing, China (CSTC, 2011BB1007) and the Genetically Modified Organisms Breeding Major Projects of China (2009ZX08009-118B).

Corresponding Authors:  LUO Ming, Tel: +86-23-68250042, Fax: +86-23-68251883, E-mail: luo0424@126.com   
About author:  YE Shu-e, E-mail: 61730797@qq.com;* These authors contributed equally to this work.

Cite this article: 

YE Shu-e, LI Fang, LI Xian-bi, HONG Qi-bin, ZHAI Yun-lan, HU Ming-yu, WEI Ting, DENG Sha-sha, PEI Yan, LUO Ming. 2015. Over-expression of GhDWF4 gene improved tomato fruit quality and accelerated fruit ripening. Journal of Integrative Agriculture, 14(10): 1980-1991.

Ali B, Hayat S, Aiman H S, Ahmad A. 2006. Effect of root applied28-homobrassinolide on the performance of Lycopersiconesculentum. Scientia Horticulturae, 110, 267-273

Altmann T. 1998. A tale of dwarfs and drugs: Brassinosteroidsto the rescue. Trends in Genetics, 14, 490-495

Anuradha S, Rao S S R. 2001. Effect of brassinosteroids onsalinity stress induced inhibition of seed germination andseedling growth of rice (Oryza sativa L.). Plant GrowthRegulation, 33, 151-153

Arteca J M, Arteca R N. 2001. Brassinosteroid inducedexaggerated growth in hydroponically grown Arabidopsisplants. Physiologia Plantarum, 112, 104-112

Asami T, Nakano T, Fujioka S. 2005. Plant brassinosteroidhormones. Vitamins & Hormones, 72, 479-504

Bishop G J, Yokota T. 2001. Plants steroid hormones,brassinosteroids: Current highlights of molecular aspectson their synthesis/metabolism, transport, perception andresponse. Plant Cell Physiology, 42, 114-120

Bai W Q, Xiao Y H, Zhao J, Song S Q, Hu L, Zeng J Y, LiX B, Hou L, Luo M, Li D M, Pei Y. 2014. Gibberellinoverproduction promotes sucrose synthase expressionand secondary cell wall deposition in cotton fibers. PLOSONE, 9, e96537.

Bishop G J. 2003. Brassinosteroid mutants of crops. Journalof Plant Growth Regulation, 22, 325-335

Bishop G J, Nomura T, Yokota T, Harrison K, Noguchi T, FujiokaS, Takatsuto S, Jones J D G, Kamiya Y. 1999. The tomatoDWARF enzyme catalyses C-6 oxidation in brassinosteroidbiosynthesis. Proceedings of the National Academy ofSciences of the United States of America, 96, 1761-1766

Bradford. 1976. A rapid and sensitive method for the quantitationof microgram quantities of protein utilizing the principle ofprotein-dye bindinn. Analytical Biochemistry, 72, 248-254

Chai Y M, Zhang Q, Tian L, Li C L, Xing Y, Qin L, Shen Y Y.2013. Brassinosteroid is involved in strawberry fruit ripening.Journal of Plant Growth Regulation, 69, 63-69

Choe S, Dilkes B P, Fujioka S, Takatsuto S, Sakurai A,Feldmann K A. 1998. The DWF4 gene of Arabidopsisencodes a cytochrome P450 that mediates multiple22α-hydroxylation steps in brassinosteroid biosynthesis.The Plant Cell, 10, 231-244

Choe S, Fujioka S, Noguchi T, Takatsuto S, Yoshida S,Feldmann K A. 2001. Overexpression of DWARF4 in thebrassinosteroid biosynthetic pathway results in increasedvegetative growth and seed yield in Arabidopsis. The PlantJournal, 26, 573-582

Clouse S D. 2002. Arabidopsis mutants reveal multiple roles forsterols in plant development. The Plant Cell, 14, 1995-2000

Clouse S D. 2011. Brassinosteroids. Arabidopsis Book, 9, e0151Clouse S D, Sasse J M. 1998. Brassinosteroids: Essentialregulators of plant growth and development. AnnualReview of Plant Physiology and Plant Molecular Biology,49, 427-451

Dhaubhadel S, Chaudhary S, Dobinson K F, Krishna P. 1999.Treatment with 24-epibrassinolide, a brassinosteroid,increases the basic thermotolerance of Brassica napus andtomato seedlings. Plant Molecular Biology, 40, 333-342

Fu F Q, Mao W H, Shi K, Zhou Y H, Asami T, Yu J Q. 2008.A role of brassinosteroids in early fruit development incucumber. Journal of Experimental Botany, 59, 2299-2308

Fujioka S, Inoue T, Takatsuto S, Yanagisawa T, Yokota T,Sakurai A. 1995. Identification of a new brassinosteroid,cathasterone, in cultured cells of catharanthus roseusas a biosynthetic precursor of teasterone. Bioscience, Biotechnology and Biochemistry, 59, 1543-1547

Fujita S, Ohnishi T, Watanabe B, Yokota T, Takatsuto S, FujiokaS, Yoshida S, Sakata K, Mizutani M. 2006. ArabidopsisCYP90B1 catalyses the early C-22 hydroxylation of C27,C28 and C29 sterols. The Plant Journal, 45, 765-774

Galpaz N, Wang Q, Menda N, Zamir D, Hirschberg J. 2008.Abscisic acid deficiency in the tomato mutant high-pigment3 leading to increased plastid number and higher fruitlycopene content. The Plant Journal, 53, 717-730

Gendron J M, Liu J S, Fan M, Bai M Y, Wenkel S, Springer PS, Barton M K, Wang Z Y. 2012. Brassinosteroids regulateorgan boundary formation in the shoot apical meristemof Arabidopsis. Proceedings of the National Academy ofSciences of the United States of America, 109, 21152-21157

Guo H, Li L, Aluru M, Aluru S, Yin Y. 2013. Mechanisms andnetworks for brassinosteroid regulated gene expression.Current Opinion in Plant Biology, 16, 545-553

Hartwig T, Chuck G S, Fujioka S, Klempien A, WeizbauerR, Potluri D P, Choe S, Johal G S, Schulz B. 2011.Brassinosteroid control of sex determination in maize.Proceedings of the National Academy of Sciences of theUnited States of America, 108, 19814-19819

Hasan S A, Irfan M, Hayat S. 2014. Response of tomatocultivars on yield and quality attributes applied with twodifferent modes of BR analogues: A comparative study.In: International Conference on Advances in Agricultural,Biological & Environmental Sciences (AABES-2014). Oct15-16, 2014, Dubai (UAE)

Hayat S, Alyemeni M N, Hasan S A. 2012. Foliar spray ofbrassinosteroid enhances yield and quality of Solanumlycopersicum under cadmium stress. Saudi Journal ofBiological Sciences, 19, 325-335

Kamuro Y, Takatsuto S. 1999. Practical application ofbrassinosteroids in agricultural fields. In: Sakurai A, YokotaT, Clouse S D, eds., Brassinosteroids: Steroidal PlantHormones. Springer-Verlag, Tokyo, Japan. pp. 223-241

Kataoka K, Yashiro Y, Habu T, Sunamoto K, Kitajima A. 2009.The addition of gibberellic acid to auxin solutions increasessugar accumulation and sink strength in developing auxininducedparthenocarpic tomato fruits. Scientia Horticulturae,123, 228-233

Khripach V A, Zhabinskii V N, de Groot A E. 2000. Twentyyears of brassinosteroids: Steroidal plant hormones warrantbetter crops for XXI century. Annals of Botany, 86, 441-447

Kim H B, Kwon M, Ryu H, Fujioka S, Takatsuto S, YoshidaS, An C S, Lee I, Hwang I, Choe S. 2006. The regulationof DWARF4 expression is likely a critical mechanism inmaintaining the homeostasis of bioactive brassinosteroidsin Arabidopsis. Plant Physiology, 140, 548-557

Koka C V, Cerny R E, Gardner R G, Noguchi T, Fujioka S,Takatsuto S, Yoshida S, Clouse S D. 2000. A putative rolefor the tomato genes DUMPY and CURL-3 in brassinosteroidbiosynthesis and response. Plant Physiology, 122, 85-98

Krishna P. 2003. Brassinosteroids-mediated stress responses.Journal of Plant Growth Regulation, 22, 289-297

Lee J M, Joung J G, McQuinn R, Chung M Y, Fei Z, TiemanD, Klee H, Giovannoni J. 2012. Combined transcriptome,genetic diversity and metabolite profiling in tomato fruitreveals that the ethylene response factor SlERF6 plays animportant role in ripening and carotenoid accumulation. ThePlant Journal, 70, 191-204

Li Zhen, Wei J, Jia C G, Wang Q M. 2010. Effect of bzr1 genetransformation on characters of fruit of tomato. ScientiaAgricultura Sinica, 43, 1-9 (in Chinese)

Lisso J, Altmann T. 2003. The effect of BRs on ripening andcomposition of the tomato fruit. Plant Biology, 7, 25-30

Lisso J, Altmann T, Müssig C. 2006. Metabolic changes in fruitsof the tomato dx mutant. Phytochemistry, 67, 2232-2238

Liu L H, Jia C G, Zhang M, Chen D L, Chen S X, Guo R F, GuoD P, Wang Q M. 2013. Ectopic expression of a BZR1-1Dtranscription factor in brassinosteroid signalling enhancescarotenoid accumulation and fruit quality attributes intomato. Plant Biotechnology Journal, 12, 105-115

Luo M, Xiao Y, Li X, Lu X, Deng W, Li D, Hou L, Hu M, Li Y,Pei Y. 2007. GhDET2, a steroid 5α-reductase, plays animportant role in cotton fiber cell initiation and elongation.The Plant Journal, 51, 419-430

Montoya T, Nomura T, Yokota T, Farrar K, Harrison K, Jones J,Kaneta T, Kamiya Y, Szekeres M, Bishop G J. 2005. Patternsof Dwarf expression and brassinosteroid accumulation intomato reveal the importance of brassinosteroid synthesisduring fruit development. The Plant Journal, 42, 262-269

Nomura T, Ueno M, Yamada Y, Takatsuto S, Takeuchi Y,Yokota T. 2007. Roles of brassinosteroids and relatedmRNAs in pea seed growth and germination. PlantPhysiology, 143, 1680-688

Ohnishi T, Nomura T, Watanabe B, Ohta D, Yokota T, MiyagawaH, Sakata K, Mizutani M. 2006a. Tomato cytochromeP450 CYP734A7 functions in brassinosteroid catabolism.Phytochemistry, 67, 1895-1906

Ohnishi T, Watanabe B, Sakata K, Mizutani M. 2006b.CYP724B2 and CYP90B3 function in the early C-22hydroxylation steps of brassinosteroid biosynthetic pathwayin tomato. Bioscience, Biotechnology, and Biochemistry,70, 2071-2080

Park S H, Morris J L, Park J E, Hirschi K D, Smith R H.2003. Efficient and genotype-independent Agrobacterium-mediated tomato transformation. Plant Physiology, 160,1253-1257

Shimada Y, Goda H, Nakamura A, Takatsuto S, FujiokaS, Yoshida S. 2003. Organ-specific expression ofbrassinosteroidbiosynthetic genes and distribution ofendogenous brassinosteroids in Arabidopsis. PlantPhysiology, 131, 287-297

Srivastava A, Handa A K. 2005. Hormonal regulation of tomatofruit development: A molecular perspective. Plant GrowthRegulation, 24, 67-82

Steber C M, McCourt P. 2001. A role for brassinosteroids ingermination in Arabidopsis. Plant Physiology, 125, 763-769

Suntornsuk L, Gritsanapun W, Nilkamhank S, Paochom A.2002. Quantitation of vitamin C content in herbal juice using direct titration. Journal of Pharmaceutical and BiomedicalAnalysis, 28, 849-855

Susila T, Amarender Reddy S, Rajkumar M, Padmaja G,RaoP V. 2012. Effects of sowing date and spraying ofbrassinosteroid on yield and fruit quality characters ofwatermelon. World Journal of Agricultural Sciences, 8,223-228

Symons G M, Davies C, Shavrukov Y, Dry I B, Reid J B,Thomas M R. 2006. Grapes on steroids. Brassinosteroidsare involved in grape berry ripening. Plant Physiology,140, 150-158

Tanabe S, Ashikari M, Fujioka S, Takatsuto S, Yoshida S, YanoM, Yoshimura A, Kitano H, Matsuoka M, Fujisawa Y, KatoH, Iwasaki Y. 2005. A novel cytochrome P450 is implicatedin brassinosteroid biosynthesis via the characterization of arice dwarf mutant, dwarf11, with reduced seed length. ThePlant Cell, 17, 776-790

Vardhini B V, Rao S S. 2002. Acceleration of ripening of tomatopericarp discs by brassinosteroids. Phytochemistry, 61,843-847

Vert G, Nemhauser J L, Geldner N, Hong F X, Chory J. 2006.Molecular mechanisms of steroid hormone signaling inplants. Annual Review of Cell and Developmental Biology,21, 177-201

Xiao Y H, Li D M, Yin M H, Li X B, Zhang M, Wang Y J, DongJ, Zhao J, Luo M, Luo X Y, Hou L, Hu L, Pei Y. 2010.Gibberellin 20-oxidase promotes initiation and elongationof cotton fibers by regulating gibberellin synthesis. Journalof Plant Physiology, 167, 829-837

Yemm E W, Willis A J. 1954. The estimation of carbohydratesin plant extracts by anthrone. Biochemical Journal, 57,508-514

Yoshimitsu Y, Tanaka K, Fukuda W, Asami T, Yoshida S,Hayashi K I, Kamiya Y, Jikumaru Y, Shigeta T, NakamuraY, Matsuo T, Okamoto S. 2011. Transcription of DWARF4plays a crucial role in auxin-regulated root elongation inaddition to brassinosteroid homeostasis in Arabidopsisthaliana. PLoS ONE, 6, e23851.

Yu J Q, Huang L F, Hu W H, Zhou Y H, Mao W H, Ye SF, Nogues S. 2004. A role for brassinosteroids in theregulation of photosynthesis in Cucumis sativus. Journalof Experimental Botany, 55, 1135-1143

Zaharah S S, Singh Z, Symons G M, Reid J B. 2012. Role ofbrassinosteroids, ethylene, abscisic acid, and indole-3-acetic acid in mango fruit ripening. Journal of Plant GrowthRegulation, 31, 363-372
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