Decapitation Crosses to Test Pollen Fertility Mutations for Defects in Stigma-style Penetration
Decapitation Crosses to Test Pollen Fertility Mutations for Defects in Stigma-style Penetration
复制标题
断头杂交测试花粉育性突变的柱头式渗透缺陷
DOI：
10.1007/978-1-0716-0672-8_3
复制
发表时间：
2020
影响因子：
2.4
通讯作者：
Harper, Jeffrey F
中科院分区：
文献类型：
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作者：
Weigand, Chyrstle;Harper, Jeffrey F
在开花植物的有性生殖过程中，花粉粒在柱头表面萌发，并穿过柱头花柱组织生长，到达子房并运送精细胞进行受精。在这里，我们概述了一种方法来测试花粉育性突变是否专门破坏花粉穿透柱头风格障碍。该方法通过手术去除柱头花柱（柱头去头）来测试是否将花粉直接转移到暴露的子房表面上显著提高突变等位基因的传递效率（TE）。为了说明这一技术，我们采用柱头断头调查的功能突变inopsisOFT 1，一个基因编码一个假定的o-岩藻糖基转移酶的分泌途径中发挥作用的损失。oft 1 - 3突变花粉表现出显着降低传输效率相比，野生型。断头杂交（这里描述）表明，柱头花柱障碍的去除减轻了传输缺陷从858倍到2.6倍，提供证据表明，大多数，但不是全部，oft 1花粉缺陷可归因于降低穿透柱头花柱障碍的能力。该方法概述了一种遗传策略，以量化突变对花粉在其到达胚珠的旅程中穿过柱头风格障碍的能力的影响。
During sexual reproduction in flowering plants, pollen grains germinate on the stigma surface and grow through the stigma-style tissue to reach the ovary and deliver sperm cells for fertilization. Here, we outline a method to test whether a pollen fertility mutation specifically disrupts pollen penetration through the stigma-style barrier. This method surgically removes the stigma-style (stigma decapitation) to test whether transferring pollen directly onto an exposed ovary surface significantly improves the transmission efficiency (TE) of a mutant allele. To illustrate this technique, we applied stigma decapitation to investigate a loss-of-function mutation inArabidopsisOFT1, a gene encoding a putative o-fucosyl transferase functioning in the secretory pathway.oft1-3mutant pollen showed a significant decrease in transmission efficiency compared to wild type. Decapitation crosses (described here) indicated that the removal of the stigma-style barrier alleviated the transmission deficiency from 858-fold to a 2.6-fold, providing evidence that most, but not all,oft1pollen deficiencies can be attributed to a reduced ability to penetrate through the stigma-style barrier. This method outlines a genetic strategy to quantify a mutation’s impact on the ability of pollen to navigate through the stigma-style barrier on its journey to the ovule.