scholarly journals Maize bioengineering with c-repeat binding factor 1 (CBF1) as a technique for desiccation toleration

2019 ◽  
Vol 3 (1) ◽  
pp. 1-10
Author(s):  
ERIC KIMANI KURIA ◽  
JESSE MACHUKA ◽  
STEVEN RUNO

Abstract. Kuria EK, Machuka J, Runo S. 2019. Maize bioengineering with c-repeat binding factor 1 (CBF1) as a technique for desiccation toleration. Trop Drylands 3: 1-10. Africa is a desiccation inclined continent leading to riskful cultivation particularly to small-scale cultivators who rely on rain-fed agriculture. Maize is the most widely cultivated main crop in Africa with more than 300 million people relying on it as their principal dietdiet fount. Desiccation causes crop fiasco, famine and poverty and this is being aggravated by climate change. There is therefore an obligation to flourish desiccation tolerablish maize. Traditional propagation techniques have been implementedcarried out in the establishment of desiccation tolerablish plants but are restricted by their requirement for labour, time and space, suggesting a limited genetic diversity within genotypes and transition of undesired traits along with the wanted ones. These restrictions are handled by utilizing these techniques along with bioengineering. Desiccation triggers a range of physiological and biochemical reactions in plants at cellular and molecular levels. These reactions include initiation of genes with several usefulness. Plant alteration for expanded desiccation toleration is generally based on the administration of either transcription and/or signaling factors or genes that directly secure plant cells contra water shortage. C-repeat binding factor (CBF) is a transcriptional factor that interacts with the desiccation responsive element (DRE), a cis-acting promoter element that governs gene expression in reaction to desiccation, brine and freezing stress. Over expression of these transcription factors, escalates stress toleration to freezing, desiccation and high brininess. In this study, three maize inbred lines and one hybrid were altered with CBF1 gene and appointed with mannose utilizing the Phosphomannose isomerase (PMI) gene. Genetic alteration was conducted through Agrobacterium tumefaciens and PCR was utilized to ascertain altered plants. Alteration frequency, alteration effectiveness and regeneration effectiveness were equated among the distinct genotypes altered. There were no remarkable dissimilarities in alteration frequency among the four maize genotypes. CML216 had the highest alteration effectiveness and regeneration effectiveness followed by A188. No alleged transgenic plants were regenerated from TL27 and A188×TL18 under the circumstances implemented on acount of their low regenerability. Further molecular analysis and desiccation stress tentatives on the expanded transgenic maize are significant prior to commercial release. Availability of desiccation tolerablish maize would bear a considerable positive collision contra famine particularly in Africa.

Plants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 758
Author(s):  
Sanjay Joshi ◽  
Christian Keller ◽  
Sharyn E. Perry

AGAMOUS-like 15 (AGL15) is a member of the MADS domain family of transcription factors (TFs) that can directly induce and repress target gene expression, and for which promotion of somatic embryogenesis (SE) is positively correlated with accumulation. An ethylene-responsive element binding factor-associated amphiphilic repression (EAR) motif of form LxLxL within the carboxyl-terminal domain of AGL15 was shown to be involved in repression of gene expression. Here, we examine whether AGL15′s ability to repress gene expression is needed to promote SE. While a form of AGL15 where the LxLxL is changed to AxAxA can still promote SE, another form with a strong transcriptional activator at the carboxy-terminal end, does not promote SE and, in fact, is detrimental to SE development. Select target genes were examined for response to the different forms of AGL15.


2000 ◽  
Vol 164 (3) ◽  
pp. R11-R16 ◽  
Author(s):  
A Takenaka ◽  
K Komori ◽  
T Morishita ◽  
SI Takahashi ◽  
T Hidaka ◽  
...  

To investigate the molecular mechanisms of increased transcription of the insulin-like growth factor-binding protein-1 (IGFBP-1) gene in dietary protein-deprived animals, the cis-acting sequence that is involved in this regulation was analyzed. We first showed that IGFBP-1 gene transcription was up-regulated by amino acid deprivation in cultured liver cell lines: H4IIE and HuH-7. Since HuH-7 cells showed a greater increase in IGFBP-1 mRNA in response to amino acid deprivation, this cell line was used in further experiments. Using a promoter function assay, we found that up-regulation of promoter activity responding to amino acid deprivation was abolished by deleting the region between -112 and -81 bp from the cap site from the gene construct. This cis-acting region includes the insulin-responsive element (IRE) and glucocorticoid responsive element (GRE) of IGFBP-1. In summary, the present observation suggests that the 32-bp (-112 to -81) in the IGFBP-1 gene 5' promoter region is involved in the induction of the IGFBP-1 gene in response to amino acid deprivation.


2017 ◽  
Vol 6 (2) ◽  
pp. 65
Author(s):  
Sitta J. ◽  
Nzuve F. M. ◽  
Olubayo F. M. ◽  
Mutinda C. ◽  
Muiru W. M. ◽  
...  

Maize (Zea mays L.) is the most widely grown staple food crop in Sub Saharan Africa (SSA) and occupies more than 33 million hectares each year. The recent outbreak and rapid spread of the Maize Lethal Necrosis (MLN) disease has emerged as a great challenge to maize production, threatening food security for the majority of households in the Eastern Africa region with yield loss estimated to be 50-90%. The disease is a result of synergistic interaction between two viruses, Sugarcane mosaic virus (SCMV) and Maize chlorotic mottle virus (MCMV). The objective of this study was to identify maize genotypes with resistance to MLN. In season one, 73 maize genotypes comprising 25 inbred lines from research institutes, 30 lines from the International Maize and Wheat Improvement Centre (CIMMYT) and 18 farmer varieties were screened for resistance to MLN. In season 2, only 48 genotypes were screened after some of the inbred lines showed complete susceptibility to MLN. These genotypes were grown in three replications in a completely randomized design in polythene bags in the greenhouse at the University of Nairobi. The plants were artificially inoculated using a mixture of SCMV and MCMV. .Weekly MLN disease severity scores using a scale of 1 to 5 (1 = highly resistant and 5 = highly susceptible) and % MLN incidence were recorded and eventually converted into Area under Disease Progress Curve (AUDPC) to give an indication of the disease intensity over time. The plants were allowed to grow to flowering stage to observe the effect of the MLN on the maize productivity. Analysis of Variance revealed wide genetic variation among the genotypes ranging from resistant to highly susceptible. In season 1, three farmer varieties namely MLR2, MLR11 and MLR13 showed resistance to MLN with a mean severity score of 2. In season 2, MLN12, MLN17, MLN18, MLN19, and MLR4 showed low MLN severity ranging from 2-3. The genotypes MLR6, MLR9, MLR16 and MLR18 showed MLN severity of 3 and early maturity traits. This study also validated the presence of MLN resistance among some CIMMYT lines depicted to show resistance in previous studies. These resistant genotypes could serve as donors in the introgression of the resistance into the adapted Kenyan maize backgrounds. This will go a long way in ensuring sustainable maize productivity while improving the livelihoods of the small-scale farmers who form the bulk of the major maize producers in Kenya.


1996 ◽  
Vol 43 (2) ◽  
pp. 369-377 ◽  
Author(s):  
J M Jankowski ◽  
P D Cannon ◽  
F Van der Hoorn ◽  
L D Wasilewska ◽  
N C Wong ◽  
...  

An in vitro transcription system from the trout testis nuclei was developed to study trout protamine gene expression. The protamine promoter contains, among others, two regulatory elements: 1) a cAMP-responsive element or CRE element (TGACGTCA) which is present in position 5' to TATA box, and 2) GC box (CCGCCC) which is present in position 3' to TATA box. The removal of the CRE-binding protein by titration (by the addition of appropriate oligonucleotides to the incubation mixture) resulted in a decrease in transcription of the protamine gene. These results were confirmed by experiments in which the pure CRE-binding factor (TPBP1) was used, as well as by those where a stimulatory effect of cAMP on protamine promoter transcription was observed. On the other hand, addition of oligonucleotides containing the GC-box sequence enhanced the protamine gene transcription indicating that the protein (Sp1 like) which binds to this sequence acts as a repressor of protamine gene expression. These results confirm the previously proposed model which suggested that the GC box played a role in negative regulation of the protamine gene expression. Involvement of some other factors in this process was also discussed.


Agronomy ◽  
2018 ◽  
Vol 8 (8) ◽  
pp. 151
Author(s):  
Cassyo de Araujo Rufino ◽  
Jucilayne Fernandes-Vieira ◽  
Jesús Martín-Gil ◽  
José Abreu Júnior ◽  
Lizandro Tavares ◽  
...  

Maize is an important food staple in many countries, and is useful in animal feed and many industrial applications. Its productivity is highly sensitive to drought stress, which may occur at any period during its growth cycle. The objective of this work was to compare the water stress influence on the performance of different maize genotypes in critical vegetative stages. Four genotypes of maize (namely a single-cross hybrid (AG 9045), a double-cross hybrid (AG 9011), a triple-cross hybrid (AG 5011), and a variety (AL Bandeirante)) were subjected to a 10-day period without irrigation in the vegetative stages that determine the number of kernel rows and the plant’s ability to take up nutrients and water (V4, V6 and V8). The impact of low water availability was assessed by analyzing plant height, height of the first ear insertion, stem diameter, yield per plant, and number of rows per ear, evincing that the yield per plant was the most sensitive parameter in all the stages. With regard to the influence of the genotype, the single-cross hybrid was demonstrated to be the most resilient to water shortage.


Plants ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 1033
Author(s):  
Chunying Wang ◽  
Tingting Lin ◽  
Mengqi Wang ◽  
Xiaoting Qi

Ethylene-responsive elements (EREs), such as the GCC box, are critical for ethylene-regulated transcription in plants. Our previous work identified a 19-bp AC-rich element (ACE) in the promoter of bean (Phaseolus vulgaris) metal response element-binding transcription factor 1 (PvMTF-1). Ethylene response factor 15 (PvERF15) directly binds ACE to enhance PvMTF-1 expression. As a novel ERF-binding element, ACE exhibits a significant difference from the GCC box. Here, we demonstrated that ACE serves as an ERE in Arabidopsis. It conferred the minimal promoter to respond to the ethylene stress and inhibition of ethylene. Moreover, the cis-acting element ACE could specifically bind the nuclear proteins in vitro. We further revealed that the first 9-bp sequence of ACE (ACEcore) is importantly required by the binding of nuclear proteins. In addition, PvERF15 and PvMTF-1 were strongly induced by ethylene in bean seedlings. Since PvERF15 activates PvMTF-1 via ACE, ACE is involved in ethylene-induced PvMTF-1 expression. Taken together, our findings provide genetic and biochemical evidence for a new ERE.


2013 ◽  
Vol 54 (9) ◽  
pp. 1415-1430 ◽  
Author(s):  
Kai Zhao ◽  
Xinjie Shen ◽  
Huazhao Yuan ◽  
Yun Liu ◽  
Xiong Liao ◽  
...  

2015 ◽  
Vol 20 (1) ◽  
Author(s):  
Qian Hu ◽  
Huili Tong ◽  
Dandan Zhao ◽  
Yunkao Cao ◽  
Weiwei Zhang ◽  
...  

AbstractThe promoter of skeletal muscle α-actin gene (ACTA1) is highly muscle specific. The core of the bovine ACTA1 promoter extends from +29 to −233, about 262 base pairs (bp), which is sufficient to activate transcription in bovine muscle satellite cells. In this study, analysis by PCR site-specific mutagenesis showed that the cis-acting element SRE (serum response element binding factor) was processed as a transcriptional activator. In order to enhance the bovine ACTA1 promoter’s activity, we used a strategy to modify it. We cloned a fragment containing three SREs from the promoter of ACTA1, and then one or two clones were linked upstream of the core promoter (262 bp) of ACTA1. One and two clones increased the activity of the ACTA1 promoter 3-fold and 10-fold, respectively, and maintained muscle tissue specificity. The modified promoter with two clones could increase the level of ACTA1 mRNA and protein 4-fold and 1.1-fold, respectively. Immunofluorescence results showed that green fluorescence of ACTA1 increased. Additionally, the number of total muscle microfilaments increased. These genetically engineered promoters might be useful for regulating gene expression in muscle cells and improving muscle mass in livestock.


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