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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >A calmodulin-binding protein from Arabidopsis has an essential role in pollen germination.
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A calmodulin-binding protein from Arabidopsis has an essential role in pollen germination.

机译：来自拟南芥的钙调蛋白结合蛋白在花粉萌发中具有重要作用。

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摘要

Calmodulin (CaM), a ubiquitous multifunctional calcium sensor in all eukaryotes, mediates calcium action by regulating the activity/function of many unrelated proteins. Although calcium and CaM are known to play a crucial role in pollen germination and pollen tube growth, the proteins that mediate their action have not been identified. We isolated three closely related CaM-binding proteins (NPG1, NPGR1, and NPGR2) from Arabidopsis. NPG1 (No Pollen Germination1) is expressed only in pollen, whereas the NPG-related proteins (NPGR1 and NPGR2) are expressed in pollen and other tissues. The bacterially expressed NPG1 bound three isoforms of Arabidopsis CaM in a calcium-dependent manner. To analyze the function of NPG1, we performed a reverse genetics screen and isolated a mutant in which NPG1 is disrupted by a T-DNA insertion. Segregation and molecular analyses of the NPG1 knockout mutant and a cross with a male sterile mutant indicate that the mutated NPG1 is not transmitted through the male gametophyte. Expression of NPG1 in the knockout mutant complemented the mutant phenotype. Analysis of pollen development in the knockout mutant by light microscopy showed normal pollen development. Pollen from NPG1 mutant in the quartet background has confirmed that NPG1 is dispensable for pollen development. However, germination studies with pollen from the mutant in the quartet background indicate that pollen carrying a mutant allele does not germinate. Our genetic, histological, and pollen germination studies with the knockout mutant line indicate that NPG1 is not necessary for microsporogenesis and gametogenesis but is essential for pollen germination.
机译：钙调蛋白（CaM）是所有真核生物中普遍存在的多功能钙传感器，通过调节许多无关蛋白的活性/功能来介导钙的作用。尽管已知钙和CaM在花粉萌发和花粉管生长中起关键作用，但尚未鉴定出介导其作用的蛋白质。我们从拟南芥中分离了三种密切相关的CaM结合蛋白（NPG1，NPGR1和NPGR2）。 NPG1（无花粉发芽1）仅在花粉中表达，而NPG相关蛋白（NPGR1和NPGR2）在花粉和其他组织中表达。细菌表达的NPG1以钙依赖的方式结合拟南芥CaM的三种同工型。为了分析NPG1的功能，我们进行了反向遗传学筛选，并分离了其中NPG1被T-DNA插入破坏的突变体。 NPG1基因敲除突变体和与雄性不育突变体的杂交的分离和分子分析表明，突变的NPG1不通过雄配子体传播。 NPG1在敲除突变体中的表达补充了突变体表型。通过光学显微镜分析敲除突变体中的花粉发育显示出正常的花粉发育。在四重奏背景下，来自NPG1突变体的花粉已证实NPG1对于花粉发育是必不可少的。但是，四元组背景中来自突变体的花粉的发芽研究表明，携带突变体等位基因的花粉不会发芽。我们的基因，组织学和花粉萌发研究表明，NPG1对于微孢子发生和配子发生不是必需的，但对于花粉萌发必不可少。

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