scholarly journals Isolation, Expression, and Promoter Analysis ofGbWRKY2: A Novel Transcription Factor Gene fromGinkgo biloba

2015 ◽  
Vol 2015 ◽  
pp. 1-17 ◽  
Author(s):  
Yong-Ling Liao ◽  
Yong-Bao Shen ◽  
Jie Chang ◽  
Wei-Wei Zhang ◽  
Shui-Yuan Cheng ◽  
...  

WRKY transcription factor is involved in multiple life activities including plant growth and development as well as biotic and abiotic responses. We identified 28WRKYgenes from transcriptome data ofGinkgo bilobaaccording to conserved WRKY domains and zinc finger structure and selected three WRKY genes, which areGbWRKY2, GbWRKY16, andGbWRKY21, for expression pattern analysis.GbWRKY2was preferentially expressed in flowers and strongly induced by methyl jasmonate. Here, we cloned the full-length cDNA and genomic DNA ofGbWRKY2. The full-length cDNA ofGbWRKY2was 1,713 bp containing a 1,014 bp open reading frame encoding a polypeptide of 337 amino acids. TheGbWRKY2genomic DNA had one intron and two exons. The deducedGbWRKY2contained one WRKY domain and one zinc finger motif.GbWRKY2was classified into Group II WRKYs. Southern blot analysis revealed thatGbWRKY2was a single copy gene inG. biloba. Manycis-acting elements related to hormone and stress responses were identified in the 1,363 bp-length 5′-flanking sequence ofGbWRKY2, including W-box, ABRE-motif, MYBCOREs, and PYRIMIDINE-boxes, revealing the molecular mechanism of upregulated expression ofGbWRKY2by hormone and stress treatments. Further functional characterizations in transiently transformed tobacco leaves allowed us to identify the region that can be considered as the minimal promoter.

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e7291 ◽  
Author(s):  
Lei Wang ◽  
Wenjing Yao ◽  
Yao Sun ◽  
Jiying Wang ◽  
Tingbo Jiang

The WRKY transcription factor family is one of the largest groups of transcription factor in plants, playing important roles in growth, development, and biotic and abiotic stress responses. Many WRKY genes have been cloned from a variety of plant species and their functions have been analyzed. However, the studies on WRKY transcription factors in tree species under abiotic stress are still not well characterized. To understand the effects of the WRKY gene in response to abiotic stress, mRNA abundances of 102 WRKY genes in Populus simonii × P. nigra were identified by RNA sequencing under normal and salt stress conditions. The expression of 23 WRKY genes varied remarkably, in a tissue-specific manner, under salt stress. Since the WRKY56 was one of the genes significantly induced by NaCl treatment, its cDNA fragment containing an open reading frame from P. simonii × P. nigra was then cloned and transferred into Arabidopsis using the floral dip method. Under salt stress, the transgenic Arabidopsis over-expressed the WRKY56 gene, showing an increase in fresh weight, germination rate, proline content, and peroxidase and superoxide dismutase activity, when compared with the wild type. In contrast, transgenic Arabidopsis displayed a decrease in malondialdehyde content under salt stress. Overall, these results indicated that the WRKY56 gene played an important role in regulating salt tolerance in transgenic Arabidopsis.


2004 ◽  
Vol 31 (1) ◽  
pp. 73 ◽  
Author(s):  
Zhihua Liao ◽  
Qiumin Tan ◽  
Yourong Chai ◽  
Kaijing Zuo ◽  
Min Chen ◽  
...  

In plants, the first committed step in the pathway for biosynthesis of isoprenoids is catalysed by 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR, EC: 1.1.1.34). Here we report for the first time the cloning of a full-length cDNA encoding HMGR (Tm–HMGR) from a taxol-producing gymnosperm, Taxus media Rehder. The full-length cDNA of Tm–HMGR (GenBank accession number: AY277740) was 2307 base pairs (bp), with a 1791-bp open reading frame (ORF) encoding a 596-amino-acid polypeptide. Bioinformatic analysis revealed that Tm–HMGR contained two trans-membrane domains and a catalytic domain, and showed high homology to other plant HMGRs. Phylogenetic analysis indicated that Tm–HMGR was more ancient than other plant HMGRs. The structural modelling showed that Tm–HMGR had the typical spatial structure of HMGRs whose catalytic domains could be folded and divided into three spatial domains, L-domain, N-domain and S-domain. Southern blot analysis revealed that Tm–HMGR belonged to a small HMGR gene family. Northern blot analysis showed that Tm–HMGR was expressed in roots, stems and needles, with higher expression in stems and needles than in roots. Functional complementation of Tm–HMGR in a HMGR-deficient mutant yeast demonstrated that Tm–HMGR mediated the biosynthesis of mevalonate and provided the general precursor for taxol biosynthesis.


2020 ◽  
Vol 32 (8) ◽  
pp. 792 ◽  
Author(s):  
Ruidong Zhang ◽  
Haitao Nie ◽  
Shulong Duan ◽  
Peng Yan ◽  
Ali Izaz ◽  
...  

Kisspeptin1 (Kiss1), a product of the Kiss1 gene, plays an important role in the regulation of reproduction in vertebrates by activating the Kiss1 receptor (Kiss1R) and its coexpression with gonadotrophin-releasing hormone (GnRH) in GnRH neurons. The purpose of this study was to clone the Kiss1 and Kiss1R genes found in the brain of Alligator sinensis and to explore their relationship with reproduction. The full-length cDNA of Kiss1 is 816bp, the open reading frame (ORF) is 417bp and the gene encodes a 138-amino acid precursor protein. The full-length cDNA of Kiss1R is 2348bp, the ORF is 1086bp and the gene encodes a 361-amino acid protein. Quantitative polymerase chain reaction showed that, except for Kiss1R expression in the hypothalamus, the expression of Kiss1 and Kiss1Rduring the reproductive period of A. sinensis was higher than that in the hypothalamus, pituitary gland and ovary during the hibernation period. The changes in GnRH2 mRNA in the hypothalamus were similar to those of GnRH1 and peaked during the reproductive period. This study confirms the existence of Kiss1 and Kiss1R in A. sinensis and the findings strongly suggest that Kiss1 and Kiss1R may participate in the regulation of GnRH secretion in the hypothalamus of alligators during the reproductive period. Furthermore, this is the first report of the full-length cDNA sequences of Kiss1 and Kiss1R in reptiles.


2011 ◽  
Vol 39 (2) ◽  
pp. 254
Author(s):  
Xiuchun DONG ◽  
Zhiyuan CHENG ◽  
Shuhan CHENG ◽  
Feng XU ◽  
Yanrong AN ◽  
...  

A full-length cDNA encoding a nuclear factor-YB (NF-YB)/HAP3/CCAAT binding factor-A (CBF-A) subunit of a CCAAT-box binding complex, designated as TaNF-YB3 was isolated from Triticum aestivum. Sequence analysis indicated that the full-length cDNA was 809 bp long, including an open reading frame (ORF) of 597 bp, which encoded a deduced polypeptide of 199 amino acids and is located in chromosome 3D. The deduced protein contained conserved structural domains and showed high identity to other plant NF-YBs. TaNF-YB3 was expressed in various organs, especially in the leaves and stamens; it was also regulated by salt, mannitol, abscisic acid, wounding, and cold. Moreover, TaNF-YB3 was down-regulated by short days and vernalization, and sensitive to the transfer of day length. It was mainly induced by light and exhibited a similar diurnal rhythmic expression pattern with the CCT-domain family gene VRN2 (TaZCCT1 and TaZCCT2), but not with CO (WCO1 and TaHd1). Overall, the results suggested that TaNF-YB3, aside from having a role in regulating day length and vernalization responses, might integrate signals from other environmental stresses to perform its functions in winter wheat adaptability and development.


2019 ◽  
Vol 110 (2) ◽  
pp. 177-184
Author(s):  
Yu Wang ◽  
Jing Nan Wang ◽  
Xue Zhun Chen ◽  
Qi Xing Hu ◽  
Qi Qing Liu ◽  
...  

Abstractp53, DNA damage regulated gene (PDRG) and apoptosis-stimulating p53 protein 2 (ASPP2) are p53-related genes which can promote apoptosis. The full-length cDNA sequence of the Px-pdrg and Px-aspp2 genes were characterized and their mRNA expression dynamics under heat stress were studied in diamondback moth (DBM) Plutella xylostella collected from Fuzhou, China. The full-length cDNA of Px-pdrg and Px-aspp2 spans 721 and 4201 bp, containing 395 and 3216 bp of the open reading frame, which encode a putative protein comprising 130 and 1072 amino acids with a calculated molecular weight of 14.58 and 118.91 kDa, respectively. As compared to 25°C, both Px-pdrg and Px-aspp2 were upregulated in chlorpyrifos-resistant (Rc) and -susceptible (Sm) strains of DBM adults and pupae under heat stress. In addition, Rc DBM showed a significantly higher expression level of Px-pdrg and Px-aspp2 in contrast to Sm DBM. The results indicate that high temperature can significantly promote apoptosis process, especially in Rc-DBM. Significant fitness cost in Rc-DBM might be associated with drastically higher transcript abundance of Px-pdrg and Px-aspp2 under the heat stress.


1991 ◽  
Vol 277 (2) ◽  
pp. 469-475 ◽  
Author(s):  
R Dumas ◽  
M Lebrun ◽  
R Douce

Acetohydroxy acid reductoisomerase (AHRI), the second enzyme in the parallel isoleucine/valine-biosynthetic pathway, catalyses an unusual two-step reaction in which the substrate, either 2-acetolactate or 2-aceto-2-hydroxybutyrate, is converted via an alkyl migration and an NADPH-dependent reduction to give 2,3-dihydroxy-3-methylbutyrate or 2,3-dihydroxy-3-methylvalerate respectively. We have isolated and characterized a full-length cDNA from a lambda gt11 spinach library encoding the complete acetohydroxy acid reductoisomerase protein precursor. The 2050-nucleotide sequence contains a 1785-nucleotide open reading frame. The derived amino acid sequence indicates that the protein precursor consists of 595 amino acid residues including a presequence peptide of 72 amino acid residues. The N-terminal sequence of the first 16 amino acid residues of the purified AHRI confirms the identity of the cDNA. The derived amino acid sequence from this open reading frame shows 23% identity with the deduced amino acid sequences of the Escherichia coli and Saccharomyces cerevisiae AHRI proteins. There are two blocks of conserved amino acid residues in these three proteins. One of these is a sequence similar to the ‘fingerprint’ region of the NAD(P)H-binding site found in a large number of NAD(P)H-dependent oxidoreductases. The other, a short sequence (Lys-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Xaa-Ser-His-Gly-Phe) containing the amino acids lysine and histidine, could well be the catalytic site of the first step of the AHRI reaction. Southern-blot analysis indicated that AHRI is encoded by a single gene per haploid genome of about 7.5 kbp containing at least four introns.


2018 ◽  
Vol 2018 ◽  
pp. 1-13 ◽  
Author(s):  
Yong-Mei Jin ◽  
Rihua Piao ◽  
Yong-Feng Yan ◽  
Mojun Chen ◽  
Ling Wang ◽  
...  

Cold stress is one of the most important abiotic stresses in rice. C2H2zinc finger proteins play important roles in response to abiotic stresses in plants. In the present study, we isolated and functionally characterized a new C2H2zinc finger protein transcription factorOsCTZFP8in rice.OsCTZFP8encodes a C2H2zinc finger protein, which contains a typical zinc finger motif, as well as a potential nuclear localization signal (NLS) and a leucine-rich region (L-box). Expression ofOsCTZFP8was differentially induced by several abiotic stresses and was strongly induced by cold stress. Subcellular localization assay and yeast one-hybrid analysis revealed that OsCTZFP8 was a nuclear protein and has transactivation activity. To characterize the function ofOsCTZFP8in rice, the full-length cDNA ofOsCTZFP8was isolated and transgenic rice with overexpression ofOsCTZFP8driven by the maize ubiquitin promoter was generated usingAgrobacterium-mediated transformation. Among 46 independent transgenic lines, 6 single-copy homozygous overexpressing lines were selected by Southern blot analysis and Basta resistance segregation assay in both T1and T2generations. Transgenic rice overexpressingOsCTZFP8exhibited cold tolerant phenotypes with significantly higher pollen fertilities and seed setting rates than nontransgenic control plants. In addition, yield per plant ofOsCTZFP8-expressing lines was significantly (p<0.01) higher than that of nontransgenic control plants under cold treatments. These results demonstrate thatOsCTZFP8was a C2H2zinc finger transcription factor that plays an important role in cold tolerance in rice.


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