Transgenic lines of Begonia maculata generated by ectopic expression of PttKN1

Biologia ◽  
2011 ◽  
Vol 66 (2) ◽  
Author(s):  
Quan-le Xu ◽  
Jiang-ling Dong ◽  
Nan Gao ◽  
Mei-yu Ruan ◽  
Hai-yan Jia ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Ectopic Expression ◽  
Class I ◽  
Wild Type Plant ◽  
Wild Type ◽  
Knox Gene ◽  
Knox Genes ◽  
Pttkn1 Gene ◽  
Transgenic Lines ◽  
First Time

AbstractKNOX (KNOTTED1-like homeobox) genes encode homeodomain-containing transcription factors which play crucial roles in meristem maintenance and proper patterning of organ initiation. PttKN1 gene, isolated from the vascular cambium of hybrid aspen (Populus tremula × P. tremuloides), is a member of class I KNOX gene family. In order to understand the roles of PttKN1 gene in meristem activity and morphogenesis as well as to explore the possibility to generate novel ornamental lines via its ectopic expression, it was introduced into the genome of Begonia maculata Raddi by Agrobacterium tumefasciens-mediated gene transformation here. Four types of transgenic plants were observed, namely coral-like (CL) type, ectopic foliole (EF) type, phyllotaxy-irregular (IP) type and cup-shaped (CS) type, which were remarkably different from corresponding wild type and were not also observed in the regenerated plantlets of wild type plant. Among these four types of transgenic plants, the phenotype of coral-like was observed for the first time in the transformants ectopically expressed KNOX genes. The observation of scanning electron microscope (SEM) showed ectopic meristems on the adaxial leaf surface of the transformants. Interestingly, the plantlets with ectopic foliole could generate new ectopic folioles from the original ectopic folioles again, and the plants regenerated from the EF-type transformants could also maintain the original morphology. The same specific RT-PCR band of the four types of transgenic plantlets showed that PttKN1 was ectopically expressed. All these data demonstrated that the ectopic expression of PttKN1 caused a series of alterations in morphology which provided possibilities producing novel ornamental lines and that PttKN1 played important roles in meristem initiation, maintenance and organogenesis events as other class I KNOX genes.

scholarly journals Over-expression of the Hybrid Aspen Homeobox PttKN1 Gene in Red Leaf Beet Induced Altered Coloration of Leaves

2015 ◽  
Vol 43 (1) ◽  
pp. 35-40
Author(s):  
Quanle XU ◽  
Mei-yu RUAN ◽  
Ying-jie TAO ◽  
Xin HU
Keyword(s):  
Transgenic Plants ◽  
Ectopic Expression ◽  
Kanamycin Resistance ◽  
Pcr Analysis ◽  
35S Promoter ◽  
Wild Type ◽  
Morphological Alterations ◽  
Pttkn1 Gene ◽  
Transgenic Lines ◽  
Morphological Modification

PttKN1 (Populus tremula × tremuloides KNOTTED1) gene belongs to the KNOXI gene family. It plays an important role in plant development, typically in meristem initiation, maintenance and organogenesis, and potentially in plant coloration. To investigate the gene functions further, it was introduced into red leaf beet by the floral dip method mediated via Agrobacterium tumefaciens. The transformants demonstrated typical phenotypes as with other PttKN1 transformants. These alterations were very different from the morphology of the wild type. Among them, morphological modification of changed color throughout the entire plant from claret of wild type to yellowish green was the highlight in those transgenic PttKN1-beet plants. The result of spraying selection showed that the PttKN1-beet plants had kanamycin resistance. PCR assay of the 35S-Promoter, NPTII and PttKN1 gene, PCR-Southern analysis of the NPTII and PttKN1 gene showed that the foreign PttKN1 gene had successfully integrated into the genome of beet plant. Furthermore, the results of RT-PCR analysis showed that the gene was ectopic expressed in transgenic plants. These data suggested that there is a correlation between the ectopic expression of PttKN1 gene and morphological alterations of beet plants. Pigment content assay showed that betaxanthins concentrations shared little difference between wild type and transgenic lines, while betacyanins content in transgenic plants was sharply decreased, indicating that the altered plant coloration of the transgenic beet plants may be caused by the changed betacyanins content. The tyrosinase study suggested that the sharply decreased of betacyanins content in transgenic plants was caused via the decreased tyrosinase level. Therefore, the reason for the altered plant coloration may be due to partial inhibition of betacyanin biosynthesis that was induced via the pleiotropic roles of PttKN1 gene.

scholarly journals Degradation of trinitrotoluene by transgenic nitroreductase in Arabidopsis plants

2018 ◽  
Vol 64 (No. 8) ◽  
pp. 379-385 ◽  
Author(s):  
Zhu Bo ◽  
Han Hongjuan ◽  
Fu Xiaoyan ◽  
Li Zhenjun ◽  
Gao Jianjie ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Transgenic Arabidopsis ◽  
Environmental Pollutant ◽  
Specific Rate ◽  
Wild Type Plant ◽  
Wild Type ◽  
Fresh Weight ◽  
Human Carcinogen ◽  
Specific Rate Constant ◽  
Nitroreductase Activity

The explosive 2,4,6-trinitrotoluene (TNT) is a highly toxic and persistent environmental pollutant. TNT is toxic to many organisms, it is known to be a potential human carcinogen, and is persistent in the environment. This study presents a system of phytoremediation by Arabidopsis plants developed on the basis of overexpression of NAD(P)H-flavin nitroreductase (NFSB) from the Sulfurimonas denitrificans DSM1251. The resulting transgenic Arabidopsis plants demonstrated significantly enhanced TNT tolerance and a strikingly higher capacity to remove TNT from their media. The highest specific rate constant of TNT disappearance rate was 1.219 and 2.297 mL/g fresh weight/h for wild type and transgenic plants, respectively. Meanwhile, the nitroreductase activity in transgenic plant was higher than wild type plant. All this indicates that transgenic plants show significantly enhanced tolerances to TNT; transgenic plants also exhibit strikingly higher capabilities of removing TNT from their media and high efficiencies of transformation.

Enhancing Biosynthesis of Flavonol Protective Biomaterials Using FLS Transgenic Plants

2017 ◽  
Vol 866 ◽  
pp. 29-32
Author(s):  
Darin Dangrit ◽  
Kanokporn Sompornpailin
Keyword(s):  
Tissue Culture ◽  
Transgenic Plants ◽  
Photosynthetic Pigment ◽  
Flavonoid Biosynthesis ◽  
Carotenoid Pigments ◽  
Chlorophyll B ◽  
Wild Type ◽  
Flavonoid Synthesis ◽  
Transgenic Lines ◽  
Tissue Culture Condition

Flavonol synthase (FLS) gene encodes an enzyme that is involved in conversion substrates into flavonols, quercetin and kaempferol. These substances are a subgroup of flavonoids which have an important role in both plant and human health. Many environmental factors such as temperature, pH and UV-A radiation have been studied and presented relationship with flavonoid synthesis. In this experiment, the combination of visible and UV-A lights was used as factors for elevating flavonoid biosynthesis of wild type (WT) plant and two lines of FLS transgenic plant under tissue culture condition. Both transgenic lines significantly enhanced the accumulation of quercetin and kaempferol substances nearly one fold higher than WT plant did. The photosynthetic pigment levels of chlorophyll A, chlorophyll B and carotenoid in transgenic lines are in the range 45.20-46.88, 16.34-17.04 and 13.63-13.46, while those of WT plants are 35.93, 13.18 and 10.55 (µg/g FW), respectively. Therefore, FLS transgenic plants containing high flavonol content showed a better in the protection photosynthetic pigments by less reductions of chlorophyll and carotenoid pigments.

scholarly journals Restoration of Wild-Type Virus by Double Recombination of Tombusvirus Mutants with a Host Transgene

1999 ◽  
Vol 12 (2) ◽  
pp. 153-162 ◽  
Author(s):  
Marise Borja ◽  
Teresa Rubio ◽  
Herman B. Scholthof ◽  
Andrew O. Jackson
Keyword(s):  
Coat Protein ◽  
Transgenic Plants ◽  
Coat Protein Gene ◽  
Protein Gene ◽  
Wild Type Virus ◽  
Type Virus ◽  
Virus Infections ◽  
Wild Type ◽  
Transgenic Lines ◽  
Double Recombination

Nicotiana benthamiana plants transformed with the coat protein gene of tomato bushy stunt virus (TBSV) failed to elicit effective virus resistance when inoculated with wild-type virus. Subsequently, R1 and R2 progeny from 13 transgenic lines were inoculated with a TBSV mutant containing a defective coat protein gene. Mild symptoms typical of those elicited in nontransformed plants infected with the TBSV mutant initially appeared. However, within 2 to 4 weeks, up to 20% of the transgenic plants sporadically began to develop the lethal syndrome characteristic of wild-type virus infections. RNA hybridization and immunoblot analyses of these plants and nontransformed N. benthamiana inoculated with virus from the transgenic lines indicated that wild-type virus had been regenerated by a double recombination event between the defective virus and the coat protein transgene. Similar results were obtained with a TBSV deletion mutant containing a nucleotide sequence marker, and with a chimeric cucumber necrosis virus (CNV) containing the defective TBSV coat protein gene. In both cases, purified virions contained wild-type TBSV RNA or CNV chimeric RNA derived by recombination with the transgenic coat protein mRNA. These results thus demonstrate that recombinant tombusviruses can arise frequently from viral genes expressed in transgenic plants.

scholarly journals Ethylene-inducible Expression of ipt Gene Produces a Dramatic Increase in Fower Bud Count in Transgenic Plants

HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 821B-821
Author(s):  
Richard J. McAvoy* ◽  
Mariya V. Khodakovskaya ◽  
Hong Liu ◽  
Yi Li
Keyword(s):  
Transgenic Plants ◽  
Vegetative State ◽  
Crop Improvement ◽  
Marked Change ◽  
Acc Oxidase ◽  
Wild Type ◽  
Flower Bud ◽  
Vegetative Development ◽  
Wide Range ◽  
Transgenic Lines

Cytokinins play an important role in regulating plant growth and development. The cytokinin gene, isopentenyl transferase (ipt), was placed under the control of the ACC oxidase promoter from the LEACO1 gene from Lycopersicon esculentum and introduced into Nicotiana tabacum (cv. Havana) and chrysanthemum (Dendranthema × grandiflorum `Iridon'). Transformants were confirmed by PCR reaction and Southern blot and analyzed for phenotypical changes under both greenhouse and growth chamber conditions. With both species, LEACO1-ipt transgenic plants displayed a wide range of vegetative and generative phenotypes. With plants growing in the vegetative state, some LEACO1-ipt transgenic lines appeared similar to the non-transgenic wild-type cultivars while other lines showed excessive lateral branch development and short internodes. With plants grown under generative conditions, several LEACO1-ipt transgenic lines showed a 2 to 10-fold increase in the number of flower buds relative to the wild-type cultivars. With chrysanthemum, dramatic increases in bud count were observed on transgenic lines that otherwise displayed a morphology similar to the non-transgenic lines. Analysis of ipt expression indicated a marked change in gene expression between the most extreme phenotypes observed in this study. LEACO1-ipt lines that express normal vegetative development but increased flower bud counts appear to have great potential for ornamental crop improvement.

scholarly journals Improved Salinity Tolerance in Carrizo Citrange Rootstock through Overexpression of Glyoxalase System Genes

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Ximena Alvarez-Gerding ◽  
Rowena Cortés-Bullemore ◽  
Consuelo Medina ◽  
Jesús L. Romero-Romero ◽  
Claudio Inostroza-Blancheteau ◽  
...  
Keyword(s):  
Salt Stress ◽  
Transgenic Plants ◽  
Stress Tolerance ◽  
Salinity Tolerance ◽  
Dry Weight ◽  
Wild Type Plant ◽  
Wild Type ◽  
Salt Stress Tolerance ◽  
Glyoxalase System ◽  
Carrizo Citrange

Citrus plants are widely cultivated around the world and, however, are one of the most salt stress sensitive crops. To improve salinity tolerance, transgenic Carrizo citrange rootstocks that overexpress glyoxalase I and glyoxalase II genes were obtained and their salt stress tolerance was evaluated. Molecular analysis showed high expression for both glyoxalase genes (BjGlyIandPgGlyII) in 5H03 and 5H04 lines. Under control conditions, transgenic and wild type plants presented normal morphology. In salinity treatments, the transgenic plants showed less yellowing, marginal burn in lower leaves and showed less than 40% of leaf damage compared with wild type plants. The transgenic plants showed a significant increase in the dry weight of shoot but there are no differences in the root and complete plant dry weight. In addition, a higher accumulation of chlorine is observed in the roots in transgenic line 5H03 but in shoot it was lower. Also, the wild type plant accumulated around 20% more chlorine in the shoot compared to roots. These results suggest that heterologous expression of glyoxalase system genes could enhance salt stress tolerance in Carrizo citrange rootstock and could be a good biotechnological approach to improve the abiotic stress tolerance in woody plant species.

Antisense RNA inhibition of pyruvate, orthophosphate dikinase and NADP malate dehydrogenase in the C4 plant Flaveria bidentis: analysis of plants with a mosaic phenotype

1999 ◽  
Vol 26 (6) ◽  
pp. 537 ◽  
Author(s):  
Anthony R. Ashton ◽  
Robert T. Furbank ◽  
Stephen J. Trevanion
Keyword(s):  
Transgenic Plants ◽  
Malate Dehydrogenase ◽  
Antisense Rna ◽  
Wild Type ◽  
Pyruvate Orthophosphate Dikinase ◽  
Orthophosphate Dikinase ◽  
Transgenic Lines ◽  
Rna Inhibition ◽  
Target Enzyme ◽  
Flaveria Bidentis

Antisense RNA suppression of either pyruvate, orthophosphate dikinase [EC 2.7.9.1] or NADP malate dehydrogenase [EC 1.1.1.82] gene expression in the C4 dicot Flaveria bidentis L. var. Kuntze produced several independent transgenic lines with leaves showing heritable, mosaic phenotypes. The appearance of these plants was highly variable, with leaves that were either predominantly green, predominantly yellow, or a mixture of the two. The yellow sectors appeared to be clonal in origin. For both sets of transgenic plants, the green and yellow sectors showed a reduction in the activity of the respective target enzyme compared to wild-type leaves. The mRNA of the target enzyme was reduced in both green and yellow sectors of leaves of both types of transformants compared to leaves from wild-type plants. The yellow sectors had decreased amounts of other photosynthetic enzymes on an area basis, but most enzyme activities and electron transport rates were similar to the green sectors on a chlorophyll basis. The mosaic phenotype could not be attributed simply to the degree of suppression of the target enzyme, because we have also obtained uniformly green plants with similar or greater enzyme suppression. The importance of this spatial variability in the effectiveness of the antisense transgenes for the analysis of transgenic plants in general is discussed.

scholarly journals Gibberellin 20-Oxidase Gene OsGA20ox3 Regulates Plant Stature and Disease Development in Rice

2013 ◽  
Vol 26 (2) ◽  
pp. 227-239 ◽  
Author(s):  
Xue Qin ◽  
Jun Hua Liu ◽  
Wen Sheng Zhao ◽  
Xu Jun Chen ◽  
Ze Jian Guo ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Critical Role ◽  
Insertion Site ◽  
Insertion Mutant ◽  
Wild Type Plant ◽  
Wild Type ◽  
Oxidase Gene ◽  
Dna Insertion ◽  
Flanking Sequences ◽  
Plant Stature

Gibberellin (GA) 20-oxidase (GA20ox) catalyses consecutive steps of oxidation in the late part of the GA biosynthetic pathway. A T-DNA insertion mutant (17S-14) in rice, with an elongated phenotype, was isolated. Analysis of the flanking sequences of the T-DNA insertion site revealed that an incomplete T-DNA integration resulted in enhanced constitutively expression of downstream OsGA20ox3 in the mutant. The accumulation of bioactive GA1 and GA4 were increased in the mutant in comparison with the wild-type plant. Transgenic plants overexpressing OsGA20ox3 showed phenotypes similar to those of the 17S-14 mutant, and the RNA interference (RNAi) lines that had decreased OsGA20ox3 expression exhibited a semidwarf phenotype. Expression of OsGA20ox3 was detected in the leaves and roots of young seedlings, immature panicles, anthers, and pollens, based on β-glucuronidase (GUS) activity staining in transgenic plants expressing the OsGA20ox3 promoter fused to the GUS gene. The OsGA20ox3 RNAi lines showed enhanced resistance against rice pathogens Magnaporthe oryzae (causing rice blast) and Xanthomonas oryzae pv. oryzae (causing bacterial blight) and increased expression of defense-related genes. Conversely, OsGA20ox3-overexpressing plants were more susceptible to these pathogens comparing with the wild-type plants. The susceptibility of wild-type plants to X. oryzae pv. oryzae was increased by exogenous application of GA3 and decreased by S-3307 treatment. Together, the results provide direct evidence for a critical role of OsGA20ox3 in regulating not only plant stature but also disease resistance in rice.

scholarly journals Seed-Specific Expression of Arabidopsis AtCYP85A2 Produces Biologically Active Brassinosteroids Such as Castasterone and Brassinolide to Improve Grain Yield and Quality in Seeds of Brachypodium Distachyon

2021 ◽  
Vol 12 ◽  
Author(s):  
Jeehee Roh ◽  
Jinyoung Moon ◽  
Ye Eun Lee ◽  
Chan Ho Park ◽  
Seong-Ki Kim
Keyword(s):  
Transgenic Plants ◽  
Grain Yield ◽  
Brachypodium Distachyon ◽  
Biologically Active ◽  
Wild Type Plant ◽  
Wild Type ◽  
Specific Expression ◽  
Yield And Quality ◽  
Grain Yield And Quality ◽  
Seed Specific Expression

Brachypodium distachyon is a monocotyledonous model plant that contains castasterone (CS) but no brassinolide (BL) as the end product of brassinosteroids (BR) biosynthesis, indicating dysfunction of BL synthase, which catalyzes the conversion of CS to BL. To increase BR activity, Arabidopsis cytochrome P450 85A2 (AtCYP85A2) encoding BR 6-oxidase/BL synthase was introduced into B. distachyon with the seed-specific promoters pBSU1, pAt5g10120, and pAt5g54000. RT-PCR analysis and GUS activity revealed that AtCYP85A2 was only expressed in the seeds of the transgenic plants pBSU1-AtCYP85A2::Bd21-3, pAt5g10120-AtCYP85A2::Bd21-3, and pAt5g54000-AtCYP85A2::Bd21-3. The crude enzyme prepared from the seeds of these three transgenic plants catalyzed the conversion of 6-deoxoCS to CS. The transgenic plants showed greater specific enzyme activity than the wild-type plant for the conversion of 6-deoxoCS to CS, indicating enhanced BR 6-oxidase activity in the transgenic plants. The enzyme solution also catalyzed the conversion of CS into BL. Additionally, BL was identified from the seeds of transgenic plants, verifying that seed-specific AtCYP85A2 encodes a functional BL synthase to increase BR activity in the seeds of transgenic Brachypodium. In comparison with wild-type Brachypodium, the transgenic plants showed better growth and development during the vegetative growing stage. The flowers of the transgenic plants were remarkably larger, resulting in increments in the number, size, and height of seeds. The total starch, protein, and lipid contents in transgenic plants were higher than those in wild-type plants, indicating that seed-specific expression of AtCYP85A2 improves both grain yield and quality in B. distachyon.

Eucalyptus grandis has at least two functional SOC1-like floral activator genes

2004 ◽  
Vol 31 (3) ◽  
pp. 225 ◽  
Author(s):  
John M. Watson ◽  
Elizabeth M. Brill
Keyword(s):  
Flowering Time ◽  
Sequence Data ◽  
Ectopic Expression ◽  
Eucalyptus Grandis ◽  
Mads Box ◽  
Wild Type ◽  
Mads Box Genes ◽  
Nucleotide Sequence Data ◽  
Transgenic Lines ◽  
Young Leaves

In a search for Eucalyptus grandis Hill ex Maiden MADS-box genes involved in floral initiation and development, we isolated two cDNAs (EgrMADS 3 and EgrMADS 4), which are functional orthologues of the Arabidopsis thaliana (L.) Heynh. floral activator gene SOC1. These two E. grandis genes are equally and most-actively expressed in vegetative tissues such as apical shoots and young leaves. The two genes are less actively, but differentially expressed in roots and unopened flowers. Ectopic expression of EgrMADS 3 or EgrMADS 4 in the late-flowering A. thaliana Ler soc1 (agl20) derivative complemented the phenotype of this mutant, and some of these transgenic lines flowered significantly earlier than the wild-type Ler ecotype. Overexpression of EgrMADS 3 or EgrMADS 4 in A. thaliana ecotype Columbia accelerated flowering time under short-day conditions. However, under the same conditions the flowering time of A. thaliana ecotype C24 was altered by the ectopic expression of EgrMADS 3, but not that of EgrMADS 4.The nucleotide sequence data reported will appear in the EMBL and GenBank Nucleotide Databases under the accession numbers AY263807 (EgrMADS 3) and AY263808 (EgrMADS 4).

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