Detection of Weak Expression of SOLO DANCERS in the Male Germline Using CYCLIN-DEPENDENT KINASE A1 Coding Sequence
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Detection of Weak Expression of SOLO DANCERS in the Male Germline Using CYCLIN‑DEPENDENT KINASE A1 Coding Sequence Yanru Zhen1 · Jilei Huang2 · Xiaojing Chen1 · Chuxiong Zhuang1,3 · Yufei Hu1 Received: 14 April 2019 / Accepted: 14 November 2019 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Male germline specification is a crucial step in double fertilization in flowering plants. Determining which genes are activated in the male germline, and how those genes are regulated, is essential to understand double fertilization. However, transcription activities in the male germline may be not easy to detect due to low-level expression of some genes and technical difficulties of isolating male germline cells. Here, through a series of gene reporter assays in Arabidopsis, we showed a weak male germline expression pattern of the SOLO DANCERS (SDS) gene, which was confirmed by RT-PCR. Compared to directly fusing the SDS sequence with GFP, adding the coding sequences of other genes such as CDKA1 can greatly enhance the detection of the male germline expression pattern of SDS. We found that SDS expression in the male germline is activated by a novel pathway that differs from the well-known DUO1 regulon. We also developed an SDS-based fluorescence reporter to analyze posttranscriptional regulation in the male germline. Our data suggest that stable gene products of CDKA1 and others may enhance the sensitivity of gene reporters, and the male germline may use diverse pathways to activate gene expression. Keywords SDS · CDKA1 · Fluorescence reporter · Male germline · Arabidopsis
Introduction Flowering plants reproduce through double fertilization. For successful double fertilization, a microspore cell divides asymmetrically into a generative cell and a vegetative cell. The generative cell further divides into two sperm cells, which finally fuse with the egg or central cell in the embryo sac (Berger and Twell 2011). The male germline, which Yanru Zhen and Jilei Huang have contributed equally to this publication. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00344-019-10061-8) contains supplementary material, which is available to authorized users. * Yufei Hu [email protected] 1
College of Life Sciences, South China Agricultural University, Guangzhou 510642, China
2
Instrumental Analysis and Research Center, South China Agricultural University, Guangzhou 510642, China
3
State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangzhou 510642, China
includes generative cells and sperm cells, is highly specialized when compared to vegetative cells, which develop into pollen tubes. How flowering plants regulate gene expression to specify the fate of the male germline is a key question in sexual reproduction. In recent years, several groups have obtained large datasets concerning which genes are expressed in male germline cells. Centrifugation-based or cell-sorting technologies were used to purify genera
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