Skip to main content
SpringerLink
Log in

Isolation of intact mesophyll protoplasts from the leaves of higher plants for photosynthetic studies

  • Published:
Proceedings / Indian Academy of Sciences

Abstract

From a survey of 104 herbaceous species from the Madurai Kamaraj University Campus collected for isolating intact mesophyll protoplasts 36 species belonging to C3, C4 and CAM photosynthetic pathways were found to yield between 15–85% of protoplasts. Species which easily yielded intact mesophyll cells by mild mechanical grinding were not amenable for protoplast isolation. The protoplasts of CAM species were the largest and those of C4 were the smallest in size. The isolated mesophyll protoplasts were photosynthetically active in terms of CO2 fixation and O2 evolution. Potassium ferricyanide was ineffective as Hill oxidant while dichlorophenolindophenol induced high Hill activity with the isolated protoplasts. The protoplasts maintained their photosynthetic stability up to 12 hr after isolation.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Anderson J M, Woo K C and Boardman N K 1972 Deficiency of photosystem II in agranal bundle sheath chloroplasts ofSorghum bicolor andZea mays; inProceedings of 2nd International Congress on Photosynthesis eds G Forti, M Avron and A Melandri (The Hague) pp 613–619

  • Aron D I 1949 Copper enzymes in isolated chloroplasts. Polyphenol oxidase inBeta vulgaris;Plant Physiol. 24 1–15

    Article  Google Scholar 

  • Aoki S and Takebe I 1969 Infection of tobacco mesophyll protoplasts by tobacco mosaic virus ribonucleic acid;Virology 39 439–448

    Article  PubMed  CAS  Google Scholar 

  • Bajaj Y P S 1977 Protoplast isolation culture and somatic hybridization; inApplied and fundamental aspects of plant cell tissue and organ culture. eds J Reinert and Y P S Bajaj (Berlin Heidelberg, New York: Springer Verlag) ch 4 pp 467–496

    Google Scholar 

  • Carlson P S 1973 The use of protoplasts for genetic research;Proc. Nat. Acad. Sci. 70 598–602

    Article  PubMed  CAS  Google Scholar 

  • Cocking E C 1972 Plant cell protoplasts—isolation and development;Ann. Rev. Plant Physiol 23 29–50

    Article  CAS  Google Scholar 

  • Davey M R and Cocking E C 1972 Uptake of bacteria by isolated higher plant protoplasts;Nature (London)239 455–456

    Article  Google Scholar 

  • Edwards G E and Black C C 1971 Isolation of mesophyll cells and bundle sheath cells fromDigitaria sanguinalis (L.) Scop. leaves and a scanning microscopy study of the internal leaf cell morphology;Plant Physiol. 47 149–156

    Article  PubMed  Google Scholar 

  • Evans P K, Keates A G and Cocking E C 1972 Isolation of protoplasts from cereal leaves;Planta 104 178–181

    Article  CAS  Google Scholar 

  • Giles K L 1974 Complementation by protoplast fusion using mutant strain of maize;Plant Cell Physiol. 15 281–285

    CAS  Google Scholar 

  • Gnanam A and Kulandaivelu G 1969 Photosynthetic studies with leaf cell suspension from higher plants;Plant Physiol. 44 1451–1456

    Article  PubMed  CAS  Google Scholar 

  • Gutierrez M, Kanai R, Huber S C, Ku S B and Edwards G E 1974 Photosynthesis in mesophyll protoplasts and bundle sheath cells of various types of C4 plants. I. Carboxylases and CO2 fixation studies;Z. Pflanzenphysiol. 72 305–319

    CAS  Google Scholar 

  • Hall M D and Cocking E C 1974 The response of isolatedAvena coleoptile protoplasts to indole 3-acetic acid;Protoplasma 19 225–234

    Article  Google Scholar 

  • Huber S C and Edwards G E 1975 C4 photosynthesis: Light dependent CO2 fixation by mesophyll cells, protoplasts and protoplast extracts ofDigitaria sanguinalis;Plant Physiol. 55 835–844

    Article  PubMed  CAS  Google Scholar 

  • Jensen R G, Franki R I B and Zaitlin M 1971 Metabolism of separated leaf cells. I. Preparation of photosynthetically active cells from tobacco;Plant Physiol. 48 9–13

    Article  PubMed  CAS  Google Scholar 

  • Kanai R and Edwards G E 1973a Separation of mesophyll protoplasts and bundle shealth cells from maize leaves for photosynthetic studies;Plant Physiol. 51 1135–1137

    Article  Google Scholar 

  • Kanai R and Edwards G E 1973b Purification of enzymatically isolated mesophyll protoplasts from C3, C4 and Crassulacean acid metabolism plants using an aqueous dextran polyethyene glycol two phase system;Plant Physiol. 52 484–490

    Article  PubMed  CAS  Google Scholar 

  • Ku S B, Gutierrez M, Kanai R and Edwards G E 1974 Photosynthesis in mesophyll protoplasts and bundle sheath cells of various types of C4 plants. II. Chlorophyll and Hill reaction studies;Z. Pflanzenphysiol. 72 320–337

    CAS  Google Scholar 

  • Lilley R McC, Fitzegerald M P, Rienits K G and Walker D A 1975 Criteria of intactness and the photosynthetic activity of spinach chloroplast preparations;New Phytol. 75 1–10

    Article  CAS  Google Scholar 

  • Nishimura M and Akazawa T 1975 Photosynthetic activities of spinach leaf protoplasts;Plant Physiol. 55 712–716

    Article  PubMed  Google Scholar 

  • Okuno T and Furusawa I 1977 A simple method for isolation of intact mesophyll protoplasts from cereal leaves;Plant Cell Physiol. 18 1357–1362

    Google Scholar 

  • Otsuki Y and Takebe I 1969 Isolation of intact mesophyll cells and their protoplasts from higher plants;Plant Cell Physiol. 10 917–921

    Google Scholar 

  • Power J B and Cocking E C 1970 Isolation of leaf protoplasts; Macromolecule uptake and growth substance response;J. Exp. Bot. 21 64–70

    Article  CAS  Google Scholar 

  • Rajendrudu G, Madhusudana Rao I, Raghavendra A S and Rama Das V S 1979 Isolation of intact mesophyll cells from the leaves of higher plants;Proc. Indian Acad. Sci. 88B 143–154

    Google Scholar 

  • Rathnam C K M and Edwards G E 1976 Protoplasts as a tool for isolating functional chloroplasts from leaves;Plant Cell Physiol. 17 177–186

    CAS  Google Scholar 

  • Ruesink A W 1971 Protoplasts of plant cells; inMethods in Enzymology ed. A San Pietro (London-New York: Academic Press) pp 197–209

    Google Scholar 

  • Takebe I, Otsuki Y and Aoki S 1968 Isolation of tobacco mesophyll cells in intact and active state;Plant Cell Physiol. 9 115–1924

    Google Scholar 

  • Vernon and Shaw E R 1969 Oxidation of 1,5-diphenyl-carbazide as a measure of photosystem 2 activity in subchloroplast fragments;Biochem. Biophys. Res. Commun. 36 878–884

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Author notes

  1. K P Chellappan

    Present address: Department of Botany, Annamalai University, 608 101, Annamalainagar

Authors and Affiliations

  1. Department of Plant Sciences, School of Biological Sciences, Madurai Kamaraj University, 625 021, Madurai

    K P Chellappan & A Gnanam

Authors
  1. K P Chellappan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A Gnanam
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chellappan, K.P., Gnanam, A. Isolation of intact mesophyll protoplasts from the leaves of higher plants for photosynthetic studies. Proc. Indian Acad. Sci. 89, 79–90 (1980). https://doi.org/10.1007/BF03046152

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF03046152

Keywords

Search

Navigation