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KMID : 0368420220650010001
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Journal of Plant Biology
2022 Volume.65 No. 1 p.1 ~ p.10
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A Method for a Rapid and Accurate Detection of Phloem Sieve Element Development: Its Application and Scalability
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Kim Hyou-Jin
Lee Ji-Young
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Abstract
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Vascular system plays an important role in plant growth and survival under dynamic environmental conditions. As part of vascular tissues, the phloem consists of phloem sieve elements (SEs) and companion cells (CCs). In the root apical meristem of Arabidopsis thaliana, CCs are generated by the asymmetric cell divisions (ACDs) of procambial cells adjoining SEs and pericycle cells. The proto- and metaphloem SEs are originated from sequential ACDs of a SE-procambium precursor and a SE precursor. To study the development of vascular tissues, researchers have mainly relied on histological methods that take days with multiple steps. While investigating molecular mechanisms underlying phloem SE development, we developed a rapid and reliable method that allows for the detection of differentiating phloem SEs. This protocol employs monoclonal anti-phloem sieve element (RS6) antibody, which was selected from hybridomas raised against SEs isolated from tissue cultures of California shield leaf, Streptanthus tortuosus. RS6 is known to recognize a SE-specific early nodulin-like protein, SE-ENOD, which is specifically localized to the plasma membrane of differentiating and mature SEs. This protocol enables quantitative assessment of phloem SEs on a large scale for a broad range of species beyond A. thaliana.
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KEYWORD
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Phloem sieve element, Immunostaining, SE-ENOD, RS6 antibody, Vibratome, Brassicaceae, Eudicot
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