Transient β-glucuronidase expression in Scots pine tissues derived from mature trees

1994 ◽  
Vol 24 (10) ◽  
pp. 2006-2011 ◽  
Author(s):  
Tuija Aronen ◽  
Hely Häggman ◽  
Anja Hohtola
Keyword(s):  
Transient Expression ◽  
Scots Pine ◽  
Growth Regulator ◽  
Genetic Regulation ◽  
Gus Expression ◽  
Expression Level ◽  
Suspension Cells ◽  
Mature Trees ◽  
Gene Construct ◽  
Bud Growth

Vegetative buds and bud-derived calli and suspension cells from 5- to 50-year-old Scots pines (Pinussylvestris L.) were used as targets for biolistic transformation. The gene construct used in the experiments was 35S CaMV–β-glucuronidase (GUS). The highest average level of transient GUS expression was found in suspension cells: 1229 ± 359 (mean ± SE) expressing cells per million. Transient expression was found in 35 of 44 (79%) tree genotypes studied. The expression level in buds and in calli was low: one or two spots per expressing bud. Growth-regulator pretreatment (BAP and 2,4-D) increased the number of GUS-expressing buds significantly. The high transient expression level in suspensions and the high percentage of GUS-expressing genotypes suggest that mature Scots pine tissues have potential for further transformation and genetic regulation studies.

Seasonal changes in the transient expression of a 35S CaMV-GUS gene construct introduced into Scots pine buds

1995 ◽  
Vol 15 (1) ◽  
pp. 65-70 ◽  
Author(s):  
T. Aronen ◽  
A. Hohtola ◽  
H. Laukkanen ◽  
H. Haggman
Keyword(s):  
Transient Expression ◽  
Scots Pine ◽  
Seasonal Changes ◽  
Gus Gene ◽  
Gene Construct

scholarly journals Establishment of Parameters for Agrobacterium Transformation of Asparagus Embryogenic Suspension Culture Cells

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 878B-878
Author(s):  
Roger A. May ◽  
Kenneth C. Sink
Keyword(s):  
Suspension Culture ◽  
Transient Expression ◽  
Gus Expression ◽  
Gus Gene ◽  
Suspension Cells ◽  
Embryogenic Cells ◽  
Culture Cells ◽  
Embryogenic Suspension ◽  
Embryogenic Suspension Culture ◽  
Embryogenic Suspension Cultures

Conditions for Agrobacterium transformation of asparagus embryogenic suspension cells were investigated using an intron-containing GUS gene in pCNL56 to detect transformation events. Embryogenic suspension cultures of Rutgers 22 were maintained on LS medium with 5 mg NAA/liter, subcultured weekly, and used 5 days thereafter. For initial experiments, cells were inoculated at 5 x 108 cfu/ml for 15 min and cocultivated for 4 days on LS medium with 10 g Phytagel/liter (EM1 medium). Subsequently, the cells were transferred to EM1 with 200 mg Timentin/liter and tested for GUS expression after a total of 6 days. The effect of acetosyringone (AS) on four A. tumefaciens strains was tested. With or without AS induction, EHA105 and C58C1(pMP90) produced a significantly greater number of GUS foci on embryogenic cells than C58C1(pGV2260) and LBA4404. Transient expression was very low from cells inoculated with C58C1(pGV2260) and nonexistent from LBA4404. AS induced significantly more GUS foci from EHA105 and C58C1(pMP90) than from the same noninduced bacteria; however, it had no effect on C58C1(pGV2260). Upon repeating the experiment using EHA105 and C58C1(pMP90) only, no differences in response were observed, although AS induced more GUS foci from both strains and EHA105 outperformed C58C1. Inoculum density was investigated using induced EHA105. 5 x 107 cfu/ml significantly increased the number of GUS foci than 108, 5 x 108, or 109, although it was not statistically different from 107, which produced slightly fewer foci.

Promoter analysis in Scots pine (Pinus sylvestris) via transient expression of luciferase fusions

2012 ◽  
Vol 29 ◽  
pp. S135-S136
Author(s):  
Muhammad Zubair Rafique ◽  
Tanja Paasela ◽  
Kean-Jin Lim ◽  
Hany El Sayed ◽  
Teemu H. Teeri
Keyword(s):  
Pinus Sylvestris ◽  
Transient Expression ◽  
Scots Pine ◽  
Promoter Analysis

scholarly journals Phenolic compounds in Scots pine heartwood: are kelo trees a unique woody substrate?

2016 ◽  
Vol 46 (2) ◽  
pp. 225-233 ◽  
Author(s):  
Parvathy Venugopal ◽  
Riitta Julkunen-Tiitto ◽  
Kaisa Junninen ◽  
Jari Kouki
Keyword(s):  
High Performance Liquid Chromatography ◽  
Fungal Infection ◽  
Scots Pine ◽  
High Performance ◽  
Phenolic Composition ◽  
Mature Trees ◽  
Fire Scars ◽  
Substrate Quality ◽  
Fungal Decomposition

Deadwood quality can be a highly significant factor in determining the occurrence of deadwood-dependent organisms such as saproxylic fungi. Rare deadwood substrates that are produced only after a lengthy senescence such as kelo trees may have unique deadwood qualities. Using high-performance liquid chromatography, we compared the phenolic composition of six types of Scots pine (Pinus sylvestris L.) substrates: living mature trees with no fungal sporocarps, living mature trees with Phellinus pini sporocarps, fallen non-kelo trees, soon-to-be kelo (standing), standing kelo, and fallen kelo. The fungal-infected living trees and fallen kelos were found to have more similarities in their phenolic composition when compared with the living and fallen trees and the standing kelos, which gets further pronounced with increasing decay. The results also highlight the uniqueness of the fungal-infected living trees and the fallen kelos and illustrate a possible correlation between fungal infection and the heartwood phenolic composition of Scots pine. However, it remains unclear to what extent the differences in phenolic compositions are caused by fungal infection and fungal decomposition. We also observed a previously undocumented correlation between the phenolic groups and fire scars on the trunks of the trees. The variation in substrate quality warrants further consideration when deadwood restoration activities are planned, as the quality of the deadwood could be as equally important as the quantity.

scholarly journals Hylobius species (Coleoptera: Curculionidae) from Siberia and the distribution patterns of adults feeding in Scots pine stands

2000 ◽  
Vol 11 (1) ◽  
pp. 57-66
Author(s):  
Andrei Gourov
Keyword(s):  
Scots Pine ◽  
Distribution Patterns ◽  
Steppe Zone ◽  
Southern Taiga ◽  
Forest Steppe ◽  
Taiga Zone ◽  
Mature Trees ◽  
Additional Food ◽  
Pine Stands ◽  
Scots Pine Stands

Seven Hylobius species are known from Siberia, while the occurrence of three of them is doubtful. Information available about the supplementary feeding of adult weevils is reviewed. It is likely that H. abietis and H. albosparsus are the only species of economic significance in this territory. In the northern forest-steppe zone, adult weevils usually prefer to feed separately on isolated or border young trees under the canopy of light Scots pine stands, but avoid the undercanopy regeneration in the dense stands. In the southern taiga zone, adults are abundant on cutover areas where they feed in clumps of young regeneration. In fresh clearcuts, up to 100% of juvenile trees may be damaged by the feeding of weevils, whereas the damage intensity declines sharply with the distance to the clearcut. An edge effect in the distribution of adult weevils needs verification and, probably, is time-dependent. Inside the stands, the crowns of mature trees may constitute an additional food niche for adults if the usual sources and preferred environmental conditions are not available.

scholarly journals Phylogenetic and Expression Analysis of CENH3 and APOLLO Genes in Sexual and Apomictic Boechera Species

2021 ◽  
Author(s):  
Evgeny Bakin ◽  
Fatih Sezer ◽  
Aslıhan Özbilen ◽  
Irem Kilic ◽  
Buket Uner ◽  
...  
Keyword(s):  
Chromosome Segregation ◽  
Genetic Regulation ◽  
Single Copy ◽  
Expression Level ◽  
Egg Cell ◽  
Copy Gene ◽  
Histone H3 Variant ◽  
Before And After ◽  
Spindle Microtubules ◽  
Mitosis And Meiosis

Apomictic plants (reproducing via asexual seeds), unlike sexual individuals, avoid meiosis and egg cell fertilization. Consequently, apomixis is very important for fixing maternal genotypes in the next plant generations. Despite the progress in the study of apomixis, molecular and genetic regulation of the latter remains poorly understood. So far APOLLO (Aspartate Glutamate Aspartate Aspartate histidine exonuclease) is one of the very few described genes associated with apomixis in Boechera species. The centromere-specific histone H3 variant encoded by CENH3 gene is essential for cell division. Mutations in CENH3 disrupt chromosome segregation during mitosis and meiosis since the attachment of spindle microtubules to a mutated form of the CENH3 histone fails. This paper presents in silico characteristic of APOLLO and CENH3 genes, which may affect apomixis. Also, we characterize the structure of CENH3, study expression levels of APOLLO and CENH3 in gynoecium/siliques of the natural diploid apomictic and sexual Boechera species at the stages of before and after fertilization. While CENH3 was a single copy gene in all Boechera species, the APOLLO gene have several polymorphic alleles associated with sexual and apomictic reproduction in the Boechera genera. Expression of the APOLLO apo-allele during meiosis was upregulated in gynoecium of apomict B. divaricarpa downregulating after meiosis until 4th day after pollination (DAP). On the 5th DAP, expression in apomictic siliques increased again. In sexual B. stricta gynoecium and siliques APOLLO apo-allele did not express. Expression of the APOLLO sex-allele during and after meiosis in gynoecium of sexual plants was several times higher than that in apomictic gynoecium. However, after pollination the sex-allele was downregulated in sexual siliques to the level of apomicts and increased sharply on the 5th DAP, while in apomictic siliques it almost did not express. At the meiotic stage, the expression level of CENH3 in the gynoecium of apomicts was two times lower than that of the sexual Boechera, decreasing in both species after meiosis and keep remaining very low in siliques of both species for several days after artificial pollination until the 4th DAP, when the expression level raised in sexual B. stricta siliques exceeding 5 times the level in apomictic B. divaricarpa siliques. We also discuss polymorphism and phylogeny of the APOLLO and CENH3 genes.

Promotion of flowering in young Larixlaricina grafts by gibberellin A4/7 and root pruning

1990 ◽  
Vol 20 (9) ◽  
pp. 1448-1452 ◽  
Author(s):  
Throstur Eysteinsson ◽  
Michael S. Greenwood
Keyword(s):  
Plant Growth ◽  
Growth Regulator ◽  
Plant Growth Regulator ◽  
Shoot Elongation ◽  
Root Pruning ◽  
Mature Trees ◽  
Foliar Sprays ◽  
Female Flowering

Flowering was promoted on potted, indoor, and field-grown 3-year-old juvenile and mature Larixlaricina (Du Roi) K. Koch grafts by foliar sprays of the plant-growth regulator gibberellin A4/7 and root pruning. Biweekly gibberellin A4/7 applications of three different durations, commencing at the start of long-shoot elongation, yielded similar increases in female flowering in the greenhouse; this effect was synergistically enhanced by root pruning. Male flowering was promoted by gibberellin A4/7 + root pruning in mature but not juvenile grafts. In the field, female flowering was also promoted by the gibberellin A4/7 treatment commencing at the start of shoot extension, but gibberellin A4/7 applications starting later were not effective. Grafted scions from mature trees responded significantly better to gibberellin A4/7 + root pruning than did seedling scions.

scholarly journals Transient expression of recombinant S1 protein of Porcine Epidemic Diarrhea Virus in Nicotiana benthamiana

2021 ◽  
Vol 19 (1) ◽  
pp. 95-105
Author(s):  
Ho Thi Thuong ◽  
Le Thu Ngoc ◽  
Nguyen Thu Giang ◽  
Trinh Thai Vy ◽  
Phan Trong Hoang ◽  
...  
Keyword(s):  
Western Blot ◽  
Transient Expression ◽  
Nicotiana Benthamiana ◽  
Porcine Epidemic Diarrhea Virus ◽  
Expression Level ◽  
Total Soluble Protein ◽  
Original Form ◽  
S Protein ◽  
Diarrhea Virus ◽  
Porcine Epidemic Diarrhea

Porcine Epidemic Diarrhea (PED) is an infectious disease with high mortality especially in suckling piglets. Among the structural proteins of Porcine Epidemic Diarrhea Virus (PEDV), the S protein (including sub-domain S1 and S2), is a homotrimer protein that plays an important role in attaching the viruses to the cell receptors. In particular, the S1 protein is considered as an important sub-component in the development of effective vaccines against PEDV. In this study, for the purpose of expressing S1 in the original form of trimmer and oligomer of trimer based on S-tag and S-protein interactions, the DNA encoding for S1 protein was fused with GCN4pII or GCN4pII-Stag, was then inserted to the pRTRA cloning vector under the control of the 35S CaMV promoter. After that, the whole cassete was inserted into the pCB301 vector and transformed into Agrobacterium tumefaciens for transient expression on Nicotiana benthamiana. The expression of recombinant S1 proteins in tobacco was determined by Western blot. The results showed that the expression levels of S1 trimer and S1 trimer S-tag proteins were equal in plants, which also indicated that S-tag fusion did not affect the expression level of the S1 protein. However, the expression level of S1 proteins was relatively low, reaching 0.005% of total soluble protein. In addition, the expression of S1 trimer S-tag protein and Sprotein-tp protein by co-transformation of two A. tumefaciens strains containing corresponding vectors in plants were also determined by Western blot. This is a premise study for the development of subunit vaccines in plants that prevent the spread of PEDV.

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