技术文章

Dualex植物叶绿素仪应用:叶片不同发育阶段叶绿素酶的定位

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2022/5/7 10:50:35

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Dualex便携式智能气象-叶绿素-花青素-黄酮醇测量仪

摘要 

背景:叶绿素酶(Chlases)是植物细胞中的管家蛋白。脱植酸酶可以催化叶绿素(Chl)形成叶绿素内酯,但叶绿素酶在植物细胞中的分布仍然具有争论。本研究制备了PmCLH2抗体,并用免疫金标记技术检测了大果委陵菜(Pachira macrocarpa,Pm)幼叶、成熟叶、黄化叶和衰老叶四个发育阶段的叶绿素酶定位。结果:透射电镜观察结果表明,在叶绿体的幼龄期、成熟期和变黄期,叶绿体的膜、基粒、叶绿体的类囊体膜、细胞质和液泡等部分全面发现了叶绿素,但在细胞壁、质膜、线粒体和细胞核中没有发现。结论:在衰老期,PmChlases主要存在于液泡中,但在叶绿体中很少发现。提出了一条解释高等植物中叶绿素、叶绿素酶和叶绿体的产生和死亡的途径。

Immunogold-labelling localization of chlorophyllase at different developmental stages of Pachira macrocarpa leaves 

Abstract

Background: Chlorophyllases (Chlases) are housekeeping proteins in plant cells. The dephytylating enzymes can catalyze chlorophyll (Chl) to form chlorophyllide, but the distribution of Chlases in plant cells is still an interesting debate. In this study, antibody of PmCLH2 was made and used by immunogold-labelling technique to detect the location of Chlase of Pachira macrocarpa (Pm) leaves at four developmental stages, including young, mature, yellowing, and senesced stages. Results: The transmission electron microscopy results show that Chlases were comprehensively found in portions of chloroplast, such as the inner membrane of the envelope, grana, and the thylakoid membrane of the chloroplast, cytosol, and vacuoles at young, mature, and yellowing stages of Pm leaves, but not in the cell wall, plasma membrane, mitochondria, and nucleus. Conclusions: PmChlases were mainly detected in vacuoles at the senescent stage, but a few were found in the chloroplasts. A pathway is proposed to explain the birth and death of Chl, Chlase, and chloroplasts in higher plants.

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