The trend of changes in chlorophyll fluorescence parameters in two Vicia faba ecotype during freezing stresses

Document Type : Original Articles

Authors

1 Research Center for Plant Sciences, Ferdowsi University of Mashhad, Mashhad, Iran

2 Faculty of Agriculture Ferdowsi University of Mashhad, Mashhad, Iran

Abstract

Introduction
Freezing tolerance is an important factor determining natural geographic distribution of plant species as well as growth and yield of many crop plants. Many plants from temperate and cold climates, including several important crop species, are able to increase their freezing tolerance in response to low, nonfreezing temperatures in a process termed cold acclimation. It is generally accepted that cellular membranes are the primary targets of freezing damage in plants. So, besides whole plant survival, determination of electrolyte leakage from plant tissues after freezing and thawing, using conductivity measurements has been the most frequently used method reported in the literatures. This method mainly monitors the ability of the plasma membrane to function as a semi-permeable barrier towards intracellular ions, but the intactness of the vacuole, as the major storage compartment for inorganic ions, may also impact the measurements. Under stress conditions, the possibility of over excitation of photosystem II (PSII) increases, and this reduces the photosynthetic rate and lead to an increase in the dissipation of absorbed energy through nonradiative processes. The measurement of chlorophyll fluorescence has been used for many years as a sensitive, reliable, and rapid method to determine the effect of environmental stresses, like drought, temperature, excessive light and air pollution on green plants. Decrease of the maximal rate of the fast rise of fluorescence after exposure of leaves to chilling temperatures, correlated well with the visual symptoms of chilling injury in several species
 
Materials & Methods
 This study was carried out to measure chlorophyll fluorescence and determine the survival of two Vicia faba ecotypes (Borujerd and Neyshabur) in the fall and winter of 2015 at Research Center for Plant Sciences, Ferdowsi University of Mashhad. Treatments were arranged as factorial based on completely randomized block design with four replications. Plants were grown in pots under natural conditions until four to six leaf stage for acclimation, and then were subjected to freezing temperatures (0, -4, -8, -12, -16, -20, -24 °C) in a thermo gradient freezer. Freezer temperature was 5 °C at the beginning and reached to -24 °C, decreasing two degrees centigrade per hour. Ice nucleation active bacteria was sprayed to nucleation formation in plants at -3°C. Plants were kept for an hour in each temperature and then were transported to a cold chamber (5±2 °C) and kept for 24 hours to avoid rapid melting. Chlorophyll fluorescence was recorded with a pulse amplitude modulation fluorometer (PAM-2000, Walz, Effeltrich, Germany) before and 3, 6, 12, 24, 48 and 72 hour after freezing stress. The efficiency of excitation energy capture by open PSII reaction centers (F´v/F´m) and the quantum yield of electron transport at photosystem II (PSII), were determined. Survival percentage was determined after three weeks recovery in greenhouse condition. Data were analyzed using a two-way ANOVA model, followed by Duncanʼs test for mean comparison at 95 % confidence level by Minitab 16 program.
 
Results & Discussion
Result indicated that maximum quantum yield of PSII photochemistry (F'v/F'm) was four percent higher in Borujerd ecotype compared to Neyshabur ecotype. No significant change was found in chlorophyll fluorescence parameters with decreasing temperature from zero to -12°C but more temperature reduction caused chlorophyll fluorescence parameters to decrease in a way that the lowest mean was observed at
-24°C. Rapid reductions were found in chlorophyll fluorescence parameters such as F'v/F'm until 24 hours after freezing stress which was followed by gently slope increasing trend, but the photochemical efficiency of photosystem II, 72 hours after stress, did not return to the level before freezing stress. Survival rate was decreased as freezing temperature decreased and reached to 83% at -12°C. More temperature reduction to
-16°C led to sever decrease in survival rate in a way that no plant survival and only five percent was observed in Borujerd and Neyshabur ecotypes, respectively. High regression coefficients were found between F'v/F'm and survival rate in both ecotypes (R2=0.99 and R2=0.98 for Borujerd and Neyshabur ecotypes, respectively)
 
Conclusion
Generally, evaluation of chlorophyll fluorescence parameters of both Vicia faba ecotypes showed that chlorophyll fluorescence has a direct relationship with survival three weeks after freezing stress and can be used as an index for assessment of freezing tolerance.

Keywords


  1. Badeck, F.W., and Rizza, F. A Combined field/laboratory method for assessment of frost tolerance with freezing tests and chlorophyll fluorescence. Agronomy 5: 71-88.
  2. Baker, N.R. 2008. Chlorophyll fluorescence: a probe of photosynthesis in vivo. Annual Review of Plant Biology 59: 89-
  3. Butler, W.L. 1978. Energy distribution in the photochemical apparatus of photosynthesis. Annual Review of Plant Physiology 29: 345-
  4. Dexter, S.T. 1993. Effects of several environmental factors on hardening of plants. Plant Physiology 8: 123-139.
  5. Ehlert, B., and Hincha, D.K. 2008. Chlorophyll fluorescence imaging accurately quantifies freezing damage and cold acclimation responses in Arabidopsis leaves. Plant Methods 4:
  6. 2009. Prod Stat: Crops. FAO Statistical Databases (Faostat), Food and Agriculture Organization of the United Nations (FAO), http://faostat.fao.org.
  7. Hincha, D.K., and Schmitt, J.M. 1992. Freeze-Thaw Injury and Cryoprotection of Thylakoid M In: G.N. Somero, C.B. Osmond and C.L. Bolis (Eds). Water and Life. Springer, p. 316-337.
  8. Jurczyk, B., Krepski, T., Kosmala, A., and Rapacz, M. 2013. Different mechanisms trigger an increase in freezing tolerance in Festuca pratensis exposed to flooding stress. Environmental and Experimental Botany 93: 45-
  9. Kaplan, F., Kopka, J., Sung, D.Y., Zhao, W., Popp, M., Porat, R., and Guy, C.L. 2007. Transcript and metabolite profiling during cold acclimation of Arabidopsis reveals an intricate relationship of cold-regulated gene expression with modifications in metabolite content. Plant Journal 50: 967-981.
  10. Koscielniak, L., and Biesaga-Koscielniak, J. 2006. Photosynthesis and non-photochemical excitation quenching components of chlorophyll excitation in maize and field bean during chilling at different photon flux density. Photosynthetica 44: 174-180.
  11. Krause, G.,and Santarius, K. 1975. Relative thermostability of the chloroplast envelope. Planta 127: 285-
  12. Krause, G.H., Grafflage, S., Rumich-Bayer, S., and Somersalo, S. 1988. Effects of freezing on plant mesophyll cells. Symposia of the Society for Experimental Biology 42: 311-327.
  13. Lindow, S.E., Arny, D.C., and Upper, C.D. 1982. Bacterial ice nucleation: a factor in frost injury to plants. Plant Physiology 70: 1084-1089.
  14. Mathur, S., Jajoo, A., Mehta, P., and Bharti, S. 2011. Analysis of elevated temperatureinduced inhibition of photosystem II using chlorophyll a fluorescence induction kinetics in wheat leaves (Triticum aestivum). Plant Biology 13: 1-
  15. Mishra, A., Heyer, A.G., and Mishra, K.B. 2014. Chlorophyll fluorescence emission can screencold tolerance of cold acclimated Arabidopsis thaliana Plant Methods 10: 38.
  16. Molina-Bravo, R., Arellano, C., Sosinski, B.R., and Fernandez, G.E. 2011. A protocol to assess heat tolerance in a segregating population of raspberry using chlorophyll fluorescence. Scientia Horticulturae 130: 524-
  17. Nezami, A., Bagheri, A., Porsa, H., Zafranieh, M., and Khamadi, N. 2011. Evaluation of cold tolerant lentil genotypes (Lens culinaris) in fall planting under supplementary irrigation. Iranian Journal of Pulses Research 1(2): 49-58. (In Persian with English Summary).
  18. Nezami, A., Khazaei, H.R., Eshghizadeh, H.R., and Riahinia, Sh. 2011. Evaluation of freezing temperature tolerance of lentil (Lens culinaris) genotypes with using chlorophyll fluorescence parameters. Agronomy Journal (Pajouhesh & Sazandegi) 99: 24-33. (In Persian with English Summary).
  19. Porsa, H., Nezami, A., Bagheri, A., and Najibnia, S. 2016. Agronomic assessment of cold tolerant chickpea (Cicer arietinum) genotypes in fall sowing at Mashhad conditions. Iranian Journal of Pulses Research 7(1): 37-53. (In Persian with English Summary).
  20. Rapacz, M., Sasal, M., and Gut, M. 2011. Chlorophyll fluorescence-based studies of frost damage and the tolerance for cold-induced photoinhibition in freezing tolerance analysis of triticale (×Triticosecale Wittmack). Journal of Agronomy and Crop Science. 197: 378–389.
  21. Sharma, D.K., Andersen, S.B., Ottosen, C.O., and Rosenqvist, E. 2012. Phenotyping of wheat cultivars for heat tolerance using chlorophyll a fluorescence. Functional Plant Biology 39: 936-
  22. Singh, A.K., Bhatt, B.P., Upadhyaya, A., Kumar, S., Sundaram, K., Singh, B.K., Chandra, N., and Bharati, R.C. 2012. Improvement of faba bean (Vicia faba L.) yield and quality through biotechnological approach: A review. African Journal of Biotechnology 11(87): 15264-15271.
  23. Vaclavik, L., Mishra, A., Mishra, K.B., and Hajslova, J. 2013. Mass spectrometry-based metabolomic fingerprinting for screening cold tolerance in Arabidopsis thaliana Analytical and Bioanalytical Chemistry 405(8): 2671-2683.
  24. Vogel, J.T., Zarka, D.G., van Buskirk, H.A., Fowler, S.G., and Thomashow, M.F. 2005. Roles of the CBF2 and ZAT12 transcription factors in configuring the low temperature transcriptome of Arabidopsis. Plant Journal 41: 195-211.
  25. Zhang, X., Wan, S., Hao, J., Hu, J., Yang, T., and Zong, X. 2016. Large-scale evaluation of pea (Pisum sativum) germplasm for cold tolerance in the field during winter in Qingdao. The Crop Journal 4(5): 377-383.
  26. Zhou, R., Yu, X., Kjær, K.H., Rosenqvist, E., Ottosen, C.O., and Wu, Z. 2015. Screening and validation of tomato genotypes under heat stress using Fv/Fm to reveal the physiological mechanism of heat tolerance. Environmental and Experimental Botany 118: 1–11.
CAPTCHA Image