scholarly journals The conserved PFT1 tandem repeat is crucial for proper flowering in Arabidopsis thaliana

2014 ◽  
Author(s):  
Pauline Rival ◽  
Maximilian O Press ◽  
Jacob Bale ◽  
Tanya Grancharova ◽  
Soledad F Undurraga ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Transgenic Plants ◽  
Tandem Repeat ◽  
Allelic Diversity ◽  
Selective Constraint ◽  
Dependent Manner ◽  
Functional Requirements ◽  
Loss Of Function ◽  
Wild Type ◽  
Genomic Regions

It is widely appreciated that short tandem repeat (STR) variation underlies substantial phenotypic variation in organisms. Some propose that the high mutation rates of STRs in functional genomic regions facilitate evolutionary adaptation. Despite their high mutation rate, some STRs show little to no variation in populations. One such STR occurs in the Arabidopsis thaliana gene PFT1 (MED25), where it encodes an interrupted polyglutamine tract. Though the PFT1 STR is large (~270 bp), and thus expected to be extremely variable, it shows only minuscule variation across A. thaliana strains. We hypothesized that the PFT1 STR is under selective constraint, due to previously undescribed roles in PFT1 function. We investigated this hypothesis using plants expressing transgenic PFT1 constructs with either an endogenous STR or with synthetic STRs of varying length. Transgenic plants carrying the endogenous PFT1 STR generally performed best across adult PFT1-dependent traits. In stark contrast, transgenic plants carrying a PFT1 transgene lacking the STR entirely phenocopied a pft1 loss-of-function mutant for flowering time phenotypes, and were generally hypomorphic for other traits, establishing the functional importance of this domain. Transgenic plants carrying various synthetic constructs occupied the phenotypic space between wild-type and pft1-loss-of-function mutants. By varying PFT1 STR length, we discovered that PFT1 can act as either an activator or repressor of flowering in a photoperiod-dependent manner. We conclude that the PFT1 STR is constrained to its approximate wild-type length by its various functional requirements. Our study implies that there is strong selection on STRs not only to generate allelic diversity, but also to maintain certain lengths pursuant to optimal molecular function.

scholarly journals First genome edited poinsettias: targeted mutagenesis of flavonoid 3′-hydroxylase using CRISPR/Cas9 results in a colour shift

2021 ◽  
Author(s):  
Daria Nitarska ◽  
Robert Boehm ◽  
Thomas Debener ◽  
Rares Calin Lucaciu ◽  
Heidi Halbwirth
Keyword(s):  
Transgenic Plants ◽  
Genome Editing ◽  
Ornamental Plants ◽  
Targeted Mutagenesis ◽  
Cloning And Expression ◽  
Euphorbia Pulcherrima ◽  
Loss Of Function ◽  
Wild Type ◽  
Promising Tool ◽  
Reddish Orange

AbstractThe CRISPR/Cas9 system is a remarkably promising tool for targeted gene mutagenesis, and becoming ever more popular for modification of ornamental plants. In this study we performed the knockout of flavonoid 3′-hydroxylase (F3′H) with application of CRISPR/Cas9 in the red flowering poinsettia (Euphorbia pulcherrima) cultivar ‘Christmas Eve’, in order to obtain plants with orange bract colour, which accumulate prevalently pelargonidin. F3′H is an enzyme that is necessary for formation of cyanidin type anthocyanins, which are responsible for the red colour of poinsettia bracts. Even though F3′H was not completely inactivated, the bract colour of transgenic plants changed from vivid red (RHS 45B) to vivid reddish orange (RHS 33A), and cyanidin levels decreased significantly compared with the wild type. In the genetically modified plants, an increased ratio of pelargonidin to cyanidin was observed. By cloning and expression of mutated proteins, the lack of F3′H activity was confirmed. This confirms that a loss of function mutation in the poinsettia F3′H gene is sufficient for obtaining poinsettia with orange bract colour. This is the first report of successful use of CRISPR/Cas9 for genome editing in poinsettia.

scholarly journals Disruption of the ARF transcriptional activator DMP1 facilitates cell immortalization, Ras transformation, and tumorigenesis

2000 ◽  
Vol 14 (14) ◽  
pp. 1797-1809 ◽  
Author(s):  
Kazushi Inoue ◽  
Renren Wen ◽  
Jerold E. Rehg ◽  
Masashi Adachi ◽  
John L. Cleveland ◽  
...  
Keyword(s):  
Cell Lines ◽  
Interleukin 2 ◽  
Embryonic Stem ◽  
Suppressor Gene ◽  
Null Mouse ◽  
Dependent Manner ◽  
Loss Of Function ◽  
Wild Type ◽  
Functional Protein ◽  
Early Passage

The DMP1 transcription factor induces the ARF tumor suppressor gene in mouse fibroblasts, leading to cell cycle arrest in a p53-dependent manner. We disrupted sequences encoding the DNA-binding domain of DMP1 in mouse embryonic stem cells and derived animals lacking the functional protein. DMP1-null animals are small at birth, and males develop more slowly than their wild-type littermates. Some adult animals exhibit seizures and/or obstuctive uropathy, each of unknown cause. The growth of explantedDMP1-null mouse embryo fibroblasts (MEFs) is progressively retarded as cells are passaged in culture on defined transfer protocols; but, unlike the behavior of normal cells, p19ARF, Mdm2, and p53 levels remain relatively low and DMP1-null MEFs do not senesce. Whereas the establishment of cell lines from MEFs is usually always accompanied by either p53 or ARF loss of function, continuously passaged DMP1-null cells readily give rise to established 3T3 and 3T9 cell lines that retain wild-type ARFand functional p53 genes. Early-passage DMP1-null cells, like MEFs from either ARF-null or p53-null mice, can be morphologically transformed by oncogenic Ha-Ras (Val-12) alone. Splenic lymphocytes harvested from both DMP1-null andARF-null mice exhibit enhanced proliferative responses in long-term cultures when stimulated to divide with antibody to CD3 and interleukin-2. Although only 1 of 40 DMP1-null animals spontaneously developed a tumor in the first year of life, neonatal treatment with dimethylbenzanthracene or ionizing radiation induced tumors of various histologic types that were not observed in similarly treated DMP1+/+ animals. Karyotypic analyses of MEFs and lymphomas from DMP1-null animals revealed pseudodiploid chromosome numbers, consistent with the retention of wild-type p53. Together, these data suggest that ARF function is compromised, but not eliminated, in animals lacking functional DMP1.

Synergistic Derepression of Gibberellin Signaling by Removing RGA and GAI Function in Arabidopsis thaliana

Genetics ◽  
2001 ◽  
Vol 159 (2) ◽  
pp. 777-785
Author(s):  
Alyssa Dill ◽  
Tai-ping Sun
Keyword(s):  
Arabidopsis Thaliana ◽  
Feedback Inhibition ◽  
Stem Elongation ◽  
Dominant Role ◽  
Signal Transduction Pathway ◽  
Loss Of Function ◽  
Wild Type ◽  
Ga Signaling ◽  
Mutant Phenotypes ◽  
Redundant Functions

Abstract RGA and GAI are negative regulators of the gibberellin (GA) signal transduction pathway in Arabidopsis thaliana. These genes may have partially redundant functions because they are highly homologous, and plants containing single null mutations at these loci are phenotypically similar to wild type. Previously, rga loss-of-function mutations were shown to partially suppress defects of the GA-deficient ga1-3 mutant. Phenotypes rescued include abaxial trichome initiation, rosette radius, flowering time, stem elongation, and apical dominance. Here we present work showing that the rga-24 and gai-t6 null mutations have a synergistic effect on plant growth. Although gai-t6 alone has little effect, when combined with rga-24, they completely rescued the above defects of ga1-3 to wild-type or GA-overdose phenotype. However, seed germination and flower development defects were not restored. Additionally, rga-24 and rga-24/gai-t6 but not gai-t6 alone caused increased feedback inhibition of expression of a GA biosynthetic gene in both the ga1-3 and wild-type backgrounds. These results demonstrate that RGA and GAI have partially redundant functions in maintaining the repressive state of the GA-signaling pathway, but RGA plays a more dominant role than GAI. Removing both RGA and GAI function allows for complete derepression of many aspects of GA signaling.

scholarly journals PUCHI Regulates Giant Cell Morphology During Root-Knot Nematode Infection in Arabidopsis thaliana

2021 ◽  
Vol 12 ◽  
Author(s):  
Reira Suzuki ◽  
Mizuki Yamada ◽  
Takumi Higaki ◽  
Mitsuhiro Aida ◽  
Minoru Kubo ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Cell Wall ◽  
Genetic Regulation ◽  
Three Dimensional ◽  
Acid Synthesis ◽  
Giant Cells ◽  
Loss Of Function ◽  
Wild Type ◽  
Root Knot Nematode ◽  
In The Wild

Parasitic root-knot nematodes transform the host’s vascular cells into permanent feeding giant cells (GCs) to withdraw nutrients from the host plants. GCs are multinucleated metabolically active cells with distinctive cell wall structures; however, the genetic regulation of GC formation is largely unknown. In this study, the functions of the Arabidopsis thaliana transcription factor PUCHI during GC development were investigated. PUCHI expression was shown to be induced in early developing galls, suggesting the importance of the PUCHI gene in gall formation. Despite the puchi mutant not differing significantly from the wild type in nematode invasion and reproduction rates, puchi GC cell walls appeared to be thicker and lobate when compared to the wild type, while the cell membrane sometimes formed invaginations. In three-dimensional (3D) reconstructions of puchi GCs, they appeared to be more irregularly shaped than those in the wild type, with noticeable cell-surface protrusions and folds. Interestingly, the loss-of-function mutant of 3-KETOACYL-COA SYNTHASE 1 showed GC morphology and cell wall defects similar to those of the puchi mutant, suggesting that PUCHI may regulate GC development via very long chain fatty acid synthesis.

scholarly journals Identification of an arabinopyranosyltransferase fromPhyscomitrella patensinvolved in the synthesis of the hemicellulose xyloglucan

2017 ◽  
Author(s):  
Lei Zhu ◽  
Murali Dama ◽  
Markus Pauly
Keyword(s):  
Arabidopsis Thaliana ◽  
Transgenic Plants ◽  
Linkage Analysis ◽  
Double Mutant ◽  
Physcomitrella Patens ◽  
Proper Function ◽  
Side Chain ◽  
Glycosidic Linkage ◽  
Nmr Analysis ◽  
Wild Type

AbstractThe hemicellulose xyloglucan consists of a backbone of a β-1,4 glucan substituted with xylosyl moieties and many other, diverse sidechains that are important for its proper function. Many, but not all glycosyltransferases involved in the biosynthesis of xyloglucan have been identified. Here, we report the identification of an hitherto elusive xyloglucan:arabinopyranosyltransferase. This glycosyltransferase was isolated from the mossPhyscomitrella patens, where it acts as aXyloglucan “D”-side-chainTransferase (XDT). Heterologous expression ofXDTin theArabidopsis thalianadouble mutantmur3.1 xlt2, where xyloglucan consists of a xylosylated glucan without further glycosyl substituents, results in the production of the arabinopyranose-containing “D” side chain as characterized by oligosaccharide mass profiling, glycosidic linkage analysis, and NMR analysis. In addition, expression of a relatedPhyscomitrellaglycosyltransferase hortholog ofXLT2leads to the production of the galactose-containing “L” side chain. The presence of the “D” and “L” xyloglucan side chains inPpXDT mur3.1 xlt2andPpXLT2 mur3.1 xlt2transgenic plants, respectively, rescue the dwarfed phenotype of untransformedmur3.1 xlt2mutants to nearly wild-type height. Expression ofPpXDTandPpXLT2in the Arabidopsismur3.1 xlt2mutant also enhanced root growth.

The role of SORBITOL DEHYDROGENASE in Arabidopsis thaliana

2012 ◽  
Vol 39 (6) ◽  
pp. 462 ◽  
Author(s):  
Marta Nosarzewski ◽  
A. Bruce Downie ◽  
Benhong Wu ◽  
Douglas D. Archbold
Keyword(s):  
Arabidopsis Thaliana ◽  
Drought Stress ◽  
Sorbitol Dehydrogenase ◽  
Primary Role ◽  
Dependent Manner ◽  
Plant Leaves ◽  
Wild Type ◽  
Concentration Dependent Manner ◽  
Knockout Mutants

SORBITOL DEHYDROGENASE (SDH, EC 1.1.1.14) catalyses the interconversion of polyols and ketoses (e.g. sorbitol ↔ fructose). Using two independent Arabidopsis thaliana (L.) Heynh. sdh knockout mutants, we show that SDH (At5g51970) plays a primary role in sorbitol metabolism as well as an unexpected role in ribitol metabolism. Sorbitol content increased in both wild-type (WT) and mutant plant leaves during drought stress, but mutants showed a dramatically different phenotype, dying even if rewatered. The lack of functional SDH in mutant plants was accompanied by accumulation of foliar sorbitol and at least 10-fold more ribitol, neither of which decreased in mutant plants after rewatering. In addition, mutant plants were uniquely sensitive to ribitol in a concentration-dependent manner, which either prevented them from completing seed germination or inhibited seedling development, effects not observed with other polyols or with ribitol-treated WT plants. Ribitol catabolism may occur solely through SDH in A. thaliana, though at only 30% the rate of that for sorbitol. The results indicate a role for SDH in metabolism of sorbitol to fructose and in ribitol conversion to ribulose in A. thaliana during recovery from drought stress.

scholarly journals PLATZ2 negatively regulates salt tolerance in Arabidopsis seedlings by directly suppressing the expression of the CBL4/SOS3 and CBL10/SCaBP8 genes

2020 ◽  
Vol 71 (18) ◽  
pp. 5589-5602
Author(s):  
Shasha Liu ◽  
Rui Yang ◽  
Miao Liu ◽  
Shizhong Zhang ◽  
Kang Yan ◽  
...  
Keyword(s):  
Salt Stress ◽  
Salt Tolerance ◽  
Transgenic Plants ◽  
Loss Of Function ◽  
Wild Type ◽  
Salt Stress Response ◽  
In The Wild ◽  
Plant Salt Tolerance

Abstract Although the salt overly sensitive (SOS) pathway plays essential roles in conferring salt tolerance in Arabidopsis thaliana, the regulatory mechanism underlying SOS gene expression remains largely unclear. In this study, AtPLATZ2 was found to function as a direct transcriptional suppressor of CBL4/SOS3 and CBL10/SCaBP8 in the Arabidopsis salt stress response. Compared with wild-type plants, transgenic plants constitutively overexpressing AtPLATZ2 exhibited increased sensitivity to salt stress. Loss of function of PLATZ2 had no observed salt stress phenotype in Arabidopsis, while the double mutant of PLATZ2 and PLATZ7 led to weaker salt stress tolerance than wild-type plants. Overexpression of AtPLATZ2 in transgenic plants decreased the expression of CBL4/SOS3 and CBL10/SCaBP8 under both normal and saline conditions. AtPLATZ2 directly bound to A/T-rich sequences in the CBL4/SOS3 and CBL10/SCaBP8 promoters in vitro and in vivo, and inhibited CBL4/SOS3 promoter activity in the plant leaves. The salt sensitivity of #11 plants constitutively overexpressing AtPLATZ2 was restored by the overexpression of CBL4/SOS3 and CBL10/SCaBP8. Salt stress-induced Na+ accumulation in both the shoots and roots was more exaggerated in AtPLATZ2-overexpressing plants than in the wild type. The salt stress-induced Na+ accumulation in #11 seedlings was also rescued by the overexpression of CBL4/SOS3 and CBL10/SCaBP8. Furthermore, the transcription of AtPLATZ2 was induced in response to salt stress. Collectively, these results suggest that AtPLATZ2 suppresses plant salt tolerance by directly inhibiting CBL4/SOS3 and CBL10/SCaBP8, and functions redundantly with PLATZ7.

Tolerance to, and Uptake and Degradation of 2,4,6-Trinitrotoluene (TNT) are Enhanced by the Expression of a Bacterial Nitroreductase Gene in Arabidopsis thaliana

2005 ◽  
Vol 60 (3-4) ◽  
pp. 272-278 ◽  
Author(s):  
Mami Kurumata ◽  
Misa Takahashi ◽  
Atsushi Sakamoto ◽  
Juan L. Ramos ◽  
Ales Nepovim ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Transgenic Plants ◽  
Degradation Products ◽  
Plant Genome ◽  
Pcr Analysis ◽  
Wild Type ◽  
In Planta ◽  
Gene Encoding ◽  
Plant Body ◽  
Wide Range

Abstract Arabidopsis thaliana was transformed with a gene encoding a nitroreductase (NTR, E.C. 1.6.99.7) with activity against a wide range of nitroaromatic compounds. The gene was transferred from Escherichia coli by an Agrobacterium-mediated in planta method. The ob­tained seeds were sowed to produce T1 plants, and they were assayed for the integration of the transgene in the plant genome. Transgenic plants that were positive with the PCR analysis were self-pollinated to produce T2 generation plants. Seven lines obtained were assayed for the NTR activity. While the noil-transformed wild-type plants showed no detectable NTR activity, the enzyme activity of the transgenic plant lines was approx. 20 times higher. Using the line with the highest NTR activity, the phytoremediation characteristics of plants against 2,4,6-trinitrotoluene (TNT) was investigated. While the wild-type plants did not grow in the presence of 0.1 mᴍ TNT, the transgenic plants grew almost normally in this condition. The uptake of TNT by seedlings of transgenic plants increased by 7 to 8 times when they were floated on TNT solution. HPLC analysis showed that the peak due to TNT taken up into plant body was much smaller in the transgenic plants as compared with that of the wild type, and that a number of peaks attributable to the degradation products of TNT, including 4-amino-2,6-dinitrotoluene, were detected in the extract from the transgenic plants. This indi­cates that the expression of bacterial NTR improved the capability of plants to degrade TNT.

A Study of Arabidopsis Cold Stress Tolerance Improvement via AthHOS1-Targeting HOS1-AmiRNA Approach

2021 ◽  
Author(s):  
Marzieh Karimi ◽  
Behrouz Shiran ◽  
Mohammad Rabei ◽  
Hossein Fallahi ◽  
Bojana Banović Đeri
Keyword(s):  
Gene Expression ◽  
Arabidopsis Thaliana ◽  
Circadian Rhythm ◽  
Comparative Analysis ◽  
Low Temperature ◽  
Transgenic Plants ◽  
Cold Stress ◽  
Stress Tolerance ◽  
Low Temperatures ◽  
Wild Type

Abstract In this study the artificial microRNAs (amiRNAs) technology targeting HOS1 gene was tested for its applicability for the improvement of cold stress tolerance in Landsberg-0 (Ler-0) ecotype of Arabidopsis thaliana. The chosen approach was designed to suppress AtHOS1 gene expression through the overexpression of amiRNA-HOS1. The effect of AtHOS1-amiRNA overexpression to transgenic plants’ response to cold stress was determined by Real Time PCR. The expression levels of amiRNA and its target, AtHOS1 gene, were observed in 3-week old seedlings of T3 generation and in wild-type plants after 6h, 12h, 24h, 48h and 96h of their exposure to cold stress (4ºC). Comparative analysis revealed that AtHOS1-amiRNA negatively regulated AtHOS1 in transgenic plants upon plants lengthen exposure (for 48h and 96h) to low temperature (Pearson’s correlation coefficient of -0.407; P < 0.05). Even though prolonged cold stress caused extended up regulation of AtHOS1 in wild type plants, in transgenic plants AtHOS1-amiRNA suppression disturbed expected AtHOS1 circadian rhythm by preventing further AtHOS1 up regulation. Moreover, transgenic plants showed AtHOS1 down regulation 96h after the cold stress onset, due to sufficient overexpression of AtHOS1-amiRNA, which allowed cold signaling amplification in transgenic plants. As a result of that, cold-acclimated transformed plants displayed 17% higher freezing tolerance (-1°C to -8°C) in comparison to wild type plants, demonstrating the success of chosen approach in improving Arabidopsis tolerance to low temperatures, at least in Ler-0 ecotype.

Sign in / Sign up

Export Citation Format

Share Document