POWRE: Characterization of a Novel ATPase

POWRE：新型 ATP 酶的表征

• 批准号: 0074617
• 负责人: Mary Bisson
• 金额: $ 7.47万
• 依托单位: SUNY at Buffalo
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-15 至 2003-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0074617&HistoricalAwards=false
• 关键词: POWRE; Characterization; Novel; ATPase

All plant cells must actively export sodium from the cytoplasm. Accumulation of sodium in the cytoplasm results in inhibition of a number of enzyme systems, and is a major cause of damage due to salinity, whether natural or due to irrigation of agricultural fields. A knowledge of the mechanisms of Na+ transport is therefore not only important in general understanding of the function of plants, but also enhances our ability to develop crop plants which are more resistant to salinization. This project will use two species of Charophyte algae that differ in their ability to tolerate saline conditions. Re-moval of Na+ from the cytoplasm by export across the plasma mem-brane is important in achieving salt tolerance, and there is evidence that an ATPase may be involved in this transport, including biophysical evidence that a Na+/H+ antiport is not sufficient to maintain the Na+ gradients measured, immunological evidence of an unusual P-type ATPase in salt-cultured algae, and molecular evidence for an unusual P-type ATPase in the salt-tolerant algae. The objectives of the proposed research are To isolate and sequence a full-length clone for an unusual ATPase from the salt-tolerant species, Chara longifolia. To use northern blot analysis to determine expression of this gene in C. longifolia adapted to saline and freshwater media, and undergoing adaptation to salt stress. To determine whether the salt-sensitive species, C. corallina, has a similar gene and, if so, under what conditions it is expressed. Long-term goals include determining the transport function of the ATPase gene either by expres-sion in yeast mutants lacking Na+ or H+ export capability or by overexpression in Chara by micro-injection of mRNA into the internode. Ultimately, information based on this research could be used to alter the salt tolerance of Chara and other plants by genetic manipulation to enhance sodium export from the cytoplasm. This research will be performed under the POWRE grant program because it will aid the author in remaining competitive in the field of salinity studies in plants. Bio-physical techniques have been successfully employed in the past to investigate important ques-tions in this field, but molecular aspects of the project must be developed in order to remain competitive.

所有植物细胞都必须主动从细胞质中输出钠。 细胞质中钠的积累会抑制许多酶系统，并且是盐度造成损害的主要原因，无论是自然盐度还是农田灌溉所致。 因此，了解 Na+ 运输机制不仅对于一般理解植物功能很重要，而且还可以增强我们开发更能抵抗盐碱化的作物的能力。 该项目将使用两种轮藻，它们耐受盐水条件的能力不同。 通过质膜输出将 Na+ 从细胞质中去除对于实现耐盐性非常重要，并且有证据表明 ATP 酶可能参与这种转运，包括 Na+/H+ 反向转运不足以维持测量的 Na+ 梯度的生物物理证据、盐培养藻类中不寻常的 P 型 ATP 酶的免疫学证据以及异常 P 型 ATP 酶的分子证据。 耐盐藻类中的 P 型 ATP 酶。 拟议研究的目标是从耐盐物种 Chara longifolia 中分离出一个不寻常 ATP 酶的全长克隆并对其进行测序。 使用 Northern blot 分析来确定该基因在适应盐水和淡水介质以及适应盐胁迫的 C. longifolia 中的表达。 确定对盐敏感的物种 C. Corallina 是否具有相似的基因，如果有，它在什么条件下表达。长期目标包括通过在缺乏 Na+ 或 H+ 输出能力的酵母突变体中表达或通过将 mRNA 微注射到节间在 Chara 中过度表达来确定 ATPase 基因的转运功能。 最终，基于这项研究的信息可用于通过基因操作来改变轮藻和其他植物的耐盐性，以增强细胞质中的钠输出。 这项研究将在 POWRE 资助计划下进行，因为它将帮助作者在植物盐度研究领域保持竞争力。 过去，生物物理技术已成功用于研究该领域的重要问题，但该项目的分子方面必须得到发展才能保持竞争力。