Cloning and characterization of aCOBRA-like gene expressedde novoduring maize germination

2003 ◽  
Vol 13 (3) ◽  
pp. 209-217 ◽  
Author(s):  
Felipe Cruz-García ◽  
Alberto Gómez ◽  
José Juan Zúñiga ◽  
Javier Plasencia ◽  
Jorge M. Vázquez-Ramos
Keyword(s):  
Cell Expansion ◽  
Developmental Stages ◽  
Mrna Differential Display ◽  
Reading Frame ◽  
Early Germination ◽  
Embryo Axes ◽  
Leaf Tissues ◽  
Root Tissues ◽  
Tissues Expression

AbstractThe search for germination-specific genes has been a laborious and unrewarding task, since many of the genes expressed during germination are also expressed in embryogenesis or in other developmental stages. By using mRNA differential display of transcript populations from maize (Zea maysL.) embryo axes, germinated for different times with or without a previous osmopriming treatment, a 682 bp cDNA was isolated that was present only after 24 h germination, and absent during osmopriming or during early germination. Screening of a cDNA library using the 682 bp probe yielded a 1554 bp cDNA that contained an open reading frame coding for 436 amino acids. This gene, referred to asZmAA9-24, was expressed in root tissues, but was not detected in shoot or leaf tissues. Expression ofZmAA9-24occurred earlier during germination (by 15 h) if embryo axes were imbibed in the presence of cytokinins or if seeds were previously osmoprimed. The predicted protein sequence ofZmAA9-24is 39.6% identical to the product of the recently identifiedArabidopsisgeneCOBRA(54.5% in the central region), which appears to participate in the regulation of cell expansion, particularly in roots, and belongs to the glycosylphosphatidylinositol (GPI)-anchored protein family.ZmAA9-24expression might be regulated by both cell expansion and the cell cycle, processes that have a central role during seed germination.

Cloning and characterization of a 2,3-oxidosqualene cyclase from Eleutherococcus trifoliatus

Holzforschung ◽  
2013 ◽  
Vol 67 (4) ◽  
pp. 463-471 ◽  
Author(s):  
Li-Ting Ma ◽  
Sheng-Yang Wang ◽  
Yen-Hsueh Tseng ◽  
Yi-Ru Lee ◽  
Fang-Hua Chu
Keyword(s):  
Physiological Role ◽  
Active Site Residues ◽  
Reading Frame ◽  
Oxidosqualene Cyclases ◽  
Oxidosqualene Cyclase ◽  
Acid Protein ◽  
Leaf Tissues ◽  
New Perspective ◽  
Amino Acid Protein

Abstract The 2,3-oxidosqualene cyclases (OSCs) are a family of enzymes that have an important role in plant triterpene biosynthesis. In this study, an OSC gene designed EtLUS from Eleutherococcus trifoliatus has been cloned. EtLUS includes a 2292-bp open reading frame and encodes a 763-amino acid protein. EtLUS has an MLCYCR motif, which is conserved in lupeol synthases. Comparison of active-site residues and gene expression in yeast showed that EtLUS synthesizes lupeol. However, EtLUS has the highest sequence identity with β-amyrin synthases from Araliaceae rather than lupeol synthases, adding new perspective to the evolution of the OSCs of Araliaceae. Furthermore, EtLUS is upregulated in leaf tissues under methyl jasmonate treatment, which can be interpreted that lupeol and its derivatives play an ecological and physiological role in plant defense against pathogens and insect herbivores.

scholarly journals Characterization of cDNA Encoding Resveratrol Synthase and Accumulation of Resveratrol in Tartary Buckwheat

2013 ◽  
Vol 8 (11) ◽  
pp. 1934578X1300801 ◽  
Author(s):  
Yeon Bok Kim ◽  
Aye Aye Thwe ◽  
YeJi Kim ◽  
Sun Kyung Yeo ◽  
Chanhui Lee ◽  
...  
Keyword(s):  
Developmental Stages ◽  
Light Condition ◽  
Tartary Buckwheat ◽  
Dark Condition ◽  
Fagopyrum Tataricum ◽  
Reading Frame ◽  
Expression Levels ◽  
Resveratrol Synthase ◽  
Key Enzyme

Resveratrol synthase (RS) is the key enzyme for biosynthesis of resveratrol which come from coumaroyl-coenzyme A (CoA) and malonyl-CoA. Here, we report the cloning and characterization of a RS gene and accumulation of resveratrol in tartary buckwheat ( Fagopyrum tataricum). FtRS was composed of 1173 bp open reading frame and 390 amino acid residues and had a theoretical molecular weight and isoelectric point value of 43.70 kDa and 6.24, respectively. The FtRS expression levels were examined in sprouts and different organs of two tartary buckwheat cultivars, Hokkai T8 (T8) and Hokkai T10 (T10). FtRS transcript levels and resveratrol contents were higher under the dark condition compared with light condition. The expression levels of different organs of T10 was not observed significant variations compared to different organs of T8. Interestingly, resveratrol was detected in the sprouts developmental stages, but no resveratrol could not detect in any other organs of both T8 and T10. Therefore, we suggest that the resveratrol content in tartary buckwheat sprouts may be attributed mainly to the dark condition. The characterization of FtRS will be helpful for better understanding of the resveratrol biosynthesis in tartary buckwheat.

scholarly journals Molecular Cloning and Characterization of Genes Expressed during Early Tomato (Lycopersicon esculentum Mill.) Fruit Development by mRNA Differential Display

1996 ◽  
Vol 121 (1) ◽  
pp. 52-56 ◽  
Author(s):  
Denise M. Tieman ◽  
Avtar K. Handa
Keyword(s):  
Differential Display ◽  
Fruit Development ◽  
Cell Expansion ◽  
Tomato Fruit ◽  
Expression Patterns ◽  
Mrna Differential Display ◽  
Differentially Expressed ◽  
Threonine Deaminase ◽  
Primer Sets ◽  
Root Tissues

The growth of tomato fruit is the result of cell division early in development followed by cell expansion until the onset of ripening. We have utilized the mRNA differential display technique to clone genes differentially expressed in 10- and 20-day-old tomato fruit, when most fruit cells are undergoing a transition to growth by cell expansion. Of 1753 total bands observed using 30 independent primer sets, 31 differential display bands were obtained only in either 10-or 20-day - old fruit RNAs. Seven differentially expressed bands from 10-day-old fruit RNAs and six from 20-day-old fruit RNAs were cloned and characterized by sequence analysis and mRNA expression patterns in developing fruit, leaf and root tissues. Two clones had sequence similarities to 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase or threonine deaminase genes, while the remaining clones did not correspond to previously characterized genes. Steady state levels of mRNAs corresponding to seven clones were upregulated between 10 and 20 days of fruit development, while two clones were downregulated during growth and ripening. Most clones also hybridize to mRNA species present in leaf and root tissues. Collectively, these results suggest a transition in gene expression between 10- and 20-day-old fruit development.

scholarly journals Comprehensive analysis and identification of drought-responsive candidate NAC genes in three semi-arid tropics (SAT) legume crops

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Sadhana Singh ◽  
Himabindu Kudapa ◽  
Vanika Garg ◽  
Rajeev K. Varshney
Keyword(s):  
Drought Stress ◽  
Candidate Genes ◽  
Developmental Stages ◽  
Biological Activities ◽  
Expression Patterns ◽  
Genome Wide ◽  
Legume Crops ◽  
Root Tissues ◽  
Semi Arid ◽  
Comprehensive Study

Abstract Background Chickpea, pigeonpea, and groundnut are the primary legume crops of semi-arid tropics (SAT) and their global productivity is severely affected by drought stress. The plant-specific NAC (NAM - no apical meristem, ATAF - Arabidopsis transcription activation factor, and CUC - cup-shaped cotyledon) transcription factor family is known to be involved in majority of abiotic stresses, especially in the drought stress tolerance mechanism. Despite the knowledge available regarding NAC function, not much information is available on NAC genes in SAT legume crops. Results In this study, genome-wide NAC proteins – 72, 96, and 166 have been identified from the genomes of chickpea, pigeonpea, and groundnut, respectively, and later grouped into 10 clusters in chickpea and pigeonpea, while 12 clusters in groundnut. Phylogeny with well-known stress-responsive NACs in Arabidopsis thaliana, Oryza sativa (rice), Medicago truncatula, and Glycine max (soybean) enabled prediction of putative stress-responsive NACs in chickpea (22), pigeonpea (31), and groundnut (33). Transcriptome data revealed putative stress-responsive NACs at various developmental stages that showed differential expression patterns in the different tissues studied. Quantitative real-time PCR (qRT-PCR) was performed to validate the expression patterns of selected stress-responsive, Ca_NAC (Cicer arietinum - 14), Cc_NAC (Cajanus cajan - 15), and Ah_NAC (Arachis hypogaea - 14) genes using drought-stressed and well-watered root tissues from two contrasting drought-responsive genotypes of each of the three legumes. Based on expression analysis, Ca_06899, Ca_18090, Ca_22941, Ca_04337, Ca_04069, Ca_04233, Ca_12660, Ca_16379, Ca_16946, and Ca_21186; Cc_26125, Cc_43030, Cc_43785, Cc_43786, Cc_22429, and Cc_22430; Ah_ann1.G1V3KR.2, Ah_ann1.MI72XM.2, Ah_ann1.V0X4SV.1, Ah_ann1.FU1JML.2, and Ah_ann1.8AKD3R.1 were identified as potential drought stress-responsive candidate genes. Conclusion As NAC genes are known to play role in several physiological and biological activities, a more comprehensive study on genome-wide identification and expression analyses of the NAC proteins have been carried out in chickpea, pigeonpea and groundnut. We have identified a total of 21 potential drought-responsive NAC genes in these legumes. These genes displayed correlation between gene expression, transcriptional regulation, and better tolerance against drought. The identified candidate genes, after validation, may serve as a useful resource for molecular breeding for drought tolerance in the SAT legume crops.

scholarly journals Characterization of full-length transcriptome in Saccharum officinarum and molecular insights into tiller development

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Haifeng Yan ◽  
Huiwen Zhou ◽  
Hanmin Luo ◽  
Yegeng Fan ◽  
Zhongfeng Zhou ◽  
...  
Keyword(s):  
Carbon Fixation ◽  
Developmental Stages ◽  
Genomic Data ◽  
Saccharum Officinarum ◽  
Full Length ◽  
Rna Seq ◽  
Pyruvate Phosphate Dikinase ◽  
Sugarcane Yield ◽  
Tiller Bud

Abstract Background Although extensive breeding efforts are ongoing in sugarcane (Saccharum officinarum L.), the average yield is far below the theoretical potential. Tillering is an important component of sugarcane yield, however, the molecular mechanism underlying tiller development is still elusive. The limited genomic data in sugarcane, particularly due to its complex and large genome, has hindered in-depth molecular studies. Results Herein, we generated full-length (FL) transcriptome from developing leaf and tiller bud samples based on PacBio Iso-Seq. In addition, we performed RNA-seq from tiller bud samples at three developmental stages (T0, T1 and T2) to uncover key genes and biological pathways involved in sugarcane tiller development. In total, 30,360 and 20,088 high-quality non-redundant isoforms were identified in leaf and tiller bud samples, respectively, representing 41,109 unique isoforms in sugarcane. Likewise, we identified 1063 and 1037 alternative splicing events identified in leaf and tiller bud samples, respectively. We predicted the presence of coding sequence for 40,343 isoforms, 98% of which was successfully annotated. Comparison with previous FL transcriptomes in sugarcane revealed 2963 unreported isoforms. In addition, we characterized 14,946 SSRs from 11,700 transcripts and 310 lncRNAs. By integrating RNA-seq with the FL transcriptome, 468 and 57 differentially expressed genes (DEG) were identified in T1vsT0 and T2vsT0, respectively. Strong up-regulation of several pyruvate phosphate dikinase and phosphoenolpyruvate carboxylase genes suggests enhanced carbon fixation and protein synthesis to facilitate tiller growth. Similarly, up-regulation of linoleate 9S-lipoxygenase and lipoxygenase genes in the linoleic acid metabolism pathway suggests high synthesis of key oxylipins involved in tiller growth and development. Conclusions Collectively, we have enriched the genomic data available in sugarcane and provided candidate genes for manipulating tiller formation and development, towards productivity enhancement in sugarcane.

scholarly journals Cloning, Characterization, and RNA Interference Effect of the UDP-N-Acetylglucosamine Pyrophosphorylase Gene in Cnaphalocrocis medinalis

Genes ◽  
2021 ◽  
Vol 12 (4) ◽  
pp. 464
Author(s):  
Yuan-Jin Zhou ◽  
Juan Du ◽  
Shang-Wei Li ◽  
Muhammad Shakeel ◽  
Jia-Jing Li ◽  
...  
Keyword(s):  
Developmental Disorders ◽  
Developmental Stages ◽  
Phylogenetic Analyses ◽  
Digital Pcr ◽  
Cnaphalocrocis Medinalis ◽  
Chitin Synthesis ◽  
Reading Frame ◽  
Rnai Technology ◽  
Glyphodes Pyloalis ◽  
Key Enzyme

The rice leaf folder, Cnaphalocrocis medinalis is a major pest of rice and is difficult to control. UDP-N-acetylglucosamine pyrophosphorylase (UAP) is a key enzyme in the chitin synthesis pathway in insects. In this study, the UAP gene from C. medinalis (CmUAP) was cloned and characterized. The cDNA of CmUAP is 1788 bp in length, containing an open reading frame of 1464 nucleotides that encodes 487 amino acids. Homology and phylogenetic analyses of the predicted protein indicated that CmUAP shared 91.79%, 87.89%, and 82.75% identities with UAPs of Glyphodes pyloalis, Ostrinia furnacalis, and Heortia vitessoides, respectively. Expression pattern analyses by droplet digital PCR demonstrated that CmUAP was expressed at all developmental stages and in 12 tissues of C. medinalis adults. Silencing of CmUAP by injection of double-stranded RNA specific to CmUAP caused death, slow growth, reduced feeding and excretion, and weight loss in C. medinalis larvae; meanwhile, severe developmental disorders were observed. The findings suggest that CmUAP is essential for the growth and development of C. medinalis, and that targeting the CmUAP gene through RNAi technology can be used for biological control of this insect.

scholarly journals Antigenic Characterization of ORF2 and ORF3 Proteins of Hepatitis E Virus (HEV)

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1385
Author(s):  
Giulia Pezzoni ◽  
Lidia Stercoli ◽  
Eleonora Pegoiani ◽  
Emiliana Brocchi
Keyword(s):  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Recombinant Antigen ◽  
Open Reading Frame ◽  
Reading Frame ◽  
E Coli ◽  
Antigenic Characterization ◽  
Antigenic Properties ◽  
Open Reading Frame 2

To evaluate the antigenic properties of Hepatitis E Virus (HEV) Open Reading Frame 2 and 3 (ORF2 and ORF3) codified proteins, we expressed different portions of ORF2 and the entire ORF3 in E. coli, a truncated ORF2, was also expressed in baculovirus. A panel of 37 monoclonal antibodies (MAbs) was raised against ORF2 (1–660 amino acids) and MAbs were mapped and characterized using the ORF2 expressed portions. Selected HEV positive and negative swine sera were used to evaluate ORF2 and ORF3 antigens’ immunogenicity. The MAbs were clustered in six groups identifying six antigenic regions along the ORF2. Only MAbs binding to the sixth ORF2 antigenic region (394–608 aa) were found to compete with HEV positive sera and efficiently catch the recombinant antigen expressed in baculovirus. The ORF2 portion from 394–608 aa demonstrated to include most immunogenic epitopes with 85% of HEV positive swine sera reacting against the region from 461–544 aa. Only 5% of the selected HEV sera reacted against the ORF3 antigen.

scholarly journals Molecular cloning and analysis of l(1)ogre, a locus of Drosophila melanogaster with prominent effects on the postembryonic development of the central nervous system.

Genetics ◽  
1990 ◽  
Vol 126 (4) ◽  
pp. 1033-1044 ◽  
Author(s):  
T Watanabe ◽  
D R Kankel
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Postembryonic Development ◽  
P Element ◽  
Reading Frame ◽  
Isolation And Characterization ◽  
The Central Nervous System ◽  
Highly Hydrophobic ◽  
Conceptual Translation

Abstract Previous genetic studies have shown that wild-type function of the l(1)ogre (lethal (1) optic ganglion reduced) locus is essential for the generation and/or maintenance of the postembryonic neuroblasts including those from which the optic lobe is descended. In the present study molecular isolation and characterization of the l(1)ogre locus was carried out to study the structure and expression of this gene in order to gain information about the nature of l(1)ogre function and its relevance to the development of the central nervous system. About 70 kilobases (kb) of genomic DNA were isolated that spanned the region where l(1)ogre was known to reside. Southern analysis of a l(1)ogre mutation and subsequent P element-mediated DNA transformation mapped the l(1)ogre+ function within a genomic fragment of 12.5 kb. Northern analyses showed that a 2.9-kb message transcribed from this 12.5-kb region represented l(1)ogre. A 2.15-kb portion of a corresponding cDNA clone was sequenced. An open reading frame (ORF) of 1,086 base paris was found, and a protein sequence of 362 amino acids with one highly hydrophobic segment was deduced from conceptual translation of this ORF.

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