Investigating the Phenophysiological Traits of Transgenic Rapeseed Lines (Brassica napus) with aroA Gene Harboring a Point Mutation of Proline 101 to Serine (P101S) under Glyphosate Herbicide Treatment

Document Type : Original Article

Authors

1 Department of Agronomy and Plant Breeding, Kermanshah Branch, Islamic Azad University, Kermanshah, Iran

2 Agricultural Biotechnology Department, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

Abstract

One of the most effective ways to breed glyphosate-resistant plants is to reduce the affinity of glyphosate to the EPSPS enzyme by manipulating the aroA gene. In this study, P101S mutation was induced in the aroA of E. coli and the mutant gene was cloned in the pUC18 plasmid. It was transferred by Agrobacterium tumefaciens to the rapeseed. To investigate the phenophysiological traits in transgenic rapeseed lines under different glyphosate treatments (0, 1.2, 2.4, 4.8, 9.6, 19.2, 38.4, 76.8 and 153.6 mM), the seeds of T2 generation transgenic plants were studied in greenhouse conditions in a factorial experiment. The results showed that the control (no herbicide) had the lowest amount of days to flowering (32.5 days), partial water pressure (14.92kPa), active photosynthetic radiation (393.4 mmol m−2 s−1), and leaf surface temperature (28.880°C). The concentration of 2.4 mM had the highest stomatal conductance (0.573 mol m−2 s−1) and photosynthesis rate (14.07 μmol m−2 s−1). The lowest value of stomatal resistance was related to the concentration of 4.8 mM (246.9 mmol m−2 s−1), and the lowest value of stomatal conductance was associated with 9.6 mM (0.047 mol m−2 s−1). The highest rate of CO2 source (416.3 ppm) and active photosynthetic radiation (576.9 mmol m−2 s−1) was seen in the 19.2 mM. The 38.4 mM had the highest number of days to flowering (48.1 days) and leaf surface temperature (38.748°C) and the lowest amount of CO2 source (386ppm). The 76.8 mM had the highest stomatal resistance (330.2 mmol m−2 s−1) and the lowest photosynthesis rate (1.36 μmol m−2 s−1). The highest partial pressure of the water source was related to 153.6 mM (19.07 kPa). In summary, different concentrations of herbicides exhibit varying degrees of phenophysiological traits, and desired traits can be improved based on these concentrations.

Graphical Abstract

Investigating the Phenophysiological Traits of Transgenic Rapeseed Lines (Brassica napus) with aroA Gene Harboring a Point Mutation of Proline 101 to Serine (P101S) under Glyphosate Herbicide Treatment

Highlights

  • The study investigated the phenophysiological traits of transgenic rapeseed lines with a P101S mutation in the aroA gene under different glyphosate treatments.
  • Results showed variations in days to flowering, stomatal conductance, photosynthesis rate, stomatal resistance, CO2 levels, and leaf surface temperature across different glyphosate concentrations.
  • The control group (no herbicide) had the lowest days of flowering, partial water pressure, active photosynthetic radiation, and leaf surface temperature.
  • A concentration of 2.4 mM exhibited the highest stomatal conductance and photosynthesis rate.
  • Findings suggest that varying herbicide concentrations can influence phenotypic traits in transgenic rapeseed lines, providing insights for trait improvement based on herbicide levels.

Keywords

Main Subjects

References

Amrhein N., Johänning D., Schab J., Schulz A. 1983. Biochemical basis for glyphosate‐tolerance in a bacterium and a plant tissue culture. FEBS Letters 157(1): 191-196. https://doi.org/10.1016/0014-5793(83)81143-0
Asaduzzaman M., Pratley J.E., Luckett D., Lemerle D., Wu H. 2020. Weed management in canola (Brassica napus L): A review of current constraints and future strategies for Australia. Archives of Agronomy and Soil Science 66(4): 427-444. https://doi.org/10.1080/03650340.2019.1624726
Barzan Z., Dehdari M., Amiri Fahliani R. 2015. Study of genetic diversity in rapeseed (Brassica napus L.) genotypes using microsatellite markers. Agricultural Biotechnology Journal 7(1): 29-42.
Bhatt P., Joshi T., Bhatt K., Zhang W., Huang Y., Chen S. 2021. Binding interaction of glyphosate with glyphosate oxidoreductase and C–P lyase: Molecular docking and molecular dynamics simulation studies. Journal of Hazardous Materials 409: 124927. https://doi.org/10.1016/j.jhazmat.2020.124927 
Chmielewska A., Kozłowska M., Rachwał D., Wnukowski P., Amarowicz R., Nebesny E., Rosicka-Kaczmarek J. 2021. Canola/rapeseed protein–nutritional value, functionality and food application: a review. Critical Reviews in Food Science and Nutrition 61(22): 3836-3856. https://doi.org/10.1080/10408398.2020.1809342
Devine M.D., Shukla A. 2000. Altered target sites as a mechanism of herbicide resistance. Crop Protection 19(8-10): 881-889. https://doi.org/10.1016/S0261-2194(00)00123-X
Duke S.O. 2021. Glyphosate: uses other than in glyphosate-resistant crops, mode of action, degradation in plants, and effects on non-target plants and agricultural microbes. Reviews of Environmental Contamination and Toxicology 255: 1-65. https://doi.org/10.1007/398_2020_53
Gaba S., Gabriel E., Chadœuf J., Bonneu F., Bretagnolle V. 2016. Herbicides do not ensure for higher wheat yield but eliminate rare plant species. Scientific Reports 6: 30112. https://doi.org/10.1038/srep30112
Gomes M.P., Le Manac’h S.G., Moingt M., Smedbol E., Paquet S., Labrecque M., Lucotte M., Juneau P. 2016. Impact of phosphate on glyphosate uptake and toxicity in willow. Journal of Hazardous Materials 304: 269-279. https://doi.org/10.1016/j.jhazmat.2015.10.043
Griffin S.L., Chekan J.R., Lira J.M., Robinson A.E., Yerkes C.N., Siehl D.L., Wright T.R., Nair S.K., Cicchillo R.M. 2021. Characterization of a glyphosate-tolerant enzyme from Streptomyces svecius: a distinct class of 5-enolpyruvylshikimate-3-phosphate synthases. Journal of Agricultural and Food Chemistry 69(17): 5096-5104. https://doi.org/10.1021/acs.jafc.1c00439
Kahrizi D. 2014. Reduction of EPSP synthase in transgenic wild turnip (Brassica rapa) weed via suppression of aroA. Molecular Biology Reports 41(12): 8177-8184. https://doi.org/10.1007/s11033-014-3718-0
Kahrizi D., Salmanian A.H. 2008. Substitution of Ala183Thr in aro A product of E. coli (k12) and transformation of rapeseed (Brassica napus L.) with altered gene confers tolerance to Roundup. Transgenic Plant Journal 2(2): 170-175. http://www.globalsciencebooks.info/Online/GSBOnline/images/0812/TPJ_2(2)/TPJ_2(2)170-175o.pdf
Kahrizi D., Salmanian A.H., Afshari A., Moieni A., Mousavi A. 2007. Simultaneous substitution of Gly96 to Ala and Ala183 to Thr in 5-enolpyruvylshikimate-3-phosphate synthase gene of E. coli (k12) and transformation of rapeseed (Brassica napus L.) in order to make tolerance to glyphosate. Plant Cell Reports 26(1): 95-104. https://doi.org/10.1007/s00299-006-0208-4
Leino L., Tall T., Helander M., Saloniemi I., Saikkonen K., Ruuskanen S., Puigbo P. 2021. Classification of the glyphosate target enzyme (5-enolpyruvylshikimate-3-phosphate synthase) for assessing sensitivity of organisms to the herbicide. Journal of Hazardous Materials 408: 124556. https://doi.org/10.1016/j.jhazmat.2020.124556
Lemerle D., Luckett D.J., Wu H., Widderick M.J. 2017. Agronomic interventions for weed management in canola (Brassica napus L.) - A review. Crop Protection 95: 69-73. https://doi.org/10.1016/j.cropro.2016.07.007
Palma-Bautista C., Vázquez-Garcia J.G., López-Valencia G., Domínguez-Valenzuela J.A., Barro F., De Prado R. 2023. Reduced glyphosate movement and mutation of the EPSPS gene (Pro106Ser) endow resistance in conyza canadensis harvested in mexico. Journal of Agricultural and Food Chemistry 71(11): 4477-4487. https://doi.org/10.1021/acs.jafc.2c07833
Pan L., Yu Q., Wang J., Han H., Mao L., Nyporko A., Maguza A., Fan L., Bai L., Powles S. 2021. An ABCC-type transporter endowing glyphosate resistance in plants. Proceedings of the National Academy of Sciences 118(16): e2100136118. https://doi.org/10.1073/pnas.2100136118
Roeintan A., Safavi S.M., Kahrizi D. 2022. Rapeseed transformation with aroA bacterial gene containing P101S mutation confers glyphosate resistance. Biochemical Genetics 60(3): 953-968. https://doi.org/10.1007/s10528-021-10136-w
Tang T., Chen G., Liu F., Bu C., Liu L., Zhao X. 2019. Effects of transgenic glufosinate-tolerant rapeseed (Brassica napus L.) and the associated herbicide application on rhizospheric bacterial communities. Physiological and Molecular Plant Pathology 106: 246-252. https://doi.org/10.1016/j.pmpp.2019.03.004
Agrotechniques in Industrial Crops

Articles in Press, Accepted Manuscript
Available Online from 29 November 2024

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