Temperature-mediated shifts in chlorophyll biosynthesis in leaves of chlorophyll b-lacking rice (Oryza sativa L.)


  • Khiem Minh NGUYEN Biodiversity Research Center, Academia Sinica, Nangang, Taipei 115; Biodiversity Program, Taiwan International Graduate Program, Academia Sinica and National Taiwan Normal University, Taipei 115; Department of Life Science, National Taiwan Normal University, Taipei 106; Faculty of Applied Sciences, Ton Duc Thang University, Ho Chi Minh City 700000 (TW)
  • Zhi-Wei YANG Taoyuan District Agricultural Research and Extension Station, Council of Agriculture, Taoyuan 327 (TW)
  • Tin-Han SHIH Biodiversity Research Center, Academia Sinica, Nangang, Taipei 115 (TW)
  • Szu-Hsien LIN Biodiversity Research Center, Academia Sinica, Nangang, Taipei 115 (TW)
  • Jun-Wei LIN Biodiversity Research Center, Academia Sinica, Nangang, Taipei 115 (TW)
  • Hoang Chinh NGUYEN Faculty of Applied Sciences, Ton Duc Thang University, Ho Chi Minh City 700000 (VN)
  • Chi-Ming YANG Biodiversity Research Center, Academia Sinica, Nangang, Taipei 115; Biodiversity Program, Taiwan International Graduate Program, Academia Sinica and National Taiwan Normal University, Taipei 115 (TW)




chlorophyll b-lacking mutant, grana, photosynthesis, temperature sensitivity


Extreme temperatures have become a threat to crop yields. To maintain plant growth and yield, chlorophyll (Chl) biosynthesis plays a crucial role in adaptation to temperature stress. This study investigated the influence of temperature on the biosynthesis and characteristics of pigments (Chl a, Chl b, and carotenoids) in the leaves of Chl b-lacking mutant rice (Chlorina 1, ch1) and wild-type rice (Norin No.8, wt). The ch1 showed thinner stacked grana caused by a decrease in thylakoid membranes per granum at 15 °C, whereas the destacked grana were observed at 35 °C after 12 h incubation. However, the grana are stacked normally, along with the absence of Chl b, and a significantly decreased amount of Chl a in both wt and ch1 were observed after heat stress exposure, demonstrating that light-harvesting complex II proteins are involved in grana stacking. Ch1 was sensitive to 15 °C during the first 4 h of incubation but it subsequently adapted to the cold environment. In addition, there were no significant differences in the photosynthesis between wt and ch1 after 12 h incubation at 35 °C. Differentially expressed gene (DEGs) analysis revealed that GluRS expression decreased, which resulted in a decline in Chl biosynthesis in wt and ch1 at 35 °C. At 8 h and 12 h, there were no significant differences in the expression of DEGs involved in Chl biosynthesis and degradation between wt and ch1 at 15 °C. ALAD expression in wt and ch1 at 15 °C decreased until it was undetectable. These findings suggested that ch1 may adapt to temperatures ranging from 15 °C to 35 °C.


Allakhverdiev SI, Kreslavski VD, Klimov VV, Los DA, Carpentier R, Mohanty P (2008). Heat stress: an overview of molecular responses in photosynthesis. Photosynthesis Research 98:541. https://doi.org/10.1007/s11120-008-9331-0

Allen DJ, Ort DR (2001). Impacts of chilling temperatures on photosynthesis in warm-climate plants. Trends in Plant Science 6:36-42. https://doi.org/10.1016/S1360-1385(00)01808-2

Allen KD, Duysen ME, Staehelin LA (1988). Biogenesis of thylakoid membranes is controlled by light intensity in the conditional chlorophyll b-deficient CD3 mutant of wheat. The Journal of Cell Biology 107:907. http://jcb.rupress.org/content/107/3/907.abstract

Aro EM, Virgin I, Andersson B (1993). Photoinhibition of photosystem II. Inactivation, protein damage and turnover. Biochimica et Biophysica Acta (BBA) - Bioenergetics 1143:113-134. https://doi.org/https://doi.org/10.1016/0005-2728(93)90134-2

Bellemare G, Bartlett SG, Chua NH (1982). Biosynthesis of chlorophyll a/b-binding polypeptides in wild type and the chlorina f2 mutant of Barley. Journal of Biological Chemistry 257:7762-7767. https://doi.org/10.1016/S0021-9258(18)34446-6

Berry J, Bjorkman O (1980). Photosynthetic response and adaptation to temperature in higher plants. Annual Review of Plant Physiology 31:491-543. https://doi.org/10.1146/annurev.pp.31.060180.002423

Bilska A, Sowiński P (2010). Closure of plasmodesmata in maize (Zea mays) at low temperature: a new mechanism for inhibition of photosynthesis. Annals of Botany 106:675-686. https://doi.org/10.1093/aob/mcq169

BukhovNG, Carpentier R (2000). Heterogeneity of photosystem II reaction centers as influenced by heat treatment of barley leaves. Physiologia Plantarum 110:279-285. https://doi.org/10.1034/j.1399-3054.2000.110219.x

Chaves MM, Flexas J, Pinheiro C (2009). Photosynthesis under drought and salt stress: regulation mechanisms from whole plant to cell. Annals of Botany 103:551-560. https://doi.org/10.1093/aob/mcn125

Cheng XX, Yu M, Zhang N, Zhou ZQ, Xu QT, Mei FZ, Qu LH (2016.) Reactive oxygen species regulate programmed cell death progress of endosperm in winter wheat (Triticum aestivum L.) under waterlogging. Protoplasma 253:311-327. https://doi.org/10.1007/s00709-015-0811-8

Cruz RPda, Sperotto RA, Cargnelutti D, Adamski JM, de FreitasTerra T, Fett JP (2013). Avoiding damage and achieving cold tolerance in rice plants. Food and Energy Security 2:96-119. https://doi.org/https://doi.org/10.1002/fes3.25

Ding Y, Shi Y, Yang S (2020). Molecular regulation of plant responses to environmental temperatures. Molecular Plant 13:544-564. https://doi.org/https://doi.org/10.1016/j.molp.2020.02.004

Eckhardt U, Grimm B, Hörtensteiner S (2004). Recent advances in chlorophyll biosynthesis and breakdown in higher plants. Plant Molecular Biology 56:1-14. https://doi.org/10.1007/s11103-004-2331-3

Fahad S, Adnan M, Hassan S, Saud S, Hussain S, Wu C, Huang J (2018). Rice responses and tolerance to high temperature. In: Advances in Rice Research for Abiotic Stress Tolerance. Woodhead Publishing, pp 201-224. https://doi.org/10.1016/B978-0-12-814332-2.00010-1

Fromme P, Melkozernov A, Jordan P, Krauss N (2003). Structure and function of photosystem I: interaction with its soluble electron carriers and external antenna systems. FEBS Letters 555:40-44. https://doi.org/10.1016/S0014-5793(03)01124-4

Gounaris K, Brain ARR, Quinn P, Williams WP (1984). Structural reorganisation of chloroplast thylakoid membranes in response to heat-stress. Biochimica et Biophysica Acta (BBA) - Bioenergetics 766:198-208. https://doi.org/10.1016/0005-2728(84)90232-9

Gounaris K, Brain APR, Quinn PJ, Williams WP (1983). Structural and functional changes associated with heat-induced phase-separations of non-bilayer lipids in chloroplast thylakoid membranes. FEBS Letters 153:47-52. https://doi.org/https://doi.org/10.1016/0014-5793(83)80117-3

Greene BA, Allred DR, Morishige DT, Staehelin LA (1988). Hierarchical response of light harvesting chlorophyll-proteins in a light-sensitive chlorophyll b-deficient mutant of maize. Plant Physiology 87:357-364.http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1054757/

Hassan I (1999). Effects of O3 and drought stress on growth, yield and physiology of tomatoes (Lycopersicon esculentum Mill. Cv Baladey). Gartenbauwiessenschaft 76:122-135.

Havaux M, Tardy F (1997). Thermostability and photostability of photosystem ii in leaves of the chlorina-f2 barley mutant deficient in light-harvesting chlorophyll a/b protein complexes. Plant Physiology 113:913-923. https://doi.org/10.1104/pp.113.3.913

Havaux M, Tardy F (1996). Temperature-dependent adjustment of the thermal stability of photosystem II in vivo: possible involvement of xanthophyll-cycle pigments. Planta 198:324-333. https://doi.org/10.1007/BF00620047

HopkinsWG, Hayden DB, Neuffer MG (1980). A light-sensitive mutant in maize (Zea mays L.) I. Chlorophyll, chlorophyll-protein and ultrastructural studies. Zeitschrift Für Pflanzenphysiologie 99:417-426. https://doi.org/https://doi.org/10.1016/S0044-328X(80)80157-7

Horie T (2019). Global warming and rice production in Asia: modeling, impact prediction and adaptation. Proceedings of the Japan Academy, Series B, 95:211-245. https://doi.org/10.2183/pjab.95.016

Hu S, Ding Y, Zhu C (2020). Sensitivity and responses of chloroplasts to heat stress in plants. Frontiers in Plant Science 11:375. https://doi.org/10.3389/fpls.2020.00375

Huang J, Qin F, Zang G, Kang Z, Zou H, Hu F, Yue C, Li X, Wang G (2013). Mutation of OsDET1 increases chlorophyll content in rice. Plant Science 210: 241-249. https://doi.org/10.1016/j.plantsci.2013.06.003

Huang S, Liu Z, Li D, Yao R, Meng Q, Feng H (2014). Screening of Chinese cabbage mutants produced by 60Co γ-ray mutagenesis of isolated microspore cultures. Plant Breeding 133:480-488. https://doi.org/10.1111/pbr.12166

Kadam NN, Xiao G, Melgar RJ, Bahuguna RN, Quinones C, Tamilselvan A, Jagadish KSV (2014). Agronomic and physiological responses to high temperature, drought, and elevated CO2 interactions in cereals. Advances in Agronomy 127:111-156. https://doi.org/10.1016/B978-0-12-800131-8.00003-0

Klimov V, Baranov S, Allakhverdiev S (1998). Bicarbonate protects the donor side of photosystem II against photoinhibition and thermoinactivation. FEBS Letters 418:243-246. https://doi.org/10.1016/S0014-5793(97)01392-6

Lai YC, Wang SY, Gao HY, Nguyen KM, Nguyen CH, Shih MC, Lin KH (2016). Physicochemical properties of starches and expression and activity of starch biosynthesis-related genes in sweet potatoes. Food Chemistry 199:556-564. https://doi.org/10.1016/j.foodchem.2015.12.053

Landi M, Zivcak M, Sytar O, Brestic M, Allakhverdiev SI (2020). Plasticity of photosynthetic processes and the accumulation of secondary metabolites in plants in response to monochromatic light environments: A review. Biochimica et Biophysica Acta (BBA) - Bioenergetics 1861:14813. https://doi.org/10.1016/j.bbabio.2019.148131

Lawlor DW, Tezara W (2009) Causes of decreased photosynthetic rate and metabolic capacity in water-deficient leaf cells: a critical evaluation of mechanisms and integration of processes. Annals of Botany 103:561–579. https://doi.org/10.1093/aob/mcn244

Lin YH, Pan KY, Hung CH, Huang HE, Chen CL, Feng TY, Huang LF (2013). Overexpression of ferredoxin, PETF, enhances tolerance to heat stress in Chlamydomonas reinhardtii. International Journal of Molecular Sciences 14:20913-20929. https://doi.org/10.3390/ijms141020913

Liu R, Dong X, Gu W, Yu L, Jin W, Qu Y, Li W (2016). Variation in the phenotypic features and transcripts of thermo-sensitive leaf-color mutant induced by carbon ion beam in Green wandering jew (Tradescantia fluminensis). Scientia Horticulturae 213:303-313. https://doi.org/https://doi.org/10.1016/j.scienta.2016.11.001

Liu W, Fu Y, Hu G, Si H, Zhu L, Wu C, Sun Z (2007). Identification and fine mapping of a thermo-sensitive chlorophyll deficient mutant in rice (Oryza sativa L.). Planta 226:785-795. https://doi.org/10.1007/s00425-007-0525-z

Markwell J, Osterman JC (1992). Occurrence of temperature-sensitive phenotypic plasticity in chlorophyll-deficient mutants of Arabidopsis thaliana L. Plant Physiology 98:392-394. https://doi.org/10.1104/pp.98.1.392

Markwell JP, DankoSJ, Bauwe H, Osterman J, Gorz HJ, Haskins FA (1986). A temperature-sensitive chlorophyll b-deficient mutant of sweetclover (Melilotus alba). Plant Physiology 81:329-334. https://doi.org/10.1104/pp.81.2.329

Masuda T, Fujita Y (2008) Regulation and evolution of chlorophyll metabolism. Photochemical & Photobiological Sciences 7:1131-1149. https://doi.org/10.1039/B807210H

Michel H, Tellenbach M, Boschetti A (1983). A chlorophyll b-less mutant of Chlamydomonas reinhardii lacking in the light-harvesting chlorophyll ab-protein complex but not in its apoproteins. Biochimica et Biophysica Acta (BBA) - Bioenergetics 725:417-424. https://doi.org/10.1016/0005-2728(83)90182-2

Mohanty P, Allakhverdiev S, Murata N (2007). Application of low temperature during photoinhibition allows characterization of individual steps in photodamage and repair of photosystem II. Photosynthesis Research 94:217-224. https://doi.org/10.1007/s11120-007-9184-y

Murakami Y, Tsuyama M, Kobayashi Y, Kodama H, Iba K (2000). Trienoic fatty acids and plant tolerance of high temperature. Science 287:476-479. https://doi.org/10.1126/science.287.5452.476

Murata N, Takahashi S, Nishiyama Y,Allakhverdiev SI (2007). Photoinhibition of photosystem II under environmental stress. Biochimica et Biophysica Acta (BBA) - Bioenergetics 1767:414-421. https://doi.org/https://doi.org/10.1016/j.bbabio.2006.11.019

Nakashima K, Tran LP, Van Nguyen D, Fujita M, Maruyama K, Todaka D, Ito Y, Hayashi N, Shinozaki K, Yamaguchi-Shinozaki K (2007). Functional analysis of a NAC‐type transcription factor OsNAC6 involved in abiotic and biotic stress‐responsive gene expression in rice. Plant Journal 51:617-630. https://doi.org/10.1111/j.1365-313X.2007.03168.x

Nakatani HY, Baliga V (1985). A clover mutant lacking the chlorophyll a- and b-containing protein antenna complexes. Biochemical and Biophysical Research Communications 131:182-189. https://doi.org/https://doi.org/10.1016/0006-291X(85)91787-5

Nguyen MK, Shih TH, Lin SH, Huang WD, Yang CM (2020). Transcription analysis of chlorophyll biosynthesis in wildtype and chlorophyll b-lacking rice (Oryza sativa L.). Photosynthetica 58:702-711. https://doi.org/10.32615/ps.2020.022

Nishiyama Y, Allakhverdiev SI, Murata N (2005). Inhibition of the repair of Photosystem II by oxidative stress in cyanobacteria. Photosynthesis Research 84:1-7. https://doi.org/10.1007/s11120-004-6434-0

Ouijja A, Farineau N, Cantrel C, Guillot-Salomon T (1988). Biochemical analysis and photosynthetic activity of chloroplasts and Photosystem II particles from a barley mutant lacking chlorophyll b. Biochimica et Biophysica Acta (BBA) - Bioenergetics 932:97-106. https://doi.org/https://doi.org/10.1016/0005-2728(88)90143-0

PetrovK, Lyubov V, Dudareva L, Nokhsorov V, Perk A, Chepalov V, Zulfugarov I (2016). The role of plant fatty acids in regulation of the adaptation of organisms to the cold climate in cryolithic zone of Yakutia. Journal of Life Science 26:519-530. https://doi.org/10.5352/JLS.2016.26.5.519

Poorter H (2004). Physiological plant ecology. Annals of Botany 93:616-617. https://doi.org/10.1093/aob/mch084

Qiu Z, Kang S, He L, Zhao J, Zhang S, Hu J, Zhu L (2018). The newly identified heat-stress sensitive albino 1 gene affects chloroplast development in rice. Plant Science 267:168-179. https://doi.org/https://doi.org/10.1016/j.plantsci.2017.11.015

RaisonJK, Roberts JKM, Berry JA (1982). Correlations between the thermal stability of chloroplast (thylakoid) membranes and the composition and fluidity of their polar lipids upon acclimation of the higher plant, Nerium oleander, to growth temperature. Biomembranes 688:218-228. https://doi.org/https://doi.org/10.1016/0005-2736(82)90597-1

RazaA, Razzaq A, Mehmood SS, ZouX, Zhang X, Lv Y, Xu J (2019). Impact of climate change on crops adaptation and strategies to tackle its outcome: A review. Plants 8:34. https://doi.org/10.3390/plants8020034

Rüdiger W (1997). Chlorophyll metabolism: From outer space down to the molecular level. Phytochemistry 46:1151-1167. https://doi.org/10.1016/S0031-9422(97)80003-9

Salvucci ME, Crafts-Brandner SJ (2004). Relationship between the heat tolerance of photosynthesis and the thermal stability of rubisco activase in plants from contrasting thermal environments. Plant Physiology 134:1460-1470. https://doi.org/10.1104/pp.103.038323

Sassa S (1982). Delta-Aminolevulinic acid dehydratase assay. Enzyme 28:133-145. https://doi.org/10.1159/000459097

Semenova GA (2004). Structural reorganization of thylakoid systems in response to heat treatment. Photosynthetica 42:521-527. https://doi.org/10.1007/S11099-005-0008-z

Shangguan Z, Shao M, Dyckmans J (1999). Interaction of osmotic adjustment and photosynthesis in winter wheat under soil drought. Journal of Plant Physiology 154:753-758.https://doi.org/https://doi.org/10.1016/S0176-1617(99)80254-5

Sharkey TD (2005). Effects of moderate heat stress on photosynthesis: importance of thylakoid reactions, rubisco deactivation, reactive oxygen species, and thermotolerance provided by isoprene. Plant, Cell & Environment 28:269-277. https://doi.org/10.1111/j.1365-3040.2005.01324.x

Soda N, Gupta BK, Anwar K, Sharan A, Govindjee, Singla-Pareek SL, Pareek A (2018). Rice intermediate filament, OsIF, stabilizes photosynthetic machinery and yield under salinity and heat stress. Scientific Reports 8:4072. https://doi.org/10.1038/s41598-018-22131-0

Strzałka K, Kostecka-Gugała A, Latowski D (2003). Carotenoids and environmental stress in plants: significance of carotenoid-mediated modulation of membrane physical properties. Russian Journal of Plant Physiology 50:168-173. https://doi.org/10.1023/A:1022960828050

Terao T, Yamashita A, Katoh S (1985). Chlorophyll b-deficient mutants of rice. 1. Absorption and fluorescence spectra and chlorophyll a/b ratios. Plant and Cell Physiology 26:1361-1367. https://doi.org/10.1093/oxfordjournals.pcp.a077036

Verkamp E, Jahn M, Jahn D, Kumar A, Soll D (1992). Glutamyl-tRNA reductase from Escherichia coli and Synechocystis 6803: Gene structure and expression. Journal of Biological Chemistry 267:8275-8280. https://doi.org/10.1016/S0021-9258(18)42438-6

Voitsekhovskaja O V, Tyutereva E V (2015). Chlorophyll b in angiosperms: Functions in photosynthesis, signaling and ontogenetic regulation. Journal of Plant Physiology 189:51-64. https://doi.org/10.1016/j.jplph.2015.09.013

Wahid A, Gelani S, Ashraf M, Foolad MR (2007). Heat tolerance in plants: an overview. Environmental and Experimental Botany 61:199-223. https://doi.org/https://doi.org/10.1016/j.envexpbot.2007.05.011

Wood WHJ, Barnett SFH, Flannery S, Hunter CN, Johnson MP (2019). Dynamic thylakoid stacking is regulated by LHCII phosphorylation but not its interaction with PSI. Plant Physiology 180:2152-2166. https://doi.org/10.1104/pp.19.00503

Xie G, Kato H, Sasaki K, Imai R (2009). A cold-induced thioredoxin h of rice, OsTrx23, negatively regulates kinase activities of OsMPK3 and OsMPK6 in vitro. FEBS Letters 583:2734-2738. https://doi.org/10.1016/j.febslet.2009.07.057

Xin C, Hou R, Wu F, Zhao Y, Xiao H, Si W, Guo J (2015). Analysis of cytosine methylation status in potato by methylation-sensitive amplified polymorphisms under low-temperature stress. Journal of Plant Biology 58:383-390. https://doi.org/10.1007/s12374-015-0316-1

Yamamoto Y, Aminaka R, Yoshioka M, Khatoon M, Komayama K, Takenaka D, Yamamoto Y (2008). Quality control of photosystem II: impact of light and heat stresses. Photosynthesis Research 98:589-608. https://doi.org/10.1007/s11120-008-9372-4

YamaneY, Kashino Y, Koike H, Satoh K (1998). Effects of high temperatures on the photosynthetic systems in spinach: oxygen-evolving activities, fluorescence characteristics and the denaturation process. Photosynthesis Research 57:51-59.https://doi.org/10.1023/A:1006019102619

Yang CM, Osterman JC, Markwell J (1990). Temperature sensitivity as a general phenomenon in a collection of chlorophyll-deficient mutants of sweetclover (Melilotus alba). Biochemical Genetics 28:31-40. https://doi.org/10.1007/BF00554819

Yang HY, Xia XW, Fang W, Fu Y, An MM, Zhou MB (2015a). Identification of genes involved in spontaneous leaf color variation in Pseudosasa japonica. Genetics and Molecular Research 14:11827-11840. http://dx.doi.org/10.4238/2015.October.2.16

Yang Y, Chen X, Xu B, Li Y, Ma Y, Wang G (2015b). Phenotype and transcriptome analysis reveals chloroplast development and pigment biosynthesis together influenced the leaf color formation in mutants of Anthurium andraeanum ‘Sonate’. Frontiers in Plant Science 6:139.https://doi.org/10.3389/fpls.2015.00139

Yordanov I, Velikova V, Tsonev T (1999). Influence of drought, high temperature, and carbamide cytokinin 4-pu-30 on photosynthetic activity of bean plants. 1. Changes in chlorophyll fluorescence quenching. Photosynthetica 37:447-457. https://doi.org/10.1023/A:1007163928253

Yu HD, Yang XF, Chen ST, Wang YT, Li JK, Shen Q, Guo FQ (2012). Downregulation of chloroplast RPS1 negatively modulates nuclear heat-responsive expression of HsfA2 and its target genes in Arabidopsis. PLoS Genetics 8:e1002669. https://doi.org/10.1371/journal.pgen.1002669

Zhao A, Fang Y, Chen X, Zhao S, Dong W, Lin Y, Gong W, Liu L (2014). Crystal structure of Arabidopsis glutamyl-tRNA reductase in complex with its stimulator protein. Proceedings of the National Academy of Science 111:6630-6635. https://doi.org/10.1073/pnas.1400166111

Zhao X, Nishimura Y, Fukumoto Y, Li J (2011). Effect of high temperature on active oxygen species, senescence and photosynthetic properties in cucumber leaves. Environmental and Experimental Botany 70:212-216. https://doi.org/10.1016/j.envexpbot.2010.09.005

Zhu JK (2016). Abiotic stress signaling and responses in plants. Cell 167:313-324. https://doi.org/10.1016/j.cell.2016.08.029




How to Cite

NGUYEN, K. M., YANG, Z.-W., SHIH, T.-H., LIN, S.-H., LIN, J.-W., NGUYEN, H. C., & YANG, C.-M. (2021). Temperature-mediated shifts in chlorophyll biosynthesis in leaves of chlorophyll b-lacking rice (Oryza sativa L.). Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 49(2), 12306. https://doi.org/10.15835/nbha49212306



Research Articles
DOI: 10.15835/nbha49212306

Most read articles by the same author(s)