Home – Home – Jornals – Siberian Journal of Forest Science 2016 number 5

Main results of allozyme-geographical study of the polymorphism, differentiation and origin of natural Pinus sylvestris L. populations in 6phylogeographical regions - Middle Siberian Tableland, Lena-Angara Tableland, South Siberia Mountains, Near-Baikalia, Western Trans-Baikalia and Northern Mongolia - have been briefly analyzed and generalized. A relative homogeneity of the population polymorphism in these regions has been revealed, with the exception of marginal ones (Tura, Ulan-Bator). Genetic distances of M. Nei (1978, D_N78) between the extremely remote populations in the investigated regions reach the level of the geographical race, but they are several time lesser in the regions’ limits - not more of the genosystmatic level of the middle differentiated local populations ( D_N78 = 0.010-0.012). The geographical group of the South Siberia Mountains stand apart distinctly ( D_N78 = 0.018±0.004) in the genogeographical structure of P. sylvestris on the basis of Nei`s genetic distances analysis and of the genosystematic scale (Sannikov, Petrova, 2012). Besides the Selenga population group (in the rank of local population, D_N78 = 0.013), also as well the Angara-Yenisei and North-Baikal population group (in the rank of subpopulation, D_N78 = 0.004-0.008) stand apart on the rest of the territory. Gradient-analysis of the genetic distances revealed the borders between the phylogeographical groups of pine populations on the Lena-Angara and Central-Siberian plateaus, on the one hand, in the South Siberia Mountains, Near-Baikal and Trans-Baikalia, on the other hand, and an important role of l. Baikal as a barrier of the migration and differentiation of the populations. As an analysis result of minimal genetic distances between 25 P. sylvestris populations in the south «non-glacial» zone and the group of 8 populations in the «glacial» zone 4 main refugiums (South-Near-Baikal, South-Ural, Upper-Vitim and Upper-Olekma) and several secondary refugiums have been revealed.

In the territory of steppe zone of Ob’-Irtysh interfluve it was studied the differentiation of scots pine populations of belt forests with use of allozyme’s, phene’s and morphometric markers. Here allocated 2 forest-seed zoning and border between them practically coincides with border between Siberian and Kulunda pine subspecies on L. F. Pravdin’s classification. Allozyme’s analysis didn’t reveal significant differentiation of stands, except for rare differences between some of them and a slight trend for decrease in effective number of alleles in the more southern populations. The analysis of a molecular variance (AMOVA) also shows absence of differentiation of populations from different forest-seed zoning. Differentiation of populations by using phenes (coloring of seeds, type of cone’s apophysis) and high-inherited morphometric characters (index form of cones and weight of 1000 seeds) was more effective at comparing populations on both levels - between and within forest-seed zoning. It allowed revealing of reliable distinctions between populations in almost 82 % of cases of total number of the compared couples of populations, thus extent of differentiation using the allozyme’s markers is almost three times lower. The assessment of population structure of pine in tape forest of Altai region, which is carried out with application of a complex of markers, indicates between-populations heterogeneity in this part of area. The received results confirm the expediency of complex researches of population structure of forest-forming species and the necessity of more precise definition of forest-seed zoning of a scots pine in the studied territory.

Most productive populations of Siberian stone pine trees were revealed as a result of many years study of the selection structure of the Siberian stone pine forests in the Republic of Altai. Based on the analysis of archive materials and ground inventory of the selection and genetic objects, the information about their quantity and preservation is provided. Currently, in the Republic of Altai, 320 elite trees have beenregistered, and on this basis 60.9 ha of forest seed plantations and 13.7 ha clone archives have been established. In the period of 2005-2014, the number of elite trees was reduced from 438 to 320. Experimental elite tree crops not presented. According to the inventory, as of 2014, at forest seed plantations and at the archives of clones grow 4.837 grafted trees of 282 clones, 139 of whichare progenies of the elite trees, selected for seed production, 124 - for resin production, and 19 - for the productivity and quality of stem wood. All clonal facilities are concentrated on breeding nursery of the Teletskoe forestry district area of ​​about 75 ha. Unique clonal objects of the Siberian stone pine in Teletskoe forestry district play a crucial role in the preservation, study and rational use of the gene pool of this tree species and require continuous monitoring of their condition and protection. Visual forest pathology inspections and molecular genetic diagnostics revealed that on clonal plantations of the Siberian stone pine, progressive needle disease developed, caused by new for Western Siberia an extremely dangerous pathogen - Dothistroma septosporum (Dorog.) M. Morelet. In this connection it is necessary to conduct appropriate sanitary measures and to carry out additional studies of population genetic features and pathways for this pathogen. The priority activities have been outlined to further development of the unified genetic and breeding complex of the Siberian stone pine in the Republic of Altai.

Three native populations of P. sylvestris, ssp. lapponica (Alakurtti, Gridino, Pjaozero) and six native populations of P. s., ssp . sylvestris (Voinitsa, Maslozero, Vodlozero, Zaonezshye, Kivach, Sortavala) along with one artificial population - Petrozavodsk seed orchard have been studied using four nuclear microsatellite primers (Spac11.8, Spac12.5, PtTX2123, PtTX2146) to find out the peculiarities of Scotch pine intraspecific diversity in Karelia. In total, 66 alleles were found. All four loci turned out to be polymorphic in all populations. The observed heterozigocity level of all studied Karelian populations was lower than expected, which is evidence of heterozygotes deficiency in Karelian pine populations. Overall, the investigated P. sylvestris populations can be described as having a high level of the genetic variability especially compared to results obtained earlier by isozyme analyses. The Petrozavodsk seed orchard ought to be mentioned as having one of the highest levels of the genetic variability, which gives evidences in favor of good state regarding representativeness of Scotch pine gene pool here. No substantial differences in genetic structure and diversity levels have been found for ss . lapponica populations vs . ss. sylvestris ones. AMOVA analysis showed that despite the significant differences between pine populations, considering both the allele composition and the genetic diversity, the major part of the variety (90 %) was allocated inside group. The investigated populations were subdivided into two clusters using UPGMA technique characterized by substantial Nei distance ( D_N = 0.273). The former one included the majority of Karelian pine populations ( D_N = 0.030-0.082) along with the Alakurtti population from Murmansk region. The latter cluster consisted ofthe most isolated «Kivach» and «Zaonezshye» populations, which had the lowest levels of genetic diversity, and the «Petrozavodsk seed orchard».

A comparative analysis of variability of morphological characteristics of Populus nigra L., Populus laurifolia Ledeb. (Salicaceae) and their natural hybrids in the flood-plain of Tom’ river (Kemerovo Oblast) revealed traits for identification of the natural hybrids. The most important of them are: the ribbing of the long shoots, types of short shoots of crown, location generative buds on the shoots, the number of bud’s glumes of the generative buds, and the shape of the leaf blade. The morphometric characteristics of leaves should be used to clarify the nature of the hybrid individuals. In female hybrids, the reliable diagnostic feature is the number of box cusps. Qualitative and quantitative characteristics of hybrids for the most part are of intermediate character, but the differentiation of shoots crown and location generative buds hybrids closer to the Populus laurifolia . Values morphometric attributes of leaves at hybrids in most cases authentically differ from ones P. nigra and P. laurifolia , and combinations of these differences in each hybrid model are individual. Hybrids don’t form separate populations, grow by single trees or form small clones in a flood-plain of Tom’ river. Occurrence of hybrids depends on stage of their development and increases in areas with significant anthropogenic pressure.

Provenance trials of Pinus sibirica Du Tour and Pinus koraiensis Siebold et Zucc. (here after «studied species») were studied. In our study we assessed the growth parameters as well as anatomical and morphological parameters of the studied species corresponding to different provenances of their testing in the south of the Krasnoyarsk and Khabarovsk territories.We determined that the growth rate of trees corresponding to different provenances is determined not only by the inherited characteristics, but also by adaptation. At both experimental regions the offspring of trees corresponding to local provenance are clearly better adapted. Nevertheless, at Krasnoyarsk Krai provenance trials, we found that the phenotypic indicators and degree of preservation of the offspring of two Korean pine corresponding to Obluchensky and Chuguevsky provenances are at the same level as for the local Siberian pine. Tree rings widths have been measured for the Siberian pine corresponding to different provenances at both plantations. We conclude that at the Ermakovskoe plantation there is a positive impact of the environmental conditions on tree-ring width for Korean pine corresponding to different provenances, and in Khabarovsk Krai there is a negative impact of the environmental conditions on tree-ring width for the Siberian pine corresponding to different provenances.

The paper presents new data on 28-30-year-olds testing of triploid poplar hybrids (2 n = 3 x = 57), obtained using in hybridization artificially obtained unreduced diploid (2 n ) pollen (induced by high temperature or colchicines treatment) in Voronezh region. The growth dynamics, preservation and timber productivity of 26 triploid and diploid (2 n = 2 x = 38) clones of Populus alba L., P. balsamifera L., P. nigra L. and intersectional hybrids ( Populus deltoides Marsh. × P. balsamifera L.) were studied. It was revealed that a preservation of triploid hybrids of white poplar at 28-year-old age was higher than diploid hybrids. The greatest loss of plants of white poplar was observed during the first three years after planting of cutting seedlings in the nursery. In comparison, major loss of plants of P. balsamifera L., P. nigra L. and intersectional hybrids ( P. deltoides Marsh. × P. balsamifera L.) was observed after the 2010 drought. Especially high plant mortality was observed in P. balsamifera clone (preservation of 40 %) and to a lesser extent in poplar «Robusta-236» (preservation of 60 %). There were no statistically significant differences in growth between groups of diploid and triploid white poplar clones. Only part of the artificially produced autotriploid hybrids (40 %) were characterized by fast growth and productivity. It is suggested that the heterosis effect in triploids associated with participation of high heterozygous 2 n gametes in hybridization. Such were formed on the basis of a synapsis of chromosomes and loss of the I-st meiotic division. Six best triploid and diploid poplar hybrids have been selected as promising for Voronezh region.

In this report the results of individual and clone variability of the number of cones on Korean pine Pinus koraiensis Siebold et Zucc. grafting trees on experimental seed plantation (up to 21 years) in Khekhtsirskii Forestry unit of Khabarovsk region are presented. The maximum characteristics of seed bearing were proper for clones of trees at the age of 170-195 with a large number of the previous year sprouts and the average annual growth in the range of 0.12-0.15 m in height and 0.24-0.29 cm in diameter. The trees over 200 years have weak vitality grafts. That’s why the age of Korean pine seed productivity breeding should be limited by this age. The coefficients of variation of the clone seed bearing characteristics amount to 35-56 % during the first decade, later - up to 154 %. The variability of the whole population is the same. Sometimes, the Rate of Relative Total Seed Productivity (RRTSP) differs ten or more times in two trees of one clone, while the best clones exceed the worst ones 2-6 times. As consistent with a single-factor analysis of variance, clone differences in the numbers of cones are not significant for P = 0.05. This characteristic heritability coefficient ( H ²) has not exceeded 0.08 during of all dates of the observations, including the twenty first years after the seed plantation creation. It is recommended to organize protection and enclosure of the most valuable plots, to optimize the graft and nursing technology for the grafted seed plantations. It is necessary to prepare the graft cuttings only from productive trees of productive clone before genetic certification.

Fast height growth selection usually considered as the basic type of Scotch pine breeding strategy. Due to this fact height growth of pine clones usually is taken as the first key feature. In the view of good seed production there is only one indicator that could be adopted as having summarized effect reflecting the real contribution of a clone to the whole seed orchard yield. This is an average number of full seeds per tree (ramet). This trait is the second most important one after the height growth ability feature. The way of two-trait breeding program with the aforesaid features seemed to be the most suitable for Scotch pine. Main theses of 4-stages complex assessment procedure have been formulated concerning Scotch pine vegetative progenies grown at seed orchards. It was stated that this system of breeding and genetic assessment is based on correlations found for habitus and reproductive features of Scotch pine clones along with the height growth ability of their seed progenies. In the first stage pine clones meeting the criteria (≥ X + 0.5σ _x ) regarding height growth are to be selected. In the second stage, clones with hard tapering stem and crown formed by long and thick branches have to be discarded. At this stage, such feature as «mean diameter of three thickest limbs at the height of 1.5-2.0 m» became the key factor. Its mean value for selected clones must not be by 1-2 % higher than for general clone set. In the third stage a complex of reproductive features for clones that have passed the previous stages are taken into account. The trait «average number of full seeds per ramet» is the key feature in this stage according to the criteria (≥ X ). The fourth stage implies that progeny trials with open-pollinated progenies ought to be laid down to calculate the general and specific combining ability. It has been concluded that high intensive selection of clones grown at the I-stage seed orchards promotes a substantial genetic gain expected at the subsequent breeding stages. Seven clones (10 % of the total) have been selected using the two-trait breeding approach. As a result, habitus features of the selected clones became much better. The height and stem diameter increased by 7.6 and 2.6 % and crown width along with branch diameter did the same by 2.1 and 1.2 %, respectively. At the same time seed yield increased by 43.0 %.

The program is based on 30 years of research by the author for breeding softwood for plantation cultivation and modeling of the course of growth of trees and forest stands. Scope of work presented progeny plus trees and conventional trees of Scots pine Рinus sylvestris L. and spruce Finnish ( Рicea  fennica (Regel) Кom.) from 16 populations of the Perm region total of 1435 families under the age of 23 years. In modeling the course, studies covered the development of stands of spruce growing on 349 plots. It was found that the problem of different types of progeny can not be solved by increasing the term of the test and on the basis of management of development of the stand test cultures by certain models, with constant regulation of their density. The summary of data on the growth of progeny in different density test cultures found that the offspring «remembers» competitive and edaphic conditions which formed the parent tree and grows best in them. Therefore, a selection of plus trees in plantations-analogues plantation crops, ie in exactly the same conditions in which we are going to grow their offspring,is based on earlier estimates of growth of offspring at the age of 4-8 years. At the same time selection of candidates for the varieties happening after a series of stepwise tests several seed yield with consistent reduction in the number of families from 500-1000 to ≈ 70. Reliability selection of candidates will be 70-80 %. The speed and reliability of the best selection progeny can be significantly increased by using chemical marker needles. The program consists of 10 stages, by 3-4 times reduces the cost and time for selection of industrial varieties of Scots pine and spruce Finnish. They can be reduced even more by using chemical marker needles typical for families with rapid growth.

Verification results of one corollary of developing theory of the authors about the coniferous hydrochory (Sannikov, Sannikova, 2007) - a hypothesis of genetic integration of Scots pine Pinus sylvestris L. populations in the direction of rivers flow - have been cited. The average genetic Nei`s distance (Nei, 1978; 0.0024 ± 0.0015) between the 8 adjacent P. sylvestris populations on the transect along all the Ob valley (Lake Teletskoje-Barnaul-Tomsk-Surgut-Khanty-Mansiisk-Geologicheskii-Berezovo-Synja), that is twice time less as between the 14 populations on the Ob`s watersheds with the adjacent rivers (Yenisei, Irtysh, Pur, Konda, Tavda, Nadym) (0.0049 ± 0.0009), has been determined on the basis of comparative allozyme analysis (Nei, 1978; 0.0024 ± 0.0015). Corresponding average gradient of the genetic distances between the population located along the Ob river-bed is also 70 % less (0.66 ± 0.43), than on the transects directed across the watersheds (1.40 ± 0.43). Probably, a lesser genetic distance and its gradients, which revealed between the Scots pine populations along river-bed Ob in comparison with ones on the transects directed across the watersheds, depended on greater rate of its seed hydrochory (up to 800 km/1000 yr) in comparison with the anemochory on the watersheds. As a whole, a hypothesis about more speed (prioritic) hydrochoric dispersal and relative genetic integration of Scots pine populations along the rivers flow in comparison with anemochoric dispersal on the watersheds has been corroborated on the basis of results generalization of the present and preceded research.

The drought resistance of trees in the Scots pine Pinus sylvestris L. populations was studied under forest steppe conditions of southern Siberia. We found large differences amongthe treesin the time and rate of needles dehydration. In the populations of the more arid growing conditions (Shira, Balgazyn) the rate of loss 50 % of the water was three to four times lower than in the population of the more favorable conditions for growth (Minusinsk). It has been established that the variability of water-holding capacity of needles from individual trees in populations varies from high to very high levels. On the contrary, for the water absorption capacity of needles characteristically has variability, as within population and between them. Is marked a great assessment reliability of water holding capacity of the needles under the pooled analysis of absolute and relative indicators of dynamics of the needles degradration. We investigated the correlation of needles’ water retention signs with a height and heterozygosity of trees. It was found that under more favorable conditions of the growth the large part of sample are the trees with a direct connection between heterozygosity and drought resistance of tree and in the worst conditions - with a reverse. The correlations of water-holding capacity of needles with the height of the tree are ambiguous: in the Minusinsk sample, the most of drought-resistant trees are characterized by better growth, in Balgazyn population - conversely. Some dwarf individuals from the Balgazyn and Shira populations in terms of drought tolerance are at same level as the typical trees, among the less drought-resistant trees found as dwarfs, and typical trees. It was concluded that there are trees in populations with different strategies to adaptation to the moisture deficit.

Endogenous, individual and ecological-geographic variation of common juniper Juniperus communis L. in 10 coenopupulations in Cis-Urals and the South Urals by 17 quantitative morphological traits of generative and vegetative organswere studied. Endogenous variation of most traits of generative organs is characterized by a low and very low level, individual variability is close or some exceeding to intra-individual one. Parameters of vegetative organs are more variable both at endogenous sample level and at individual one. Ecological-geographic variability of many traits some yields or corresponds to the level of endogenous and individual variability. However, separate traits (mass of 1000 cones, number of seeds per cone, length of annual increment of III order shoots, needle length and width) vary more between coenopopulations than in the limits of ones. Some traits are connect by correlation dependencies with natural-climatic factors of locations - geographic latitude and to a lesser degree with longitude and altitude, sum of active temperatures and amount of precipitation. The obtained data indicate the significant morphological diversity of individuals within coenopopulations as well as phenotypic specifics of coenopopulations and two main inhabited regions - Cis-Urals and the South Urals. The presented material is the base for further analysis of population structure of common juniper in the region, development of measures for species gene pool preservation, fulfillment of breeding selection of individuals and coenopopulations by one or another economically valuable trait (large-coned, with the most portion of pulp in cones, long-needled and others).

Allozyme polymorphism of Siberian stone pine Pinus sibirica Du Tour has been studied in 14 artificial stands in the South Urals and Bashkir Cis-Urals on the base of 7 gene-enzyme system analysis. The following values of genetic diversity are determined: mean number of alleles per locus ( A ) constitutes 1.69±0.08; portion of polymorphic loci ( P ₉₅) - 50.0 %; the average expected heterozygosity ( H_e ) - 0.121±0.015; the average observed heterozygosity ( H_o ) - 0.127±0.017.The level of genetic variability in artificial stands of Siberian stone pine in the region is somewhat inferior to that in natural populations of the species. The highest genotype heterozygosity is determined in high-productive 110 year-old artificial stand in the South Urals (Beloretsky-2 site), and also in Ufimsky and Mishkinsky sites in Bashkir Cis-Urals. The lowest heterozygosity values are revealed in Birsky and Tuimazinsky sites characterized by the weakened vital state of individuals. In total we can speak about the maintenance of essential part of the species’ genetic polymorphism under introduction, especially in some stands. Genetic similarity of the studied stands is shown: inter-sample component of the total genetic diversity ( F_ST ) constitutes 2.2 %, the average Nei’s genetic distance ( D ) - 0.0033±0.00023, that is also typical of natural populations of Siberian stone pine in the species range. The obtained data about the genetic variability level of artificial stands in a complex with forestry characteristics give evidence of the successful species introduction in the region and the necessity of resumption of works on Siberian stone pine culture establishment in an industrial scale.

In this work was tested method of DNA-diagnostic for identification and further localization of pest holes of Heterobasidion annosum (Fr.) Bref s.l. Heterobasidion annosum is dangerous enemy for coniferous trees because they have no effective natural resistance against this pathogen. Therefore it can easily spread in stand and be the cause of its death. The most aggressive and rate of spreading of pathogen require appropriate events foridentifying and mapping of infectious holes. Identification of H. annosum by fruit bodies is usually difficult because of the rarity of their formation. Furthermore, this method cannot allow determination of the hidden infectious borders in the dying stand. For example, in the clone bank of Pinus sylvestris from Ozersky forestry during long time attempts todeterminecauses of trees’ death were unsuccessful because fruit bodies weren’t found there and the experiments for induction of fruit bodies also yielded nothing. We have applied the method of DNA-analysis and identified that the cause of trees’ death was H. annosum . The rapidity and accuracy are advantages of DNA-diagnosiswhile the conventional methods are time-consuming. However, DNA-markers aren’t a panacea without a complete description of the stand infected by pathogen and the events aimed against it. This is a preliminary study where we wanted to demonstrate applicability of the method and began the first stage for further detail localization of pestholes.