Evaluation of gene expression, enzymatic and biochemical activity of leaves in native and foreign genotypes of bread wheat before and after anthesis

Document Type : Original Article


1 Department of Plant Breeding and Biotechnology, Gorgan University of Agricultural Sciences and Natural Resources , Gorgan, Iran

2 Department of plant breeding and biotechnology, Gorgan University of Agricultural Sciences and Natural Resources, Iran.



This experiment was carried out in Golestan province, in a randomized complete block design (RCBD) with four replicates. Wheat genotypes were included N-91-9, N-91-17, Tiregan, Nogal, Euclide, Agorazado, Lucullus, and Antonius. Sampling from Flag leaf and other leaves at 10 stages included 7DBA (Day before anthesis), 3DBA, Anthesis, 3DAA (Day after anthesis), 7DAA, 11DAA, 15DAA, 19DAA, 23DAA, and 27DAA was done and carried to laboratory in liquid nitrogen and stored at -80 °C. Part of the leaf samples was carried with a paper pocket and incubated in the oven at 80 °C for 48 hours. Biochemical traits such as chlorophyll content, net photosynthetic rate, sucrose content, sucrose synthase (SS) and sucrose phosphate synthase (SPS) content, grain yield, and photosynthetic genes expression included RbcL, RbcS, and Rca was measured in all stages. The result had shown that flag leaf in the post-anthesis stages has a significant role in photosynthesis and sucrose production in the plant and cultivars with higher chlorophyll content also had higher photosynthesis. Also, in genotypes with higher SS enzyme activity, more sucrose is synthesized and due to the role of SPS, increasing its amount is involved in the transfer of sucrose from the leaves to other organs. The RbcL, RbcS, and Rca genes had higher expression in cultivars with higher photosynthesis than the control (Tiregan). Among the studied genotypes, the results showed that Nogal cultivar and line N-91-17 had superior in terms of grain yield and other measured traits and can be recommended for use in future research.


Chen, D. Q., S. W. Wang, B.L. Xiong, B. B. Cao and X. P. Deng. 2015. Carbon/Nitrogen imbalance associated with drought-induced leaf senescence in Sorghum bicolor. PloS One. 10 (8): e0137026.
Douglas, C. D., M. K. Tsung and C. F. Frederick. 1988. Enzymes of sucrose and hexose metabolism in development kernels of two inbreeds of maize. Plant Physiology. 86: 1013–1019.
Ezat Ahmadi, M., Gh. Noormohammadi, M. Ghodsi and M. Kafi. 2012. Evaluation of drought tolerance and use of stem stocks of promising bread wheat genotypes under different moisture stress and photosynthetic conditions. Iranian Journal of Field Crops Research. 9(4): 758-769.
Fahad, S., A. A. Bajwa, U. Nazir, S. A. Anjum, A. Farooq, A. Zohaib, S. Sadia, W. Nasim, S. Adkins, S. Saud, M. Z. Ihsan, H. Alharby, C. Wu, D. Wang and J. Huang. 2017. Crop Production under Drought and Heat Stress: Plant Responses and Management Options. Front. Plant Sci. 8:1147. doi: 10.3389/fpls.2017.01147.
Golabadi, M., P. Golkar and B. Bahari. 2015. Remobilization assay of dry matter from different shoot organs under drought stress in wheat (Triticum aestivum L.). Agronomy Research. 13(5):1202-1214.
Julius, B.T., K. A. Leach, T. M. Tran, R. A. Mertz and M. D. Braun. 2017. Sugar Transporters in Plants: New Insights and Discoveries. Plant Cell Physiol. 58(9): 1442–1460. doi:10.1093/pcp/pcx090.
Koch, K. 2004. Sucrose metabolism: regulatory mechanisms and pivotal roles in sugar sensing and plant development. Curr. Opin. Plant Biol. 7: 235–246.
Kumar, R. R., S. Goswami, K. Singh, K. Dubey, S. Singh, R. Sharma, N. Verma, Y. K. Kala, G. K. Rai, M. Grover, D. C. Mishra, B. Singh, H. Pathak, V. Chinnusamy, A. Rai and S. Praveen. 2016. Identification of Putative Rubisco Activase (TaRca1) The Catalytic Chaperone Regulating Carbon Assimilatory Pathway in Wheat (Triticum aestivum) under the Heat Stress. Front. Plant Sci. 7:986. doi: 10.3389/fpls.2016.00986.
Lan, J., B. Zhang, H. Zhou, Y. Liu, H. Li. 2003. Comparison on photosynthetic rate and interrelated characteristics of winter wheat varieties from different eras. Journal of Shenyang Agricultural University. 34(1):12-15.
Lemoine, R., S. La Camera, R. Atanassova, F. Dédaldéchamp, T. Allario, N. Pourtau, J. L. Bonnemain, M. Laloi, P. Coutos-Thévenot, L. Maurousset, M. Faucher, C. Girousse, P. Lemonnier, J. Parrilla and M. Durand. 2013. Source-to-sink transport of sugar and regulation by environmental factors. Plant Science. 4: 1-21. doi: 10.3389/fpls.2013.00272.
Li, X and C. Wang. 2013. Physiological and metabolic enzymes activity changes in transgenic rice plants with increased phosphoenolpyruvate carboxylase activity during the flowering stage. Acta Physiologiae Plantarum. 35:1503–1512.
 Liu, C., Y. Liu, Y. Lu, Y. Liao, J. Nie, X. Yuan and F. Chen. 2019. Use of a leaf chlorophyll content index to improve the prediction of above-ground biomass and productivity. PeerJ 6:e6240 DOI 10.7717/peerj.6240.
Ludewig, F and U.I. Flu¨gge. 2013. Role of metabolite transporters in source-sink carbon allocation. Front Plant Sci. 4:231.
Maillard, A., S. Diquélou, V. Billard, P. Laîné, M. Garnica, M. Prudent, J. M. Garcia-Mina, J.C. Yvin and A. Ourry. 2015. Leaf mineral nutrient remobilization during leaf senescence and modulation by nutrient deficiency. Front. Plant Sci. 6:317. doi: 10.3389/fpls.2015.00317.
Miron, D and A. A. Schaffer. 1991. Sucrose Phosphate Synthase, Sucrose Synthase, and Invertase Activities in Developing Fruit of Lycopersicon esculentum Mill. and the Sucrose Accumulating Lycopersicon hirsutum Humb. and Bonpl. Plant Physiology. 95: 623-627.
Navabpour, S. 2011. Molecular study and expression of photosynthetic genes and aging process in flag leaf and other leaves in barley. Journal of Production and Processing of Crops and Horticultural Products. 2(4): 13-24.
Pfaf, M. W. 2001. A new mathematical model for relative quantification in real-time RT–PCR. Nucleic Acids Res. 29(9): e45–e45.
Porra, R. J., W. A. Thompson and P. E. Kriedmann. 1989. Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. Biochemistry Biophysics Acta. 975:384-394.
Roca, M., K. Chen and A. Pérez-Gálvez. 2016. Chlorophylls. Handbook on Natural Pigments in Food and Beverages. 125–158. doi:10.1016/b978-0-08-100371-8.00006-3.
Ruan, Y. L. 2014. Sucrose metabolism: gateway to diverse carbon use and sugar signaling. Annu. Rev. Plant Biol. 65: 33–67.
Shi, H., B. Wang, P. Yang, Y. Li and F. Miao. 2016. Differences in Sugar Accumulation and Mobilization between Sequential and Non Sequential Senescence Wheat Cultivars under Natural and Drought Conditions. PLoS ONE 11(11): e0166155. doi: 10.1371/ journal. pone.0166155.
Stein, O and D. Granot. 2019. An Overview of Sucrose Synthases in Plants. Front. Plant Sci. 10:95. doi: 10.3389/fpls.2019.00095.
Suzuki, Y and A. Makino. 2012. Availability of Rubisco small subunit up-regulates the transcript levels of large subunit for stoichiometric assembly of its holoenzyme in rice. Plant Physiology. 160, 533–540.
Suzuki, Y and A. Makino. 2013. Translational downregulation of RBCL is operative in the coordinated expression of Rubisco genes in senescent leaves in rice. Journal of Experimental Botany. 64(4): 1145–1152.
Suzuki, Y., T. Miyamoto, R. Yoshizawa, T. Mae, A. Makino. 2009. Rubisco content and photosynthesis of leaves at different positions in transgenic rice with an overexpression of RBCS. Plant Cell and Environment. 32, 417–427.
Vitlin Gruber, A and L. Feiz. 2018. Rubisco Assembly in the Chloroplast. Front. Mol. Biosci. 5:24. doi: 10.3389/fmolb.2018.00024.
Wang, B., M. Ma, H. Lu, Q. Meng, G. Li and X. Yang. 2015. Photosynthesis, sucrose metabolism, and starch accumulation in two NILs of winter wheat. Photosynth Res. DOI 10.1007/s11120-015-0126-9.
Zhang, S. W., C. F. Wang, F. Miao, C. J. Zhou, Y. H. Yao and G.X. LI. 2012. Photosynthetic characteristics and its significance of topmost three leaves at fruiting stage in wheat with presenile flag leaf. Acta agronomica sinica. 38(12): 2258–2266. doi:10.3724/SP.J.1006.2012.02258.
Zhao, L., Y. Xia, X. Y. Wu, J. H. M. Schippers and H.C. Jing. 2018. Phenotypic Analysis and Molecular Markers of Leaf Senescence. Plant Senescence. 35–48. doi:10.1007/978-1-4939-7672-0_3.