Open Access

ARTICLE

A Simple Clear Technique in Observing Vascular Development of Grape Ovary

by Teng Fei¹, Youmei Li¹, Bo Li², Zhaosen Xie^1,*

1 College of Horticulture and Garden, Yangzhou University, Yangzhou, 225009, China
2 Shandong Academy of Grape, Shandong Academy of Agricultural Sciences, Jinan, 250199, China

* Corresponding Author: Zhaosen Xie. Email:

Phyton-International Journal of Experimental Botany 2023, 92(7), 2117-2132. https://doi.org/10.32604/phyton.2023.028208

Received 05 December 2022; Accepted 27 March 2023; Issue published 29 May 2023

Abstract

The vascular system of the grapevine (Vitis vinifera L.) flower is a channel for transporting water and nutrients to the ovary. It plays an important role in the development of the ovary and fertilization through pollination. However, the vascular bundles in the flower are so tiny that they are difficult to sample and observe by traditional slicing techniques. In this study, ‘Summer Black’ grape flowers were selected as the test materials, and the tissue samples were treated by the optical clearing technique. After simple compaction, the structure and development of the vasculature were observed by common microscopy, fluorescence microscopy and laser confocal microscopy. The results showed that the transparency effects of 3% NaOH and a saturated trichloroacetaldehyde composite agreed well with the observations of the vascular structure and the developmental process of the flower in different periods. Moreover, the samples after optical clearing could be reconstructed in 3D, which helped us know more about its development and function. According to these observations, the vasculature of the ‘Summer Black’ flower can be divided into ovule vascular bundles, peripheral vascular bundles and central vascular bundles. The peripheral vascular bundles were composed of the first-order vascular bundles and the inferior vascular bundles which branched from the superior vascular bundles. These bundles branched in different directions with no discernible pattern. The two different branching methods were as follows. First, the inferior vascular bundle was directly connected to a superior vascular bundle. Secondly, some of the superior vascular bundles bent in different ways, forming the inferior vascular bundle connecting the superior vascular bundles by a metamorphosed vessel with a triangular shape. In a comparison of the developmental changes in various periods, the growth of vascular bundles at each period was directly proportional to the growth of the flower. Laser confocal scanning was used to explore the three-dimensional morphology of the peripheral vascular bundle and showed that the peripheral vascular bundle of grapes was not completely parallel to the flower’s epidermal cells. As a result, the optical clearing technique was convenient and authentic compared with the traditional slicing operation for tiny flower organs. With these advantages according to the observations, this study provides a feasible technique and useful information for the study of vascular bundle development in grape flower organs.

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Keywords

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