High-throughput chlorophyll fluorescence screening of Setaria viridis for mutants with altered CO2 compensation points
Robert A. Coe A B D , Jolly Chatterjee A * , Kelvin Acebron C * , Jacqueline Dionora A * , Reychelle Mogul A , HsiangChun Lin A , Xiaojia Yin A , Anindya Bandyopadhyay A , Xavier R. R. Sirault D , Robert T. Furbank B and W. Paul Quick A E F
+ Author Affiliations
- Author Affiliations
A C4 Rice Centre, International Rice Research Institute (IRRI), Los Baños, Philippines.
B ARC Centre of Excellence for Translational Photosynthesis, Research School of Biology, Australian National University, GPO Box 1500, Canberra, ACT 2601, Australia.
C IBG-2, Forschungszentrum Jülich (FZJ), Jülich, 52425 Jülich, Germany.
D CSIRO Agriculture Flagship, High Resolution Plant Phenomics GPO Box 1500, Canberra, ACT 2601, Australia.
E Department of Animal and Plant Sciences, University of Sheffield, Sheffield, S10 2TN, UK.
F Corresponding author. Email: w.p.quick@irri.org
Functional Plant Biology 45(10) 1017-1025 https://doi.org/10.1071/FP17322
Submitted: 17 November 2017 Accepted: 27 March 2018 Published: 8 May 2018
Abstract
To assist with efforts to engineer a C4 photosynthetic pathway into rice, forward-genetic approaches are being used to identify the genes modulating key C4 traits. Currently, a major challenge is how to screen for a variety of different traits in a high-throughput manner. Here we describe a method for identifying C4 mutant plants with increased CO2 compensation points. This is used as a signature for decreased photosynthetic efficiency associated with a loss of C4 function. By exposing plants to a CO2 concentration close to the CO2 compensation point of a wild-type plant, individuals can be identified from measurements of chlorophyll a fluorescence. We use this method to screen a mutant population of the C4 monocot Setaria viridis (L.) P.Beauv. generated using N-nitroso-N-methylurea (NMU). Mutants were identified at a frequency of 1 per 157 lines screened. Forty-six candidate lines were identified and one line with a heritable homozygous phenotype selected for further characterisation. The CO2 compensation point of this mutant was increased to a value similar to that of C3 rice. Photosynthesis and growth was significantly reduced under ambient conditions. These data indicate that the screen was capable of identifying mutants with decreased photosynthetic efficiency. Characterisation and next-generation sequencing of all the mutants identified in this screen may lead to the discovery of novel genes underpinning C4 photosynthesis. These can be used to engineer a C4 photosynthetic pathway into rice.
Additional keywords: C4 photosynthesis, C4 rice, forward genetics, high-throughput phenomics.
References
Aro EM, Virgin I, Andersson B (1993) Photoinhibition of photosystem II. Inactivation, protein damage and turnover. Biochimica et Biophysica Acta 1143, 113–134.| Photoinhibition of photosystem II. Inactivation, protein damage and turnover.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK3sXms1Kgtb0%3D&md5=ddb1e54e441e49a1c8c64b5cb30cfd6aCAS |
Badger MR, Fallahi H, Kaines S, Takahashi S (2009) Chlorophyll fluorescence screening of Arabidopsis thaliana for CO2 sensitive photorespiration and photoinhibition mutants. Functional Plant Biology 36, 867–873.
| Chlorophyll fluorescence screening of Arabidopsis thaliana for CO2 sensitive photorespiration and photoinhibition mutants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXhtlOgs7rP&md5=67d3a9fad1ab06ffe7e43f271ffb9c59CAS |
Belgio E, Kapitonova E, Chmeliov J, Duffy CDP, Ungerer P, Valkunas L, Ruban AV (2014) Economic photoprotection in photosystem II that retains a complete light-harvesting system with slow energy traps. Nature Communications 5, 8–16.
| Economic photoprotection in photosystem II that retains a complete light-harvesting system with slow energy traps.Crossref | GoogleScholarGoogle Scholar |
Björkman O, Demmig B (1987) Photon yield of O2 evolution and chlorophyll fluorescence characteristics at 77 K among vascular plants of diverse origins. Planta 170, 489–504.
| Photon yield of O2 evolution and chlorophyll fluorescence characteristics at 77 K among vascular plants of diverse origins.Crossref | GoogleScholarGoogle Scholar |
Blackwell RD, Murray AJ, Lea PJ, Kendall AC, Hall NP, Turner JC, Wallsgrove RM (1988) The value of mutants unable to carry out photorespiration. Photosynthesis Research 16, 155–176.
| The value of mutants unable to carry out photorespiration.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC2czlvFOjuw%3D%3D&md5=77101031eb471f5b6a876293b2e707e3CAS |
Cousins AB, Baroli I, Badger MR, Ivakov A, Lea PJ, Leegood RC, von Caemmerer S (2007) The role of phosphoenolpyruvate carboxylase during C4 photosynthetic isotope exchange and stomatal conductance. Plant Physiology
| The role of phosphoenolpyruvate carboxylase during C4 photosynthetic isotope exchange and stomatal conductance.Crossref | GoogleScholarGoogle Scholar |
Demmig-Adams B, Adams WW (2006) Photoprotection in an ecological context: the remarkable complexity of thermal energy dissipation. New Phytologist 172, 11–21.
| Photoprotection in an ecological context: the remarkable complexity of thermal energy dissipation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XhtVyisrnE&md5=447b9a274ed00d820a3b2589f949dda9CAS |
Dever LV, Balckwell RD, Fullwood NJ, Lacuesta M, Leegood RC, Onek LA, Pearson M, Lea PJ (1995) The isolation and characterisation of mutant of the C4 photosynthetic pathway. Journal of Experimental Botany 46, 1363–1376.
| The isolation and characterisation of mutant of the C4 photosynthetic pathway.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXovFans7w%3D&md5=c4309af0d10a7d14313707effcf19bbcCAS |
Feldman AB, Murchie EH, Leung H, Baraoidan M, Coe RA, Yu S-M, Lo S-F, Quick WP (2014) Increasing leaf vein density of mutagenesis: laying the foundations for C4 rice. PLoS One 9, e94947
| Increasing leaf vein density of mutagenesis: laying the foundations for C4 rice.Crossref | GoogleScholarGoogle Scholar |
Furbank RT, Walker DA (1986) Chlorophyll a fluorescence as a quantitative probe of photosynthesis: effects of CO2 concentration during gas transients on chlorophyll fluorescence in spinach leaves. New Phytologist 104, 207–213.
| Chlorophyll a fluorescence as a quantitative probe of photosynthesis: effects of CO2 concentration during gas transients on chlorophyll fluorescence in spinach leaves.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL28Xmt1ejs78%3D&md5=134a233ba2d1836c43016cf6d725e0d1CAS |
Furbank RT, Chitty JA, von Caemmer S, Jenkins CLD (1996) Antisense RNA inhibition of RbcS gene expression reduced rubsico level and photosynthesis in the C4 plant Flaveria bidentis. 111, 725–734.
Furbank RT, von Caemmerer S, Sheehy J, Edwards G (2009) C4 rice: a challenge for plant phenomics. Functional Plant Biology 36, 845–856.
| C4 rice: a challenge for plant phenomics.Crossref | GoogleScholarGoogle Scholar |
Genty B, Briantais J-M, Baker NR (1989) The relationship between the quantum yield of photosynthetic electron transport and quenching of chlorophyll fluorescence. Biochimica et Biophysica Acta. G, General Subjects 990, 87–92.
| The relationship between the quantum yield of photosynthetic electron transport and quenching of chlorophyll fluorescence.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL1MXhsFWntL4%3D&md5=9d9e270bbaccee4721105a2ded548bc0CAS |
Hanson DT, Franklin LA, Samuelsson G, Badger MR (2003) The Chlamydomonas reinhardtii cia3 mutant lacking a thylakoid lumen-localized carbonic anhydrase is limited by CO2 supply to Rubisco and not photosystem II function in vivo. Plant Physiology 132, 2267–2275.
| The Chlamydomonas reinhardtii cia3 mutant lacking a thylakoid lumen-localized carbonic anhydrase is limited by CO2 supply to Rubisco and not photosystem II function in vivo.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXmsVanurc%3D&md5=1838477e7aaef48c0364caffb8b97c0fCAS |
Hibberd JM, Sheehy JE, Langdale JA (2008) Using C4 photosynthesis to increase the yield of rice-rationale and feasibility. Current Opinion in Plant Biology 11, 228–231.
| Using C4 photosynthesis to increase the yield of rice-rationale and feasibility.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXktFCmsLs%3D&md5=210e2ca773dc84880662e3c4c51ef59dCAS |
Horton P, Ruban A (2005) Molecular design of the photosystem II light-harvesting antenna: photosynthesis and photoprotection. Journal of Experimental Botany 56, 365–373.
| Molecular design of the photosystem II light-harvesting antenna: photosynthesis and photoprotection.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXovVymtg%3D%3D&md5=0eda3e64c398f3d81e4acdb929634774CAS |
Ishida S, Uebayashi N, Tazoe Y, Ikeuci M, Homma K, Sato F, Endo T (2014) Diurnal and developmental changes in energy allocation of absorbed light and PSII in field-grown rice. Plant & Cell Physiology 55, 171–182.
| Diurnal and developmental changes in energy allocation of absorbed light and PSII in field-grown rice.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXlvFKhtQ%3D%3D&md5=2bd9d41470487472f7ef4be5deaff292CAS |
Kajala K, Covshoff S, Karki S, Woodfield H, Trolley BJ, Dionora MJA, Mogul RT, Mabilangan AE, Danila FR, Hibberd JM, Quick WP (2011) Strategies for engineering a two-cell C4 photosynthetic pathway into rice. Journal of Experimental Botany 62, 3001–3010.
| Strategies for engineering a two-cell C4 photosynthetic pathway into rice.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXnsFWjtLo%3D&md5=7d5b61b4809739678fe8e07257196f2eCAS |
Laing WA, Ogren WL, Hageman RH (1974) Regulation of soybean net photosynthetic CO2 fixation by the interaction of CO2, O2 and ribulose-1,5 diphosphate carboxylase. Plant Physiology 54, 678–685.
| Regulation of soybean net photosynthetic CO2 fixation by the interaction of CO2, O2 and ribulose-1,5 diphosphate carboxylase.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaE2MXktVygtg%3D%3D&md5=7350fc0ce90fee2c56582590c15d8ea4CAS |
Laisk A, Edwards GE (1998) Oxygen and electron flow in C4 photosynthesis: Mehler reaction, photorespiration and CO2 concentration in the bundle sheath. Planta 205, 632–645.
| Oxygen and electron flow in C4 photosynthesis: Mehler reaction, photorespiration and CO2 concentration in the bundle sheath.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1cXksFGmsL4%3D&md5=a733897791089ba535c456e7059e56f7CAS |
Long SP, Humphries S, Falkowski PG (1994) Photoinhibition of photosynthesis in nature. Annual Review of Plant Physiology and Plant Molecular Biology 45, 633–662.
| Photoinhibition of photosynthesis in nature.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2cXlt12rsLg%3D&md5=4143401a648eeb3c6f1ef67cdf6e4503CAS |
Ludwig M, von Caemmerer S, Price GD, Badger MR, Furbank RT (1998) Expression of tobacco carbonic anhydrase in the C4 dicot Flaveria bidentis leads to increased leakiness of the bundle sheath ad a defective CO2-concentrating mechanism. Plant Physiology 117, 1071–1081.
| Expression of tobacco carbonic anhydrase in the C4 dicot Flaveria bidentis leads to increased leakiness of the bundle sheath ad a defective CO2-concentrating mechanism.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1cXkvVyqtrw%3D&md5=2a33758779938f0a66ca74556fd628f6CAS |
Meurer J, Meierhoff K, Westhoff P (1996) Isolation of high-chlorophyll- fluorescence mutants of Arabidopsis thaliana and their characterization by spectroscopy, immunoblotting and northern hybridisation. Planta 198, 385–396.
| Isolation of high-chlorophyll- fluorescence mutants of Arabidopsis thaliana and their characterization by spectroscopy, immunoblotting and northern hybridisation.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK28XkvVeqtg%3D%3D&md5=4d300db244207ee2a07380c4dfbed550CAS |
Niyogi KK, Bjorkman O, Grossman AR (1997) Chlamydomonas xanthophyll cycle mutants identified by video imaging of chlorophyll fluorescence quenching. The Plant Cell 9, 1369–1380.
| Chlamydomonas xanthophyll cycle mutants identified by video imaging of chlorophyll fluorescence quenching.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2sXlvVKgsLo%3D&md5=7ccd3979b28923dc5560c94f391bb560CAS |
Peisker M (1974) A model describing the influence of oxygen on photosynthetic carboxylation. Photosynthetica 8, 47–50.
Quick WP, Stitt M (1989) An examination of factors contributing to non-photochemical quenching of chlorophyll fluorescence in barley leaves. Biochimica et Biophysica Acta 977, 287–296.
| An examination of factors contributing to non-photochemical quenching of chlorophyll fluorescence in barley leaves.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK3cXnsVSlug%3D%3D&md5=cd25f5172fbbbc9d27c1e98da5a72a23CAS |
Rizal G, Thakur V, Dionora J, Karki S, Wanchana S, Acebron K, Larazo N, Garcia R, Mabilangan A, Montecillo F, Danila F, Mogul R, Pablico P, Leung H, Langdale JA, Sheehy J, Kelly S, Quick WP (2015) Two forward genetic screen for vein density mutants in sorghum converge on a cytochrome P450 gene in the brassinosteroid pathway. The Plant Journal 84, 257–266.
| Two forward genetic screen for vein density mutants in sorghum converge on a cytochrome P450 gene in the brassinosteroid pathway.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2MXhs1Wqu73J&md5=9aee8d1b802c7f01f02813cca584d014CAS |
Rizal G, Karki S, Thakur V, Wanachana S, Alonso-Cantabrana H, Dionora J, Sheehy JE, Furbank R, von Caemmerer S, Quick WP (2017) A sorghum (Sorghum bicolor) mutant with altered carbon isotope ratio. PLoS One 12, e0179567
| A sorghum (Sorghum bicolor) mutant with altered carbon isotope ratio.Crossref | GoogleScholarGoogle Scholar |
Shikanai T, Munekage Y, Shimizu K, Endo T, Hashimoto T (1999) Identification and characterization of Arabidopsis mutants with reduced quenching of chlorophyll fluorescence. Plant & Cell Physiology 40, 1134–1142.
| Identification and characterization of Arabidopsis mutants with reduced quenching of chlorophyll fluorescence.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXns1eitrs%3D&md5=dd75571ab5f7940b1c42b81622a3c3c6CAS |
Somerville CR (1986) Analysis of photosynthesis with mutants of higher plants and algae. Annual Review of Plant Physiology and Plant Molecular Biology 37, 467–507.
| Analysis of photosynthesis with mutants of higher plants and algae.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL28XksFOrtL8%3D&md5=a3858d7ee97b47d0b4f0ea02801aa120CAS |
Somerville CR (2001) An early Arabidopsis demonstration. Resolving a few issues concerning photorespiration. Plant Physiology 125, 20–24.
| An early Arabidopsis demonstration. Resolving a few issues concerning photorespiration.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXjslymt78%3D&md5=2af897a68892465433c49941979c7975CAS |
Vogan PJ, Frohlich MW, Sage RF (2007) The functional significance of C3-C4 intermediate traits in Heliotropium L. (Boraginaceae): gas exchange perspectives. Plant, Cell & Environment 30, 1337–1345.
| The functional significance of C3-C4 intermediate traits in Heliotropium L. (Boraginaceae): gas exchange perspectives.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXhtFCgu7jN&md5=e949f551e9bd42f0f0cc974a6eb9089aCAS |
von Caemmerer S (2000) ‘Biochemical models of leaf photosynthesis. Vol. 2.’ (CSIRO Publishing: Melbourne)
von Caemmerer S, Furbank RT (2003) The C4 pathway: an efficient CO2 pump. Photosynthesis Research 77, 191–207.
| The C4 pathway: an efficient CO2 pump.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXnsF2mtLw%3D&md5=5b08cbf56af85398d4501659db8a490fCAS |
von Caemmerer S, Millgate A, Farquhar GD, Furbank RT (1997) Reduction of ribulose-1,5-bisphosphate carboxylase/oxygenase by antisense RNA in the C4 plant Flaveria bidentis leads to reduced assimilation rates and increased carbon isotope discrimination. Plant Physiology 113, 469–477.
| Reduction of ribulose-1,5-bisphosphate carboxylase/oxygenase by antisense RNA in the C4 plant Flaveria bidentis leads to reduced assimilation rates and increased carbon isotope discrimination.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2sXht1Cktrg%3D&md5=54ad9e337f869c1c2a1f1ec23eb98becCAS |
Wang Q, Zhang Q, Fan D, Lu C (2006) Photosynthetic light and CO2 utilization and C4 traits of two novel super-rice hybrids. Journal of Plant Physiology 163, 529–537.
| Photosynthetic light and CO2 utilization and C4 traits of two novel super-rice hybrids.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XjtVeksrw%3D&md5=df4a5ced7e89598da9c171ba1bd9747eCAS |
