《植物生理学报》 2018, 54(2): 237-246

组织透明法观察葡萄叶片生长过程中气孔与叶脉形态结构特征变化

谢兆森^1,*, 杜鸿儒¹, 李建宝¹, Bhaskar Bondada²

¹河北科技师范学院园艺科技学院, 河北秦皇岛066600; ²华盛顿州立大学葡萄栽培与酿造科研中心, 美国华盛顿州里奇兰市 99354

通信作者：谢兆森；E-mail: xiezhaosen@sina.com

摘　要：

本文以‘摩尔多瓦’葡萄叶片为试验材料, 采用组织透明法观察了葡萄叶片生长过程中叶片表皮细胞、气孔和叶脉形态结构的变化,测定与气孔和叶脉功能相关的生理指标变化, 比较了组织透明法、指甲油法和撕取法在观察葡萄叶片气孔上的实验效果。结果表明: 组织透明法能够清晰观察到葡萄叶片生长过程中叶片表皮细胞、气孔和叶脉形态结构的变化。相对于指甲油法、撕取法, 组织透明法操作简单, 且能保持组织完整性, 提升观察叶片细胞结构的实验效果。从叶片生长过程中气孔形态结构变化规律来看, 葡萄嫩叶齿尖存在大量的大气孔, 而叶片中部只观察到中央大气孔和正在发育的气孔保卫细胞母细胞。随着叶片生长, 叶片气孔保卫细胞母细胞逐渐发育形成成熟气孔, 叶片末端叶脉也是随叶片生长而生长, 叶脉密度逐渐增加, 提高了树体往叶片供水的效率。从与气孔和叶脉功能相关的各项生理指标变化来看, 气孔导度随着气孔逐渐形成和成熟而逐渐升高, 但叶片含水量和水势下降, 有利于拉升水分和养分从根系往地上树体各器官运输, 满足整个植株生长发育的需要。

关键词：葡萄; 组织透明; 叶片; 气孔; 叶脉

收稿：2017-09-06   修定：2018-01-15

资助：国家自然科学基金项目(31470104)和河北省自然科学基金(C2018407036)。

Morphological and structural changes of stomata and leaf veins during growth of grape leaves using tissue clearing technique

XIE Zhao-Sen^1,*, DU Hong-Ru¹, LI Jian-Bao¹, Bhaskar Bondada²

¹College of Horticulture Science and Technology, Hebei Normal University of Science and Technology, Qinhuangdao, Hebei 066600, China; ²Washington State University, Viticulture and Wine Science Center, Richland, WA 99354, USA

Corresponding author: XIE Zhao-Sen; E-mail: xiezhaosen@sina.com

Abstract:

In this study, ‘Moldova’ grapevine (Vitis vinifera) leaves were cleared to examine leaf epidermal cells, stomata, and minor vein system. These features were compared with other techniques such as leaf imprints using nail polish and tearing/peeling of leaf epidermis. Among these techniques, the tissue clearing technique emerged as the best way of studying leaf morphological features as it is easy and provides clear images of the highest resolution. For instance, it revealed numerous large stomates scattered throughout the abaxial surfaces of dentate margins of young leaves as opposed to few large stomates centrally localized along with guard mother cells in the lamina. As the leaves developed, the guard mother cells transformed into mature stomata and the minor vein system increased its vein density. These collectively increased stomatal conductance but the water content and water potential declined explaining the paradigm that plants use most water to transpire and only a small quantity to sustain growth.

Key words: grape (Vitis vinifera); tissue clearing; leaf; stomata; vein