Improvement of biomass production in transgenic Melia azedarach L. plants by the expression of a GA20-oxidase gene

Improvement of biomass production in transgenic Melia azedarach L. plants by the expression of a GA20-oxidase gene

Melia azedarach L. is a multipurpose timber tree that is important in forestry plantations. Genetic modification to improvethe growth rate and wood quality is important to produce new cultivars of M. azedarach. In this study, the effects of AtGA20ox geneexpression on plant growth, wood formation, and biomass production in transgenic M. azedarach plants were investigated. AtGA20oxwas inserted into the binary vector pBI101 under the control of the xylem specific promoter CAD4 and transferred to M. azedarach viaAgrobacterium-mediated method. The integration and expression of the transgenes were confirmed using PCR, RT-PCR, and Southernblotting. The specificity of CAD4 in the xylem was indicated by the higher expression of the AtGA20ox gene in the stem tissues comparedto the leaves. The larger stem diameter resulted from higher xylem cell numbers and a wider xylem zone in all the transgenic plants. Inaddition, the transgenic plants also grew more quickly. Consequently, transgenic M. azedarach increased its biomass production 2- to4-fold compared to the wild-type plants at 90 days under greenhouse conditions. Our results suggest that the expression of ectopicAtGA20ox under the control of a xylem-specific promoter is an appropriate method to improve plant growth, biomass production, andwood formation in M. azedarach and other forestry plants.

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  • Ayano M, Kani T, Kojima M, Sakakibara H, Kitaoka T et al. (2014). Gibberellin biosynthesis and signal transduction is essential for internode elongation in deepwater rice. Plant Cell Environment 37: 2313-2324.
  • Barakat A, Bagniewska-Zadworna A, Choi A, Plakkat U, DiLoreto DS et al. (2009). The cinnamyl alcohol dehydrogenase gene family in Populus: phylogeny, organization, and expression. BMC Plant Biology 9: 26.
  • Biemelt S (2004). Impact of altered gibberellin metabolism on biomass accumulation, lignin biosynthesis, and photosynthesis in transgenic tobacco plants. Plant Physiology 135: 254-265.
  • Carrera E, Bou J, García-Martínez JL, Prat S (2000). Changes in GA 20-oxidase gene expression strongly affect stem length, tuber induction and tuber yield of potato plants. Plant Journal 22: 247-256.
  • Coles JP, Phillips AL, Croker SJ, Garía-Lepe R, Lewis MJ et al. (1999). Modification of gibberellin production and plant development in Arabidopsis by sense and antisense expression of gibberellin 20-oxidase genes. Plant Journal 17: 547-556.
  • Dayan J (2016). Gibberellin transport. Annual Plant Reviews 49: 95- 120.
  • Dayan J, Schwarzkopf M, Avni A, Aloni R (2010). Enhancing plant growth and fiber production by silencing GA 2-oxidase. Plant Biotechnology Journal 8: 425-435.
  • Dellaporta SL, Wood J, Hicks JB (1983). A plant DNA minipreparation: version II. Plant Molecular Biology Reporter 1: 19-21.
  • Do PT, De Tar JR, Lee H, Folta MK, Zhang ZJ (2016). Expression of ZmGA20ox cDNA alters plant morphology and increase biomass production of switchgrass (Panicum virgatum L.). Plant Biotechnology Journal 14: 1532-1540.
  • Dong DX, Thang BV, Giang HV, Son LV, Ha CH (2011). Transformation of gibberellin 20-oxidase encoded gene to paradise tree (Melia azedarach L.) via Agrobacterium tumefaciens. Vietnam Journal of Biotechnology 9: 217-222 (in Vietnamese with an abstract in English).
  • Eriksson ME, Israelsson M, Olsson O, Moritz T (2000). Increased gibberellin biosynthesis in transgenic trees promotes growth, biomass production and xylem fiber length. Nature Biotechnology 18: 784-788.
  • Fagoaga C, Tadeo FR, Iglesias DJ, Huerta L, Lliso I et al. (2007). Engineering of gibberellin levels in citrus by sense and antisense overexpression of a GA 20-oxidase gene modifies plant architecture. Journal of Experimental Botany 58: 1407- 1420.
  • FAO (2004). Forest Genetic Resources Working Paper FGR/59E. Rome, Italy: FAO.
  • Feuillet C, Lauvergeat V, Deswarte C, Pilate G, Boudet A et al. (1995). Tissue- and cell-specific expression of a cinnamyl alcohol dehydrogenase promoter in transgenic poplar plants. Plant Molecular Biology 27: 651-667.
  • García-Hurtado N, Carrera E, Ruiz-Rivero O, López-Gresa MP, Hedden P et al. (2012). The characterization of transgenic tomato overexpressing gibberellin 20-oxidase reveals induction of parthenocarpic fruit growth, higher yield, and alteration of the gibberellin biosynthetic pathway. Journal of Experimental Botany 63: 5803-5813.
  • Hedden P, Kamiya Y (1997). Gibberellin biosynthesis: enzymes, genes and their regulation. Annual Review of Plant Physiology and Plant Molecular Biology 48: 431-460.
  • Hedden P, Thomas SG (2012). Gibberellin biosynthesis and its regulation. Biochemical Journal 444: 11-25.
  • Huang RC, Tadera K, Yagi F, Minami Y, Okamura H et al. (1996). Limonoids from Melia azedarach. Phytochemistry 43: 581-583.
  • Jeon HW, Cho JS, Park EJ, Han KH, Choi YI et al. (2015). Developing xylem-preferential expression of PdGA201, a gibberellin 20-oxidase 1 from Pinus densiflora, improves woody biomass production in a hybrid poplar. Plant Biotechnology Journal 14: 1161-1170.
  • Ko JH, Kim HT, Hwang ID, Han KH (2012). Tissue-type-specific transcriptome analysis identifies developing xylem-specific promoters in poplar. Plant Biotechnology Journal 10: 587-596.
  • Mauriat M, Petterle A, Bellini C, Moritz T (2014). Gibberellins inhibit adventitious rooting in hybrid aspen and Arabidopsis by affecting auxin transport. Plant Journal 78: 372-384.
  • Ngo VT, Jiang X, Ha VH, Nguyen TH, Ho VG (2010) Vector construction and transformation of 4CL1 gene into Chinaberrytree (Melia azedarach L.). VNU Journal of Science, Natural Sciences and Technology 26: 205-210.
  • Nhung NH, Thao BP, Doai NV, Phong NV, Anh LTV et al. (2017). Transformation of GA20-oxidase gene under regulation of xylem-specific promoter CAD4 in tobacco (Nicotiana tabacum). VNU Journal of Science 33: 77-83 (in Vietnamese with an abstract in English).
  • Nirsatmanto A, Gyokusen K (2007) Genetic transformation of Melia azedarach L., using Agrobacterium mediated transformation. Indonesian Journal of Forestry Research 4 (1): 1-8.
  • Olszewski N, Sun TP, Gubler F (2002). Gibberellin signaling: biosynthesis, catabolism, and response pathways. Plant Cell 14: 61-80.
  • Ragni L, Nieminen K, Pacheco-Villalobos D, Sibout R, Schwechheimer C et al. (2011). Mobile gibberellin directly stimulates Arabidopsis hypocotyl xylem expansion. Plant Cell 23: 1322-1336.
  • Rubae AY (2009). The potential uses of Melia azedarach L. as pesticidal and medicinal plant, review. American-Eurasian Journal of Sustainable Agriculture 3 (2): 185-194.
  • Sharma N, Sharma N (1995). Effect of leaf powder/extract of neem and Persian lilac on post-harvest diseases of tomato fruit. National Academy Science Letters 18: 11-14.
  • Vidal AM, Gisbert C, Talon M, Primo-Millo E, Lopez-Diaz I et al. (2001). The ectopic overexpression of a citrus gibberellin 20-oxidase enhances the non-13-hydroxylation pathway of gibberellin biosynthesis and induces an extremely elongated phenotype in tobacco. Physiologia Plantarum 112: 251-260.
  • Voorend W, Nelissen H, Vanholme R, De Vliegher A, Breusegem FV (2015). Overexpression of GA20-oxidase1 impacts plant height, biomass allocation and saccharification efficiency in maize. Plant Biotechnology Journal 14: 997-1007.
  • Xu YL, Li L, Wu K, Peeters AJ, Gage DA et al. (1995). The GA5 locus of Arabidopsis thaliana encodes a multifunctional gibberellin 20-oxidase: molecular cloning and functional expression. Proceedings of the National Academy of Sciences of the United States of America 92: 6640-6644.
  • Yamaguchi S (2008). Gibberellin metabolism and its regulation. Annual Review of Plant Biology 59: 225-251.