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:: Volume 6, Issue 2 (Fall and Winter 2021) ::
FOP 2021, 6(2): 147-164 Back to browse issues page
Effect of the type of inoculation medium on chimeric chitinase gene transfer to lisianthus (Eustoma grandiflorum [Raf.] Shinn) for resistance to the fungal disease Fusarium solani
Mohammad Mehdi Fakhraei , AliReza Motallebi Azar * , Hassan Salehi , Nasser Mahna , Mostafa Motallebi
University of Tabriz
Abstract:   (2699 Views)
Lisianthus is in the top ten flowers in the world and in 2017, it was the fifth largest exporter and seller of ornamental plants in the world. Resistance to fungal diseases, especially Fusarium, is one of the most important breeding objectives for lisianthus. Little work has been done on the modification of resistance to fungal diseases in this ornamental plants. Therefore, gene transfer of chimeric chitinase to lisianthus will be an important contribution to achieve this important goal. The transfer and expression of chitinase gene in plants show a high level of resistance to fungal infections and a delay in the onset of symptoms when exposed to fungal pathogens. The researchers showed that the activity of chimeric chitinase in destroying fungal walls is significantly different from that of Chit42 chitinase. The aim of this study was to achieve resistance to Fusarium solani in lisianthus by transfer of chimeric chitinase gene using Agrobacterium tumefactions. In this study, the effect of different culture media, MS, ½MS and LB, two pH values of 5.2 and 5.8 and 30 and 15 mg L-1 sucrose and 30 mg L-1 maltose in the inoculation medium were investigated. The results showed that the B treatment with MS inoculation medium containing 30 g L-1 maltose with a pH of 5.2 gave the best response in gene transfer to lisianthus and the presence of a more stringent selection medium containing 100 mg L-1 of kanamycin was superior to the other treatments with an average regeneration of 11.13 plantlets per leaflet. The results of this study show that MS medium is more efficient than LB medium for gene transfer to lisianthus. Lowering the pH and changing the carbohydrate source from sucrose to maltose in the inoculum medium increased the transformation efficiency. In this study, 471 plantlets were generated from 45 leaf explants during gene transfer of chimeric chitinase to lisianthus in a more stringent selection medium and out of 21 randomly selected plantlets and lines, 10 lines were selected by PCR and 8 lines responded positively to bioassay test. This is the first report of chimeric chitinase gene transfer in lisianthus.
Keywords: Agrobacterium, Bioassay, Fusarium, Fungal disease resistance, Transgenic line
Full-Text [PDF 1857 kb]   (316 Downloads)    
Type of Study: Research | Subject: Special
Received: 2021/12/11 | Accepted: 2021/12/22 | Published: 2022/03/7
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58. Bradley, J. M., Deroles, S. C., Boase, M. R., Bloor, S., Swinny, E., Davies, K. M. (1999). Variation in the ability of the maize Lc regulatory gene to upregulate flavonoid biosynthesis in heterologous systems. Plant Science, 140(1), 31-39. [DOI:10.1016/S0168-9452(98)00200-3]
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61. Clough, S. J., Bent, A. F. (1998). Floral dip: A simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant Journal, 16(6), 735-743. https://doi.org/10.1046/j.1365-313x.1998.00343.x [DOI:10.1046/j.1365-313X.1998.00343.x]
62. Curtis, I. S., Nam, H. G. (2001). Transgenic radish (Raphanus sativus L. longipinnatus Bailey) by floral-dip method - Plant development and surfactant are important in optimizing transformation efficiency. Transgenic Research, 10(4), 363-371. [DOI:10.1023/A:1016600517293]
63. Fakhraei, M. M., Arab, M., Panahi, M. E. shariat. (2014a). Effect of cold, heat and chemical stresses on the induction of androgenesis in lisianthus (Eustoma grandiflorum). Journal of Applied Crop Breeding, 2(1), 1-12 (In Persian).
64. Fakhraei, M. M., Arab, M., Panahi, M. E. shariat. (2014b). Effect of cultivar, growth regulators and light during incubation on induction of haploid in lisianthus (Eustoma grandiflorum) through microspore culture. Journal of Crop Production and Processing, 4(12), 171-178 (In Persian).
65. Fan, Y., Fang, W., Guo, S., Pei, X., Zhang, Y., Xiao, Y., Li, D., Jin, K., Bidochka, M. J., Pei, Y. (2007). Increased insect virulence in Beauveria bassiana strains overexpressing an engineered chitinase. Applied and Environmental Microbiology, 73(1), 295-302. [DOI:10.1128/AEM.01974-06]
66. Gao, N., Shen, W., Cao, Y., Su, Y., Shi, W. (2009). Influence of bacterial density during preculture on Agrobacterium-mediated transformation of tomato. Plant Cell, Tissue and Organ Culture, 98(3), 321-330. [DOI:10.1007/s11240-009-9566-2]
67. Gilardi, G., Gullino, M. . (2006). Ornamentali - Prime osservazioni sulla suscettibilita a Fusarium oxysporum f. sp. eustomae di diverse cultivar di lisianthus (Eustoma grandiflorum). Informatore Fitopatologico, 56, 30-33.
68. Grison, R., Grezes-Besset, B., Schneider, M., Lucante, N., Olsen, L., Leguay, J. J., Toppan, A. (1996). Field tolerance to fungal pathogens of Brassica napus constitutively expressing a chimeric chitinase gene. Nature Biotechnology, 14(5), 643-646. [DOI:10.1038/nbt0596-643]
69. Guan, C., Liu, X., Song, X., Wang, G., Ji, J., Jin, C. (2014). Overexpression of a peroxiredoxin Q gene, SsPrxQ, in Eustoma grandiflorum Shinn enhances its tolerance to salt and high light intensity. Molecular Breeding, 33(3), 657-667. [DOI:10.1007/s11032-013-9982-1]
70. Hahm, Y.-I. (1998). Occurrence of Fusarium wilt on lisianthus (Eustoma grandiflorum) caused by Fusarium oxysporum f. sp. eustomae. Korean Journal of Plant Pathology, 14(2), 188-190.
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Fakhraei M M, Motallebi Azar A, Salehi H, mahna N, Motallebi M. Effect of the type of inoculation medium on chimeric chitinase gene transfer to lisianthus (Eustoma grandiflorum [Raf.] Shinn) for resistance to the fungal disease Fusarium solani. FOP 2021; 6 (2) :147-164
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گل و گیاهان زینتی Flower and Ornamental Plants
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