scholarly journals The ferroxidase LPR5 functions in the maintenance of phosphate homeostasis and is required for normal growth and development of rice

2020 ◽  
Vol 71 (16) ◽  
pp. 4828-4842
Author(s):  
Hao Ai ◽  
Yue Cao ◽  
Ajay Jain ◽  
Xiaowen Wang ◽  
Zhi Hu ◽  
...  
Keyword(s):  
Growth And Development ◽  
Xylem Sap ◽  
Sufficient Conditions ◽  
Normal Growth ◽  
Yield Potential ◽  
Specific Expression ◽  
Gus Reporter ◽  
Ferroxidase Activity ◽  
Vitro Analysis

Abstract Members of the Low Phosphate Root (LPR) family have been identified in rice (Oryza sativa) and expression analyses have been conducted. Here, we investigated the functions of one of the five members in rice, LPR5. qRT-PCR and promoter–GUS reporter analyses indicated that under Pi-sufficient conditions OsLPR5 was highly expressed in the roots, and specific expression occurred in the leaf collars and nodes, and its expression was increased under Pi-deficient conditions. In vitro analysis of the purified OsLPR5 protein showed that it exhibited ferroxidase activity. Overexpression of OsLPR5 triggered higher ferroxidase activity, and elevated concentrations of Fe(III) in the xylem sap and of total Fe in the roots and shoots. Transient expression of OsLPR5 in Nicotiana benthamiana provided evidence of its subcellular localization to the cell wall and endoplasmic reticulum. Knockout mutation in OsLPR5 by means of CRISPR-Cas9 resulted in adverse effects on Pi translocation, on the relative expression of Cis-NATOsPHO1;2, and on several morphological traits, including root development and yield potential. Our results indicate that ferroxidase-dependent OsLPR5 has both a broad-spectrum influence on growth and development in rice as well as affecting a subset of physiological and molecular traits that govern Pi homeostasis.

scholarly journals Methylation of Histone H3 Lysine 36 Is Required for Normal Development in Neurospora crassa

2005 ◽  
Vol 4 (8) ◽  
pp. 1455-1464 ◽  
Author(s):  
Keyur K. Adhvaryu ◽  
Stephanie A. Morris ◽  
Brian D. Strahl ◽  
Eric U. Selker
Keyword(s):  
Neurospora Crassa ◽  
Growth And Development ◽  
Histone H3 ◽  
Normal Growth ◽  
Wild Type ◽  
Set Domain ◽  
Amino Terminal ◽  
Histone H3 Gene ◽  
Type Gene

ABSTRACT The SET domain is an evolutionarily conserved domain found predominantly in histone methyltransferases (HMTs). The Neurospora crassa genome includes nine SET domain genes (set-1 through set-9) in addition to dim-5, which encodes a histone H3 lysine 9 HMT required for DNA methylation. We demonstrate that Neurospora set-2 encodes a histone H3 lysine 36 (K36) methyltransferase and that it is essential for normal growth and development. We used repeat induced point mutation to make a set-2 mutant (set-2 RIP1 ) with multiple nonsense mutations. Western analyses revealed that the mutant lacks SET-2 protein and K36 methylation. An amino-terminal fragment that includes the AWS, SET, and post-SET domains of SET-2 proved sufficient for K36 HMT activity in vitro. Nucleosomes were better substrates than free histones. The set-2 RIP1 mutant grows slowly, conidiates poorly, and is female sterile. Introducing the wild-type gene into the mutant complemented the defects, confirming that they resulted from loss of set-2 function. We replaced the wild-type histone H3 gene (hH3) with an allele producing a Lys to Leu substitution at position 36 and found that this hH3 K36L mutant phenocopied the set-2 RIP1 mutant, confirming that the observed defects in growth and development result from inability to methylate K36 of H3. Finally, we used chromatin immunoprecipitation to demonstrate that actively transcribed genes in Neurospora crassa are enriched for H3 methylated at lysines 4 and 36. Taken together, our results suggest that methylation of K36 in Neurospora crassa is essential for normal growth and development.

scholarly journals FOREIGN GENE EXPRESSION IN TRANSGENIC CRANBERRY PLANTS

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 621d-621
Author(s):  
Rod Serres ◽  
David Russell ◽  
Dan Mahr ◽  
Brent McCown
Keyword(s):  
Normal Growth ◽  
Foreign Gene ◽  
Foreign Gene Expression ◽  
Specific Expression ◽  
Genome Expression ◽  
Feeding Trials ◽  
Blackheaded Fireworm ◽  
Foreign Genes ◽  
Lepidopteran Resistance

Genetically transformed Vaccinium macrocarpon `Stevens' and `Pilgrim' plants have been obtained using electric discharge particle acceleration. Three foreign genes, kan encoding a selectable marker, gus a reporter gene, and B.t.k. conferring lepidopteran resistance, were incorporated into the genome. Expression of kan was assayed by culturing shoots in vitro on media with several concentrations of kanamycin. Expression among transformed clones (transclones) varied from high resistance (normal growth at 300 mg/L kan) to no resistance. Histochemical analyses for gus expression revealed variability among transclones. Some transclones exhibited no gus expression, others had consistent area-specific expression while others displayed random expression. In preliminary feeding trials with blackheaded fireworm larvae, B.t.k. expression was found to be ineffective at controlling insect development. We have recovered plants transformed with a different promoter driving the B.t.k. gene in an effort to enhance expression.

In vitro analysis of rat surfactant protein A gene expression

1996 ◽  
Vol 270 (4) ◽  
pp. L504-L516
Author(s):  
K. J. Stuempfle ◽  
M. Koptides ◽  
P. G. Quinn ◽  
J. Floros
Keyword(s):  
Protein Interactions ◽  
Protein Complexes ◽  
Protein A ◽  
Surfactant Protein ◽  
In Vitro Transcription ◽  
Proximal Promoter ◽  
Specific Expression ◽  
Thyroid Transcription Factor 1 ◽  
Vitro Analysis

Previous findings suggest that the rat proximal promoter segment (rPPS) of the SP-A gene is important in the regulation of the lung-specific expression of the gene. In this report, two regions within the rPPS containing thyroid transcription factor-1 (TTF-1) binding sites are identified that form strong lung- and thyroid- specific DNA-protein complexes. These regions bind nuclear polypeptides with similar apparent molecular mass to TTF-1, suggesting that TTF-1 binds to these regions. Two regions within the rPPS that form weaker lung-specific DNA-protein complexes are also identified. The transcription start site is mapped, and a functional analysis shows that the sequences of the 5' flanking region are sufficient to support in vitro transcription but are not sufficient to reproduce tissue-specific expression. Together, these results show that lung- and thyroid-specific, as well as lung-specific, DNA-protein interactions occur within the rPPS but are not sufficient for the lung-specific expression of the rat SP-A gene to be duplicated in vitro.

FACTORS CONTROLLING EMBRYO GROWTH AND DEVELOPMENT IN BARLEY (HORDEUM VULGARE L.)

1962 ◽  
Vol 40 (11) ◽  
pp. 1515-1523 ◽  
Author(s):  
L. J. La Croix ◽  
J. Naylor ◽  
E. N. Larter
Keyword(s):  
Energy Source ◽  
Cell Division ◽  
Growth And Development ◽  
Normal Growth ◽  
Growth Stimulation ◽  
Hordeum Vulgare L ◽  
Cell Extension ◽  
Single Leaf ◽  
Tetraploid Level

Normal growth and development of barley proembryos occurred in excised intact florets, but not in ovaries from which lemmas and paleas were removed. The lemmas and paleas of the barley floret apparently contained a substance (referred to as "hull factor") which inhibited cell extension and stimulated cell division in the embryo. Evidence was obtained that this stimulation was not due to the provision of a simple energy source such as sucrose. In the absence of the "hull factor", ovaries cultured in vitro were found to contain embryos having nuclei in which the DNA content was equivalent to the tetraploid level, while mitosis was almost completely lacking. A similar embryo growth stimulation was obtained when a single leaf was left on an excised barley spike where lemmas and paleas were removed.

The native structure of the eukaryotic ribosome

1983 ◽  
Vol 41 ◽  
pp. 432-433
Author(s):  
R.A. Milligan ◽  
P.N.T. Unwin
Keyword(s):  
Ribosomal Proteins ◽  
Crystal Form ◽  
Native Structure ◽  
X Ray Diffraction ◽  
Eukaryotic Ribosome ◽  
Vitro Analysis ◽  
In Vitro Analysis ◽  
Resolution Structure ◽  
Stable Unit

A detailed understanding of the mechanism of protein synthesis will ultimately depend on knowledge of the native structure of the ribosome. Towards this end we have investigated the low resolution structure of the eukaryotic ribosome embedded in frozen buffer, making use of a system in which the ribosomes crystallize naturally.The ribosomes in the cells of early chicken embryos form crystalline arrays when the embryos are cooled at 4°C. We have developed methods to isolate the stable unit of these arrays, the ribosome tetramer, and have determined conditions for the growth of two-dimensional crystals in vitro, Analysis of the proteins in the crystals by 2-D gel electrophoresis demonstrates the presence of all ribosomal proteins normally found in polysomes. There are in addition, four proteins which may facilitate crystallization. The crystals are built from two oppositely facing P4 layers and the predominant crystal form, accounting for >80% of the crystals, has the tetragonal space group P4212, X-ray diffraction of crystal pellets demonstrates that crystalline order extends to ~ 60Å.

1163: Radial Dilation of Ureteral Balloons: A Comparative in-Vitro Analysis

2005 ◽  
Vol 173 (4S) ◽  
pp. 315-316
Author(s):  
Kari Hendlin ◽  
Brynn Lund ◽  
Manoj Monga
Keyword(s):  
Vitro Analysis ◽  
In Vitro Analysis

Role of the 807 C/T Polymorphism of the α2 Gene in Platelet GP Ia Collagen Receptor Expression and Function

1999 ◽  
Vol 81 (06) ◽  
pp. 951-956 ◽  
Author(s):  
J. Corral ◽  
R. González-Conejero ◽  
J. Rivera ◽  
F. Ortuño ◽  
P. Aparicio ◽  
...  
Keyword(s):  
Venous Thrombosis ◽  
Cell Types ◽  
Receptor Expression ◽  
Case Control Studies ◽  
Platelet Surface ◽  
Vitro Analysis ◽  
And Function ◽  
In Vitro Analysis

SummaryThe variability of the platelet GP Ia/IIa density has been associated with the 807 C/T polymorphism (Phe 224) of the GP Ia gene in American Caucasian population. We have investigated the genotype and allelic frequencies of this polymorphism in Spanish Caucasians. The T allele was found in 35% of the 284 blood donors analyzed. We confirmed in 159 healthy subjects a significant association between the 807 C/T polymorphism and the platelet GP Ia density. The T allele correlated with high number of GP Ia molecules on platelet surface. In addition, we observed a similar association of this polymorphism with the expression of this protein in other blood cell types. The platelet responsiveness to collagen was determined by “in vitro” analysis of the platelet activation and aggregation response. We found no significant differences in these functional platelet parameters according to the 807 C/T genotype. Finally, results from 3 case/control studies involving 302 consecutive patients (101 with coronary heart disease, 104 with cerebrovascular disease and 97 with deep venous thrombosis) determined that the 807 C/T polymorphism of the GP Ia gene does not represent a risk factor for arterial or venous thrombosis.

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