scholarly journals Expression of a Rice Chlorophyll a/b Binding Protein Promoter in Sweetpotato

2007 ◽  
Vol 132 (4) ◽  
pp. 551-556 ◽  
Author(s):  
Guo-qing Song ◽  
Hideo Honda ◽  
Ken-ichi Yamaguchi
Keyword(s):  
Chlorophyll A ◽  
Ipomoea Batatas ◽  
Binding Protein ◽  
Gus Expression ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Target Tissue ◽  
Storage Roots ◽  
Specific Expression ◽  
Pests And Diseases

Leaves are usually the target tissue for expressing transgenes conferring resistances to herbicides, pests, and diseases. To achieve leaf-specific expression, a light-harvest chlorophyll a/b binding protein (CAB) of photosystem-II (CAB2) promoter (CAB2-p) from rice (Oryza sativa L.) and the cauliflower mosaic virus 35S promoter were fused to the β-glucuronidase (GUS) reporter and subsequently evaluated in transgenic sweetpotato [Ipomoea batatas L. (Lam.)]. The 35S promoter-directed GUS activities varied from 46.0 to 61.2 nmol 4-methyl-umbelliferyl-β-D-glucuronide (4-MU) per minute per milligram of protein in leaf, stem, primary, and storage roots. In contrast, the CAB2-p directed an uneven distribution of GUS activities (4-MU at 1.1 to 12.6 nmol·min−1·mg−1 protein); GUS activity in mature leaves was ≈12-fold as high as that in storage roots. In addition, GUS assay in leaf tissues revealed that CAB2-p enabled a developmentally controlled and light-regulated GUS expression. These results indicate that the rice CAB2-p could be used to drive leaf-specific expression of linked genes in sweetpotato.

scholarly journals Overexpression of the Golden SNP-Carrying Orange Gene Enhances Carotenoid Accumulation and Heat Stress Tolerance in Sweetpotato Plants

Antioxidants ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 51
Author(s):  
So-Eun Kim ◽  
Chan-Ju Lee ◽  
Sul-U Park ◽  
Ye-Hoon Lim ◽  
Woo Sung Park ◽  
...  
Keyword(s):  
Heat Stress ◽  
Stress Tolerance ◽  
Ipomoea Batatas ◽  
Radical Scavenging Activity ◽  
Radical Scavenging ◽  
Cauliflower Mosaic Virus ◽  
Carotenoid Content ◽  
35S Promoter ◽  
Storage Roots ◽  
Heat Stress Tolerance

Carotenoids function as photosynthetic accessory pigments, antioxidants, and vitamin A precursors. We recently showed that transgenic sweetpotato calli overexpressing the mutant sweetpotato (Ipomoea batatas [L.] Lam) Orange gene (IbOr-R96H), which carries a single nucleotide polymorphism responsible for Arg to His substitution at amino acid position 96, exhibited dramatically higher carotenoid content and abiotic stress tolerance than calli overexpressing the wild-type IbOr gene (IbOr-WT). In this study, we generated transgenic sweetpotato plants overexpressing IbOr-R96H under the control of the cauliflower mosaic virus (CaMV) 35S promoter via Agrobacterium-mediated transformation. The total carotenoid contents of IbOr-R96H storage roots (light-orange flesh) and IbOr-WT storage roots (light-yellow flesh) were 5.4–19.6 and 3.2-fold higher, respectively, than those of non-transgenic (NT) storage roots (white flesh). The β-carotene content of IbOr-R96H storage roots was up to 186.2-fold higher than that of NT storage roots. In addition, IbOr-R96H plants showed greater tolerance to heat stress (47 °C) than NT and IbOr-WT plants, possibly because of higher DPPH radical scavenging activity and ABA contents. These results indicate that IbOr-R96H is a promising strategy for developing new sweetpotato cultivars with improved carotenoid contents and heat stress tolerance.

scholarly journals Identification of the Core Pollen-Specific Regulation in the Rice OsSUT3 Promoter

2020 ◽  
Vol 21 (6) ◽  
pp. 1909
Author(s):  
Dandan Li ◽  
Rucong Xu ◽  
Dong Lv ◽  
Chunlong Zhang ◽  
Hong Yang ◽  
...  
Keyword(s):  
Pollen Development ◽  
Ectopic Expression ◽  
Gus Expression ◽  
35S Promoter ◽  
Crop Breeding ◽  
Specific Expression ◽  
Regulatory Motifs ◽  
Gus Reporter ◽  
Specific Regulation ◽  
Promoter Deletions

The regulatory mechanisms of pollen development have potential value for applications in agriculture, such as better understanding plant reproductive regularity. Pollen-specific promoters are of vital importance for the ectopic expression of functional genes associated with pollen development in plants. However, there is a limited number of successful applications using pollen-specific promoters in genetic engineering for crop breeding and hybrid generation. Our previous work led to the identification and isolation of the OsSUT3 promoter from rice. In this study, to analyze the effects of different putative regulatory motifs in the OsSUT3 promoter, a series of promoter deletions were fused to a GUS reporter gene and then stably introduced into rice and Arabidopsis. Histochemical GUS analysis of transgenic plants revealed that p385 (from −385 to −1) specifically mediated maximal GUS expression in pollen tissues. The S region (from −385 to −203) was the key region for controlling the pollen-specific expression of a downstream gene. The E1 (−967 to −606), E2 (−202 to −120), and E3 (−119 to −1) regions enhanced ectopic promoter activity to different degrees. Moreover, the p385 promoter could alter the expression pattern of the 35S promoter and improve its activity when they were fused together. In summary, the p385 promoter, a short and high-activity promoter, can function to drive pollen-specific expression of transgenes in monocotyledon and dicotyledon transformation experiments.

scholarly journals MOLECULAR CHARACTERIZATION AND TRANSIENT EXPRESSION OF MAIZE SUS1 PROMOTER

HortScience ◽  
1994 ◽  
Vol 29 (4) ◽  
pp. 253f-254
Author(s):  
Xiao-Fang Huang ◽  
Binh Nguyen-Quoc ◽  
Serge Yelle
Keyword(s):  
Transient Expression ◽  
Gus Expression ◽  
35S Promoter ◽  
Plant Tissues ◽  
Upstream Sequence ◽  
Specific Expression ◽  
Gene Sequence Analysis ◽  
Sequence Elements ◽  
Young Leaves

Sucrose synthase (SS) is one of the key enzymes in plant carbohydrate metabolism. In maize, this enzyme is encoded by two genes, Sh1 and Sus1. We have isolated and determined the 5'-upstream sequence of maize Sus1 gene and compared it with the corresponding sequence in Sh1 gene. Sequence analysis revealed that there was a weak homology between the two promoters and no common sequence elements were found. To understand the differential regulation of the expression of the two genes, we constructed chimeric GUS fusions using the two promoters of SS genes. By using the biolistic system, we delivered these constructs into various plant tissues, and their transient expression was studied. Our results showed that the two promoters of SS genes directed tissue-specific expression in the same way that the two genes are expressed in vivo. The effectiveness of the expression of the constructs was recorded by counting the total blue expression units (blue spots) per shot and by fluorometric assays. High levels of GUS activity were detected in the immature embryos, young coleoptiles, and heterotrophic young leaves bombarded with the Sus–GUS construct. More than 100 expression units were observed in these tissues. Compared with the transient expression of the 35S promoter in the same tissue, Sus promoter activity was twice as high. Strong Sus–GUS expression was also detected in the aleurone cells of developing kernels. In contrast, the Sh-GUS construct was expressed only in the endosperm with an activity twice as high as that of Sus–GUS and 35S–GUS in the same tissue. The results will be discussed in terms of the physiological roles of the two SS isozymes in plant tissues.

Research note: The 35S promoter is not constitutively expressed in the transgenic tropical actinorhizal tree Casuarina glauca

2002 ◽  
Vol 29 (5) ◽  
pp. 649 ◽  
Author(s):  
Aziz Smouni ◽  
Laurent Laplaze ◽  
Didier Bogusz ◽  
Fathia Guermache ◽  
Florence Auguy ◽  
...  
Keyword(s):  
Vascular Tissue ◽  
Gus Expression ◽  
Cauliflower Mosaic Virus ◽  
Camv 35S Promoter ◽  
35S Promoter ◽  
Casuarina Glauca ◽  
Camv 35S ◽  
Highly Active ◽  
One Year

The tropical nitrogen-fixing tree, Casuarina glauca Sieb. ex Spreng. was genetically transformed using Agrobacterium tumefaciens C58C1(pGV2260; pBIN19GUSINT). We report on the expression pattern conferred by the cauliflower mosaic virus (CaMV) 35S promoter in transgenic C. glauca plants grown in vitro, and for one year in a greenhouse. Histochemical assays in shoots from in vitro plants revealed β-glucuronidase (GUS) staining in apical and axillary buds, and in nearly all tissues near the base of the stem. In roots, the CaMV 35S drove strong GUS expression in the apex and vascular tissue. In 1-year old plants grown in a greenhouse, the CaMV 35S promoter was highly active, except in peripheral suberized tissues. Transgenic C. glauca plants were nodulated by the actinomycete Frankia. Histochemical assays on vibratome sections of transgenic nodules demonstrated intense GUS activity in the vascular bundle, the phellogen, and in strands of uninfected cells filled with polyphenols. GUS expression was undetectable in Frankia-infected cells.

Agrobacterium-mediated transformation of Amaranthus hypochondriacus: light- and tissue-specific expression of a pea chlorophyll a/b-binding protein promoter

1997 ◽  
Vol 16 (12) ◽  
pp. 847-852 ◽  
Author(s):  
A. E. Jofre-Garfias ◽  
N. Villegas-Sepúlveda ◽  
J. L. Cabrera-Ponce ◽  
R. M. Adame-Alvarez ◽  
L. Herrera-Estrella ◽  
...  
Keyword(s):  
Chlorophyll A ◽  
Binding Protein ◽  
Specific Expression ◽  
Tissue Specific ◽  
Agrobacterium Mediated Transformation ◽  
Amaranthus Hypochondriacus ◽  
Tissue Specific Expression

scholarly journals Factors Affecting Transient Gene Expression in Apple Cotyledons following Particle Bombardment

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 617a-617
Author(s):  
Hong Y. Yang ◽  
Schuyler S. Korban
Keyword(s):  
Gene Transfer ◽  
Transient Expression ◽  
Particle Bombardment ◽  
Transient Gene Expression ◽  
Gus Expression ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Tungsten Particle ◽  
Factors Affecting ◽  
Efficient Gene

Developing an efficient gene transfer system for apple (Malus ×domestica L.) remains a major objective in genetic engineering efforts of this fruit crop. Transient expression of the uidA gene coding for β-glucuronidase (GUS) and driven by the cauliflower mosaic virus 35S promoter (CaMV35S) has been induced in apple cotyledonary explants of mature seeds by tungsten particle bombardment using the Particle Inflow Gun (PIG). Several factors that affect transient expression of the GUS gene in apple cotyledons were investigated. The gene transfer efficiency was monitored by recording the number of blue spots observed on explants two days following bombardment. Precultivation of cotyledons for 18 hours before bombardment significantly increased the number of blue foci. Of the three different precipitation methods tested including water, 25% PEG, and 60% glycerol, the latter was the most effective for coating DNA onto tungsten particles. Washing DNA-coated tungsten particles with 70% ethanol and resuspending in 100% ethanol significantly enhanced gene delivery to cotyledons. The amount of particles used for each bombardment also influenced GUS expression. About 0.5 mg of particles per shot resulted in the highest number of blue foci. Using larger quantity of particles (i.e., 2 mg) drastically decreased GUS expression probably due to the toxicity of tungsten particles.

scholarly journals Mesorhizobium loti Increases Root-Specific Expression of a Calcium-Binding Protein Homologue Identified by Promoter Tagging in Lotus japonicus

2000 ◽  
Vol 13 (6) ◽  
pp. 606-616 ◽  
Author(s):  
K. Judith Webb ◽  
Leif Skøt ◽  
Margaret N. Nicholson ◽  
Bodil Jørgensen ◽  
Sue Mizen
Keyword(s):  
Calcium Binding ◽  
Binding Protein ◽  
Lotus Japonicus ◽  
Root Hairs ◽  
Gus Expression ◽  
Calcium Binding Protein ◽  
Gus Activity ◽  
Specific Expression ◽  
Mesorhizobium Loti ◽  
Dna Insertion

A promoter tagging program in the legume Lotus japonicus was initiated to identify plant genes involved in the nitrogen-fixing symbiosis between legumes and rhizobia. Seven transformed plant lines expressing the promoterless reporter gene uidA (β-glucuronidase; GUS) specifically in roots and/or nodules were identified. Four of these expressed GUS in the roots only after inoculation with nodule-forming Mesorhizobium loti. In one line (T90), GUS activity was found in the root epidermis, including root hairs. During seedling growth, GUS expression gradually became focused in developing nodules and disappeared from root tissue. No GUS activity was detected when a non-nodulating mutant of M. loti was used to inoculate the plants. The T-DNA insertion in this plant line was located 1.3 kb upstream of a putative coding sequence with strong homology to calcium-binding proteins. Four motifs were identified, which were very similar to the “EF hands” in calmodulin-related proteins, each binding one Ca2+. We have named the gene LjCbp1 (calcium-binding protein). Northern (RNA) analyses showed that this gene is expressed specifically in roots of L. japonicus. Expression was reduced in roots inoculated with non-nodulating M. loti mutants and in progeny homozygous for the T-DNA insertion, suggesting a link between the T-DNA insertion and this gene.

Field performance and heavy metal concentrations of transgenic flue-cured tobacco expressing a mammalian metallothionein-β-glucuronidase gene fusion

Genome ◽  
1993 ◽  
Vol 36 (2) ◽  
pp. 255-260 ◽  
Author(s):  
J. E. Brandle ◽  
H. Labbe ◽  
J. Hattori ◽  
B. L. Miki
Keyword(s):  
Heavy Metal ◽  
Gene Fusion ◽  
Metal Tolerance ◽  
Heavy Metal Tolerance ◽  
Leaf Tissue ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Specific Expression ◽  
High Background ◽  
Transgenic Lines

Cadmium (Cd) is a nonessential heavy metal that can cause acute and chronic illness in humans. Some plant species such as tobacco (Nicotiana tabacum L.) tend to accumulate high levels of Cd in leaf tissue, the consumed portion of the plant. Tissue-specific expression of mammalian metallothionein has been suggested as a means of partitioning Cd in nonconsumed portions of transgenic plants. The purpose of the experiment reported here was to evaluate Cd concentration and agronomic performance of four field-grown transgenic tobacco lines harbouring a metallothionein-β-glucuronidase (MG) gene fusion driven by the constitutive 35S promoter of cauliflower mosaic virus. The trial was grown in a region of Canada known to have high background levels of Cd. The agronomic evaluation showed that some of the transgenic lines were equal to, while others performed more poorly than, the untransformed control for yield, days to flower, and leaf number. Gene expression measured by β-glucuronidase activity showed that all of the transgenic lines expressed the MG gene in the upper portion of the plant. One line did not express the MG gene in the roots. Cd levels in the leaf tissue of transformed lines were not significantly different from the untransformed control.Key words: cadmium, β-glucuronidase, transformation, heavy metal tolerance.

scholarly journals Expression of a Defense-Related 3-Hydroxy-3-Methylglutaryl CoA Reductase Gene in Response to Parasitization by Orobanche spp.

1998 ◽  
Vol 11 (6) ◽  
pp. 530-536 ◽  
Author(s):  
James H. Westwood ◽  
Xueshu Yu ◽  
Chester L. Foy ◽  
Carole L. Cramer
Keyword(s):  
Gene Promoter ◽  
Attachment Site ◽  
Gus Expression ◽  
Cauliflower Mosaic Virus ◽  
35S Promoter ◽  
Gus Reporter ◽  
Host Root ◽  
Reductase Gene ◽  
Defense Related Gene ◽  
Coa Reductase

Orobanche spp. are angiosperms that live parasitically on the roots of other plants, and are capable of significantly reducing the yield and quality of their crop hosts. We have demonstrated that parasitization by Orobanche induces expression of hmg2, a defense-related isogene of 3-hydroxy-3-methylglutaryl CoA reductase (HMGR) in tobacco. Transgenic tobacco plants expressing a construct containing 2.3 kb of the tomato hmg2 gene promoter fused to the β-glucuronidase (GUS) reporter gene were parasitized by O. aegyptiaca. Expression of the hmg2:GUS construct was detected within 1 day following penetration of the host root by the O. aegyptiaca radicle and was localized to the region immediately around the site of parasite invasion. This expression continued and intensified over the course of O. aegyptiaca development. In addition, the hmg2:GUS expression was induced by secondary parasitization, where secondary roots of O. aegyptiaca contacted the host root at a distance from the primary attachment site. This GUS expression was specific to plants containing the hmg2:GUS construct, and was not observed in control plants transformed with a construct of the cauliflower mosaic virus 35S promoter fused to the GUS gene. These results indicate that Orobanche parasitization initiates rapid and sustained induction of a defense-related gene in the host root.

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