A new monoclonal antibody against rhamnogalacturonan II and its application to immunocytochemical detection of rhamnogalacturonan II in Arabidopsis roots
A new monoclonal antibody against rhamnogalacturonan II and its application to immunocytochemical detection of rhamnogalacturonan II in Arabidopsis roots
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DOI：
10.1080/09168451.2018.1485479
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发表时间：
2018-06
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影响因子：
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通讯作者：
Ye Zhou;Masaru Kobayashi;T. Awano;T. Matoh;K. Takabe
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作者：
Ye Zhou;Masaru Kobayashi;T. Awano;T. Matoh;K. Takabe
鼠李糖半乳糖醛酸聚糖II（RG-II）是存在于植物初生细胞壁中的一个果胶大分子区域。RG-II可以作为硼酸盐-RG-II复合物（B-RG-II）从细胞壁中溶解，其中两个RG-II片段通过硼酸二酯键交联。在此，制备了抗B-RG-II的兔单克隆抗体，其识别B-RG-II和RG-II单体而无需硼酸酯交联。一个结构未知的果胶片段也被抗体识别，但既不是homogalacturonan也不是rhamnogalacturonan I被识别。使用该抗体进行拟南芥根尖细胞的免疫电镜分析。在发育中的细胞板和细胞壁中检测到信号，其在纵向壁中比在横向壁中更密集。这些结果与我们先前在悬浮培养的烟草细胞中获得的结果一致，证实了RG-II在其组装的早期阶段存在于细胞板中。缩略语：B：硼; B-RG-II：硼酸根-RG-Ⅱ络合物; ELISA：酶联免疫吸附试验; IgG：免疫球蛋白G; mBSA：甲基化牛血清白蛋白; PGA：聚半乳糖醛酸; PLL：聚赖氨酸; RG-I：鼠李糖半乳糖醛酸聚糖I; RG-II：利用兔单克隆抗体对拟南芥根细胞中的鼠李半乳糖醛酸聚糖II进行免疫金标记。在细胞壁（A）和细胞板（B）中都检测到信号。
ABSTRACT Rhamnogalacturonan II (RG-II) is a region of pectin macromolecules that is present in plant primary cell walls. RG-II can be solubilized from cell walls as a borate-RG-II complex (B-RG-II), where two RG-II fragments are cross-linked via a borate diester linkage. Here, a rabbit monoclonal antibody against B-RG-II was prepared, which recognized both B-RG-II and RG-II monomers without borate ester-crosslinking. A pectic fragment with unknown structure was also recognized by the antibody, but neither homogalacturonan nor rhamnogalacturonan I was recognized. Immunoelectron microscopic analyses of Arabidopsis root tip cells were performed using this antibody. The signal was detected in developing cell plates and cell walls, which were denser in longitudinal walls than in transverse walls. These results coincide with our previous results obtained in suspension cultured tobacco cells, confirming that RG-II is present in cell plates at an early stage of their assembly. Abbreviations: B: boron; B-RG-II: borate-RG-II complex; ELISA: enzyme-linked immunosorbent assay; IgG: immunoglobulin G; mBSA: methylated bovine serum albumin; PGA: polygalacturonic acid; PLL: poly-l-lysine; RG-I: rhamnogalacturonan I; RG-II: rhamnogalacturonan II Graphical Abstract Immunogold labeling of rhamnogalacturonan II in Arabidopsis root cells using rabbit monoclonal antibody. Signals were detected in both cell walls (A) and cell plates (B).