A new original "cryostabilization" technique was applied to different animal tissues and organs. In this study, we apply this technique also to vegetal tissues (ripening and ripe fleshy fruits, buds and embryos). The proposed method preserves good tissue morphology and intracellular substances, and avoids loss of epicuticular waxes; moreover it maintains in situ some enzymatic activities. Being some samples very difficult to handle the original protocol had to be modified process. The correct orientation in embedding of samples is an important requirement which is not easy to achieve when dealing with small fragments of tissues. To obtain slices oriented correctly, Vitis vinifera L. somatic and zygotic embryos, very little in size, and Corylus avellana L. leaves, are pre-included in agar 1%. This pre-inclusion in agar either facilitates the sample manipulation either allows to achieve a satisfactory precision in slicing. In a study on apricot buds (Prunus armeniaca L.), vacuum was used to facilitate reagent penetration, in spite of tissue hardness and impermeability. Observations were performed on slices cut with glass blades and stained with different histological and histochemical dyes. In Vitis vinifera the method allowed to distinguish normal and anomalous embryo development in different stages. In Corylus avellana different thickness of leave lacunose and palisade tissues could be put in relation with sun or shadow conditions of growth. In apricot buds the morphology of meristematic tissues is well preserved and starch reserves could be visualized, up the starting of hydrolysis at the inception of the new vegetative season.

Application of an innovative technique for histological and histochemical study of different vegetal tissues

DORE, Bruno Emilio;
2009-01-01

Abstract

A new original "cryostabilization" technique was applied to different animal tissues and organs. In this study, we apply this technique also to vegetal tissues (ripening and ripe fleshy fruits, buds and embryos). The proposed method preserves good tissue morphology and intracellular substances, and avoids loss of epicuticular waxes; moreover it maintains in situ some enzymatic activities. Being some samples very difficult to handle the original protocol had to be modified process. The correct orientation in embedding of samples is an important requirement which is not easy to achieve when dealing with small fragments of tissues. To obtain slices oriented correctly, Vitis vinifera L. somatic and zygotic embryos, very little in size, and Corylus avellana L. leaves, are pre-included in agar 1%. This pre-inclusion in agar either facilitates the sample manipulation either allows to achieve a satisfactory precision in slicing. In a study on apricot buds (Prunus armeniaca L.), vacuum was used to facilitate reagent penetration, in spite of tissue hardness and impermeability. Observations were performed on slices cut with glass blades and stained with different histological and histochemical dyes. In Vitis vinifera the method allowed to distinguish normal and anomalous embryo development in different stages. In Corylus avellana different thickness of leave lacunose and palisade tissues could be put in relation with sun or shadow conditions of growth. In apricot buds the morphology of meristematic tissues is well preserved and starch reserves could be visualized, up the starting of hydrolysis at the inception of the new vegetative season.
2009
LXXXII
72
75
cryostabilization; embedding; polar resin (GMA); agar; fruit trees
DORE B.; PONSO A.; VALLANIA R.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2318/57903
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