Histochemical detection of acetogenins and storage molecules in the endosperm of Annona macroprophyllata Donn Sm. seeds

  • G. Laguna Hernández | glagher@ciencias.unam.mx Universidad Nacional Autónoma de México, Mexico.
  • A.E. Brechú-Franco Universidad Nacional Autónoma de México, Mexico.
  • I. De la Cruz-Chacón Universidad de Ciencias y Artes de Chiapas, Mexico.
  • A.R. González-Esquinca Universidad de Ciencias y Artes de Chiapas, Mexico.

Abstract

Acetogenins (ACGs) are bioactive compounds with cytotoxic properties in different cell lines. They are antitumoural, antiparasitic, antimalarial, insecticidal, antimicrobial, antifungal and antibacterial. These secondary metabolites function in plant defence and are found in specific organelles and specific cells, thereby preventing toxicity to the plant itself and permitting site-specific defence. The aim of this work was to histochemically determine the in situ localisation of ACGs in the endosperm of Annona macroprophyllata seeds using Kedde’s reagent. Additionally, the colocalisation of ACGs with other storage molecules was analysed. The seeds were analysed after 6 and 10 days of imbibition, when 1 or 2 cm of the radicle had emerged and metabolism was fully established. The seeds were then transversally cut in half at the midline and processed using different histological and histochemical techniques. Positive reactions with Kedde’s reagent were only observed in fresh, unfixed sections that were preserved in water, and staining was found only in the large cells (the idioblasts) at the periphery of the endosperm. The ACGs’ positive reaction with Sudan III corroborated their lipid nature. Paraffin sections stained with Naphthol Blue Black showed reactions in the endosperm parenchyma cells and stained the proteoplasts blue, indicating that they might correspond to storage sites for albumin-like proteins. Lugol’s iodine, which is similar in chemical composition to Wagner’s reagent, caused a golden brown reaction product in the cytoplasm of the idioblasts, which may indicate the presence of alkaloids. Based on these results, we propose that Kedde’s reagent is an appropriate histochemical stain for detecting ACGs in situ in idioblasts and that idioblasts store ACGs and probably alkaloids. ACGs that are located in idioblasts found in restricted, peripheral areas of the endosperm could serve as a barrier that protects the seeds against insects and pathogen attack.

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Author Biographies

G. Laguna Hernández, Universidad Nacional Autónoma de México

Ecología y Recursos Naturales, 

Profesor Titular A de Tiempo Completo.

A.E. Brechú-Franco, Universidad Nacional Autónoma de México

Ecología y Recursos Naturales

Profesor Titular A de Tiempo Completo

I. De la Cruz-Chacón, Universidad de Ciencias y Artes de Chiapas

Instituto de Ciencias Biológicas

Profesor Tiempo Completo Asociado

A.R. González-Esquinca, Universidad de Ciencias y Artes de Chiapas

Instituto de Ciencias Biológicas

Investigadora Titular C

Published
2015-07-09
Section
Technical Notes
Supporting Agencies
Universidad Nacional Autónoma de México, Universidad de Ciencias y Artes de Chiapas
Keywords:
Annona, Kedde reagent, acetogenin histochemistry, storage reserves histochemistry, acetogenin idioblast.
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How to Cite
Laguna Hernández, G., Brechú-Franco, A., De la Cruz-ChacónI., & González-Esquinca, A. (2015). Histochemical detection of acetogenins and storage molecules in the endosperm of Annona macroprophyllata Donn Sm. seeds. European Journal of Histochemistry, 59(3). https://doi.org/10.4081/ejh.2015.2502