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Brassinosteroids promote photosynthesis and growth by enhancing activation of Rubisco and expression of photosynthetic genes in Cucumis sativus

Abstract

Brassinosteroids (BRs) are a new group of plant growth substances that promote plant growth and productivity. We showed in this study that improved growth of cucumber (Cucumis sativus) plants after treatment with 24-epibrassinolide (EBR), an active BR, was associated with increased CO2 assimilation and quantum yield of PSII (ΦPSII). Treatment of brassinazole (Brz), a specific inhibitor for BR biosynthesis, reduced plant growth and at the same time decreased CO2 assimilation and ΦPSII. Thus, the growth-promoting activity of BRs can be, at least partly, attributed to enhanced plant photosynthesis. To understand how BRs enhance photosynthesis, we have analyzed the effects of EBR and Brz on a number of photosynthetic parameters and their affecting factors, including the contents and activity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco). Northern and Western blotting demonstrated that EBR upregulated, while Brz downregulated, the expressions of rbcL, rbcS and other photosynthetic genes. In addition, EBR had a positive effect on the activation of Rubisco based on increased maximum Rubisco carboxylation rates (V c,max), total Rubisco activity and, to a greater extent, initial Rubisco activity. The accumulation patterns of Rubisco activase (RCA) based on immunogold-labeling experiments suggested a role of RCA in BR-regulated activation state of Rubisco. Enhanced expression of genes encoding other Calvin cycle genes after EBR treatment may also play a positive role in RuBP regeneration (J max), thereby increasing maximum carboxylation rate of Rubisco (V c,max). Thus, BRs promote photosynthesis and growth by positively regulating synthesis and activation of a variety of photosynthetic enzymes including Rubisco in cucumber.

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Abbreviations

A sat :

Light-saturated rate of CO2 assimilation

BRs:

Brassinosteroids

Brz:

Brassinazole

Chl:

Chlorophyll

C i :

Intercellular CO2 concentration

EBR:

24-Epibrassinolide

FBP:

Fructose-1,6-bisphosphate

FBPA:

Fructose-1,6-bisphosphate aldolase

FBPase:

Fructose-1,6-bisphosphatase

Fru6P:

Fructose 6-phosphate

Fv/Fm:

Maximum quantum yield of PSII

Fv′/Fm′:

Efficiency of energy capture by open PSII

Gs:

Stomatal conductance

J max :

Maximum RuBP regeneration rates

PRK:

Ribulose-5-phosphate kinase

qP:

Photochemical quenching coefficient

rbcL:

Rubisco large subunit gene

rbcS:

Rubisco small subunit gene

rca :

Rubisco activase gene

Rubisco:

Ribulose-1,5-bisphosphate carboxylase/oxygenase

RuBP:

Ribulose-1,5-bisphosphate

RuPE:

Ribulose phosphate epimerase

SBP:

Sedoheptulose-1,7-bisphosphate

SBPase:

Sedoheptulose-1,7-bisphosphatase

Sed7P:

Sedoheptulose 7-phosphate

TPI:

Triose-3-phosphate isomerase

V c,max :

Maximum Rubisco carboxylation rates

ΦPSII :

Quantum yield of PSII

References

  1. Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399

  2. Arnon DI (1949) Copper enzymes in isolated chloroplasts: polyphenol oxidase in Beta vulgaris. Plant Physiol 24:1–15

  3. Asami T, Min YK, Nagata N, Yamagishi K, Takatsuto S, Fujioka S, Murofushi N, Yamaguchi I, Yoshida S (2000) Characterization of brassinazole, a triazole-type brassinosteroid biosynthesis inhibitor. Plant Physiol 123:93–99

  4. Asami T, Mizutani M, Fujioka S, Goda H, Min YK, Shimada Y, Nakano T, Takatsuto S, Matsuyama T, Nagata N, Sakata K, Yoshida S (2001) Selective interaction of triazole derivatives with DWF4, a cytochrome P450 monooxygenase of the brassinosteroid biosynthesis pathway, correlates with brassinosteroid deficiency in planta. J Biol Chem 276:25687–25691

  5. Bajguz A, Asami T (2004) Effects of brassinazole, an inhibitor of brassinosteroid biosynthesis, on light- and dark-grown Chlorella vulgaris. Planta 218:869–877

  6. Bancos S, Nomura T, Sato T, Molnar G, Bishop GJ, Koncz C, Yokota T, Nagy F, Szekeres M (2002) Regulation of transcript levels of the Arabidopsis cytochrome P450 genes involved in brassinosteroid biosynthesis. Plant Physiol 130:504–513

  7. Bishop GJ (2007) Refining the plant steroid hormone biosynthesis pathway. Trends Plant Sci 12:377–380

  8. Bishop GJ, Koncz C (2002) Brassinosteroids and plant steroid hormone signaling. Plant Cell 14(Suppl.):S97–S110

  9. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254

  10. Buysse J, Merckx R (1993) An improved colorimetric method to quantify sugar content of plant tissue. J Exp Bot 44:1627–1629

  11. Catterou M, Dubois F, Schaller H, Aubanelle L, Vilcot B, Sangwan-Norreel BS, Sangwan RS (2001) Brassinosteroids, microtubules and cell elongation in Arabidopsis thaliana. I. Molecular, cellular and physiological characterization of the Arabidopsis bul1 mutant, defective in the 7-sterol-C5-desaturation step leading to brassinosteroid biosynthesis. Planta 212:659–672

  12. Choe SW, Dilkes BP, Fujioka S, Takatsuto S, Sakurai A, Feldmann KA (1998) The DWF4 gene of Arabidopsis encodes a cytochrome P450 that mediates multiple 22 α-hydroxylation steps in brassinosteroid biosynthesis. Plant Cell 10:231–243

  13. Choe S, Tanaka A, Noguchi T, Fujioka S, Takatsuto S, Ross AS, Tax FE, Yoshida S, Feldmann KA (2000) Lesions in the sterol Δ7 reductase gene of Arabidopsis cause dwarfism due to a block in brassinosteroid biosynthesis. Plant J 21:431–443

  14. Choe S, Fujioka S, Noguchi T, Takatsuto S, Yoshida S, Feldmann KA (2001) Overexpression of DWARF4 in the brassinosteroid biosynthetic pathway results in increased vegetative growth and seed yield in Arabidopsis. Plant J 26:573–582

  15. Clouse SD, Sasse JM (1998) Brassinosteroids: essential regulators of plant growth and development. Annu Rev Plant Physiol Plant Mol Biol 49:427–451

  16. Dhaubhadel S, Browning KS, Gallie DR, Krishna P (2002) Brassinosteroid functions to protect the translational machinery and heat-shock protein synthesis following thermal stress. Plant J 29:681–691

  17. Ethier GJ, Livingston NJ (2004) On the need to incorporate sensitivity to CO2 transfer conductance into the Farquhar-von Caemmerer-Berry leaf photosynthesis model. Plant Cell Environ 27:137–153

  18. Genty B, Briantais JM, Baker NR (1989) The relationship between the quantum yield of photosynthetic electron transport and quenching of chlorophyll fluorescence. Biochim Biophys Acta 990:87–92

  19. Grove MD, Spencer GF, Rohwedder WK, Mandava N, Worley JF, Warthen JD Jr, Steffens GL, Flippen-Anderson JL, Cook JC Jr (1979) Brassinolide, a plant growth-promoting steroid isolated from Brassica napus pollen. Nature 281:216–217

  20. Hammond E, Andrews T, Mott K, Woodrow I (1998) Regulation of Rubisco activation in antisense plants of tobacco containing reduced levels of Rubisco activase. Plant J 14:101–110

  21. Harrison EP, Olcer H, Lloyd JC, Long SP, Raines CA (2001) Small decreases in SBPase cause a linear decline in the apparent RuBP regeneration rate, but do not affect Rubisco carboxylation capacity. J Exp Bot 52:1779–1784

  22. Hayat S, Ahmad A, Mobin M, Hussain A, Fariduddin Q (2000) Photosynthetic rate, growth, and yield of mustard plants sprayed with 28-homobrassinolide. Photosynthetica 38:469–471

  23. Horton P, Ruban AV, Walters RG (1996) Regulation of light harvesting in green plants. Annu Rev Plant Physiol Plant Mol Biol 47:655–684

  24. Hu YX, Bao F, Li JY (2000) Promotive effect of brassinosteroids on cell division involves a distinct CycD3-induction pathway in Arabidopsis. Plant J 24:693–701

  25. Irving LJ, Robinson D (2006) A dynamic model of Rubisco turnover in cereal leaves. New Phytol 169:493–504

  26. Jin SH, Hong J, Li XQ, Jiang DA (2006) Antisense inhibition of Rubisco activase increases Rubisco contents and alters the proportion of Rubisco activase in strom and thylakiods in chloroplast of rice leaves. Ann Bot 97:736–744

  27. Khripach V, Zhabinskii V, De Groot A (2000) Twenty years of brassinosteroids: steroidal plant hormones warrant better crops for the XXI century. Ann Bot 86:441–447

  28. Koßmann J, Sonnewald U, Willmitzer L (1994) Reduction of the chloroplastic fructose-1, 6-bisphosphatase in transgenic potato plants impairs photosynthesis and plant growth. Plant J 6:637–650

  29. Krishna P (2003) Brassinosteroid-mediated stress responses. J Plant Growth Regul 22:289–297

  30. Laemmli UK (1970) Cleavage of structural proteins during the assembly of head of bacteriophage T4. Nature 227:680–685

  31. Li JM, Nagpal P, Vitart V, McMorris TC, Chory J (1996) A role for brassinosteroids in light-dependent development of Arabidopsis. Science 272:398–401

  32. Lilley RM, Walker DA (1974) An improved spectrophotometric assay for ribulose-bisphosphate carboxylase. Biochim Biophys Acta 358:226–229

  33. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 25:402–408

  34. Long SP, Zhu X, Naidu SL, Ort DR (2006) Can improvement in photosynthesis increase crop yields? Plant Cell Environ 29:315–330

  35. Mate CJ, von Caemmerer S, Evans JR, Hudson GS, Andrews TJ (1996) The relationship between CO2-assimilation rate, Rubisco carbamylation and Rubisco activase content in activase-deficient transgenic tobacco suggests a simple model of activase action. Planta 198:604–613

  36. Müssig C, Fischer S, Altmann T (2002) Brassinosteroid-regulated gene expression. Plant Physiol 129:1241–1251

  37. Nakashita H, Yasuda M, Nitta T, Asami T, Fujioka S, Arai Y, Sekimata K, Takatsuto S, Yamaguchi I, Yoshida S (2003) Brassinosteroid functions in a broad range of disease resistance in tobacco and rice. Plant J 33:887–898

  38. Neill S, Desikan R, Hancock J (2002) Hydrogen peroxide signalling. Curr Opin Plant Biol 5:388–395

  39. Noguchi T, Fujioka S, Takatsuto S, Sakurai A, Yoshida S, Li JM, Chory J (1999) Arabidopsis det2 is defective in the conversion of (24R)-24-methylcholest-4-en--3-one to (24R)-24-methyl-5α-cholestan-3-one in brassinosteroid biosynthesis. Plant Physiol 120:833–839

  40. Oswald O, Martin T, Dominy PJ, Graham IA (2001) Plastid redox state and sugars: interactive regulators of nuclear-encoded photosynthetic gene expression. Proc Natl Acad Sci USA 98:2047–2052

  41. Paul M, Pellny T (2003) Carbon metabolite feedback regulation of leaf photosynthesis and development. J Exp Bot 54:539–547

  42. Pfannschmidt T, Allen JF, Oelmüller R (2001) Principles of redox control in photosynthesis gene expression. Physiol Plant 112:1–9

  43. Portis AR, Li C, Wang D, Salvucci ME (2008) Regulation of Rubisco activase and its interaction with Rubisco. J Exp Bot 59:1597–1604

  44. Salvucci ME, Portis AR Jr, Ogren WL (1985) A soluble chloroplast protein catalyzes ribulosebisphosphate carboxylase/oxygenase activation in vivo. Photosynth Res 7:193–201

  45. Salvucci ME, Portis AR Jr, Ogren WL (1986) Light and CO2 response of ribulose-1, 5-bisphosphate carboxylase/oxygenase activation in Arabidopsis leaves. Plant Physiol 80:655–659

  46. Sasse JM (1997) Recent progress in brassinosteroid research. Physiol Plant 100:696–701

  47. Schlüter U, Köpke D, Altmann T, Müssig C (2002) Analysis of carbohydrate metabolism of CPD antisense plants and the brassinosteroids-deficient cbb1 mutant. Plant Cell Environ 25:783–791

  48. Sharkey TD, Savitch LV, Butz ND (1991) Photometric method for routine determination of kcat and carbamylation of Rubisco. Photosynth Res 28:41–48

  49. Strand A, Zrenner R, Trevanion S, Stitt M, Gustafsson P, Gardeström P (2000) Decreased expression of two key enzymes in the sucrose biosynthesis pathway, cytosolic fructose-1, 6-bisphosphatase and sucrose phosphate synthase, has remarkably different consequences for photosynthetic carbon metabolism in transgenic Arabidopsis thaliana. Plant J 23:759–770

  50. Streusand VJ, Portis AR (1987) Rubisco activase mediates ATP-dependent activation of ribulose bisphosphate carboxylase. Plant Physiol 85:152–154

  51. Suzuki Y, Makino A, Mae T (2001) Changes in the turnover of Rubisco and levels of mRNAs of rbcL and rbcS in rice leaves from emergence to senescence. Plant Cell Environ 24:1353–1360

  52. Szekeres M, Németh K, Koncz-Kálmán Z, Mathur J, Kauschmann A, Altmann T, Rédei GP, Nagy F, Schell J, Koncz C (1996) Brassinosteroids rescue the deficiency of CYP90, a cytochrome P450, controlling cell elongation and de-etiolation in Arabidopsis. Cell 85:171–182

  53. Tanaka K, Asami T, Yoshida S, Nakamura Y, Matsuo T, Okamoto S (2005) Brassinosteroid homeostasis in Arabidopsis is ensured by feedback expressions of multiple genes involved in its metabolism. Plant Physiol 138:1117–1125

  54. van Kooten O, Snel JFH (1990) The use of chlorophyll fluorescence nomenclature in plant stress physiology. Photosynth Res 25:147–150

  55. von Caemmerer S, Farquhar GD (1981) Some relationships between the biochemistry of photosynthesis and the gas exchange of leaves. Planta 153:367–387

  56. Wu CY, Trieu A, Radhakrishnan P, Kwok SF, Harris S, Zhang K, Wang JL, Wan JM, Zhai HQ, Takatsuto S, Matsumoto S, Fujioka S, Feldmann KA, Pennell RI (2008) Brassinosteroids regulate grain filling in rice. Plant Cell 20:2130–2145

  57. Wullschleger SD (1993) Biochemical limitations to carbon assimilation in C3 plants: a retrospective analysis of the A/Ci curves from 109 species. J Exp Bot 44:907–920

  58. Xia XJ, Wang YJ, Zhou YH, Tao Y, Mao WH, Shi K, Asami T, Chen Z, Yu JQ (2009) Reactive oxygen species are involved in brassinosteroids-induced stress tolerance in Cucumis sativus. Plant Physiol 150:801–814

  59. Yokota T (1997) The structure, biosynthesis and function of brassinosteroids. Trends Plant Sci 2:137–143

  60. Yu JQ, Huang LF, Hu WH, Zhou YH, Mao WH, Ye SF, Nogués S (2004) A role for brassinosteroids in the regulation of photosynthesis in Cucumis sativus. J Exp Bot 55:1135–1143

  61. Zhang N, Kallis RP, Ewy RG, Portis AR (2002) Light modulation of Rubisco in Arabidopsis requires a capacity for redox regulation of the larger Rubisco activase isoform. Proc Natl Acad Sci USA 99:3330–3334

  62. Zhou YH, Yu JQ, Huang LF, Nogués S (2004) The relationship between CO2 assimilation, photosynthetic electron transport and water–water cycle in chill-exposed cucumber leaves under low light and subsequent recovery. Plant Cell Environ 27:1503–1514

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Acknowledgments

This work was supported by the National Basic Research Program of China (2009CB119000), National Natural Science Foundation of China (3050344; 30671428) and the Program for Promotion of Basic Research Activities for Innovative Bioscience (PROBRAIN). We thank Dr J Hong for help with the immunogold-labeling experiment.

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Correspondence to Jing-Quan Yu.

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Xia, X., Huang, L., Zhou, Y. et al. Brassinosteroids promote photosynthesis and growth by enhancing activation of Rubisco and expression of photosynthetic genes in Cucumis sativus . Planta 230, 1185 (2009). https://doi.org/10.1007/s00425-009-1016-1

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Keywords

  • Cucumber (Cucumis sativus)
  • Immunogold labeling
  • Productivity
  • Rubisco activase
  • Sugar metabolism