Dualex植物葉綠素儀應(yīng)用：葉片不同發(fā)育階段葉綠素酶的定位

Dualex便攜式智能氣象-葉綠素-花青素-黃酮醇測量儀

摘要 

背景：葉綠素酶（Chlases）是植物細胞中的管家蛋白。脫植酸酶可以催化葉綠素（Chl）形成葉綠素內(nèi)酯，但葉綠素酶在植物細胞中的分布仍然具有爭論。本研究制備了PmCLH2抗體，并用免疫金標記技術(shù)檢測了大果委陵菜（Pachira macrocarpa，Pm）幼葉、成熟葉、黃化葉和衰老葉四個發(fā)育階段的葉綠素酶定位。結(jié)果：透射電鏡觀察結(jié)果表明，在葉綠體的幼齡期、成熟期和變黃期，葉綠體的膜、基粒、葉綠體的類囊體膜、細胞質(zhì)和液泡等部分全面發(fā)現(xiàn)了葉綠素，但在細胞壁、質(zhì)膜、線粒體和細胞核中沒有發(fā)現(xiàn)。結(jié)論：在衰老期，PmChlases主要存在于液泡中，但在葉綠體中很少發(fā)現(xiàn)。提出了一條解釋高等植物中葉綠素、葉綠素酶和葉綠體的產(chǎn)生和死亡的途徑。

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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