Research Article

Effect of Elevated CO2 Concentration On Four Populus by the Fast Fluorescence Rise OJIP  

Fanjuan Meng1 , Ruyu Xie1 , Mu Peng1 , Tao Wang1 , Fachun Guan2
1 College of Life Science, Northeast Forestry University, Harbin 150040, China
2 Tibet Agricultural and Animal Husbandry University, Linzhi 860000, China
Author    Correspondence author
International Journal of Molecular Evolution and Biodiversity, 2019, Vol. 9, No. 1   doi: 10.5376/ijmeb.2019.09.0001
Received: 08 Jan., 2019    Accepted: 10 Mar., 2019    Published: 23 May, 2019
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Preferred citation for this article:

Xie R.Y. , Peng M., Wang T., Meng F.J.and Guan F.C., 2019, Effect of elevated CO2 concentration on four populus by the fast fluorescence rise OJIP, International Journal of Molecular Evolution and Biodiversity, 9(1): 1-8 (doi: 10.5376/ijmeb.2019.09.0001)





Increased atmospheric carbon dioxide (CO2) can influence the stability of ecosystems in the world. The chlorophyll fluorescence technique was considered as an effective tool to evaluate the photosynthetic levels on many plant species. In this study, we analyzed the state of PSII on four kinds of populus (Populus L.) (populus X, Pop ulusdeltoides × cathayana, Poplus alba ′Berdinensis′ L, Populus euramerican ′N3016′ × Populus ussuriensis) under high CO2 condition. The results show that the PSII performance was negatively influenced by CO2 stress. High CO2 stress resulted in down-regulation of Fm, φPo (=Fv/Fm), ψEo, φEo and PItotal in four kinds of populus. And a significant decrease in the P-step level of the fluorescence transients OJIP curves of four populus species after 7 days of treatment was observed. Therefore, a fast decrease of the P-step level indicated there was main change to fluorescence transients. These results indicated that changes of PSII attribute to the degradation of antenna pigment and inhibition of the electron transport at the acceptor side of PSII.

Populus L.; Atmospheric carbon dioxide (CO2); Photosynthetic performance; OJIP


Environmental changes for emissions of greenhouse gasses have influenced the stability of ecosystems in the world (IPCC, 2007). Accordingly, the increased atmospheric CO2 concentration (EC) has become the most important environmental issues in the world. Especially, human activities lead to increased CO2 concentration in the atmosphere, which is expected to reach 700 μL/L by the middle of the next century (Amthor, 2010; Griffin et al., 2001; Long et al., 2004; Peters and Hertwich, 2008). Various researches showed EC can effect the development, growth and yield of plant. In addition, EC can increase photosynthetic rates. In the result, high photosynthetic rates will improve growth of plants. For these reasons, there is a large of reports on the physiological responses of plants to EC, however, little studies have given account of the response of woody plants to EC. In this study, here, we tested the effects of EC on the growth and photosynthetic characteristics of four populus (Populus L.).


Poplars (Populus L.) are most widespread and fastest growing tree species, and their wood can be used for construction, paper, and fossil fuels. Poplars are known to be sensitive to various environmental stresses (Cui et al., 1999; Jiang et al., 2007). To date, the effects of EC on physiology and growth of poplars have been studied. For instance, EC treatments did not bring changes in structure of wood of birch (Kostiainen et al., 2008). EC also reduced tree height and photosynthetic pigment contents of Populus alba × glandulosa. Generally, the photosynthetic ability was reported to a reliable indirect indicator of tree performances under different environmental stress. However, there is few knowledge on photosynthetic responses on different Populus species to EC (Verhoeven et al., 1999; Jiang et al., 2002). Several past studies have found chlorophyll a fluorescence technique to be suitable as a stress indicator and for monitoring physiological changes in the plant. The chlorophyll a fluorescence OJIP transient (OJIP transient) technique was suitable.


Due to simple, non-destructive, and rapid characteristics, this technique has been used to evaluate the photosynthetic levels on many plant species (Strasser et al., 2000; Strasser et al., 2004; Zushi et al., 2012; Guo and Tan, 2015). The OJIP transient is defined by the O, J, I, and P steps, which correspond to the redox states of the photosystem (PS II and PS I) (Strasser et al., 2000; Strasser et al., 2004). Indeed, several studies have been conducted on OJIP transients in salt-stressed plants (Guo and Tan, 2015). For example, in salt-stressed wheat leaves, OJIP transients and the calculated JIP parameters can be used as sensitive methods to assess the stress level and to identify the action sites of salt stress (Mehta et al., 2010). It has also been shown that in salt-stressed canola (Brassica napus) leaves (100 mM and 200 mM NaCl), salt stress does not significantly affect the values of JIP parameters related to the electron donor site in PSII; However, the values of the JIP parameters that are related to the electron acceptor site significantly decrease with an increasing concentration of salt (Jafarinia and Shariati, 2012).


In our study, we have examined photosynthetic responses of four Populus spp. species to EC by chlorophyll a fluorescence OJIP transient (OJIP transient) technique. Due to simple, non-destructive, and rapid characteristics, this technique has been used to evaluate the photosynthetic levels on many plant species. Indeed, this method can provides valuable information to the redox states of the photosystem (PS II and PS I) (Šesták et al., 2001; Force et al., 2003; Jiang et al., 2003; Heerden et al., 2004; Albert et al., 2005; Schansker et al., 2005; An, 2006; Ilík et al., 2006; Strauss et al., 2007; Tóth et al., 2007; Chen et al., 2008; Strasserf et al., 2008; Yordanov et al., 2008). PSⅡ is very sensitive to environmental stresses (Verhoeven et al., 1999; Jiang et al., 2002). However, a study of comparing influences of EC on PSII behaviors of woody species is still lack. Therefore, a detailed comparison of high CO2-induced changes in PSII photochemistry in different Populus spp. species was carried out by OJIP technique. In this study, we analyzed the transient fluorescence using JIP-test on four Populus species (populus X, Populus deltoides × cathayana , Poplus alba ′Berdinensis′ L , Populus euramericana ′N3016′ × Populus ussuriensis) under atmospheric CO2 (1500 ±50 μmol/mol). Our objectives were to determine that whether different Populus species vary in their PSⅡ traits under EC.


1 Results           

1.1 OJIP curves on four populus

In OJIP curves of leaves, each step exhibited a different response to EC for four populus species (Figure 1A-D). All O-J-I-P transients were normalized at the O step and P step. Effects of EC on JIP parameter values were different, and specific changes were observed in some populus species. For example, relative fluorescence intensity was decreased for populus X at the 7th day after treatment, especially, the fluorescence intensity from I- to P- step was inhibited (Figure 1A). Conversely, relative fluorescence intensity of Populus deltoides × cathayana was increased and relative fluorescence intensity was also increased from J- to I- step (Figure 1B). However, relative fluorescence intensity from J- to I- step of Populus alba ′Berdinensis′ L was similar between control and treatment (Figure 1C). For Populus euramericana ′N3016′ × Populus ussuriensis, an increase in relative fluorescence intensity was also observed from I- to P- step (Figure 1D).


1.2 Fluorescence parameters on four populus  

The values (treated and control) of the JIP test parameters are shown in Table 1 and Table 2. No significant changes were observed in the minimum fluorescence (Fo), fluorescence at the J-step (Fj) and fluorescence at the I-step (Fi). In contrast, drastic decreases in F maximum fluorescence (Fm) values of four populus species were recorded after 7 days of CO2 treatment (Table 2). Similarly, all populus species showed significant decrease in maximum quantum yield of PSII primary photochemistry (φPo), the efficiency on the energy of a trapped exciton converting into electron transport beyond QA- (ψEo), the quantum yield of electron transport beyond QA (φEo) and total performance index per absorption basis (PItotal) after treatment. Especially, PItotal value sharply decreased after treatment. On the other hand, the values of relative variable fluorescence at the J-step (Vj) and relative variable fluorescence at the I-step (Vi) on treated-plants were higher than those of controls. The value in ABS/RC of treated plants is higher than those of control plants. However, effects of EC on Populus deltoides × cathayana did not show specific changes in CO2-treated plants compared control plants (Table 2).



Figure 1 The relative fluorescence transients in fully dark-adapted Populus X leaves (A), Populus deltoides × cathayana leaves (B), Populus alba Berdinensis′ L. leaves (C) and Populus euramericana N3016× Populus ussuriensis leaves (D) under CO2 stress and the control condition at 7 days


2 Discussion

In this study, the shapes of chlorophyll a fluorescence transient (O-J-I-P) were markedly different in four populus species after high CO2 treatment, however the values of Fv/Fm showed similar decreasing tendency (Table 2). Thus, there is the heterogeneous behavior of PSII in populus leaves different from Fv/Fm. Generally, the level of photochemical reaction can be evaluated according to the chlorophyll fluorescence intensity. And Fv/Fm can also used to describe the trapping efficiency of the absorbed light, which can reduce primary quinone electron acceptor of PSII (QA) (And and Weis, 2003). Therefore, the different fluorescence transients of leaves under high CO2 levels indicated that the photochemical reactions were different for different populus species, although there were similar decreasing Fv/Fm (reflecting trapping efficiencies of the absorbed light). In contrast, previous studies showed the Fv/Fm ratio reflects the photochemical efficiency of PSII (Rachoski et al., 2015). Various environmental stress can lead to inhibition of photosynthetic efficiency, accordingly, affecting state of the photosynthetic apparatus.


For O-J-I-P, point O can represent the fluorescence of PS Ⅱ action center, if all of the electron acceptor (QA, AB, PQ, etc.) are fully open in the maximum oxidation state. The fluorescence intensity of point O is connected with the content of the antenna pigment and the activity of action center. Point J reflects the rate of reduction of QA, which is connected with reaction centre pigments, light-harvesting pigment and the state of QA and QB. If the electrons transfer from QA to QB is restricted, the value of point J will rise (Ainsworth and Long, 2005). Here, higher J value in the leaves was observed. In addition, point I reflects the heterogeneity of PQ, the electron acceptor state in point I mainly is related to QA- and QB2-. Furthermore, the structure and function of PSⅡ complexes and the size of the PQ library is attributed to appearing time of point P. In this study, high CO2 level caused a significant decrease in the P-step level of the fluorescence transients OJIP curves of four populus species after 7 days of treatment. Therefore, a fast decrease of the P-step level indicated that there was main change to fluorescence transients. Generally, the “P” level was connected with the process of the electron transportation from QB to PQ, and it can mark concentrations of both QA- QB2- and PQH (Stirbet et al., 1998; Hill et al., 2004). Additionally, no significant changes in Fo, Fj and Fi of different populus species were observed after high CO2 treatment. Generally, high CO2 stress can lead to decrease of Fm, which reflects maximum fluorescence intensity at the P-step (Li et al., 2005). Therefore, this result suggested that the Fm may be a good marker of PSII vitality under high CO2 stress.


The PSII switches from the process of converting light energy into biochemical energy storage to the energy conversion process that transforms absorbed light energy into heat dissipation (Thach et al., 2007). One of PItotal is used to analyze the response of the plant’s PSII. Accordingly, it is sensitive to changes in either antenna properties, trapping efficiency or electron transport beyond QA (Oukarroum et al., 2007). In this study, we found that PSII performance can be represented using PItotal. In general, PItotal is also influenced by changes in antenna, RC, electron transport and end-acceptor reduction dependent parameters. Thus, PItotal integrates the response of RC/ABS, TRo/ABS(=Fv/Fm), ETo/TRo(=[1−Vj]) and REo/ETo(=[1 − Vi]/[1−Vj]).



Table 1 Formulae and explanation the technical data of the OJIP curves and the selected JIP-test parameters used in this study



Table 2 The fluorescence parameters on four Populus species under CO2 stress

Note: Values of the mean ± standard error for the un-manipulated control are given. The parameters are, minimum fluorescence at 20 s, Fo; fluorescence intensity at 2 ms, Fj; fluorescence intensity at 30 ms, Fi; maximum fluorescence, Fm; relative variable fluorescence at the J-step, Vj; relative variable fluorescence at the I-step, Vi; measure of the average total absorbance per active PSII RC, ABS/RC; maximum quantum yield of primary photochemistry, φPo(=TRo/ABS); probability that a trapped exciton moves an electron into the electron transport chain beyond QA-, ETo/TRo; total performance index per absorption basis, PItotal (see also Table 1). Results are presented as mean of five individual measurements


In summary, inhibition of PSII under high CO2 stress has involved in changes of Fm and PItotal. In other words, these two parameters can be used to evaluate the state of PSII. These results also indicated that changes of PSII attribute to the degradation of antenna pigment and inhibition of the electron transport at the acceptor side of PSII.


3 Materials and Methods

3.1 Plant material and stress treatment

The experiments were carried out in College of Life Science, Northeast Forestry University from Harbin, Heilongjiang Province. Four Populus species (populus X, Populus deltoides × cathayana, Populus alba Berdinensis′ and Populus euramericana ′N3016′ × Populus ussuriensis) were used as experimental materials. The cuttings were grown in plastic pots (60 cm in length, 25 cm in breadth and 15cm in depth) filled with 1.5kg of soil and sand (2:1). There are 10 cuttings in each pot. Potted plants were grown in the conditions: day/night air temperature, 28/22°C; photoperiod, 12h; relative humidity, 65-85%. One Chamber was kept at a CO2 concentration (average±SD) of 370 ± 15 μmol/mol (control), while another chamber was treated with an elevated CO2 concentration of 1500 ±50μmol/mol (treatment). At the 7th day after treatment, we examined the effect of EC on every variety of poplar using JIP-test. Ten biological replicates were repeated.


3.2 Chlorophyll a fluorescence transient measurement and the JIP-test

The OJIP transients were measured using fully expanded leaves. All measurements were carried out between 13:00 and 15:00 in May, 2016. After the plants had been exposed to high and normal CO2 concentration for 7 d, respectively, all leaves were exposed to a 30-min dark condition, before determination of OJIP transients. Accordingly, OJIP transients were measured using a portable chlorophyll a fluorometer (Hansatech Instruments, Ltd., King’s Lynn Norfolk, UK). In addition, we calculated values of several JIP parameters according to the JIP-test equations based on the methods of Strasser et al., Strasser et al., Tsimilli-Michael and Strasser (Strasser et al., 2004; Strasser et al., 2010; Tsimilli-Michael and Strasser, 2008).


In general, there are four important inflection points on the fluorescence fast dynamic curve, namely the O - J - I - P turning point. They are: 1) O point shows that the system (PSI) release amount of the fluorescent light, which can be used as the light of the PSI and efficiency, the fluorescence intensity is at 0.02 ms (F0). 2) Fluorescence intensity at 2ms is called Fj, J inflection point, which can reflect the reaction in the accumulation of the electron acceptor QA- to QB-, the oxidation of the Q. 3) Fluorescence intensity at 30ms is called Fi. 4)Maximum fluorescence intensity is Fm.


3.3 Date analysis

Each experiment was conducted at least ten times independently. All data presented were mean values of each treatment.


Authors’ contributions

Ruyu Xie, Mu Peng and Fanjuan Meng performed and designed the experiments; Tao Wang, Fanjuan Meng and Fachun Guan performed and analyzed the data.



This project was supported by the National Natural Science Foundation of China (31660552), the Fundamental Research Funds for the Central Universities (2572018CG05) and college students' innovation training program (201810225133).



Ainsworth E.A., and Long S.P., 2005, What have we learned from 15 years of free-air CO2 enrichment (face)? A meta-analytic review of the responses of photosynthesis, canopy properties and plant production to rising CO2, New Phytologist, 165(2): 351


Albert K.R., Mikkelsen T.N., and Ro-Poulsen H., 2005, Effects of ambient versus reduced UV-B radiation on high arctic Salix arctica, assessed by measurements and calculations of chlorophyll a, fluorescence parameters from fluorescence transients, Physiologia Plantarum, 124(2): 208–226


Amthor J.S., 2010, Terrestrial higher-plant response to increasing atmospheric [CO2] in relation to the global carbon cycle, Global Change Biology, 1(4): 243-274


An L., 2006, The polyphasic chlorophyll a fluorescence rise measured under high intensity of exciting light, Functional Plant Biology, 33(1): 9-30


And G.H.K., and Weis E., 2003, Chlorophyll fluorescence and photosynthesis: the basics, annual review of plant physiology, 42(42): 313-349


Chen L.S., Li P., and Cheng L., 2008, Effects of high temperature coupled with high light on the balance between photooxidation and photoprotection in the sun-exposed peel of apple, Planta, 228(5): 745-56


Cui C., Huang C., Yu G., Cui Y., and Zhao J., 1999, Research of nursery growth rule of populus xiaohei planting by cutting, Journal Ofence of Teachers College & University


Force L., Critchley C., and Rensen J.J.S.V., 2003, New fluorescence parameters for monitoring photosynthesis in plants, Photosynthesis Research, 78(1): 17-33


Griffin D.W., Garrison V.H., Herman J.R., and Shinn E.A., 2001, African desert dust in the Caribbean atmosphere: Microbiology and public health, Aerobiologia, 17(3): 203-213


Heerden P.D.R.V., Strasser R.J., and Krüger G.H.J., 2004, Reduction of dark chilling stress in N 2 -fixing soybean by nitrate as indicated by chlorophyll a, fluorescence kinetics, Physiologia Plantarum, 121(2): 239


Hill R., Larkum A.W., Frankart C., Kühl M., and Ralph P.J., 2004, Loss of functional photosystem II reaction centres in zooxanthellae of corals exposed to bleaching conditions: using fluorescence rise kinetics, Photosynthesis Research, 82(1): 59-72



Ilík P., Schansker G., Kotabová E., Váczi P., Strasser R.J., and Barták M., 2006, A dip in the chlorophyll fluorescence induction at 0.2-2 s in Trebouxia-possessing lichens reflects a fast reoxidation of photosystem I, A comparison with higher plants, Biochimica et biophysica acta, 1757(1): 12


IPCC, 2007, Intergovernmental panel on climate change, Climatic change 2007: the physical science basis, Geneva


Jiang C.D., Gao H.Y., and Zou Q., 2002, Characteristics of photosynthetic apparatus in Mn-starved maize leaves, Photosynthetica, 40(2): 209-213


Jiang C.D., Gao H.Y., and Zou Q., 2003, Changes of donor and acceptor side in photosystem 2 complex induced by iron deficiency in attached soybean and maize leaves, Photosynthetica, 41(2): 267-271


Jiang Z.H., Wang X.Q., Fei B.H., Ren H.Q., and Liu X.E., 2007, Effect of stand and tree attributes on growth and wood quality characteristics from a spacing trial with Populus xiaohei, Annals of Forest Science, 64(8): 807-814


Kostiainen K., Kaakinen S., Warsta E., Kubiske M.E., and Nelson N.D., 2008, Wood properties of trembling aspen and paper birch after 5 years of exposure to elevated concentrations of CO2 and O3, Tree Physiology, 28(5): 805-813


Li P.M., Gao H.Y., and Strasser R.J., 2005, Application of the fast chlorophyll fluorescence induction dynamics analysis in photosynthesis study., Acta Photophysiologica Sinica, 31(6): 559


Long S.P., Ainsworth E.A., Rogers A., and Ort D.R., 2004, Rising atmospheric carbon dioxide: plants face the future, Annual review of plant biology, 2004: 591-628



Oukarroum A., Madidi S.E., Schansker G., and Strasser R.J. 2007, Probing the responses of barley cultivars (Hordeum vulgare L.) by chlorophyll a, fluorescence OLKJIP under drought stress and re-watering, Environmental & Experimental Botany, 60(3): 438-446


Peters G.P., and Hertwich E.G., 2008, CO2 embodied in international trade with implications for global climate policy, Environmental Science & Technology, 42(5): 1401-1407


Rachoski M., Gazquez A., Calzadilla P., Bezus R., Rodriguez A., Ruiz O., Menendez A., and Maiale S., 2015, Chlorophyll fluorescence and lipid peroxidation changes in rice somaclonal lines subjected to salt stress, Acta Physiologiae Plantarum, 37(6): 1-12


Schansker G., Kissimon J., and Kovács L., 2005, Biophysical studies of photosystem II-related recovery processes after a heat pulse in barley seedlings (Hordeum vulgare L.), J. Plant Physiol., Journal of Plant Physiology, 162(2): 181


Šesták Z., 2001, Probing photosynthesis, mechanisms, regulation and adaptation, Photosynthetica, 39(1): 10


Stirbet A., Strasser B.J., Strasser R.J., and Govindjee., 1998, Chlorophyll a fluorescence Induction in Higher Plants: Modelling and Numerical Simulation, Journal of Theoretical Biology, 193(1): 131-151


Strasser R.J., Tsimillimichael M., and Srivastava A., 2004, Analysis of the Chlorophyll a Fluorescence Transient, Chlorophyll a Fluorescence, Springer Netherlands, 2004: 321-362


Strasser R.J., Tsimilli-Michael M., Qiang S., and Goltsev V., 2010, Simultaneous in vivo recording of prompt and delayed fluorescence and 820-nm reflection changes during drying and after rehydration of the resurrection plant Haberlea rhodopensis, Biochim Biophys Acta, 1797(6–7): 1313-1326


Strasserf R.J., Srivastava A., and Govindjee., 2008, Polyphasic chlorophyll a fluorescence transient in plants and cyanobacteria. Photochem Photobiol, Photochemistry & Photobiology, 61(1):32-42


Strauss A.J., Ghj K., Strasser R.J., and van Heerden P.D., 2007, The role of low soil temperature in the inhibition of growth and PSII function during dark chilling in soybean genotypes of contrasting tolerance, Physiologia Plantarum, 131(1): 89–105


Thach L.B., Shapcott A., Schmidt S., and Critchley C., 2007, The OJIP fast fluorescence rise characterizes Graptophyllum species and their stress responses., Photosynthesis Research, 94(2): 423-436


Tóth S.Z., Schansker G., Garab G., and Strasser R.J., 2007, Photosynthetic electron transport activity in heat-treated barley leaves: the role of internal alternative electron donors to photosystem II, Biochimica Et Biophysica Acta, 1767(4): 295-305


Tsimilli-Michael M., and Strasser R.J., 2008, In vivo Assessment of Stress Impact on Plant’s Vitality: Applications in Detecting and Evaluating the Beneficial Role of Mycorrhization on Host Plants, Mycorrhiza, Springer Berlin Heidelberg, pp.679-703


Verhoeven A.S., Adams W.W., Demmigadams B., Croce R., and Bassi R., 1999, Xanthophyll cycle pigment localization and dynamics during exposure to low temperatures and light stress in vinca major, Plant Physiology, 120(3): 727



Yordanov I., Goltsev V., Stefanov D., Chernev P., Zaharieva I., Kirova M., Gecheva V., and Strasser R.J., 2008, Preservation of photosynthetic electron transport from senescence-induced inactivation in primary leaves after decapitation and defoliation of bean plants, Journal of Plant Physiology, 165(18): 1954

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