scholarly journals Isolation and Functional Characterization of Two SHORT VEGETATIVE PHASE Homologous Genes from Mango

2021 ◽  
Vol 22 (18) ◽  
pp. 9802
Author(s):  
Xiao Mo ◽  
Cong Luo ◽  
Haixia Yu ◽  
Jinwen Chen ◽  
Yuan Liu ◽  
...  

The SHORT VEGETATIVE PHASE (SVP) gene is a transcription factor that integrates flowering signals and plays an important role in the regulation of flowering time in many plants. In this study, two full-length cDNA sequences of SVP homologous genes—MiSVP1 and MiSVP2—were obtained from ‘SiJiMi’ mango. Sequence analysis showed that the MiSVPs had typical MADS-box domains and were highly conserved between each other. The analysis of expression patterns showed that the MiSVPs were expressed during flower development and highly expressed in vegetative tissues, with low expression in flowers/buds. The MiSVPs could responded to low temperature, NaCl, and PEG treatment. Subcellular localization revealed that MiSVP1 and MiSVP2 were localized in the nucleus. Transformation of Arabidopsis revealed that overexpression of MiSVP1 delayed flowering time, overexpression of MiSVP2 accelerated flowering time, and neither MiSVP1 nor MiSVP2 had an effect on the number of rosette leaves. Overexpression of MiSVP1 increased the expression of AtFLC and decreased the expression of AtFT and AtSOC1, and overexpression of MiSVP2 increased the expression levels of AtSOC1 and AtFT and decreased the expression levels of AtFLC. Point-to-point and bimolecular fluorescence complementation (BiFC) assays showed that MiSVP1 and MiSVP2 could interact with SEP1-1, SOC1D, and AP1-2. These results suggest that MiSVP1 and MiSVP2 may play a significant roles in the flowering process of mango.

2018 ◽  
Vol 143 (2) ◽  
pp. 101-109
Author(s):  
Jiaqi Hu ◽  
Hye-Ji Kim ◽  
Houbin Chen ◽  
Biyan Zhou

Short vegetative phase (SVP), a MADS-domain transcription factor, was shown to act as a repressor of flowering in arabidopsis (Arabidopsis thaliana). Although the role of SVPs in flowering is well characterized in the model plant arabidopsis, little is known in evergreen woody litchi (Litchi chinensis). In this study, three litchi SVP homologs (LcSVP1, LcSVP2, and LcSVP3) were cloned, and the bioinformatic analysis of the LcSVPs was carried out to identify their molecular characteristics. Their expression patterns in the apical meristem (AM) during the transition from vegetative to reproductive phase were studied under natural flowering inductive conditions. Also, brassinosteroid (BR) treatment under low temperature conditions was performed to elucidate the role of LcSVPs in the BR-regulated flowering. The results showed that LcSVPs belonged to the MADS superfamily. LcSVP relative expression levels in AMs of the early- and late-flowering cultivars showed decreasing trends with the transition from vegetative to reproductive growth. Under low temperature condition, relative expression levels of LcSVP1, LcSVP2, and LcSVP3 in AMs or panicle primordia showed decreasing trends, whereas those in the AMs of the BR-treated trees remained at relatively high levels. Relative expression analysis of the litchi homolog, flowering locus t 1 (LcFT1), showed that the BR-treated leaves had lower relative expression level than nontreated control leaves. The findings suggest that LcSVPs act as repressors involved in flowering in natural conditions and the BR-regulated flowering.


2020 ◽  
Author(s):  
Cuiyu Liu ◽  
Yujie Zhao ◽  
Xueqing Zhao ◽  
Jianmei Dong ◽  
Zhaohe Yuan

Abstract Backgrounds: Pomegranate (Punica granatum L.) is an important commercial fruit tree, with moderate tolerance to salinity. The balance of Cl− and other anions in pomegranate tissues are affected by salinity, however, the accumulation patterns of anions are poorly understood. The chloride channel (CLC) gene family is involved in conducting Cl−, NO3−, HCO3− and I−, but its characteristics have not been reported on pomegranate.Results: In this study, we identified seven PgCLC genes, consisting of four antiporters and three channels, based on the presence of the gating glutamate (E) and the proton glutamate (E). Phylogenetic analysis revealed that seven PgCLCs were divided into two clades, with clade I containing the typical conserved regions GxGIPE (I), GKxGPxxH (II) and PxxGxLF (III), whereas clade II not. Multiple sequence alignment revealed that PgCLC-B had a P [proline, Pro] residue in region I, which was suspected to be a NO3–/H+ exchanger, while PgCLC-C1, PgCLC-C2, PgCLC-D and PgCLC-G contained a S [serine, Ser] residue, with a high affinity to Cl−. We determined the content of Cl−, NO3−, H2PO4−, and SO42− in pomegranate tissues after 18 days of salt treatments (0, 100, 200 and 300 mM NaCl). Compared with control, the Cl− content increased sharply in pomegranate tissues. Salinity inhibited the uptake of NO3− and SO42− , but accelerated H2PO4− uptake. The results of real-time reverse transcription PCR (qRT-PCR) revealed that PgCLC genes had tissue-specific expression patterns. The high expression levels of three antiporters PgCLC-C1, PgCLC-C2 and PgCLC-D in leaves might be contributed to sequestrating Cl− into the vacuoles. However, the low expression levels of PgCLCs in roots might be associated with the exclusion of Cl− from root cells. Also, the up-regulated PgCLC-B in leaves indicated that more NO3− was transported into leaves to mitigate the nitrogen deficiency.Conclusions: Our findings suggested that the PgCLC genes played important roles in balancing of Cl− and NO3− in pomegranate tissues under salt stress. This study establishes a theoretical foundation for the further functional characterization of CLC genes in pomegranate.


Development ◽  
2020 ◽  
Vol 148 (1) ◽  
pp. dev193870
Author(s):  
Hendry Susila ◽  
Zeeshan Nasim ◽  
Katarzyna Gawarecka ◽  
Ji-Yul Jung ◽  
Suhyun Jin ◽  
...  

ABSTRACTPHOSPHORYLETHANOLAMINE CYTIDYLYLTRANSFERASE 1 (PECT1) regulates phosphatidylethanolamine biosynthesis and controls the phosphatidylethanolamine:phosphatidylcholine ratio in Arabidopsis thaliana. Previous studies have suggested that PECT1 regulates flowering time by modulating the interaction between phosphatidylcholine and FLOWERING LOCUS T (FT), a florigen, in the shoot apical meristem (SAM). Here, we show that knockdown of PECT1 by artificial microRNA in the SAM (pFD::amiR-PECT1) accelerated flowering under inductive and even non-inductive conditions, in which FT transcription is almost absent, and in ft-10 twin sister of ft-1 double mutants under both conditions. Transcriptome analyses suggested that PECT1 affects flowering by regulating SHORT VEGETATIVE PHASE (SVP) and GIBBERELLIN 20 OXIDASE 2 (GA20ox2). SVP misexpression in the SAM suppressed the early flowering of pFD::amiR-PECT1 plants. pFD::amiR-PECT1 plants showed increased gibberellin (GA) levels in the SAM, concomitant with the reduction of REPRESSOR OF GA1-3 levels. Consistent with this, GA treatment had little effect on flowering time of pFD::amiR-PECT1 plants and the GA antagonist paclobutrazol strongly affected flowering in these plants. Together, these results suggest that PECT1 also regulates flowering time through a florigen-independent pathway, modulating SVP expression and thus regulating GA production.


2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Cuiyu Liu ◽  
Yujie Zhao ◽  
Xueqing Zhao ◽  
Jianmei Dong ◽  
Zhaohe Yuan

Abstract Backgrounds Pomegranate (Punica granatum L.) is an important commercial fruit tree, with moderate tolerance to salinity. The balance of Cl− and other anions in pomegranate tissues are affected by salinity, however, the accumulation patterns of anions are poorly understood. The chloride channel (CLC) gene family is involved in conducting Cl−, NO3−, HCO3− and I−, but its characteristics have not been reported on pomegranate. Results In this study, we identified seven PgCLC genes, consisting of four antiporters and three channels, based on the presence of the gating glutamate (E) and the proton glutamate (E). Phylogenetic analysis revealed that seven PgCLCs were divided into two clades, with clade I containing the typical conserved regions GxGIPE (I), GKxGPxxH (II) and PxxGxLF (III), whereas clade II not. Multiple sequence alignment revealed that PgCLC-B had a P [proline, Pro] residue in region I, which was suspected to be a NO3−/H+ exchanger, while PgCLC-C1, PgCLC-C2, PgCLC-D and PgCLC-G contained a S [serine, Ser] residue, with a high affinity to Cl−. We determined the content of Cl−, NO3−, H2PO4−, and SO42− in pomegranate tissues after 18 days of salt treatments (0, 100, 200 and 300 mM NaCl). Compared with control, the Cl− content increased sharply in pomegranate tissues. Salinity inhibited the uptake of NO3− and SO42−, but accelerated H2PO4− uptake. The results of real-time reverse transcription PCR (qRT-PCR) revealed that PgCLC genes had tissue-specific expression patterns. The high expression levels of three antiporters PgCLC-C1, PgCLC-C2 and PgCLC-D in leaves might be contributed to sequestrating Cl− into the vacuoles. However, the low expression levels of PgCLCs in roots might be associated with the exclusion of Cl− from root cells. Also, the up-regulated PgCLC-B in leaves indicated that more NO3− was transported into leaves to mitigate the nitrogen deficiency. Conclusions Our findings suggested that the PgCLC genes played important roles in balancing of Cl− and NO3− in pomegranate tissues under salt stress. This study established a theoretical foundation for the further functional characterization of the CLC genes in pomegranate.


2021 ◽  
Author(s):  
Xiaohong Zhang ◽  
Zhongying Ren ◽  
Genhai Hu ◽  
Hengling Wei ◽  
Shuli Fan ◽  
...  

Abstract The AP1/FUL transcription factors are important for floral development, but the underlying molecular mechanisms remain unclear. In this study, we cloned and identified two AP1/FUL-like genes, GhAP1.1 and GhFUL2, in upland cotton, which is a commonly cultivated economically valuable crop. Sequence alignments and phylogenetic analyses indicated GhAP1.1 and GhFUL2, which are encoded by genes in the AP1/FUL clade, have conserved N-terminals, but diverse C-terminal domains. A quantitative real-time PCR analysis revealed that GhAP1.1 and GhFUL2 were expressed in the flower and root, and had the opposite expression patterns during different shoot apical meristem stages. The upregulated expression of GhAP1.1 in Arabidopsis and the silencing of GhAP1.1 did not induce significant changes to the flowering time or floral organs, but the transcript levels of the florigen FT gene and the AP1 homolog GhMADS42 increased. The overexpression of GhFUL2 in Arabidopsis delayed flowering and promoted bolting by decreasing the FT and LFY expression levels. Silencing GhFUL2 in cotton dramatically increased the expression of GhFT and GhMADS42 and promoted flowering. Additionally, yeast two-hybrid and bimolecular fluorescence complementation assays indicated that GhAP1.1 can interact with the SVP homolog GhSVP1, whereas GhFUL2 can form heterodimers with SEP1, SEP4 homologs, and GhSVP1. Therefore, we proved that the functional divergence of GhAP1.1 and GhFUL2, which involved changes in sequences and expression patterns, influenced the regulation of cotton flower development.


2020 ◽  
Author(s):  
Cuiyu Liu ◽  
Yujie Zhao ◽  
Xueqing Zhao ◽  
Jianmei Dong ◽  
Zhaohe Yuan

Abstract BackgroundsPomegranate (Punica granatum L.) is an important commercial fruit tree, with moderate tolerance to salinity. The balance of Cl− and other anions in pomegranate tissues are affected by salinity, however, the accumulation patterns of anions are poorly understood. The chloride channel (CLC) gene family is involved in conducting Cl−, NO3−, HCO3− and I−, but its characteristics have not been reported on pomegranate.ResultsIn this study, we identified seven PgCLC genes, consisting of four antiporters and three channels, based on the presence of the gating glutamate (E) and the proton glutamate (E). Phylogenetic analysis revealed that seven PgCLCs were divided into two clades, with clade I containing the typical conserved regions GxGIPE (I), GKxGPxxH (II) and PxxGxLF (III), whereas clade II not. Multiple sequence alignment revealed that PgCLC-B had a P [proline, Pro] residue in region I, which is suspected to be a NO3–/H+ exchanger, while PgCLC-C1, PgCLC-C2, PgCLC-D and PgCLC-G contained a S [serine, Ser] residue, with a high affinity to Cl−. We were determined the content of Cl−, NO3−, H2PO4−, and SO42− in pomegranate tissues after 18 days of salt treatments (0, 100, 200 and 300 mM NaCl). Compared with control, the Cl− content increased sharply in tissues and was ranked as leaf > stem > root. The uptake of NO3− and SO42− was inhibited by high salinity, while that of H2PO4− increased. The results of real-time reverse transcription PCR (qRT-PCR) revealed that PgCLC genes had tissue-specific expression patterns. The high expression levels of PgCLC-C1, PgCLC-C2 and PgCLC-D in leaves suggested they played roles in sequestrating Cl− into the vacuoles. However, the low expression levels of PgCLCs in roots might be contributed to the exclusion of Cl− from root cells. Also, the non-significantly changed concentration of NO3− in leaves and the up-regulated PgCLC-B indicated an acceleration of transporting NO3− into leaves to mitigate the nitrogen deficiency.ConclusionsOur findings suggested that PgCLC genes played important roles in balance of Cl− and NO3− in pomegranate tissues under salt stress. This study establishes a theoretical foundation for the further functional characterization of CLC genes in pomegranate.


2020 ◽  
Author(s):  
Cuiyu Liu ◽  
Yujie Zhao ◽  
Xueqing Zhao ◽  
Jianmei Dong ◽  
Zhaohe Yuan

Abstract Backgrounds: Pomegranate (Punica granatum L.) is an important commercial fruit tree, with moderate tolerance to salinity. The balance of Cl− and other anions in pomegranate tissues are affected by salinity, however, the accumulation patterns of anions are poorly understood. The chloride channel (CLC) gene family is involved in conducting Cl−, NO3−, HCO3− and I−, but its characteristics have not been reported on pomegranate.Results: In this study, we identified seven PgCLC genes, consisting of four antiporters and three channels, based on the presence of the gating glutamate (E) and the proton glutamate (E). Phylogenetic analysis revealed that seven PgCLCs were divided into two clades, with clade I containing the typical conserved regions GxGIPE (I), GKxGPxxH (II) and PxxGxLF (III), whereas clade II not. Multiple sequence alignment revealed that PgCLC-B had a P [proline, Pro] residue in region I, which was suspected to be a NO3–/H+ exchanger, while PgCLC-C1, PgCLC-C2, PgCLC-D and PgCLC-G contained a S [serine, Ser] residue, with a high affinity to Cl−. We determined the content of Cl−, NO3−, H2PO4−, and SO42− in pomegranate tissues after 18 days of salt treatments (0, 100, 200 and 300 mM NaCl). Compared with control, the Cl− content increased sharply in pomegranate tissues. Salinity inhibited the uptake of NO3− and SO42− , but accelerated H2PO4− uptake. The results of real-time reverse transcription PCR (qRT-PCR) revealed that PgCLC genes had tissue-specific expression patterns. The high expression levels of three antiporters PgCLC-C1, PgCLC-C2 and PgCLC-D in leaves might be contributed to sequestrating Cl− into the vacuoles. However, the low expression levels of PgCLCs in roots might be associated with the exclusion of Cl− from root cells. Also, the up-regulated PgCLC-B in leaves indicated that more NO3− was transported into leaves to mitigate the nitrogen deficiency.Conclusions: Our findings suggested that the PgCLC genes played important roles in balancing of Cl− and NO3− in pomegranate tissues under salt stress. This study established a theoretical foundation for the further functional characterization of CLC genes in pomegranate.


PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e8249
Author(s):  
Huifeng Li ◽  
Kun Ran ◽  
Qinglong Dong ◽  
Qiang Zhao ◽  
Song Shi

Background NAC transcription factors play important roles in the regulation of plant growth, development, abiotic and biotic stress responses. The transcriptional level of MdNACs in different tissues and under various biotic and abiotic stress treatments was determined to provide a solid foundation for studying the function of MdNACs. Methods Thirty-two full-length cDNA sequences of Md NACs were isolated by homologous comparison and RT-PCR confirmation, and the obtained cDNA sequences and the deduced amino acid sequences were analyzed with bioinformatics methods. The prediction of subcellular locations of MdNAC proteins was performed using CELLO v.2.5, PSORT, and SoftBerry ProtComp 9.0. Expression levels of MdNACs were detected in 16 different tissues using an array. Expression patterns of MdNACs were detected in response to Alternaria alternata apple pathotype (AAAP) infection using RNA-seq, and the expression of MdNACs was analyzed under NaCl and mannitol treatments using RT-qPCR. Results The sequencing results produced 32 cDNAs (designated as MdNAC24-39, MdNAC54-65, and MdNAC67-70 with GenBank accession No. MG099861–MG099876, MG099891–MG099902, and MG099904–MG099907, respectively). Phylogenetic analysis revealed that MdNAC34 belonged to the ATAF group, MdNAC63 belonged to the AtNAC3 group, MdNAC24, MdNAC26-30, MdNAC32-33, MdNAC35, MdNAC37-39, MdNAC56-57, MdNAC59-62, MdNAC64-65, and MdNAC67-70 belonged to the NAM group, and MdNAC25, MdNAC36, MdNAC54-55, and MdNAC58 belonged to the VND group. Predictions of subcellular localization showed that MdNAC24-27, MdNAC29-30, MdNAC33-37, MdNAC39, MdNAC54-65, and MdNAC67-70 proteins were located in the nucleus, MdNAC28 proteins were located in the cytoplasm, MdNAC31-32 proteins were located in the nucleus and cytoplasm, and MdNAC38 proteins were located in the nucleus and plasma membrane. Array results indicated that 32 MdNACs were expressed in all examined tissues at various expression levels. RNA-seq results showed that expression levels of MdNAC26-28, MdNAC33-34, MdNAC60, MdNAC62-65, and MdNAC68 were induced, but MdNAC24, MdNAC32, and MdNAC58 were down-regulated in response to AAAP infection. Under salt treatment, MdNAC24, MdNAC27, MdNAC29, MdNAC34, MdNAC37, MdNAC39, MdNAC54, MdNAC59, and MdNAC63 transcription levels were induced. Under mannitol treatment, MdNAC32 and MdNAC54 transcription levels were induced, but MdNAC24, MdNAC28, MdNAC30, MdNAC33, MdNAC35, MdNAC37, MdNAC55, MdNAC56, MdNAC58, and MdNAC59 were down-regulated. Taken together, the results indicated that the cloned MdNAC genes were expressed constitutively in all examined tissues. These genes were up-regulated or down-regulated in response to AAAP infection and to salt or mannitol, which suggested they may be involved in the regulation of growth, development, and stress response in apple.


2021 ◽  
Author(s):  
Xiaohong Zhang ◽  
Zhongying Ren ◽  
Genhai Hu ◽  
Hengling Wei ◽  
Shuli Fan ◽  
...  

Abstract The AP1/FUL transcription factors are important for floral development, but the underlying molecular mechanisms remain unclear. In this study, we cloned and identified two AP1/FUL-like genes, GhAP1.1 and GhFUL2, in upland cotton, which is a commonly cultivated economically valuable crop. Sequence alignments and phylogenetic analyses indicated GhAP1.1 and GhFUL2, which are encoded by genes in the AP1/FUL clade, have conserved N-terminals, but diverse C-terminal domains. A quantitative real-time PCR analysis revealed that GhAP1.1 and GhFUL2 were expressed in the flower and root, and had the opposite expression patterns during different shoot apical meristem stages. The upregulated expression of GhAP1.1 in Arabidopsis and the silencing of GhAP1.1 did not induce significant changes to the flowering time or floral organs, but the transcript levels of the florigen FT gene and the AP1 homolog GhMADS42 increased. The overexpression of GhFUL2 in Arabidopsis delayed flowering and promoted bolting by decreasing the FT and LFY expression levels. Silencing GhFUL2 in cotton dramatically increased the expression of GhFT and GhMADS42 and promoted flowering. Additionally, yeast two-hybrid and bimolecular fluorescence complementation assays indicated that GhAP1.1 can interact with the SVP homolog GhSVP2.2, whereas GhFUL2 can form heterodimers with GhSEP3/GhSEP4 homologs, and GhSVP2.2. Therefore, we proved that the functional divergence of GhAP1.1 and GhFUL2, which involved changes in sequences and expression patterns, influenced the regulation of cotton flower development and plant architecture.


2020 ◽  
Vol 52 (3) ◽  
pp. 244-251
Author(s):  
Joon Ki Hong ◽  
Sang-Ryeol Park ◽  
Eun Jung Suh ◽  
Jihee Park ◽  
Yeon-Hee Lee

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