scholarly journals A Small Natural Light-induced Bidirectional Promoter of Rapeseed (Brassica Napus)

2021 ◽  
Author(s):  
Ruijia Zhu ◽  
Li Zhang ◽  
Tao Wei ◽  
Xuefei Jiang ◽  
Maolin Wang
Keyword(s):  
Brassica Napus ◽  
Transcription Initiation ◽  
Molecular Breeding ◽  
Leaf Tissue ◽  
Bidirectional Promoter ◽  
Histochemical Staining ◽  
Sequence Length ◽  
Homeodomain Protein ◽  
Genes Expression ◽  
Application Potential

Abstract A bidirectional promoter is an intergenic region located between a pair of adjacent and oppositely transcribed genes (‘head-to-head’ genes) that concurrently promote both genes expression. In the genome of Brassica napus, we identified a bidirectional promoter (265bp long), named PBn265. PBn265 was located between the transcription initiation codons (ATG) of two genes that encode the homeodomain protein SHH2, and chloroplast GROUP II intron splicing factor CFM3. Its bidirectional promoting activity was verified by transient expression of Nicotiana benthamiana leaf tissue via Agrobacterium-mediated transformation of PBn265-F::EGFP and PBn265-R::mCherry. The expression of both reporter genes, EGFP linked to one end and GUS on the other end of the PBn265 sequence, was observed in the various tissues of the transgenic Arabidopsis thaliana using histochemical staining and fluorescence microscopy. Furthermore, we also found that the promoting activity of this sequence was regulated by illumination. Considering its short sequence length and light inducible regulation, this promoter likely has application potential in bioengineering and agricultural molecular breeding.

A rapid effect of applied brassinolide on abscisic acid concentrations in Brassica napus leaf tissue subjected to short-term heat stress

2008 ◽  
Vol 55 (3) ◽  
pp. 165-167 ◽  
Author(s):  
Leonid V. Kurepin ◽  
Mirwais M. Qaderi ◽  
Thomas G. Back ◽  
David M. Reid ◽  
Richard P. Pharis
Keyword(s):  
Abscisic Acid ◽  
Heat Stress ◽  
Brassica Napus ◽  
Leaf Tissue ◽  
Short Term ◽  
Rapid Effect

scholarly journals Molecular Breeding of Transgenic Rice Plants Expressing a Bacterial Chlorocatechol Dioxygenase Gene

2002 ◽  
Vol 68 (8) ◽  
pp. 4061-4066 ◽  
Author(s):  
Masami Shimizu ◽  
Tetsuya Kimura ◽  
Takayoshi Koyama ◽  
Katsuhisa Suzuki ◽  
Naoto Ogawa ◽  
...  
Keyword(s):  
Transgenic Rice ◽  
Molecular Breeding ◽  
Ralstonia Eutropha ◽  
Leaf Tissue ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Gene Encoding ◽  
Rice Plants ◽  
Transgenic Rice Plants ◽  
Dioxygenase Gene

ABSTRACT The cbnA gene encoding the chlorocatechol dioxygenase gene from Ralstonia eutropha NH9 was introduced into rice plants. The cbnA gene was expressed in transgenic rice plants under the control of a modified cauliflower mosaic virus 35S promoter. Western blot analysis using anti-CbnA protein indicated that the cbnA gene was expressed in leaf tissue, roots, culms, and seeds. Transgenic rice calluses expressing the cbnA gene converted 3-chlorocatechol to 2-chloromucote efficiently. Growth and morphology of the transgenic rice plants expressing the cbnA gene were not distinguished from those of control rice plants harboring only a Ti binary vector. It is thus possible to breed transgenic plants that degrade chloroaromatic compounds in soil and surface water.

scholarly journals Cloning and characterization of the DIR1 promoter from Eucommia ulmoides Oliv and its response to hormonal and abiotic stress.

2021 ◽  
Author(s):  
Li Ziyun ◽  
Li Biao ◽  
Zhao yichen ◽  
Zhao Degang
Keyword(s):  
Transgenic Tobacco ◽  
Transcription Initiation ◽  
Core Promoter ◽  
Promoter Sequence ◽  
Inhibitory Effect ◽  
Initiation Site ◽  
Histochemical Staining ◽  
Eucommia Ulmoides ◽  
Tobacco Seedlings ◽  
High Concentrations

Abstract The lignans of Eucommia ulmoides have been extensively studied and shown to have a dual mechanism of regulating blood pressure. Studies have shown that DIR1 is a key gene in the biosynthetic pathway of lignans in Eucommia ulmoides Oliv. In this study, a 2000 bp upstream promoter sequence was cloned, and part of the sequence (1495 bp) and its 5'-end truncated segment were constructed into the pCAMBIA1391Z expression plasmid upstream of ß-glucuronidase (GUS). Agrobacterium-mediated genetic transformation produced stable transgenic tobacco lines. The results showed that although both full-length and truncated promoters could initiate GUS expression, their levels were influenced by the degree of deletion at the 5' end. GUS histochemical staining showed that the core promoter region was located in the region containing the transcription initiation site (TIS) within 212 bp. In addition, the DIR1 promoter responded to environmental and hormonal stressors, such as jasmonic acid (MeJA), abscisic acid (ABA), D-mannitol (drought mimic), and high concentrations of NaCl. In transgenic tobacco seedlings, MeJA, D-mannitol, and ABA could activate the DIR1 promoter, whereas high concentrations of NaCl could inhibit it. In E. ulmoides Oliv seedlings, MeJA, NaCl, and D-mannitol activated the DIR1 promoter, whereas ABA had an inhibitory effect. In summary, our findings provide a theoretical basis for the use of the DIR1 promoter in plant genetic engineering, indicating its potential. Our study also presents novel insights for lignan biosynthesis and sheds light on the mechanisms of E. ulmoides Oliv in response to stress.

scholarly journals Transcriptome sequence reveals candidate genes involving in the post-harvest hardening of trifoliate yam Dioscorea dumetorum

2021 ◽  
Author(s):  
Christian Siadjeu ◽  
Eike Mayland-Quellhorst ◽  
Sascha Laubinger ◽  
Dirk C. Albach
Keyword(s):  
Functional Analysis ◽  
Candidate Genes ◽  
Molecular Breeding ◽  
Rna Seq ◽  
Post Harvest ◽  
Genes Expression ◽  
Genes Encoding ◽  
Hypothetical Mechanism ◽  
Trifoliate Yam ◽  
Storage Ability

AbstractStorage ability of D. dumetorum is restricted by a severe phenomenon of post-harvest hardening which starts 72h after harvest and renders tubers inedible. Previous work has only focused on the biochemistry changes affecting the PHH on D. dumetorum. To the best of our knowledge nobody has identified candidate genes responsible for hardness on D. dumetorum. Here, transcriptome analysis of D. dumetorum tubers was performed, 4 months after emergence (4MAE), after harvest (AH), 3 days AH (3DAH) and 14 days AH (14DAH) on four accessions using RNA-Seq. In total between AH and 3DAH, 165, 199,128 and 61 differentially expressed genes (DEGs) were detected in Bayangam 2, Fonkouankem 1, Bangou 1 and Ibo sweet 3 respectively. Functional analysis of DEGs revealed that genes encoding for cellulose synthase A, xylan O-acetyltransferase chlorophyll a/b binding protein 1,2,3,4 and transcription factor MYBP were found predominantly and significantly up-regulated 3DAH, implying that genes were potentially involved in the post-harvest hardening. A hypothetical mechanism of this phenomenon and its regulation has been proposed. These findings provide the first comprehensive insights into genes expression in yam tubers after harvest and valuable information for molecular breeding against the post-harvest hardening. A hypothetical mechanism of this phenomenon and its regulation has been proposed. These findings provide the first comprehensive insights into genes expression in yam tubers after harvest and valuable information for molecular breeding against the post-harvest hardening.

Developmental basis of homeosis in precociously germinating Brassica napus embryos: phase change at the shoot apex

Development ◽  
1997 ◽  
Vol 124 (6) ◽  
pp. 1149-1157
Author(s):  
D.E. Fernandez
Keyword(s):  
Molecular Markers ◽  
Brassica Napus ◽  
Shoot Apex ◽  
Leaf Tissue ◽  
Precocious Germination ◽  
Organ Identity ◽  
The Status ◽  
Mode Characteristic ◽  
In Situ Hybridizations

Precociously germinating Brassica napus (oilseed rape) embryos produce extra cotyledons or chimeric organs with sectors of cotyledon and leaf tissue, rather than leaves, at the shoot apex. To investigate this phenomenon in more detail, scanning electron microscopy was used to examine the development of organ primordia at the shoot apex. In situ hybridizations with molecular markers of the embryonic phase were used to assess the status of individual cells in the shoot apex with regard to the transition between embryonic and vegetative phases. The results indicate that, under conditions that support precocious germination, primordia develop at the shoot apex in the mode characteristic of postgerminative growth, i.e. they arise sequentially in a spiral phyllotaxy. Cells in the rest of the embryo, however, can continue to express molecular markers of the embryonic phase for several weeks after the start of culture. When patterns of gene expression and the fate of individual primordia were compared, a strong correlation was found between organ identity and the status of cells in the vicinity of the meristem with regard to phase. Primordia that develop in situations where neighboring cells are in the embryonic phase always produce organs with cotyledon morphology. Primordia that develop in situations where neighboring cells have exited the embryonic phase produce leaves. Based on an examination of situations where chimeric organs are produced, I propose that short range interactions or signalling are likely to be involved in communicating information about phase to developing primordia.

Temporal and spatial distribution of BN28 during low temperature acclimation of Brassica napus cv. Cascade seedlings

1997 ◽  
Vol 75 (1) ◽  
pp. 28-35
Author(s):  
Mitchel D. de Beus ◽  
Anne M. Johnson-Flanagan ◽  
Joseph G. Boothe
Keyword(s):  
Brassica Napus ◽  
Low Temperature ◽  
Leaf Age ◽  
Temporal Distribution ◽  
Leaf Tissue ◽  
Temperature Acclimation ◽  
Ground Tissue ◽  
Temporal And Spatial Distribution ◽  
Apical Meristems ◽  
Mature Leaves

The objective of this study was to determine the spatial and temporal distribution of the peptide BN28 in Brassica napus cv. Cascade seedlings during low temperature acclimation. Immunoblots revealed that BN28 was present in leaves and shoot apical meristems of plants grown under low temperature but was absent from older tissues in the stem. Immunocytochemistry was used to determine the distribution throughout leaf and apical meristem tissues. BN28 was found throughout the apical meristems, was localized in the ground tissue and epidermis of young leaves, and was restricted to the ground tissue and guard cells in mature leaves. Differences in total accumulation were also noted, with the youngest leaves having the highest accumulation of BN28 and the quantity decreasing with leaf age. Despite these differences, plant developmental stage did not affect the accumulation of BN28 in individual leaves. Post-transcriptional controls are expected to regulate accumulation of the protein, as bn28 mRNA accumulates during acclimation in both young and mature leaves. Immunolocalization studies of BN28 in acclimated leaf tissue confirmed that BN28 is cytoplasmically localized and has no apparent weak association with organelles or other cellular membrane systems. Key words: acclimation, Brassica, development, immunocytochemistry, low temperature, protein synthesis.

scholarly journals Tissue-specific mRNA profiling of the Brassica napus-Sclerotinia sclerotiorum interaction uncovers novel regulators of plant immunity

2021 ◽  
Author(s):  
Philip Walker ◽  
Ian Girard ◽  
Shayna Giesbrecht ◽  
Steve Whyard ◽  
Dilantha Fernando ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Sclerotinia Sclerotiorum ◽  
Defense Response ◽  
Plant Immunity ◽  
Leaf Tissue ◽  
Specific Gene ◽  
Tissue Specific ◽  
Biotrophic Pathogens ◽  
Dna Insertion ◽  
Insight Into

White mold in Brassica napus (canola) is caused by the fungal pathogen Sclerotinia sclerotiorum and is responsible for significant losses in crop yield across the globe. With advances in high-throughput transcriptomics, our understanding of the B. napus defense response to S. sclerotiorum is becoming clearer; however, the response of individual tissue layers directly at the site of infection has yet to be explored. Using laser microdissection coupled with RNA sequencing, we profiled the epidermis, mesophyll and vascular leaf tissue layers in response to S. sclerotiorum. This strategy increases the number of genes detected compared to whole-leaf assessments and provides unprecedented information on tissue-specific gene expression networks in response to pathogen attack. Our findings provide novel insight into the conserved and specific roles of ontogenetically distinct leaf tissue layers in response to infection. Using bioinformatics tools, we identified several defense genes that might coordinate plant immunity responses shared across different tissue layers within the leaf. These genes were functionally characterized by challenging T-DNA insertion lines of Arabidopsis with necrotrophic, hemi-biotrophic, and biotrophic pathogens, ultimately converging on the PR5-like RECEPTOR KINASE (PRK5). Together, these data provide insight on the complexity of the B. napus defense response directly at the site of infection.

The human cut homeodomain protein represses transcription from the c-myc promoter

1994 ◽  
Vol 14 (6) ◽  
pp. 4251-4257
Author(s):  
D Dufort ◽  
A Nepveu
Keyword(s):  
Transcription Initiation ◽  
Binding Activity ◽  
Size Exclusion ◽  
Homeodomain Protein ◽  
Human Homolog ◽  
Mobility Shift ◽  
Expression Library ◽  
Nuclear Extracts ◽  
Exon 1 ◽  
Mobility Shift Assay

Studies of the c-myc promoter have shown that efficient transcription initiation at the P2 start site as well as the block to elongation of transcription require the presence of the ME1a1 protein binding site upstream of the P2 TATA box. Following fractionation by size exclusion chromatography, three protein-ME1a1 DNA complexes, a, b, and c, were detected by electrophoretic mobility shift assay. A cDNA encoding a protein present in complex c was isolated by screening of an expression library with an ME1a1 DNA probe. This cDNA was found to encode the human homolog of the Drosophila Cut homeodomain protein. The bacterially expressed human Cut (hu-Cut) protein bound to the ME1a1 site, and antibodies against hu-Cut inhibited the ME1a1 binding activity c in nuclear extracts. In cotransfection experiments, the hu-Cut protein repressed transcription from the c-myc promoter, and this repression was shown to be dependent on the presence of the ME1a1 site. Using a reporter construct with a heterologous promoter, we found that c-myc exon 1 sequences were also necessary, in addition to the ME1a1 site, for repression by Cut. Taken together, these results suggest that the human homolog of the Drosophila Cut homeodomain protein is involved in regulation of the c-myc gene.

scholarly journals A soluble transcription factor, Oct-1, is also found in the insoluble nuclear matrix and possesses silencing activity in its alanine-rich domain.

1996 ◽  
Vol 16 (8) ◽  
pp. 4366-4377 ◽  
Author(s):  
M K Kim ◽  
L A Lesoon-Wood ◽  
B D Weintraub ◽  
J H Chung
Keyword(s):  
Nuclear Matrix ◽  
Transcription Initiation ◽  
Initiation Site ◽  
Homeodomain Protein ◽  
Mobility Shift ◽  
Rich Domain ◽  
Gel Mobility Shift ◽  
Silencer Element

Expression of the human thyrotropin beta (hTSHbeta) gene is restricted to thyrotrophs, at least in part, by silencing. Using transient-transfection assays, we have localized a silencer element to a region between -128 and -480 bp upstream of the transcription initiation site. The silencing activity was overcome in a thyrotroph-specific manner by an unknown enhancer located in the sequences at -approximately 10000 to -1200 bp. The ubiquitous POU homeodomain protein Oct-1 recognized the A/T-rich silencer element at multiple sites in gel mobility shift assays and in vitro footprinting analyses. The silencing activity of Oct-1 was localized in its C-terminal alanine-rich domain, suggesting that Oct-1 plays a role in silencing of the hTSHbeta promoter. Further, a significant fraction of Oct-1 was shown to be associated with the nuclear matrix, and the hTSHbeta silencer region was tethered to a nuclear matrix of human cells in vivo, suggesting a possible role of the Oct-1-hTSHbeta silencer region interaction in chromatin organization.

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