Multiplication of hybrid aspen (Populus tremula L. x P. tremuloides Michx.) from cuttings

The primary aim of the present study was to find an efficient and simple method of vegetative propagation for producing large numbers of hybrid aspen (Populus tremuloides L. x P. tremula Michx.) plants for forest plantations. The key objectives were to investigate the main physiological factors that affect the ability of cuttings to regenerate and to determine whether these factors could be manipulated by different growth conditions. In addition, clonal variation in traits related to propagation success was examined. According to our results, with the stem cutting method, depending on the clone, it is possible to obtain only 1−8 plants from one stock plant per year. With the root cutting method the corresponding values for two-year-old stock plants are 81−207 plants. The difference in number of cuttings between one- and two-year-old stock plants is so pronounced that it is economically feasible to grow stock plants for two years. There is no reason to use much older stock plants as a source of cuttings, as it has been observed that rooting ability diminishes as root diameter increases. Clonal variation is the most important individual factor in propagation of hybrid aspen. The fact that the efficiently sprouted clones also rooted best facilitates the selection of clones for large-scale propagation. In practice, root cuttings taken from all parts of the root system of hybrid aspen were capable of producing new shoots and roots. However, for efficient rooting it is important to use roots smaller than one centimeter in diameter. Both rooting and sprouting, as well as sprouting rate, were increased by high soil temperature; in our studies the highest temperature tested (30ºC) was the best. Light accelerated the sprouting of root cuttings, but they rooted best in dark conditions. Rooting is essential because without roots the sprouted cutting cannot survive long. For aspen the criteria for clone selection are primarily fiber qualities and growth rate, but ability to regenerate efficiently is also essential. For large-scale propagation it is very important to find clones from which many cuttings per stock plant can be obtained. In light of production costs, however, it is even more important that the regeneration ability of the produced cuttings be high.

The Significance of Wild Plants in the Evolutionary Ecology of Three Major Viruses Infecting Cultivated Sweetpotato in Uganda

The studies presented in this thesis contribute to the understanding of evolutionary ecology of three major viruses threatening cultivated sweetpotato (Ipomoea batatas Lam) in East Africa: Sweet potato feathery mottle virus (SPFMV; genus Potyvirus; Potyviridae), Sweet potato chlorotic stunt virus (SPCSV; genus Crinivirus; Closteroviridae) and Sweet potato mild mottle virus (SPMMV; genus Ipomovirus; Potyviridae). The viruses were serologically detected and the positive results confirmed by RT-PCR and sequencing. SPFMV was detected in 24 wild plant species of family Convolvulacea (genera Ipomoea, Lepistemon and Hewittia), of which 19 species were new natural hosts for SPFMV. SPMMV and SPCSV were detected in wild plants belonging to 21 and 12 species (genera Ipomoea, Lepistemon and Hewittia), respectively, all of which were previously unknown to be natural hosts of these viruses. SPFMV was the most abundant virus being detected in 17% of the plants, while SPMMV and SPCSV were detected in 9.8% and 5.4% of the assessed plants, respectively. Wild plants in Uganda were infected with the East African (EA), common (C), and the ordinary (O) strains, or co-infected with the EA and the C strain of SPFMV. The viruses and virus-like diseases were more frequent in the eastern agro-ecological zone than the western and central zones, which contrasted with known incidences of these viruses in sweetpotato crops, except for northern zone where incidences were lowest in wild plants as in sweetpotato. The NIb/CP junction in SPMMV was determined experimentally which facilitated CP-based phylogenetic and evolutionary analyses of SPMMV. Isolates of all the three viruses from wild plants were genetically similar to those found in cultivated sweetpotatoes in East Africa. There was no evidence of host-driven population genetic structures suggesting frequent transmission of these viruses between their wild and cultivated hosts. The p22 RNA silencing suppressor-encoding sequence was absent in a few SPCSV isolates, but regardless of this, SPCSV isolates incited sweet potato virus disease (SPVD) in sweetpotato plants co-infected with SPFMV, indicating that p22 is redundant for synergism between SCSV and SPFMV. Molecular evolutionary analysis revealed that isolates of strain EA of SPFMV that is largely restricted geographically in East Africa experience frequent recombination in comparison to isolates of strain C that is globally distributed. Moreover, non-homologous recombination events between strains EA and C were rare, despite frequent co-infections of these strains in wild plants, suggesting purifying selection against non-homologous recombinants between these strains or that such recombinants are mostly not infectious. Recombination was detected also in the 5 - and 3 -proximal regions of the SPMMV genome providing the first evidence of recombination in genus Ipomovirus, but no recombination events were detected in the characterized genomic regions of SPCSV. Strong purifying selection was implicated on evolution of majority of amino acids of the proteins encoded by the analyzed genomic regions of SPFMV, SPMMV and SPCSV. However, positive selection was predicted on 17 amino acids distributed over the whole the coat protein (CP) in the globally distributed strain C, as compared to only 4 amino acids in the multifunctional CP N-terminus (CP-NT) of strain EA largely restricted geographically to East Africa. A few amino acid sites in the N-terminus of SPMMV P1, the p7 protein and RNA silencing suppressor proteins p22 and RNase3 of SPCSV were also submitted to positive selection. Positively selected amino acids may constitute ligand-binding domains that determine interactions with plant host and/or insect vector factors. The P1 proteinase of SPMMV (genus Ipomovirus) seems to respond to needs of adaptation, which was not observed with the helper component proteinase (HC-Pro) of SPMMV, although the HC-Pro is responsible for many important molecular interactions in genus Potyvirus. Because the centre of origin of cultivated sweetpotato is in the Americas from where the crop was dispersed to other continents in recent history (except for the Australasia and South Pacific region), it would be expected that identical viruses and their strains occur worldwide, presuming virus dispersal with the host. Apparently, this seems not to be the case with SPMMV, the strain EA of SPFMV and the strain EA of SPCSV that are largely geographically confined in East Africa where they are predominant and occur both in natural and agro-ecosystems. The geographical distribution of plant viruses is constrained more by virus-vector relations than by virus-host interactions, which in accordance of the wide range of natural host species and the geographical confinement to East Africa suggest that these viruses existed in East African wild plants before the introduction of sweetpotato. Subsequently, these studies provide compelling evidence that East Africa constitutes a cradle of SPFMV strain EA, SPCSV strain EA, and SPMMV. Therefore, sweet potato virus disease (SPVD) in East Africa may be one of the examples of damaging virus diseases resulting from exchange of viruses between introduced crops and indigenous wild plant species. Keywords: Convolvulaceae, East Africa, epidemiology, evolution, genetic variability, Ipomoea, recombination, SPCSV, SPFMV, SPMMV, selection pressure, sweetpotato, wild plant species Author s Address: Arthur K. Tugume, Department of Agricultural Sciences, Faculty of Agriculture and Forestry, University of Helsinki, Latokartanonkaari 7, P.O Box 27, FIN-00014, Helsinki, Finland. Email: tugume.arthur@helsinki.fi Author s Present Address: Arthur K. Tugume, Department of Botany, Faculty of Science, Makerere University, P.O. Box 7062, Kampala, Uganda. Email: aktugume@botany.mak.ac.ug, tugumeka@yahoo.com

Havupuiden erikoismuotojen lisäysmenetelmät

Havupuiden erikoismuotoja on käytetty koristekasveina jo vuosisatoja ympäri maailmaa. Niitä on lisätty pääsääntöisesti pistokkaista ja varttamalla. Suomessa kotimaisten metsäpuidemme erikois-muotoja on kartoitettu ja kerätty kokoelmiin järjestelmällisemmin 1960-luvulta alkaen. Taimisto-viljelijät, puutarhasuunnittelijat ja kotipuutarhurit ovat olleet enenevässä määrin kiinnostuneita näistä kotimaisista kestävistä havukasveista. Yli 90 prosenttia markkinoillamme olevista havukas-veista tuodaan ulkomailta, joten on selvää, että niiden talvenkestävyydessä on ongelmia. Tämän tutkimuksen tavoitteena oli selvittää kotimaisille erikoismuodoille sopivia lisäysmene-telmiä ja siten edistää kotimaisen havukasvituotannon mahdollisuuksia. Aineistona kokeissa oli kotimaisia erikoismuotoja metsäkuusesta (Picea abies (L.) Karsten) ja kotikatajasta (Juniperus communis L.), tavallisia metsäkuusia sekä kahdeksan ulkomaista havupuutaksonia. Lisäysmene-telmistä tutkittiin varttamista ja pistokaslisäystä ja kokeet suoritettiin Metsäntutkimuslaitoksen toimipaikoissa Lopen Haapastensyrjässä sekä Punkaharjulla. Varttamiskokeessa vertailtiin koti-maisen kuusen erikoismuotokloonien varttamisen onnistumista. Pistokaskokeissa tutkittiin geno-tyypin, emopuun iän, pistokasoksan sijainnin sekä hormonikäsittelyn vaikutusta havukasvien pis-tokkaiden juurtumiseen. Tavalliset metsäkuuset toimivat kontrolleina. Tutkimus osoitti, että varttaminen onnistui erinomaisesti kaikilla erikoismuotoklooneilla. Ovat-ko vartteet esteettisesti katsottuna koristekäyttöön sopivia, jää vielä seurattavaksi. Pistokaskokeis-sa havaittiin, että juveniilisuus vaikutti pistokkaiden juurtumiseen, mutta iäkkäistäkin puista lisää-minen onnistuu, kunhan genotyyppi on sopiva. Keskimäärin alaoksat juurtuivat paremmin kuin latvuksen yläosista otetut pistokasoksat, mutta vain yhdellä kloonilla ero oli tilastollisesti merkit-sevä. Hormonikäsittely heikensi selvästi kotimaisen kuusen ja katajan pistokkaiden juurtumista, mutta ulkomaisiin havupuulajeihin käsittelyllä ei ollut vaikutusta. Kotimaisen havukasvituotannon pohjaksi pitäisi tehdä kloonivalintaa, jossa koristearvon lisäksi otettaisiin huomioon myös kloonin lisättävyys. Taimien tuottaminen pistokkaista on selvästi edul-lisempaa kuin vartteiden tuottaminen, joskin varte kasvaa myyntikuntoon nopeammin kuin pisto-kastaimi. Pistokastaimi on kuitenkin omajuurinen ja stabiilimpi kasvutavaltaan kuin varte. Tämä korostuu etenkin kääpiömuotoja tuotettaessa.