آمار
| تعداد نشریات | 28 |
| تعداد شماره‌ها | 426 |
| تعداد مقالات | 4,164 |
| تعداد مشاهده مقاله | 6,739,393 |
| تعداد دریافت فایل اصل مقاله | 4,988,826 |
Analysis of the Effect of Chimeric Chitinase Expressed by Synthetic Promoter in T2 Generation of Transgenic Canola | ||
| Journal of Genetic Resources | ||
| دوره 9، شماره 1، اردیبهشت 2023، صفحه 48-58 اصل مقاله (632.34 K) | ||
| نوع مقاله: Research Article | ||
| شناسه دیجیتال (DOI): 10.22080/jgr.2023.24506.1336 | ||
| نویسندگان | ||
| Mehdi Moradyar؛ Mohammadreza Zamani* ؛ Mostafa Motallebi؛ Esmat Jourabchi | ||
| Department of Plant Biotechnology, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran | ||
| تاریخ دریافت: 26 مرداد 1401، تاریخ بازنگری: 26 آذر 1401، تاریخ پذیرش: 16 مهر 1402 | ||
| چکیده | ||
| The chitinase enzymes are known to play an important role in the plant defense system against phytopathogenic fungi. The effect of chimeric chitinase, which is chitinase-42 with a chitin-binding domain (ChBD), was previously analyzed in the T0 generation of the transgenic canola. In this research, three homozygous lines (pGFC3, pGFC13, and pGFC26) containing a single copy of the transgene (chimeric chitinases) on the two homologous chromosomes were selected in the T2 generation using a kanamycin-resistant marker (NPTII gene). The selected homozygous plants in T2 generation were induced by chitin as an elicitor in the greenhouse. The results of the semi-quantitative RT-PCR, chitinase enzyme activity, and growth inhibition of phytopathogenic fungi demonstrated that the synthetic inducible promoter of transgenic plants was induced by chitin. The results of chitinase activity of extracted protein from all transgenic lines containing inducible promoters showed a 3.2-5.8-fold increase in chitinase activity compared to non-induced plants. The antifungal activity of the inducibly expressed chitinase was examined on Sclerotinia sclerotiorum and Rhizoctonia solani. The results showed that fungal growth inhibition increased via elicitor treatment of the inducible promoter, 82% for S. sclerotiorum and 62% for R. solani, respectively. The result of light microscopic observation demonstrated morphological changes in hyphae and that the expressed enzyme can lyse the mycelial cell walls of R. solani. Moreover, resistance to S. sclerotiorum in the intact leaves of transgenic plants (T2) was confirmed using bioassay analysis. Based on these results, it seems that the synthetic inducible promoter containing F cis-acting element driving chimeric chitinase is suitable for increasing the resistance of the canola transgenic plant when attacked by phytopathogenic fungi. | ||
| کلیدواژه‌ها | ||
| Cis-acting element؛ Phytopathogenic fungi؛ Chitinase activity؛ Transgenic canola؛ Inducible promoter | ||
| مراجع | ||
|
Aghazadeh R, Zamani M, Motallebi M, Moradyar M, Moghadassi Jahromi Z. 2016. Co-transformation of canola by chimeric chitinase and tlp genes towards improving resistance to Sclerotinia sclerotiorum. World Microbiol Biotechnol 32(9): 1-12.
Broekaert WF, Delauré SL, De Bolle MFC, Cammue BPA. 2006. The role of ethylene in host-pathogen interactions. Annu Rev Phytopathol 44(1): 393-416.
Cazzonelli CI, Velten J. 2008. In vivo characterization of plant promoter element interaction using synthetic promoters. TransgenicRes 17(3): 437-457.
Chernin L, Ismailov Z, Haran S, Chet I. 1995. Chitinolytic Enterobacter agglomerans antagonistic to fungal plant pathogens. Appl Environ Microbiol 61(5): 1720-1726.
Chernin LS, De la Fuente L, Sobolev V, Haran S, Vorgias CE, Oppenheim AB, …, Chet I. 1997. Molecular cloning, structural analysis, and expression in Escherichia coli of a chitinase gene from Enterobacter agglomerans. Appl Environ Microbiol 63(3): 834-839.
Chet I, Inbar J. 1994. Biological control of fungal pathogens. Biotechnol. Appl. Biochem. 48(1): 37-43.
Gunaratna K.R, Balasubramanian R. 1994. Partial purification and properties of extracellular chitinase produced by Acremonium obclavatum, an antagonist to the groundnut rust, Puccinia arachidis. World J Microbiol Biotechnol 10(3): 342-345.
Gurr SJ, Rushton PJ. 2005. Engineering plants with increased disease resistance: what are we going to express? Trends Biotechnol 23(6): 275-282.
Hammond-Kosack KE, Parker JE. 2003. Deciphering plant–pathogen communication: fresh perspectives for molecular resistance breeding. Curr Opin Biotechnol 14(2): 177-193.
Harighi MJ, Motallebi M, Zamani MR. 2006. Antifungal activity of heterologous expressed chitinase 42 (Chit42) from Trichoderma atroviride PTCC5220. Iran J Biotechnol 4(2): 95-103.
Heise A, Lippok B, Kirsch C, Hahlbrock K. 2002. Two immediate-early pathogen-responsive members of the AtCMPG gene family in Arabidopsis thaliana and the W-box-containing elicitor-response element of AtCMPG1. Proc Natl Acad Sci 99(13): 9049-9054.
Joosten MHAJ, Verbakel HM, Nettekoven ME, van Leeuwen J, van der Vossen RTM, de Wit PJGM. 1995. The phytopathogenic fungus Cladosporium fulvum is not sensitive to the chitinase and β-1,3-glucanase defence proteins of its host, tomato. Physiol Mol Plant Pathol 46(1): 45-59.
Khademi M, Varasteh-Shams M, Nazarian-Firouzabadi F, Ismaili A. 2020. New recombinant antimicrobial peptides confer resistance to fungal pathogens in tobacco plants. Front Plant Sci 11: 1236. https://doi.org/10.3389/fpls.2020.01236.
Kobayash DY, Reedy RM, Bick J, Oudemans P.V. 2002. Characterization of a chitinase gene from Stenotrophomonas maltophilia strain 34S1 and its involvement in biological control. Appl Environ Microbiol 68(3): 1047-1054.
Kunz C, Sellam O, Bertheau Y. 1992. Purification and characterization of a chitinase from the hyperparasitic fungus Aphanocladium album. Physiol Mol Plant Pathol 40(2): 117-131.
Limón MC, Chacón MR, Mejías R, Delgado-Jarana J, Rincón AM, Codón AC, …, Benítez T. 2004. Increased antifungal and chitinase specific activities of Trichoderma harzianum CECT 2413 by addition of a cellulose binding domain. Appl Microbiol Biotechnol 64(5): 675-685.
Lorito M, Farkas V, Rebuffat S, Bodo B, Kubicek CP. 1996. Cell wall synthesis is a major target of mycoparasitic antagonism by Trichoderma harzianum. J Bacteriol 178(21): 6382-6385.
Lorito M, Woo SL, Fernandez IG, Colucci G, Harman GE, Pintor-Toro JA,…, Scala F. 1998. Genes from Mycoparasitic fungi as a source for improving plant resistance to fungal pathogens. Proc Natl Acad Sci 95(14): 7860-7865.
Matroodi S, Motallebi M, Zamani M, Moradyar M. 2013. Designing a new chitinase with more chitin binding and antifungal activity. World J. Microbiol. Biotechnol. 29(8): 1517-1523.
Mazarei M, Teplova I, Hajimorad MR, Stewart CN. 2008. Pathogen phytosensing: plants to report plant pathogens. Sen 8(4): 2628-2641.
Mekapogu M, Jung JA, Kwon OK, Ahn MS, Song HY, Jang S. 2021. Recent progress in enhancing fungal disease resistance in ornamental plants. Int J Mol Sci 22(15): 7956. https://doi.org/10.3390/ijms22157956.
Moradyar M, Zamani MR, Motalebi M, Aghazadeh R. 2016 a. Transformation of canola with a chimeric chitinase gene under control of the SP-FF synthetic pathogen-inducible promoter. Cell and Mol Res (Iran J Biol). 28(4): 599-611.
Moradyar M, Motallebi M, Zamani MR, Aghazadeh R. 2016 b. Pathogen-induced expression of chimeric chitinase gene containing synthetic promoter confers antifungal resistance in transgenic canola. In Vitro Cell Dev Biol 52(2): 119-129.
Neuhaus JM, Ahl-Go P, Hinz U, Flores S, Meins F. 1991. High-level expression of a tobacco chitinase gene in Nicotiana sylvestris susceptibility of transgenic plants to Cercospora nicotianae infection. Plant Mol Biol 16(1): 141-151.
Nürnberger T, Brunner F. 2002. Innate immunity in plants and animals: emerging parallels between the recognition of general elicitors and pathogen-associated molecular patterns. Curr Opin Plant Biol 5(4): 318-324.
Pan H, Wei Y, Xin F, Zhou M, Zhang S. 2006. Characterization and biocontrol ability of fusion chitinase in Escherichia coli carrying chitinase cDNA from Trichothecium roseum. Z. Naturforsch. 61(5-6): 397-404.
Qing CM, Fan L, Lei Y, Bouchez D, Tourneur C, Yan L, …, Robaglia C. 2000. Transformation of Pakchoi (Brassica rapa L. ssp. Chinensis) by Agrobacterium infiltration. Mol Breed 6(1): 67-72.
Rushton PJ, Reinstädler A, Lipka V, Lippok B, Somssich IE. 2002. Synthetic plant promoters containing defined regulatory elements provide novel insights into pathogen- and wound-induced signaling. Plant Cell Rep 14(4): 749-762.
Salinas J, Oeda K, Chua NH. 1992. Two G-box-related sequences confer different expression patterns in transgenic tobacco. Plant Cell Rep 4(12): 1485-1493.
Shokouhifar F, Zamani MR, Motallebi M. 2011 a. Expression pattern of the synthetic pathogen-inducible promoter (SynP-FF) in the transgenic canola in response to Sclerotinia sclerotiorum. Iran J Biotechnol 9(1): 1-10.
Shokouhifar F, Zamani MR, Motallebi M, Mousavi A, Malboobi MA. 2011 b. Construction and functional analysis of pathogen-inducible synthetic promoters in Brassica napus. Biol Plant 55(4): 689-695.
Tiwari S, Mishra DK, Chandrasekhar K, Singh PK, Tuli R. 2011. Expression of δ-endotoxin Cry1EC from an inducible promoter confers insect protection in peanut (Arachis hypogaea L.) plants. Pest Manag Sci 67(2): 137-145.
Venter M. 2007. Synthetic promoters: genetic control through cis engineering. Trends Plant Sci 12(3): 118-124.
Zou X, Song E, Peng A, He Y, Xu L, Lei T, …, Chen S. 2014. Activation of three pathogen-inducible promoters in transgenic citrus (Citrus sinensis Osbeck) after Xanthomonas axonopodis pv. Citri infection and wounding. Plant Cell Tissue Organ Cult 117(1): 85-98. | ||
|
آمار تعداد مشاهده مقاله: 249 تعداد دریافت فایل اصل مقاله: 262 |
||