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MES

Company: Thermo Fisher Scientific
Catalog#: BP300-100
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Quantifying the Capacity of Phloem Loading in Leaf Disks with [14C]Sucrose
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Date:
2017-12-20
[Abstract]  Phloem loading and transport of photoassimilate from photoautotrophic source leaves to heterotrophic sink organs are essential physiological processes that help the disparate organs of a plant function as a single, unified organism. We present three protocols we routinely use in combination with each other to assess (1) the relative rates of sucrose (Suc) loading into the phloem vascular system of mature leaves (this protocol), (2) the relative rates of carbon loading and transport through the phloem (Yadav et al., 2017a), and (3) the relative rates of carbon unloading into heterotrophic sink organs, specifically roots, after long-distance transport (Yadav et al., 2017b). We propose that conducting all three protocols on experimental and control plants provides a ... [摘要]  来自光合自养源的光合同化物的韧皮部装载和运输到异养宿主器官是必不可少的生理过程,其帮助植物的不同器官作为单一的统一生物体起作用。我们提出了三种方案,我们经常使用相互组合,以评估(1)蔗糖(Suc)加载到成熟叶片的韧皮部血管系统(本协议)的相对比率,(2)碳负荷和运输通过韧皮部(Yadav et al。,2017a),和(3)在长距离运输后碳卸载到异养汇器官特别是根中的相对速率(Yadav等人, / em>。,2017b)。我们建议,在实验和对照植物上进行所有三种方案提供了全植物碳分配的可靠比较,并且将与单独进行的单个方案相关联的歧义降至最低(Dasgupta等人,2014; Khadilkar 。,2016)。在该方案中,从成熟莲座叶中分离的拟南芥叶片用含有[14 C] Suc的缓冲溶液浸润。 Suc转运蛋白(SUCs或SUTs)将Suc载入韧皮部,并将多余的,卸载在叶片中的Suc洗掉。通过冻干叶盘的放射自显影显示标记的Suc加载到静脉中,并通过闪烁计数进行定量。结果表示为每单位叶盘鲜重或面积的每分钟崩解。

【背景】光合同化物从源头到宿主器官的运输对于整个植物的正常生长和维持至关重要。叶片中的韧皮部负载是将在叶肉细胞中合成的光合同化物递送至韧皮部脉管系统的伴生细胞(CC)和筛分元素(SE)。三种不同的加载机制被认可。其中两个消耗能量在CC和SEs中累积高浓度的糖,并在源叶韧皮部产生高静水压力。第一种是外源韧皮部装载,其中将Suc(和/或一些物种中的糖醇)穿过质膜从细胞壁空间(即,质外体)装载进入CC中的CC质子动力的牺牲(Giaquinta,1983)。第二种是聚合物捕获,其中Suc通过特化的胞间连丝(Botmodesmata)扩散到韧皮部,并转化为太大而不能扩散回来的寡糖(Turgeon,1996)。第三种机制是被动加载,其中叶肉细胞中的溶质浓度最高,胞间连丝为被动移入CC和SE提供了一个开放的途径(Rennie和Turgeon,2009)。 ...

In vitro CLE Peptide Bioactivity Assay on Plant Roots
Author:
Date:
2015-12-20
[Abstract]  Plant CLAVATA3/ESR (CLE)-related proteins play diverse roles in plant growth and development including regulating the development of root meristem. The mature functional forms of CLE peptides are typically 12-13 amino acids (aa) in length that are derived from the conserved C-termini of their precursor proteins. Genes encoding small secreted peptides sharing similarity to plant CLE proteins have recently been cloned from plant-parasitic nematodes, pests that infect many important crops. It is demonstrated that exogenous application of synthetic 12-14 aa CLE peptides corresponding to the CLE domain of their precursor proteins can suppress plant root growth. This protocol is to evaluate the bioactivity of CLE peptides originated from plant-parasitic nematodes by measuring the growth of ... [摘要]  植物CLAVATA3/ESR(CLE)相关蛋白质在植物生长和发育中发挥多种作用,包括调节根分生组织的发育。 CLE肽的成熟功能形式通常为12-13个氨基酸(aa)长度,其衍生自其前体蛋白的保守C-末端。 最近从植物寄生线虫,感染许多重要作物的害虫克隆了编码与植物CLE蛋白相似的小分泌肽的基因。 证明外源应用对应于其前体蛋白的CLE结构域的合成12-14aa CLE肽可以抑制植物根生长。 该协议是通过测量CLE肽处理后植物根的生长或根尖分生组织(RAM)的大小来评价源自植物寄生线虫的CLE肽的生物活性。 用于研究的植物包括拟南芥和马铃薯。

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