A novel ERF transcription factor, CmERF12, negatively influences chrysanthemum embryo development

Author(s):  
Sujuan Xu ◽  
Huizhong Hou ◽  
Ze Wu ◽  
Jingya Zhao ◽  
Fengjiao Zhang ◽  
...  

Abstract Embryo abortion often occurs during distant hybridization events. Apetala 2/ethylene-responsive factor (AP2/ERF) proteins are key transcription factor (TF) regulators of plant development and stress resistance, but their roles in hybrid embryo development are poorly understood. We isolated a novel AP2/ERF TF, CmERF12, from chrysanthemum and showed that it adversely affects embryo development during distant hybridization. Transcriptome and real-time quantitative PCR data demonstrated that CmERF12 is expressed at significantly higher levels in aborted ovaries compared with normal ovaries. CmERF12 localizes to the cell nucleus and contains a conserved EAR motif that mediates its transcription repressor function in yeast and plant cells. We generated an amiR-CmERF12 transgenic Chrysanthemum morifolium (C.m.) var. ‘Yuhualuoying’ and conducted distant hybridization with the wild-type tetraploid, Chrysanthemum nankingense (C.n.), revealing that CmERF12 knockdown significantly promoted embryo development and increased the seed setting rates during hybridization. The expression of various embryo development-related genes was up-regulated in developing ovaries from the ♀amiR-CmERF12-C.m. × ♂C.n. cross. Furthermore, CmERF12 directly interacted with CmSUF4 and significantly reduced its ability to activate its target gene CmEC1. Overall, we invented an original method to overcome plant distant hybridization barriers and unraveled the mechanism by which CmERF12 negatively affects chrysanthemum embryo development.

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Sujuan Xu ◽  
Ze Wu ◽  
Huizhong Hou ◽  
Jingya Zhao ◽  
Fengjiao Zhang ◽  
...  

AbstractDistant hybridization is widely used to develop crop cultivars, whereas the hybridization process of embryo abortion often severely reduces the sought-after breeding effect. The LEAFY COTYLEDON1 (LEC1) gene has been extensively investigated as a central regulator of seed development, but it is far less studied in crop hybridization breeding. Here we investigated the function and regulation mechanism of CmLEC1 from Chrysanthemum morifolium during its seed development in chrysanthemum hybridization. CmLEC1 encodes a nucleic protein and is specifically expressed in embryos. CmLEC1’s overexpression significantly promoted the seed-setting rate of the cross, while the rate was significantly decreased in the amiR-CmLEC1 transgenic chrysanthemum. The RNA-Seq analysis of the developing hybrid embryos revealed that regulatory genes involved in seed development, namely, CmLEA (late embryogenesis abundant protein), CmOLE (oleosin), CmSSP (seed storage protein), and CmEM (embryonic protein), were upregulated in the OE (overexpressing) lines but downregulated in the amiR lines vs. wild-type lines. Future analysis demonstrated that CmLEC1 directly activated CmLEA expression and interacted with CmC3H, and this CmLEC1–CmC3H interaction could enhance the transactivation ability of CmLEC1 for the expression of CmLEA. Further, CmLEC1 was able to induce several other key genes related to embryo development. Taken together, our results show that CmLEC1 plays a positive role in the hybrid embryo development of chrysanthemum plants, which might involve activating CmLEA’s expression and interacting with CmC3H. This may be a new pathway in the LEC1 regulatory network to promote seed development, one perhaps leading to a novel strategy to not only overcome embryo abortion during crop breeding but also increase the seed yield.


Author(s):  
Peiqi An ◽  
Qing Cao ◽  
Chen Wang ◽  
Junhun Wang ◽  
Hanguo Zhang ◽  
...  

Abstract Larix olgensis is one of the main coniferous tree species in northeastern China and has excellent timber properties and strong tolerance to stress. Thirteen HD-Zip family genes with a complete CDS region were identified on the basis of cambium transcriptome data from Larix olgensis. All 13 genes were analyzed via bioinformatics by their conserved domain protein sequence and amino acid composition, including their physicochemical properties and protein structure. The spatiotemporal expression and abiotic stress responses of these genes were analyzed by real-time quantitative PCR. The results showed that the 13 HD-Zip genes of Larix olgensis were expressed in the roots, stems, and leaves at different stages. The expression of three of these genes (LoHDZ2, LoHDZ11, LoHDZ13) was highest in nonlignified roots, indicating that they might be related to the secondary growth of Larix olgensis; in addition, three genes (LoHDZ5, LoHDZ9, LoHDZ10) were highly expressed in partially and completely lignified stems and leaves. These 13 genes were expressed specifically under drought stress. The expression of two of them (LoHDZ1, LoHDZ5) was obviously upregulated, and the expression of 6 genes (LoHDZ2, LoHDZ3, LoHDZ4, LoHDZ8, LoHDZ10, LoHDZ13) was significantly downregulated. The expression trends indicate that these genes could be involved in drought stress. The expression of all 13 genes was downregulated when the plants were treated with 0.2 M NaCl for 96 h, indicating that these genes are inhibited by salt stress. Overall, the results have significant implications for the study of the gene function of members of the LoHD-Zip transcription factor family.


2020 ◽  
Vol 32 (2) ◽  
pp. 167
Author(s):  
D. Lee ◽  
K. Choi ◽  
J. Oh ◽  
S. Kim ◽  
M. Lee ◽  
...  

Recent studies suggest that endogenous and exogenous free fatty acids play various important roles in mammalian oocyte and pre-implantation embryo development. Among fatty acids, linoleic acid (LA) has been reported to affect the apoptosis pathway via nuclear transcription factor-kappa B (NF-κB). The transcription factor NF-κB is a key modulator of apoptosis in a variety of cell types, but to date, this specific function of NF-κB has not been demonstrated in porcine pre-implantation embryos. To examine the effect of linoleic acid on invitro-produced parthenogenetic pig embryos, we treated LA by concentration (0, 10, 25, 50, and 100 µM) to identify developmental rate, NF-κB expression, and mRNA level of apoptotic-related genes. In addition, the mechanism was confirmed by examining the protein and mRNA expression of NF-kb and c-jun by immunostaining and quantitative PCR at the blastocyst stage. Linoleic acid had a positive effect on embryo development without toxicity at a certain concentration (25 µM), but toxicity was confirmed at higher (50-100μM) concentrations. Furthermore, it was confirmed that the concentration of NF-κB increased as the treatment concentration of LA increased, which was found to increase even at the concentration at which embryo development decreased. Previous studies have shown that the NF-κB pathway is involved in regulating anti- and pro-apoptotic gene expression. We also investigated the effects of LA on anti- (Bcl-xL, Mcl-1) and pro- (BAX1, TP53, Caspase3) apoptotic genes and NF-κB activation-related genes (RelA, JNK1, JNK2, IL-6) in porcine embryos. We have found that down-regulation of pro-apoptotic gene expression occurs in the LA-treated group. It was also found that Bcl-xL, one of the anti-apoptotic genes, was not affected by LA, which appears to be an effect of IL-6. In contrast, Mcl-1, an anti-apoptotic gene known not to be affected by IL-6, was found to have increased expression mRNA level in LA-treated pig embryos. Furthermore, through double-staining of apoptosis and immunocytochemistry, as the concentration of NF-kB level increases, the nuclear translocation of c-jun, the protein of which was also related with apoptosis, increased gradually depending on the LA concentration. These data could support that porcine embryo can use exogenous LA as a metabolic energy source. The data also demonstrate the important role of NF-kB in porcine early embryo development. Support was provided by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET) through the Development of High Value-Added Food Technology program funded by the Ministry of Agriculture, Food and Rural Affairs (MAFRA, 118042-03-1-HD020).


2019 ◽  
Vol 70 (21) ◽  
pp. 6229-6244
Author(s):  
Gang Li ◽  
Wenxuan Zou ◽  
Liufang Jian ◽  
Jie Qian ◽  
Jie Zhao

Abstract Embryogenesis is an essential process during seed development in higher plants. It has previously been shown that mutation of the Arabidopsis non-SMC element genes AtNSE1 or AtNSE3 leads to early embryo abortion, and their proteins can interact with each other directly. However, the crucial regions of these proteins in this interaction and how the proteins are cytologically involved in Arabidopsis embryo development are unknown. In this study, we found that the C-terminal including the Ring-like motif of AtNSE1 can interact with the N-terminal of AtNSE3, and only the Ring-like motif is essential for binding with three α motifs of AtNSE2 (homologous to AtMMS21). Using genetic assays and by analysing molecular markers of cell fate decisions (STM, WOX5, and WOX8) in mutant nse1 and nse3 embryos, we found that AtNSE1 and AtNSE3 work non-redundantly in early embryo development, and that differentiation of the apical meristem and the hypophysis fails in the mutants, which have disrupted auxin transportation and responses. However, the upper cells of the suspensor in the mutants seem to have proper embryo cell identity. Cytological examination showed that cell death occurred from the early embryo stage, and that vacuolar programmed cell death and necrosis in the nse1 and nse3 mutant embryos led to ovule abortion. Thus, AtNSE1 and AtNSE3 are essential for maintaining cell viability and growth during early embryogenesis. Our results improve our understanding of the functions of SMC5/6 complex in early embryogenesis in Arabidopsis.


Heredity ◽  
1994 ◽  
Vol 72 (2) ◽  
pp. 193-200 ◽  
Author(s):  
S Abbo ◽  
G Ladizinsky

2011 ◽  
Vol 68 (4) ◽  
pp. 583-596 ◽  
Author(s):  
Jing Lu ◽  
Hongping Ju ◽  
Guoxin Zhou ◽  
Chuanshu Zhu ◽  
Matthias Erb ◽  
...  

2015 ◽  
Vol 168 (3) ◽  
pp. 1025-1035 ◽  
Author(s):  
John Danzer ◽  
Eric Mellott ◽  
Anhthu Q. Bui ◽  
Brandon H. Le ◽  
Patrick Martin ◽  
...  

2011 ◽  
Vol 23 (2) ◽  
pp. 353 ◽  
Author(s):  
Damien B. B. P. Paris ◽  
Ewart W. Kuijk ◽  
Bernard A. J. Roelen ◽  
Tom A. E. Stout

Real-time quantitative PCR (qPCR) is invaluable for investigating changes in gene expression during early development, since it can be performed on the limited quantities of mRNA contained in individual embryos. However, the reliability of this method depends on the use of validated stably expressed reference genes for accurate data normalisation. The aim of the present study was to identify and validate a set of reference genes suitable for studying gene expression during equine embryo development. The stable expression of four carefully selected reference genes and one developmentally regulated gene was examined by qPCR in equine in vivo embryos from morula to expanded blastocyst stage. SRP14, RPL4 and PGK1 were identified by geNorm analysis as stably expressed reference genes suitable for data normalisation. RPL13A expression was less stable and changed significantly during the period of development examined, rendering it unsuitable as a reference gene. As anticipated, CDX2 expression increased significantly during embryo development, supporting its possible role in trophectoderm specification in the horse. In summary, it was demonstrated that evidence-based selection of potential reference genes can reduce the number needed to validate stable expression in an experimental system; this is particularly useful when dealing with tissues that yield small amounts of mRNA. SRP14, RPL4 and PGK1 are stable reference genes suitable for normalising expression for genes of interest during in vivo morula to expanded blastocyst development of horse embryos.


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