scholarly journals A Highly Mutable GST is Essential for Bract Colouration in Euphorbia Pulcherrima Willd. Ex Klotsch

2020 ◽  
Author(s):  
Vinicius Vilperte ◽  
Robert Boehm ◽  
Thomas Debener
Keyword(s):  
Genetic Variability ◽  
Anthocyanin Biosynthesis ◽  
Mutation Breeding ◽  
Euphorbia Pulcherrima ◽  
Wild Type ◽  
Coding Region ◽  
X Ray ◽  
Short Repeat ◽  
Irradiation Treatment ◽  
Dynamic Mutations

Abstract Background: Mutation breeding is an extraordinary tool in plant breeding to increase the genetic variability, where mutations in anthocyanin biosynthesis are targets to generate distinctive phenotypes in ornamental species. In poinsettia, ionizing radiation is routinely applied in breeding programs to obtaining a range of colours, with nearly all pink and white varieties being obtained after γ- or X-ray mutagenesis of red varieties. In the present study we performed a thorough characterization of a potential mutagenesis target gene as the main responsible for the ‘white paradox’ in poinsettiaResults: We identified a GST gene in poinsettia (Bract1) as an essential factor for the expression of anthocyanin-based red colouration of bracts, which presents a high phylogenetic similarity to known anthocyanin-related GSTs. Red poinsettia varieties and white mutants generated from these varieties by X-ray were analysed for polymorphisms related to the ‘white paradox’ in the species. A 4 bp mutation in a short repeat within the coding region of Bract1 is most likely responsible for the appearance of white phenotypes upon irradiation treatment. The polymorphism between wild-type and mutant alleles co-segregates with the phenotype in progeny from heterozygous red and white parents. Moreover, overexpression of Bract1 wild-type allele in Arabidopsis tt19 mutants restored the anthocyanin phenotype, while the Bract1 mutated allele showed to be non-functional. Conclusions: The identified repeat seems to be highly unstable, since mutated plants can be easily detected among fewer than 200 shoots derived from 10 mutated plants. Our data indicate that particular short repeat sequences, similar to microsatellite sequences or so-called dynamic mutations, might be hot spots for genetic variability. Moreover, the identification of the Bract1 mutation fills a gap on the understanding on the molecular mechanism of colour formation in poinsettia

scholarly journals A highly mutable GST is essential for bract colouration in Euphorbia pulcherrima Willd. Ex Klotsch

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Vinicius Vilperte ◽  
Robert Boehm ◽  
Thomas Debener
Keyword(s):  
Genetic Variability ◽  
Anthocyanin Biosynthesis ◽  
Mutation Breeding ◽  
Euphorbia Pulcherrima ◽  
Wild Type ◽  
Coding Region ◽  
X Ray ◽  
Short Repeat ◽  
Irradiation Treatment ◽  
Dynamic Mutations

AbstractBackgroundMutation breeding is an extraordinary tool in plant breeding to increase the genetic variability, where mutations in anthocyanin biosynthesis are targets to generate distinctive phenotypes in ornamental species. In poinsettia, ionizing radiation is routinely applied in breeding programs to obtaining a range of colours, with nearly all pink and white varieties being obtained after γ- or X-ray mutagenesis of red varieties. In the present study we performed a thorough characterization of a potential mutagenesis target gene as the main responsible for the ‘white paradox’ in poinsettia.ResultsWe identified aGSTgene in poinsettia (Bract1) as an essential factor for the expression of anthocyanin-based red colouration of bracts, which presents a high phylogenetic similarity to known anthocyanin-related GSTs. Red poinsettia varieties and white mutants generated from these varieties by X-ray were analysed for polymorphisms related to the ‘white paradox’ in the species. A 4 bp mutation in a short repeat within the coding region ofBract1is most likely responsible for the appearance of white phenotypes upon irradiation treatment. The polymorphism between wild-type and mutant alleles co-segregates with the phenotype in progeny from heterozygous red and white parents. Moreover, overexpression ofBract1wild-type allele in Arabidopsistt19mutants restored the anthocyanin phenotype, while theBract1mutated allele showed to be non-functional.ConclusionsThe identified repeat seems to be highly unstable, since mutated plants can be easily detected among fewer than 200 shoots derived from 10 mutated plants. Our data indicate that particular short repeat sequences, similar to microsatellite sequences or so-called dynamic mutations, might be hot spots for genetic variability. Moreover, the identification of theBract1mutation fills a gap on the understanding on the molecular mechanism of colour formation in poinsettia.

scholarly journals Biological effects of electron beam to target turning X-ray (EBTTX) on two freesia (Freesia hybrida) cultivars

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e10742
Author(s):  
Yi-rui Li ◽  
Ling Liu ◽  
Dan Wang ◽  
Li Chen ◽  
Hao Chen
Keyword(s):  
Electron Beam ◽  
Survival Rate ◽  
Irradiation Dose ◽  
Biological Effects ◽  
Lethal Dose ◽  
Mutation Breeding ◽  
Flower Number ◽  
X Ray ◽  
Irradiation Treatment ◽  
Flowering Rate

Electron beam to target turning X-ray (EBTTX) is an emerging irradiation technology that can potentially accelerate the breeding process of plants. The biological effects of EBTTX irradiation on the two freesia cultivars (the red freesia and the purple freesia) were investigated by establishing an irradiation-mediated mutation breeding protocol. The germination rate, survival rate, plant height, leaf number and area, root number and length of the two freesia cultivars decreased following different irradiation doses (25, 50, 75, and 100-Gy). A high irradiation dose exhibited stronger inhibition effects on these plant growth parameters, and the survival rate of the two freesia cultivars was 0.00% following the 100-Gy irradiation treatment. The median lethal dose (LD50) based on survival rates was 54.28-Gy for the red freesia and 60.11-Gy for the purple freesia. The flowering rate, flower number, and pollen vigor were significantly decreased by irradiation treatment. At 75-Gy irradiation, the flowering rate, flower number and pollen viability of the two varieties reached the minimum, exhibiting strong inhibitory effects. Meanwhile, 75-Gy irradiation significantly decreased the chlorophyll content and increased the malondialdehyde (MDA) content of the two freesia cultivars. Furthermore, as the irradiation dose increased, the changes in the micro-morphology of the leaf epidermis and pollen gradually increased according to a scanning electron microscope (SEM) analysis. These results are expected to provide useful information for the mutation breeding of different freesia cultivars and other flowering plants.

Lethal Dose (LD50) Fixation and Sensitivity of Fenugreek (Trigonellafoenum-graecumL.) to Gamma Radiation for Induction of Mutation

2017 ◽  
Vol 4 (04) ◽  
Author(s):  
ANURADHA PATEL ◽  
POONAM VERMA ◽  
SHARDA CHOUDHARY ◽  
ARVIND KUMAR VERMA
Keyword(s):  
Genetic Variability ◽  
Gamma Rays ◽  
Growth Parameters ◽  
Seedling Survival ◽  
Lethal Dose ◽  
Germination Percentage ◽  
Mutation Breeding ◽  
Vigour Index ◽  
Small Flower ◽  
Dose Dependent

Fenugreek (Trigonella foenum-graecumL.) is an annual crop, mainly used as a spiceand leafy vegetable crop in many parts of the world. Classical breeding in fenugreek is restricted due to its low genetic variability and small flower size which hamper manual emasculation and pollination. Mutation breeding is an effective way to enrich genetic variability in crop plants. An experiment was conducted to determine the lethal dose of the physical mutagen gamma rays in fenugreek. The dry seeds of fenugreek were exposed to different doses of gamma rays i.e. 150Gy, 200Gy, 250Gy, 300Gy and 350Gy. These irradiated seeds were sown in the Petri plates with non-irradiated seeds (control). As the dose of gamma rays increased, there was a decrease in germination percentage, seedling survival, root length, shoot length and vigour index. Among five doses of gamma rays, the maximum seed germination was observed at lowest dose 150Gy (93%), followed by 200Gy (83%), 250Gy (76%), 300Gy (76%) and 350Gy (64%). The seedling survival was decreased from 90% (in control) to 56% in 350Gy dose of gamma rays. The gamma rays dose of 150Gy gave stimulatory effect on seedlings growth. The growth parameters were dose dependent, as the dose of gamma rays increased from 200Gy to 350Gy. The gamma rays dose of 350Gy showed 64% seeds germination and 56% of seedlings survival. Therefore, it is concluded that the LD50 dose for fenugreek is close to 350Gy. This information would be highly useful for initiating mutation breeding programme in fenugreek

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 Treatment of Cancer and Inflammation With Low-Dose Ionizing Radiation

Dose-Response ◽  
2017 ◽  
Vol 15 (1) ◽  
pp. 155932581769753 ◽  
Author(s):  
Shuji Kojima ◽  
Mitsutoshi Tsukimoto ◽  
Noriko Shimura ◽  
Hironobu Koga ◽  
Akishisa Murata ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Ulcerative Colitis ◽  
Bone Metastasis ◽  
Low Dose ◽  
Case Reports ◽  
Low Dose Radiation ◽  
X Ray ◽  
Radiation Hormesis ◽  
Irradiation Treatment ◽  
Dose Radiation

There is considerable evidence from experimental studies in animals, as well as from clinical reports, that low-dose radiation hormesis is effective for the treatment of cancer and ulcerative colitis. In this study, we present 3 case reports that support the clinical efficacy of low-dose radiation hormesis in patients with these diseases. First, a patient with prostate cancer who had undergone surgical resection showed a subsequent increase in prostate-specific antigen (PSA). His PSA value started decreasing immediately after the start of repeated low-dose X-ray irradiation treatment and remained low thereafter. Second, a patient with prostate cancer with bone metastasis was treated with repeated low-dose X-ray irradiation. His PSA level decreased to nearly normal within 3 months after starting the treatment and remained at the low level after the end of hormesis treatment. His bone metastasis almost completely disappeared. Third, a patient with ulcerative colitis showed a slow initial response to repeated low-dose irradiation treatment using various modalities, including drinking radon-containing water, but within 8 months, his swelling and bleeding had completely disappeared. After 1 year, the number of bowel movements had become normal. Interest in the use of radiation hormesis in clinical practice is increasing, and we hope that these case reports will encourage further clinical investigations.

scholarly journals Essential Function of the Pseudorabies Virus UL36 Gene Product Is Independent of Its Interaction with the UL37 Protein

2004 ◽  
Vol 78 (21) ◽  
pp. 11879-11889 ◽  
Author(s):  
Walter Fuchs ◽  
Barbara G. Klupp ◽  
Harald Granzow ◽  
Thomas C. Mettenleiter
Keyword(s):  
Gene Product ◽  
Pseudorabies Virus ◽  
Rabbit Kidney ◽  
Tegument Protein ◽  
Wild Type ◽  
Coding Region ◽  
Ordered Structures ◽  
Ca 50 ◽  
Essential Function ◽  
Full Length Clone

ABSTRACT The large tegument protein encoded by the UL36 gene of pseudorabies virus (PrV) physically interacts with the product of the adjacent UL37 gene (B. G. Klupp, W. Fuchs, H. Granzow, R. Nixdorf, and T. C. Mettenleiter, J. Virol. 76:3065-3071, 2002). To analyze UL36 function, two PrV recombinants were generated by mutagenesis of an infectious PrV full-length clone in Escherichia coli: PrV-ΔUL36F exhibited a deletion of virtually the complete UL36 coding region, whereas PrV-UL36BSF contained two in-frame deletions of 238 codons spanning the predicted UL37 binding domain. Coimmunoprecipitation experiments confirmed that the mutated gene product of PrV-UL36BSF did not interact with the UL37 protein. Like the previously described PrV-ΔUL37 (B. G. Klupp, H. Granzow, and T. C. Mettenleiter, J. Virol. 75:8927-8936, 2001) but in contrast to PrV-ΔUL36F, PrV-UL36BSF was able to replicate in rabbit kidney (RK13) cells, although maximum virus titers were reduced ca. 50-fold and plaque diameters were reduced by ca. 45% compared to wild-type PrV. PrV-ΔUL36F was able to productively replicate after repair of the deleted gene or in a trans-complementing cell line. Electron microscopy of infected RK13 cells revealed that PrV-UL36BSF and phenotypically complemented PrV-ΔUL36F were capable of nucleocapsid formation and egress from the nucleus by primary envelopment and deenvelopment at the nuclear membrane. However, reenvelopment of nucleocapsids in the cytoplasm was blocked. Only virus-like particles without capsids were released efficiently from cells. Interestingly, cytoplasmic nucleocapsids of PrV-UL36BSF but not of PrV-ΔUL36F were found in large ordered structures similar to those which had previously been observed with PrV-ΔUL37. In summary, our results demonstrate that the interaction between the UL36 and UL37 proteins is important but not strictly essential for the formation of secondary enveloped, infectious PrV particles. Furthermore, UL36 possesses an essential function during virus replication which is independent of its ability to bind the UL37 protein.

scholarly journals Systemic Polyomavirus Genome Increase and Dissemination of Capsid-Defective Genomes in Mammary Gland Tumor-Bearing Mice

2000 ◽  
Vol 74 (15) ◽  
pp. 6975-6983 ◽  
Author(s):  
Julie J. Wirth ◽  
Li Chen ◽  
Michele M. Fluck
Keyword(s):  
Mammary Gland ◽  
Viral Genome ◽  
Wild Type Strain ◽  
Neonatal Infection ◽  
Wild Type ◽  
Coding Region ◽  
Live Virus ◽  
Viral Genomes ◽  
Mammary Gland Tumors ◽  
Low Levels

ABSTRACT BALB/c mice that developed tumors 7 to 8 months following neonatal infection by polyomavirus (PYV) wild-type strain A2 were characterized with respect to the abundance and integrity of the viral genome in the tumors and in 12 nontumorous organs. These patterns were compared to those found in tumor-free mice infected in parallel. Six mice were analyzed in detail including four sibling females with mammary gland tumors. In four of five mammary gland tumors, the viral genome had undergone a unique deletion and/or rearrangement. Three tumor-resident genomes with an apparently intact large T coding region were present in abundant levels in an unintegrated state. Two of these had undergone deletions and rearrangements involving the capsid genes and therefore lacked the capacity to produce live virus. In the comparative organ survey, the tumors harboring replication-competent genomes contained by far the highest levels of genomes of any tissue. However, the levels of PYV genomes in other organs were elevated by up to 1 to 2 orders of magnitude compared to those detected in the same organs of tumor-free mice. The genomes found in the nontumorous organs had the same rearrangements as the genomes residing in the tumors. The original wild-type genome was detected at low levels in a few organs, particularly in the kidneys. The data indicate that a systemic increase in the level of viral genomes occurred in conjunction with the induction of tumors by PYV. The results suggest two novel hypotheses: (i) that genomes may spread from the tumors to the usual PYV target tissues and (ii) that this dissemination may take place in the absence of capsids, providing an important path for a virus to escape from the immune response. This situation may offer a useful model for the spread of HPV accompanying HPV-induced oncogenesis.

scholarly journals Structural defects of a Pax8 mutant that give rise to congenital hypothyroidism

1999 ◽  
Vol 341 (1) ◽  
pp. 89-93 ◽  
Author(s):  
Gianluca TELL ◽  
Lucia PELLIZZARI ◽  
Gennaro ESPOSITO ◽  
Carlo PUCILLO ◽  
Paolo Emidio MACCHIA ◽  
...  
Keyword(s):  
Dna Sequence ◽  
Congenital Hypothyroidism ◽  
Dna Sequences ◽  
Dna Interaction ◽  
Structural Defects ◽  
Molecular Defect ◽  
Wild Type ◽  
Coding Region ◽  
Induced Fit ◽  
Helical Content

Pax proteins are transcriptional regulators that play important roles during embryogenesis. These proteins recognize specific DNA sequences via a conserved element: the paired domain (Prd domain). The low level of organized secondary structure, in the free state, is a general feature of Prd domains; however, these proteins undergo a dramatic gain in α-helical content upon interaction with DNA (‘induced fit’). Pax8 is expressed in the developing thyroid, kidney and several areas of the central nervous system. In humans, mutations of the Pax8 gene, which are mapped to the coding region of the Prd domain, give rise to congenital hypothyroidism. Here, we have investigated the molecular defects caused by a mutation in which leucine at position 62 is substituted for an arginine. Leu62 is conserved among Prd domains, and contributes towards the packing together of helices 1 and 3. The binding affinity of the Leu62Arg mutant for a specific DNA sequence (the C sequence of thyroglobulin promoter) is decreased 60-fold with respect to the wild-type Pax8 Prd domain. However, the affinities with which the wild-type and the mutant proteins bind to a non-specific DNA sequence are very similar. CD spectra demonstrate that, in the absence of DNA, both wild-type Pax8 and the Leu62Arg mutant possess a low α-helical content; however, in the Leu62Arg mutant, the gain in α-helical content upon interaction with DNA is greatly reduced with respect to the wild-type protein. Thus the molecular defect of the Leu62Arg mutant causes a reduced capability for induced fit upon DNA interaction.

scholarly journals Comparative reactions of recombinant papaya ringspot viruses with chimeric coat protein (CP) genes and wild-type viruses on CP-transgenic papaya

2001 ◽  
Vol 82 (11) ◽  
pp. 2827-2836 ◽  
Author(s):  
Chu-Hui Chiang ◽  
Ju-Jung Wang ◽  
Fuh-Jyh Jan ◽  
Shyi-Dong Yeh ◽  
Dennis Gonsalves
Keyword(s):  
Coat Protein ◽  
Sequence Similarity ◽  
Transcriptional Gene Silencing ◽  
Papaya Ringspot Virus ◽  
Wild Type ◽  
Coding Region ◽  
Transgenic Papaya ◽  
Cp Gene ◽  
Post Transcriptional Gene Silencing

Transgenic papaya cultivars SunUp and Rainbow express the coat protein (CP) gene of the mild mutant of papaya ringspot virus (PRSV) HA. Both cultivars are resistant to PRSV HA and other Hawaii isolates through homology-dependent resistance via post-transcriptional gene silencing. However, Rainbow, which is hemizygous for the CP gene, is susceptible to PRSV isolates from outside Hawaii, while the CP-homozygous SunUp is resistant to most isolates but susceptible to the YK isolate from Taiwan. To investigate the role of CP sequence similarity in overcoming the resistance of Rainbow, PRSV HA recombinants with various CP segments of the YK isolate were constructed and evaluated on Rainbow, SunUp and non-transgenic papaya. Non-transgenic papaya were severely infected by all recombinants, but Rainbow plants developed a variety of symptoms. On Rainbow, a recombinant with the entire CP gene of YK caused severe symptoms, while recombinants with only partial YK CP sequences produced a range of milder symptoms. Interestingly, a recombinant with a YK segment from the 5′ region of the CP gene caused very mild, transient symptoms, whereas recombinants with YK segments from the middle and 3′ parts of the CP gene caused prominent and lasting symptoms. SunUp was resistant to all but two recombinants, which contained the entire CP gene or the central and 3′-end regions of the CP gene and the 3′ non-coding region of YK, and the resulting symptoms were mild. It is concluded that the position of the heterologous sequences in the recombinants influences their pathogenicity on Rainbow.

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