scholarly journals Functional Characterization of the Arabidopsis Abscisic Acid Transporters NPF4.5 and NPF4.6 in Xenopus Oocytes

2020 ◽  
Vol 11 ◽  
Author(s):  
Sophie Léran ◽  
Mélanie Noguero ◽  
Claire Corratgé-Faillie ◽  
Yann Boursiac ◽  
Chantal Brachet ◽  
...  
Keyword(s):  
Abscisic Acid ◽  
Xenopus Oocytes ◽  
Functional Characterization

scholarly journals Functional characterization of Lilium lancifolium cold-responsive Zinc Finger Homeodomain (ZFHD) gene in abscisic acid and osmotic stress tolerance

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11508
Author(s):  
Yubing Yong ◽  
Yue Zhang ◽  
Yingmin Lyu
Keyword(s):  
Abscisic Acid ◽  
Transgenic Plants ◽  
Zinc Finger ◽  
Promoter Region ◽  
Salt Water ◽  
Functional Characterization ◽  
Regulatory Elements ◽  
Response Analysis ◽  
Lilium Lancifolium

Background. We have previously performed an analysis of the cold-responsive transcriptome in the mature leaves of tiger lily (Lilium lancifolium) by gene co-expression network identification. The results has revealed that a ZFHD gene, notated as encoding zinc finger homeodomain protein, may play an essential regulating role in tiger lily response to cold stress. Methods. A further investigation of the ZFHD gene (termed as LlZFHD4) responding to osmotic stresses, including cold, salt, water stresses, and abscisic acid (ABA) was performed in this study. Based on the transcriptome sequences, the coding region and 5′ promoter region of LlZFHD4 were cloned from mature tiger lily leaves. Stress response analysis was performed under continuous 4 °C, NaCl, PEG, and ABA treatments. Functional characterization of LlZFHD4 was conducted in transgenic Arabidopsis, tobacco, and yeast. Results. LlZFHD4 encodes a nuclear-localized protein consisting of 180 amino acids. The N-terminal region of LlZFHD4 has transcriptional activation activity in yeast. The 4 °C, NaCl, PEG, and ABA treatments induced the expression of LlZFHD4. Several stress- or hormone-responsive cis-acting regulatory elements (T-Box, BoxI. and ARF) and binding sites of transcription factors (MYC, DRE and W-box) were found in the core promoter region (789 bp) of LlZFHD4. Also, the GUS gene driven by LlZFHD4 promoter was up-regulated by cold, NaCl, water stresses, and ABA in Arabidopsis. Overexpression of LlZFHD4 improved cold and drought tolerance in transgenic Arabidopsis; higher survival rate and better osmotic adjustment capacity were observed in LlZFHD4 transgenic plants compared to wild type (WT) plants under 4 °C and PEG conditions. However, LlZFHD4 transgenic plants were less tolerant to salinity and more hypersensitive to ABA compared to WT plants. The transcript levels of stress- and ABA-responsive genes were much more up-regulated in LlZFHD4 transgenic Arabidopsis than WT. These results indicate LlZFHD4 is involved in ABA signaling pathway and plays a crucial role in regulating the response of tiger lily to cold, salt and water stresses.

cAMP-activated C1 conductance is expressed in Xenopus oocytes by injection of shark rectal gland mRNA

1991 ◽  
Vol 260 (3) ◽  
pp. C664-C669 ◽  
Author(s):  
S. K. Sullivan ◽  
K. Swamy ◽  
M. Field
Keyword(s):  
Xenopus Oocytes ◽  
Functional Characterization ◽  
Equilibrium Potential ◽  
Rectal Gland ◽  
Calcium Buffer ◽  
Cl Channel ◽  
Shark Rectal Gland ◽  
Cl Channels ◽  
Cl Conductance

Development of reliable expression systems for use in identification and functional characterization of proteins required for secretory Cl channel activity is key to understanding the molecular basis of cystic fibrosis (CF). Until now, heterologous expression of epithelial Cl channels had not been accomplished. We show here that Xenopus oocytes express an adenosine 3',5'-cyclic monophosphate (cAMP)-activated Cl conductance after injection of mRNA from shark rectal gland. Current through this conductance was rapidly activated by intracellular application of cAMP, reversed near the chloride equilibrium potential (ECl), blocked by the Cl channel inhibitor 5-nitro-2-(3-phenylpropylamino) benzoate, and was not affected by preincubation with the intracellular calcium buffer bis-(2-amino-5-methylphenoxy)-ethane-N,N,N',N'-tetraacetic acid tetraacetoxymethyl ester, a condition that prohibits activation of the endogenous Ca-activated Cl conductance.

scholarly journals Revisiting the functional properties of NPF6.3/NRT1.1/CHL1 in xenopus oocytes

2018 ◽  
Author(s):  
Mélanie Noguero ◽  
Sophie Léran ◽  
Eléonore Bouguyon ◽  
Chantal Brachet ◽  
Pascal Tillard ◽  
...  
Keyword(s):  
Transport Properties ◽  
Xenopus Oocytes ◽  
Nitrate Concentration ◽  
Functional Characterization ◽  
Nitrate Transport ◽  
Experimental Conditions ◽  
Point Mutant ◽  
Electrode Voltage ◽  
Nitrate Transporters

ABSTRACTWithin the Arabidopsis NPF proteins, most of the characterized nitrate transporters are low-affinity transporters, whereas the functional characterization of NPF6.3/NRT1.1 has revealed interesting transport properties: the transport of nitrate and auxin, the eletrogenicity of the nitrate transport and a dual-affinity transport behavior for nitrate depending on external nitrate concentration. However, some of these properties remained controversial and were challenged here. We functionally express WT NPF6.3/NRT1.1 and some of its mutant in Xenopus oocytes and used a combination of uptake experiments using 15N-labelled nitrate and two-electrode voltage-clamp. In our experimental conditions in xenopus oocytes, in the presence or in the absence of external chloride, NPF6.3/NRT1.1 behaves as a non-electrogenic and pure low-affinity transporter. Moreover, further functional characterization of a NPF6.3/NRT1.1 point mutant, P492L, allowed us to hypothesize that NPF6.3/NRT1.1 is regulated by internal nitrate concentration and that the internal perception site involves the P492 residue.

Functional characterization of two alternatively spliced transcripts of tomato ABSCISIC ACID INSENSITIVE3 (ABI3) gene

2013 ◽  
Vol 82 (1-2) ◽  
pp. 131-145 ◽  
Author(s):  
Yongfeng Gao ◽  
Jikai Liu ◽  
Zhiguo Zhang ◽  
Xiaochun Sun ◽  
Ni Zhang ◽  
...  
Keyword(s):  
Abscisic Acid ◽  
Functional Characterization ◽  
Alternatively Spliced ◽  
Abscisic Acid Insensitive3

scholarly journals Functional characterization of the ABF gene family in upland cotton (Gossypium hirsutum L.)

2017 ◽  
Author(s):  
Tyson C. C. Kerr ◽  
Haggag Abdel-Mageed ◽  
MiYoung Kang ◽  
Dakota Cryer ◽  
Randy D. Allen
Keyword(s):  
Abscisic Acid ◽  
Abiotic Stress ◽  
Gossypium Hirsutum ◽  
Water Deficit ◽  
Upland Cotton ◽  
Expression Patterns ◽  
Functional Characterization ◽  
Ectopic Expression ◽  
Differentially Expressed

AbstractThe AREB/ABF bZIP transcription factors play a pivotal role in abscisic acid-dependent abiotic stress-responsive gene expression. Despite the perennial damage and reduced productivity that result from water-deficit and unpredictable early season temperature fluctuations, these critical genes have not been previously examined in upland cotton (Gossypium hirsutum). Here, we report the isolation of the G. hirsutum ABF homologs, characterization of their expression patterns in response to abiotic stress treatments, and examination of their functions through heterologous ectopic expression in Arabidopsis. As expected for an allotetraploid, G. hirsutum ABF homologs are present in the genome as homeologous pairs. These genes are differentially expressed, both among the homologs and within the homeologous pairs, in response to exogenous abscisic acid (ABA) application, dehydration, and chilling temperatures. Furthermore, heterologous ectopic expression of many of the G. hirsutum ABF genes in Arabidopsis conferred increased tolerance to water deficit and osmotic stress, as well as cold tolerance, in a gene specific manner. These results indicate the G. hirsutum ABF homologs are functional in Arabidopsis and, as in other species, are likely to play an essential role in the abiotic stress response.HighlightThe Gossypium hirsutum ABF homeologs are differentially expressed in response to abiotic stress, and their ectopic expression in Arabidopsis can confer increased water deficit tolerance.

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