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Tuija Aronen

Publications and source records attributed to Tuija Aronen.

5 recordsLinked to original sources

Does lignin modification affect feeding preference or growth performance of insect herbivores in transgenic silver birch (Betula pendula Roth)?

Transgenic silver birch (Betula pendula Roth) lines were produced in order to modify lignin biosynthesis. These lines carry COMT (caffeate/5-hydroxyferulate O-methyltransferase) gene from Populus tremuloides driven by constitutive promoter 35S CaMV (cauliflower mosaic virus) or UbB1 (ubiquitin promoter from sunflower). The decreased syringyl/guaiacyl (S/G) ratio was found in stem and leaf lignin of 35S CaMV-PtCOMT transgenic silver birch lines when compared to non-transformed control or UbB1-PtCOMT lines. In controlled feeding experiments the leaves of transgenic birch lines as well as controls were fed to insect herbivores common in boreal environment, i.e., larvae of Aethalura punctulata, Cleora cinctaria and Trichopteryx carpinata (Lepidoptera: Geometridae) as well as the adults of birch leaf-feeding beetles Agelastica alni (Coleoptera: Chrysomelidae) and Phyllobius spp. (Coleoptera: Curculionidae). The feeding preferences of these herbivores differed in some cases among the tested birch lines, but these differences could not be directly associated to lignin modification. They could as well be explained by other characteristics of leaves, either natural or caused by transgene site effects. Growth performance of lepidopteran larvae fed on transgenic or control leaves did not differ significantly.

Animals↗

Vitrification, a complementary cryopreservation method for Betula pendula Roth.

Cryopreservation--the storage of plant germplasm in liquid nitrogen--provides a modern tool for the conservation of forest genetic resources. It is especially applicable for species in which their micropropagation can be initiated from mature tree buds, e.g., silver birch (Betula pendula Roth), thus enabling the conservation of specific genotypes: endangered elite trees and trees expressing rare, valuable or interesting characteristics. The aim of the present study was to develop a vitrification protocol applicable for the cryostorage of silver birch that avoids the use of expensive sophisticated freezers. The average recovery of vitrified axillary silver birch buds was 71% using a protocol that started with four-week cold hardening of bud-bearing in vitro donor shoots on modified medium under short day conditions. After cold hardening, the excised axillary buds were precultivated on medium containing 0.7 M sucrose for 24 h under the same conditions as during the cold hardening period. Following preculture, the buds were treated with loading solution containing 2M glycerol and 0.4 M sucrose for 20 min at room temperature. Finally, the buds were dehydrated with PVS2 cryoprotectant for 120 min followed by direct immersion in liquid nitrogen. According to the morphology and the RAPD profiles of regenerated plants in the greenhouse, the genetic fidelity of the vitrified birch material seems to have remained unchanged.

Betula↗

Expression of Vitreoscilla haemoglobin in hybrid aspen (Populus tremula x tremuloides).

We describe the first ever expression of Vitreoscilla haemoglobin (VHb) in an economically important boreal woody plant hybrid aspen (Populus tremula x tremuloides). VHb has mainly been expressed in biotechnologically important unicellular organisms of both prokaryotic and eukaryotic origin. VHb expression, in this study, was analysed under different greenhouse cultivation conditions and under elevated UV-B illumination. Microscope analyses of leaves grown under optimized conditions revealed significant differences both in cell structure and size when the transgenic VHb lines were compared with the control lines. VHb lines displayed a higher relative volume of mitochondria and a significantly enhanced accumulation of starch in chloroplasts, all of which pointed towards changes in cellular energy production. Under elevated UV-B illumination, the differences between VHb lines became evident. Some specific VHb lines had elevated levels of total flavonoids, individual quercetin, kaempferol- and myricetin-derivatives relative to controls and other transgenic lines. This observation may reflect the availability of extra energy resources for secondary metabolite production and possibly an enhanced protection ability of these transgenic lines against UV-B illumination. Thus, all these findings point to changes in the energy metabolism of VHb lines. In the cultivation conditions tested this observation did not, however, result in a general improvement of elongation growth.

Journal Article↗

Silver birch (Betula pendula) plants with aux and rol genes show consistent changes in morphology, xylem structure and chemistry.

The effects of Agrobacterium pRiA4 rol and aux genes, controlled by their endogenous promoters, on tree growth and wood anatomy and chemistry were studied in 5- and 7-year-old silver birch (Betula pendula Roth) plants. Southern hybridization confirmed the following rol and aux gene combinations: control plants (no genes transferred); plants with rolC and rolD genes; plants with rolA, rolB, rolC and rolD genes; and plants with rolA, rolB, rolC, rolD, aux1 and aux2 genes. Transgene mRNA was most abundant in phloem/cambium samples and in the developing xylem, whereas no expression was detected in leaves. Plants with rolC and rolD genes or with all the rol genes were significantly shorter and had smaller leaves and a more bushy growth habit than control plants or plants with both aux and rol genes. Morphological observations and wood chemistry analyses revealed that plants with rol genes produced less xylem and broke bud later than control plants or plants with both aux and rol genes. Tension wood was detected in both control and transgenic plants irrespective of their gene combination, probably as a result of greenhouse cultivation. Xylem fibers were shorter in transgenic plants than in control plants, and plants with all the rol genes were characterized by shorter vessels compared with the control plants and a smaller proportional area of vessels compared with the other groups. In addition, silver birch plants with all the rol genes had approximately a 3.3% lower concentration of total acid soluble carbohydrates than control plants. We conclude that the rolC and rolD genes induced the typical "rol-phenotype," and that this was emphasized by concomitant expression of the rolA and rolB genes and alleviated by the presence of aux1 and aux2 genes. We observed consistent phenotypic effects of rol and aux genes on the morphology, anatomy and cell wall chemistry of the plants.

Base Sequence↗

Genetic transformation of silver birch (Betula pendula) by particle bombardment.

We used in vitro callus and shoot cultures as target material for genetic transformation of silver birch (Betula pendula Roth) by particle bombardment. Cultivation of in vitro shoot cultures before particle bombardment and a long selection period, combined with a high concentration of selective agent after bombardment, led to the production of transformed plantlets that were stable, and no escapes were found among the tree lines produced. Clonal variation in transformation efficiency was found in transient expression of the beta-glucuronidase gene in callus cultures and in plantlets transformed by stable integration of the ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) and neomycin phosphotransferase (npt2) genes.

Journal Article↗