Planta

, Volume 158, Issue 6, pp 487–500 | Cite as

Phytochrome control of RNA levels in developing pea and mung-bean leaves

  • William F. Thompson
  • Marylee Everett
  • Neil O. Polans
  • Richard A. Jorgensen
  • Jeffrey D. Palmer
Article
Received:
Accepted:

Abstract

We have examined phytochrome effects on the abundance of transcripts from several nuclear and chloroplast genes in buds of dark-grown pea seedlings and primary leaves of dark-grown mung-bean seedlings. Probes for nuclear-coded RNAs were selected from a library of cDNA clones and included those corresponding to the small subunit (SS) of ribulosebisphosphate carboxylase and a chlorophyll a/b binding protein (AB). Transcripts from chloroplast genes for RuBP carboxylase large subunit (LS) and a 32,000-dalton photosystem II polypeptide (PII) were assayed with cloned fragments of the chloroplast genome. In addition, we present data on transcripts from a number of other nuclear genes of unknown function, several of which change in abundance during light-induced development. Transcript levels were measured as a proportion of total RNA by a dot blot assay in which RNA from different tissues or stages is fixed to nitrocellulose and hybridized with 32P-labeled probes prepared from cloned DNAs. Several patterns of induction can be seen. For example, although both SS and AB RNAs show positive, red/far-red reversible responses in both pea and mung bean, in pea buds the induction ratio for SS RNA is much higher than that for AB RNA, while just the reverse is true for mung-bean leaves. In addition, treatment with lowfluence red light produces full induction of the pea AB RNA, while SS RNA in the same tissue does not reach a maximum steady-state level until after about 24 h of supplementary high-intensity white light. In pea buds, chloroplast genes (LS, PII) also show clear responses to phytochrome, as measured by the steady-state levels of their RNA products. Chloroplast DNA levels (as a fraction of the total cellular DNA) show the same response pattern, which may indicate that in peas many of the light effects we see are related to a general stimulation of chloroplast development. In mung beans, the levels of plastid DNA and RNA are already quite high in the leaves of 7-d dark-grown seedlings, and light effects are much less pronounced. The results are consistent with the notion that chloroplast development is arrested at a later stage in dark-grown mung-bean leaves than in etiolated pea buds.

Key words

Chlorophyll a/b-binding protein Chloroplast DNA Phytochrome (RNA levels) RNA levels, light Pisum (light and RNA) Ribulosebisphosphate carboxylase Vigna 

Abbreviations

AB

chlorophyll a/b polypeptide of the light-harvesting complex

FR

far-red light

PII

32,000-dalton photosystem II polypeptide

R

red light

SS

small subunit and

LS

large subunit of ribulosebisphospate carboxylase (RuP2; 3-phospho-D-glycerate carboxylase [dimerizing], EC 4.1.1.39)

This is a preview of subscription content, log in to check access.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Apel, K. (1979) Phytochrome-induced appearance of mRNA activity for the apoprotein of the light-harvesting chlorophyll a/b protein of barley (Hordeum vulgare). Eur. J. Biochem. 97, 183–188Google Scholar
  2. Apel, K. (1981) The protochlorophyllide holochrome of barley (Hordeum vulgare L.): phytochrome-induced decrease of translatable mRNA coding for the NADPH: protochlorophyllide oxidoreductase. Eur. J. Biochem. 120, 89–93Google Scholar
  3. Arnon, D. (1949) Copper enzymes in isolated chloroplasts. Plant Physiol. 24, 1–15Google Scholar
  4. Bedbrook, J.R., Link, G. Coen, D.M., Bogorad, L., Rich, A. (1978) Maize plastid gene expressed during photoregulated development. Proc. Natl. Acad. Sci. USA 75, 3060–3064Google Scholar
  5. Bedbrook, J., Smith, S., Ellis, R. (1980) Molecular cloning and sequencing of cDNA encoding the precursor to the small subunit of chloroplast ribulose 1,5-bisphosphate carboxylase. Nature (London) 287, 692–697Google Scholar
  6. Biessmann, H., Craig, E.A., McCarthy, B.S. (1979) Rapid quantitation of individual RNA species in a complex population. Nucleic Acids Res. 7, 981–996Google Scholar
  7. Birnboim, H.D., Doly, J. (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 7, 1513–1523Google Scholar
  8. Bolivar, F., Rodriguez, R.L., Greene, T.J., Betlach, M.C., Heyneker, H.L., Boyer, H.W. (1977) Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2, 95–113Google Scholar
  9. Broglie, R., Bellemare, G., Bartlett, S.G., Chua, N.-H., Cashmore, A.R. (1981) Cloned DNA sequences complementary to mRNAs encoding precursors to the small subunit of ribulose-1,5-bisphosphate carboxylase and a chlorophyll a/b binding polypeptide. Proc. Natl. Acad. Sci. USA 78, 7304–7308Google Scholar
  10. Cuellar, R.E. (1982) The structure and evolution of repeated DNA sequences in Pisum sativum: a molecular characterization. Ph.D. thesis, Stanford University. Stanford, Cal., USAGoogle Scholar
  11. Dagert, M., Ehrlich, S.D. (1979) Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene 6, 23–28Google Scholar
  12. Dyer, T.A., Miller, R.H., Greenwood, A.D. (1971) Leaf nucleic acids characteristics and role in the differentiation of plastids. J. Exp. Bot. 22, 125–136Google Scholar
  13. Eaglesham, A.R.J., Ellis, R.J. (1974) Protein synthesis in chloroplast. II. Light-driven synthesis of membrane proteins by isolated pea chloroplasts. Biochim. Biophys. Acta 335, 396–407Google Scholar
  14. Edelman, M. (1981) Nucleic acids of chloroplasts and mitochondria. In: The biochemistry of plants, vol. 6, pp. 249–301, Marcus, A., ed. Academic Press, New York LondonGoogle Scholar
  15. Ellis, J.R. (1981) Chloroplst proteins: synthesis, transport, and assembly. Annu. Rev. Plant Physiol. 32, 111–137Google Scholar
  16. Everett, M., Jorgensen, R.A., Thompson, W.F. (1981) Phytochrome control of transcript abundance in developing pea leaves. Carnegie Inst. Washington Yearb. 80, 79–80Google Scholar
  17. Gallagher, T.F., Ellis, R.J. (1982) Light-stimulated transcription of genes for two chloroplast polypeptides in isolated pea leaf nuclei. EMBO J. 1, 1493–1498Google Scholar
  18. Geiser, M., Doring, H., Wostemeyer, J., Behrens, V., Tillman, E., Starlinger, P. (1980) A cDNA clone from Zea mays endosperm sucrose synthetase mRNA. Nucleic Acids Res. 8, 6175–6188Google Scholar
  19. Glisin, V., Crkvenjakov, R., Byus, C. (1974) Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry 13, 2633–2637Google Scholar
  20. Gorton, H.L., Briggs, W.R. (1980) Phytochrome responses to end-of-day irradiation in light-grown corn grown in the presence and absence of Sandoz 9789. Plant Physiol. 66, 1024–1026Google Scholar
  21. Grunstein, M.G., Hogness, D.S. (1975) Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc. Natl. Acad. Sci. USA 72, 3961–3965Google Scholar
  22. Hallick, R.B., Chelm, B.K., Gray, P.W., Orozco, E.M. Jr. (1977) Use of aurintricarboxylic acid as an inhibitor of nucleases during nucleic acid isolation. Nucleic Acids Res. 4, 3055–3064Google Scholar
  23. Heinze, H., Herzfeld, F., Kiper, M. (1980) Light-induced appearance of polysomal poly A-rich messenger RNA during greening of barley plants. Eur. J. Biochem. 111, 137–144Google Scholar
  24. Henshall, J.D., Goodwin, T.W. (1964) The effect of red and far red light on carotenoid and chlorophyll formation in pea seedlings. Photochem. Photobiol. 3, 243–247Google Scholar
  25. Hong, Y.-N., Schopfer, P. (1981) Control by phytochrome of urate oxidase and allantoinase activities during peroxisome development in the cotyledons of mustard (Sinapis alba L.) seedlings. Planta 152, 325–335Google Scholar
  26. Jorgensen, R.A., Thompson, W.F. (1980) Observing patterns of gene expression at the RNA level: the greening response in Pisum. Carnegie Inst. Washington Yearb. 79, 116–119Google Scholar
  27. Kafatos, F.C., Jones, C., Efstratiadis, A. (1979) Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure. Nucleic Acids Res. 7, 1541–1552Google Scholar
  28. Kirk, J.T.O., Tilney-Bassett, R.A.E. (1978) The plastids: their chemistry, structure, growth, and inheritance, 2nd edn. Elsevier, AmsterdamGoogle Scholar
  29. Klein, S., Schiff, J.A., Holowinsky, A.W. (1972) Events surrounding the early development of Euglena chloroplasts. II. Normal development of fine structure and the consequences of preillumination. Dev. Biol. 28, 253–273Google Scholar
  30. Link, G. (1982) Phytochrome control of plastid mRNA in mustard (Sinapis alba L.). Planta 154, 81–86Google Scholar
  31. Mandoli, D.F., Briggs, W.R. (1981) Phytochrome control of two low-irradiance responses in etiolated oat seedlings. Plant Physiol. 67, 733–739Google Scholar
  32. Maniatis, T., Jeffrey, A., Kleid, D.G. (1975) Nucleotide sequence of the rightward operator of phage λ. Proc. Natl. Acad. Sci. USA 72, 1184–1188Google Scholar
  33. McIntosh, L., Poulsen, C., Bogorad, L. (1980) Chloroplast gene sequence for the large subunit of ribulose bisphosphate carboxylase of maize. Nature (London) 288, 556–560Google Scholar
  34. Murray, M.G., Peters, D.L., Thompson, W.F. (1981) Ancient repeated sequences in the pea and mung bean genomes and implications for genome evolution. J. Mol. Evol. 17, 31–42Google Scholar
  35. Myers, J.C., Spiegelman, S. (1978) Sodium pyrophosphate inhibition of RNA·DNA hybrid degradation by reverse transcriptase. Proc. Natl. Acad. Sci. USA 75, 5329–5333Google Scholar
  36. Palmer, J.D., Thompson, W.F. (1981) Clone banks of the mung bean, pea, and spinach chloroplast genomes. Gene 15, 21–26Google Scholar
  37. Palmer, J.D., Thompson, W.F. (1982) Chloroplast DNA rearrangements are more frequent when a large inverted repeat sequence is lost. Cell 29, 537–550Google Scholar
  38. Roychoudhury, R., Jay, E., Wu, R. (1976) Terminal labeling and addition of homopolymer tracts to duplex DNA fragments by terminal deoxynucleotidyl transferase. Nucleic Acid Res. 3, 101–116Google Scholar
  39. Sasaki, Y., Ishiye, M., Sakihama, T., Kamikubo, T. (1981) Light-induced increase of mRNA activity coding for the small subunit of ribulose-1,5-bisphosphate carboxylase. J. Biol. Chem. 256, 2315–2320Google Scholar
  40. Schiff, J.A. (1978) Photocontrol of chloroplast development in Euglena. In: Chloroplast development, pp. 747–767, Akoyunolgov, G., ed. Elsevier/North Holland Biomedical Press, AmsterdamGoogle Scholar
  41. Schiff, J.A. (1980) Development, inheritance, and evolution of plastids and mitochondria. In: The biochemistry of plants, vol. 1, pp. 209–272, N.E. Tolbert, ed. Academic Press, New York LondonGoogle Scholar
  42. Schopfer, P. (1977) Phytochrome control of enzymes. Annu. Rev. Plant Physiol. 28, 223–252Google Scholar
  43. Shinozaki, K., Sasaki, Y., Sakihama, T., Kamikubo, T. (1982) Coordinate light-induction of two mRNAs, encoded in nuclei and chloroplasts, of ribulose 1,5-bisphosphate carboxylase/oxygenase. FEBS Lett. 144, 73–76Google Scholar
  44. Silflow, C.D., Hammett, J.R., Key, J.L. (1979) Sequence complexity of polyadenylated ribonucleic acid from soybean suspension culture cells. Biochemistry 18, 2725–2731Google Scholar
  45. Sims, T., Hague, D. (1981) Light-stimulated increase of translatable mRNA for phosphoenolpyruvate carboxylase in leaves of maize. J. Biol. Chem. 256, 8252–8255Google Scholar
  46. Smith, H. (1975) Phytochrome and photomorphogenesis. McGraw-Hill, MaidenheadGoogle Scholar
  47. Smith, H., Billett, E.E., Giles, A.B. (1977) The photocontrol of gene expression in higher plants. In: Regulation of enzyme synthesis and activity in plants, pp 93–127, Smith, H., ed. Academic Press, New York LondonGoogle Scholar
  48. Smith, S., Ellis, R.J. (1981) Light-stimulated accumulation of transcripts of nuclear anc chloroplast genes for ribosebisphosphate carboxylase. J. Mol. Appl. Genet. 1, 127–137Google Scholar
  49. Sullivan, D., Brisson, M., Verma, D.P. (1980) Reverse transcription of 25S soybean ribosomal RNA in the absence of exogenous primer. Biochim. Biophys. Res. Comm. 94, 144–150Google Scholar
  50. Tobin, E.M. (1978) Light regulation of specific mRNA species in Lemna gibba L. G-3. Proc. Natl. Acad. Sci. USA 75, 4749–4753Google Scholar
  51. Tobin, E.M. (1981a) White light effects on the mRNA for the light-harvesting chlorophyll a/b protein in Lemna gibba L. G-3. Plant Physiol. 67, 1078–1083Google Scholar
  52. Tobin, E.M. (1981b) Phytochrome-mediated regulation of messenger RNAs for the small subunit of ribulose 1,5 bisphosphate carboxylase and the light-harvesting chlorophyll a/b protein in Lemna gibba. Plant Mol. Biol. 1, 35–51Google Scholar
  53. Wahl, G.M., Stern, M., Stark, G.R. (1979) Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc. Natl. Acad. Sci. USA 76, 3683–3687Google Scholar

Copyright information

© Springer-Verlag 1983

Authors and Affiliations

  • William F. Thompson
    • 1
  • Marylee Everett
    • 1
  • Neil O. Polans
    • 1
  • Richard A. Jorgensen
    • 1
  • Jeffrey D. Palmer
    • 1
  1. 1.Department of Plant BiologyCarnegie Institution of WashingtonStanfordUSA

Personalised recommendations