Overcoming the difficulties in collecting apoplastic fluid from rice leaves by the infiltration-centrifugation method

Isamu Nouchi, Kentaro Hayashi, Syuntaro Hiradate, Satoru Ishikawa, Minehiko Fukuoka, Charles P. Chen, Kazuhiko Kobayashi

研究成果: Contribution to journalArticle査読

23 被引用数 (Scopus)

抄録

Physiological and biochemical studies on the leaf apoplast have been facilitated by the use of the infiltration-centrifugation technique to collect intercellular washing fluid (IWF). However, this technique has been difficult to implement in rice (Oryza sativa L.) for various reasons. We compared the collection efficiency of leaf IWF between two types of rice varieties (Indica and Japonica), as well as between rice and other species (spinach, snap bean and wheat). Although the extraction of IWF in most species took only 2-3 min, it took up to 35 min in rice. The difficulty in infiltration with rice was ascribed to the small stomatal aperture and hydrophobicity of the leaves. In this study, we have established an improved method for collecting IWF and determining the apoplastic air and water volumes in rice leaves. We have shortened the infiltration time to 8 min via the following improvements: (i) infiltration under outdoor shade in the daytime to prevent stomatal closure and a rise in temperature of the infiltration medium; (ii) soaking of leaves in a surfactant solution to decrease the leaf hydrophobicity; and (iii) continuous pressurization using a sealant injector to facilitate the infiltration. The rapid collection of IWF achieved using this technique will facilitate study of the leaf apoplast in rice.

本文言語英語
ページ(範囲)1659-1668
ページ数10
ジャーナルPlant and Cell Physiology
53
9
DOI
出版ステータス出版済み - 9 2012
外部発表はい

All Science Journal Classification (ASJC) codes

  • 生理学
  • 植物科学
  • 細胞生物学

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