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35LICHEN TRANSPLANTATION METHOD SUCCESS IN TEMPERATE FORESTS: CASE STUDIES IN RUSSIA AND LATVIA
A. Mezaka1, I.S. Ryabcev2, I.M. Ryabceva2, I.N. Urbanavichene3
1 Daugavpils University, email: anna.mezakaadu. lv JSC "ECOPROJECT", email: ryabtsev@ecopro. spb.ru 3Komarov Botanical Institute RAS email: urbanavichene@gmail.com
The article describes experiments on translocation of epiphytic lichens produced in Latvia based on published data and unpublished data in Russia. Lobaria pulmonaria was transplanted in Latvia. During the transplant, several methods of securing the thallus were used: with the help of a fishing line, fishing net and medical gauze. As a result of the experiment, 75% of the thallus recorded with a fishing line or fishing net were preserved for three years, and more, 85 % with medical bandage. Differences in the vital condition of Lobaria pulmonaria thallus recorded 3 years after transplantation between WKH and managed forest were not found. Translocation of crustoseepiphytic lichens Chaenotheca ferruginea, Graphis scripta and Pertusaria albescens was performed in Russia. Lichen thalli were transplanted with a substrate site. Several types of adhe-sives were used for fixation: carboxymethyl cellulose gel (CMC), CMC gel on a medical gauze substrate, vaccination var, PVA glue, acrylic and cyanoacrylic adhesives, mechanical fixation with stationery pins, as well as complex fixation with a graft seal with mechanical fixation. As a result of the experiment, 57% of Chaenotheca ferruginea samples, Picea abies phorophyte tree and 61% of Graphis scripta and Pertusaria albescens samples, Alnus glutinosa phorophyte tree were preserved after 2 years. Viable thalli of lichen transplants that have not lost their ability to reproduce are noted on all preserved fragments. Key words: translocations, epiphyte, crustose lichens, foliose lichens
Introduction
Economic activity on forest lands leads to the loss of biodiversity at various levels in the boreal and hemiboreal zones of European Russia (Identification and survey..., 2009). In the zone of influence of the construction of various industrial facilities, natural ecosystems are being replaced by natural and anthropogenic complexes everywhere. As a result of the operation and functioning of industrial facilities, land is seized and deforestation changes the hydrological regime of rivers, destruction and violation of the habitat conditions of flora and fauna. The study of the consequences of these impacts and the search for possible ways to minimize and compensate for damage caused to natural objects are extremely relevant today, especially if it concerns the protection of rare and endangered species listed in the Red Book of the Russian Federation (Forest Code of the Russian Federation, 2006).
Temperate forests have been impacted in considerable degree with intensive cuttings for decades all over the world. Today it is important to conserve biodiversity what has been left in temperate forests and plan long-term forestry approaches and biodiversity conservation. Among other forest dwelling organisms, biodiversity in temperate forests forms also lichens. Several lichen species are also indicators of forest continuity and indicate high biological value of the forest (Ek et al., 2002).
Lichen transplantation is an important approach in studies focused in lichen conservation. Different lichen transplantation methods have been used before (Market et al., 2001). Hanging branches with Hypogymniaphysodes (L.) Nyl. in Poland (Bialonska & Dayan, 2005), sterile medical gauze with fragments of Graphis sterlingiana E. A. Tripp & Lendemer, Hypotrachyna virginica (Hale) Hale, Lepraria lanata T0nsberg, pieces of burlap with Lepraria finkii (B. de Lesd.) R.C. Harris, monofilament loop with silicon sealant for Usnea angulata Ach. in USA (Allen, 2017), horizontal wooded stick with Usnea longissima Ach. in Sweden (Jansson et al., 2009). Lobaria pulmonaria (L.) Hoffm. was attached to the tree stem with plastic net and metal staples in Sweden (Gustafsson et al., 2013) and using bark samples (discs) with Parmelia caperata (L.) Ach. in USA (Brodo, 1961). However, most of these methods were focused on fruticose and foliose lichens, but not much on crustose lichens. This could be related to difficulties to transplant microscopical lichens and monitoring them through time. In New Zealand (Leddy et al, 2019), were replanted not only fragments of foliose lichen thallus Crocodia aurata (Ach.) Link, but also soredia placed in gauze bags. After a year's monitoring of the vitality more than half of thallus and soredia took roots and some of them started reproduction producing new soralia.
The aim of the present study was to evaluate successful methods in lichen transplantation experiments and their application both in Russia and Latvia in temperate forests.
Material and Methods
We compiled already published results of lichen transplantation experiments in Latvia in Zïlu plavas, Skrïveri district, Central Latvia in nemoral Woodland key habitat (WKH) and managed nemoral forest, where red-listed lichen Lobaria pulmonaria was transplanted (Mezaka, 2014) and unpublished data about lichen transplantation experiments in Russia.
In transplantation of Lobaria pulmonaria in Latvia were used three methods: fishing line, fishing net and medical bandage (Fig. 1) which were used to press the thallus (9 cm ) of Lobaria pulmonaria in north direction at 1.20 m on a tree. In total 20 transplants were located on 20 trees in WKH and other 20 in managed forest. The vitality of the each transplant was evaluated with one of four grades (1-high vitality; 4 - a transplant died) with digital photos taken in 2006 (beginning of transplantation) and then in 2007, 2008 and in 2009 (Mezaka, 2014).
Transplantation experiments in Russia were conducted in old-growth spruce-deciduous forests in Tosnensky district, Leningrad oblast near Lisinsky Nature Reserve (Fig. 2). Lichen species Chaenotheca ferruginea (Turner) Mig., Graphis scripta (L.) Ach. and Pertusaria albescens (Huds.) M. Choisy & Werner were used as transplants in Russia.On the chosen place there was a forest with the domination of fir trees and black alders, where Chaenotheca ferruginea was replanted on fir trunks, Graphis scripta and Pertusaria albescens - on alder trunks. During the transplantation of all three types of crustose epiphytic lichen species, a method of replantation of thallus with fragments of bark was developed. In the experiments, eight variants were used for sticking: carboxymethylcellulose (CMC) gel, graft seal (GS), aqueous
Fig. 1. Lobaria pulmonaria transplant with the medical bandage from 2008 in managed forest in Latvia.
dispersion of polyvinyl acetate (PVA), acrylic and cyanoacrylic glues, a mechanical variant with safety pins, and combined variants. In total 654 samples of Chaenotheca ferruginea; 210 samples of Graphis scripta and 80 samples of Pertusaria albescens were transplanted. In course of the study, less effective gluing methods (< 50% survivability within six months) have been excluded from the experiment.
A plot on a tree-accepter trunk was chosen to place fragments of the bark from the donor-tree in Russia. The accepter plots had the same exposition as the control plot. The dead layer of the bark was transferred from the chosen plots (for recipient plots of the third series of experiments the removal of the bark was carried out with 'planting nests' of the same size (Fig 2).
ABC
Fig. 2. Recipient plots of Chaenotheca ferruginea on the fir tree trunk: A - the first series, B - the second series, C - the third series.
During the monitoring of the translocation of crustose epiphytic lichen species following parameters were evaluated:
• preservation of transferred fragments;
• projective coverage of thallus on fragments;
• a share of necrosis and other damages;
• quantity (projective coverage) of the organs of reproduction (lichen apo-thecia and soredia spots);
• development of the species beyond the transferred bark;
• development of other epiphytic species on the removed bark (species confusedness and projective coverage).
The main part of these parameters is better to evaluate in laboratory conditions with the microscope research.
Depending on the condition of lichen thallus samples and their organs of reproduction (a share of necrosis and other damages) during the monitoring and the classification of different conditions of lichen on transferred fragments was made. Six classes of the condition were identified (Table 1).
When assessing the success of the settlement of the studied crustose lichen species (Chaenotheca ferruginea, Graphis scripta), we studied flushes with distilled water from the bark areas surrounding the samples of transplanted lichens. Preparations were prepared from the collected water samples and the number and vitality of spores, the presence of vegetative propagules of other lichen species (Lepraria spp.) were evaluated using a light microscope.
Table 1. Classes of lichen condition on the samples
Class Thallus characteristic
I Lichen thallus and the upper layer of bark without visible changes
II Shrinking of projective coverage of lichen not more than 10%. Can be seen small spots of necrosis, decreasing of the number of reproduction organs, traces of mechanical damage of fragments, small resin extraction, extraction of used glue substrate and growth of other epiphytic species.
III Shrinking of projective coverage of lichen from 10% to 25%. Decreasing of the number of reproduction organs.
IV Shrinking of projective coverage of lichen from 25 to 50%. Significant mechanical damage of the transferred fragment (rasin extraction, exfoliation, gluing, damage because of the corrosive pins).
V Shrinking of projective coverage of lichen from 50 to 75%. Significant mechanical damage of the transferred fragment more than 50% (rasin extraction, exfoliation, gluing, damage because of the corrosive pins). Disappearance of reproduction organs.
VI Shrinking of projective coverage of lichen more than 75%.
Results
In total 25% of transplants with Lobaria pulmonaria which were pressed with fishing line or plastic net fallen down, but only 15 % of transplants pressed with cotton bandage fallen down during all experiment time until 2009. The vitality of Lobaria pulmonaria did not show significant difference after 3 years between WKH and managed forest (Mezaka, 2014).
The lichen transplantation was evaluated for the fragments of the fir bark (as a percentage to the original quantity) with Chaenotheca ferruginea and for the fragments of the black alder bark with Graphis scripta and Pertusaria albecens (Table 2).
Table 2. Crustose lichen transplantation methods with different fastening options
Variants Total, % CMC CMC + gause. Graft seal PVA Mechanic. Acril Graft seal+mechanic cyanoacril
Picea abies -Chaenotheca ferruginea 57 36 43 27 84 58 32 98 88
Alnus glutinosa -Graphis scripta, Pertusaria albescens 61 - - 40 74 83 0 73 71
The best preservation of the samples in all series for all species was when PVA was used for sticking, also good preservation was marked if mechanical sticking combined mechanical and graft seal sticking and cyanoacryl were used. The worst preservation was marked when CMC (both variants) and acryl were used.
The deterioration of the vital condition of almost all the studied samples was noted in Chaenotheca ferruginea (life condition class V-II, the average life condition class of all preserved samples -1.73). At the same time, it should be noted that the presence of living lichen-object thallus is noted on all preserved transferred objects.
The vital state of Graphis scripta and Pertusaria albescens on the preserved transferred samples has practically not changed (life state class III-1, average class 1, 3).
Discussion
The cotton bandage method was found to be more successful than fishing line and plastic net methods in foliose lichen transplantation in Latvia. Our results show that probably dispersal and establishment limitation are significant predictors shaping epiphytic foliose lichen vitality in nemoral forest stands since significant differences were not found in transplant vitality between managed forest and WKH (Mezaka, 2014). Similar conclusions were addressed also in other studies (Belinchon et al., 2017). However, it is important to note that lichen establishment limitation effects should be separated from the dispersal limitation (Werth et al., 2006).
In experiments on translocation of crustose epiphytic lichens, an important step is the selection of a recipient site suitable for highly specialized species in terms of ecological properties. The most important aspects for a more successful transplantation of epiphytic crustose lichens are the following:
• environmental conditions suitable for the transplanted species (insolation, humidity, the nature of the bark surface, etc.) on the new phorophyte;
• competition with surrounding lichen species (absence of fast-growing species Lepraria);
• weather conditions (during the wet period, the glue solidification time increases, in very dry and hot - the glue melts - graft seal and acrylic).
The following reasons for the deterioration of the lichen-object thallus on the preserved samples were distinguished:
• mechanical destruction of the upper layer of the bark due to the influence of external factors (rain and wind) when placing the sample outside (above) the bark structure of the phorophyte acceptor tree (violation is characteristic for all types of lichens);
• formation of necrosis from a pin (Fig. 3) with a mechanical or complex method of fastening (because of the pin corrosion and rust ingress into the thallus (the violation is characteristic for all types);
A B
Fig. 3. Formation of necrosis of thallus around pins with a mechanical (complex) method of fixation of samples: A - a scale of spruce with Chaenotheca ferruginea; B - a fragment of the bark of the black alder with Graphis scripta.
• local necrosis of the lichen thallus occurs for a pair of Chaenoteca ferruginea - Picea obovata, when a mechanical (complex) method of fixation is used (a pin enters the resin passages and resin is released onto the surface of the sample, Fig. 4);
• in samples of the black alder bark, with a mechanical (less often complex) method of fixation, splitting of the transferred fragment of bark is often observed, eventually leading to the loss of sample sections;
Fig. 4. Resin extraction onto the surface of a spruce bowl with Chaenotheca ferruginea.
• overgrowth of aggressive species of epiphytic organisms, in particular lichens from the Lepraria sp., on the transferred samples, which suppress the development and cause the death of lichens-experimental objects (Fig. 5).
Fig. 5. Overgrowth of the transferred fragment with Chaenotheca ferruginea lichens from the Lepraria sp.
In an experiment with crustose epiphytic lichens, we tested the our hypothesis about the possibility of taking into account the dispersion of transplanted lichens on adjacent attachment points (planting) with the help of spores or soredia.
The results of this experiment will allow us to choose the most effective option for fixing bark fragments during translocation of epiphytic crustose lichens. Epiphytic lichen species with a strategy of vegetative reproduction with the help of soredia take root most quickly (Scheidegger et al., 1995; Leddy et al., 2019).
This should be taken into account planning nature protection and long-term forestry in landscape and forest stand scale. Retaining potential substrate trees, for epiphyte distribution in nemoral-managed forests adjacent to nemoral old-growth forests with species with limited dispersal, transplantation, and forest continuity could ensure the continued existence of epiphytic species, especially specialists with limited dispersal abilities in boreo-nemoral zone.
Acknowledgements
The present study in Latvia was partly funded by the European Regional Development Fund Postdoctoral research aid project "Epiphyte metapopulation dynamics in boreo-nemoral forest landscape" (Nr. 1.1.1.2/VIAA/3/19/469). The works in Russia were carried out with the financial support of Nord Stream 2 AG under the contract PO 18-5082 from 12.04.2018. The study by I.N. Urbanavichene was carried out within the framework of the institutional research project ("Herbarium collections of BIN RAS (history, conservation, investigation and replenishment)") of the Komarov Botanical Institute of the Russian Academy of Sciences ((№ AAAA-A18-118022090078-2).
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УСПЕХИ МЕТОДОВ ТРАНСПЛАНТАЦИИ ЛИШАЙНИКОВ В ЛЕСАХ УМЕРЕННОЙ ЗОНЫ: ПРИМЕРЫ В РОССИИ И ЛАТВИИ
1 11 3 А. Meжaкa , И.С. Рябцев , И.М. Рябцева , И.Н. Урбанавичене
lDaugavpils University, email: anna.mezaka a du. Iv 2JSC "ECOPROJECT", email: tyabtsev'a ecopro.spb.ru
o
Komarov Botanical Institute RAS email: urbanavichene@gmail. com
В статье описаны эксперименты по транслокации эпифитных лишайников произведенные в Латвии и России. В Латвии проводили пересадку макролишайника Lobaria pulmonaria. При пересадке использовали несколько методов закрепления таллома: при помощи лески, рыболовной сети и медицинской марли. В результате эксперимента за три года сохранилось 75% пересаженных талломов, зафиксированных при помощи лески или рыболовной сети, и только 13% талломов, зафиксированных хлопчатобумажной марлей. Различий в жизненном состоянии талломов Lobaria pulmonaria через 3 года после пересадки (2009 г.) между ключевыми лесными сообществами (WKH) и нарушенными лесами выявлено не было. В России производили пересадку накипных эпифитных лишайников Chaenotheca ferruginea, Graphis scripta and Pertusaria albescens. Пересадку талломов лишайников производили с участком субстрата (чешуйками или кусочками коры). Для фиксации применяли несколько вариантов адгезивов: гель карбоксиметилцеллюлозы (КМЦ), гель КМЦ на подложке из медицинской марли, прививочный вар, клей ПВА, акриловый и цианакриловый клеи, механическая фиксация при помощи канцелярских булавок, а также комплексную фиксацию садовым варом с булавками. В результате эксперимента через 2 года сохранилось 57% образцов Chaenotheca ferruginea (форофит Picea abies) и 61% образцов Graphis scripta and Pertusaria albescens -(форофит Alnus glutinosa). На всех сохранившихся фрагментах отмечаются жизнеспособные талломы лишайников-объектов, не потерявшие способности к размножению. Ключевые слова: транслокация, эпифитные лишайники, накипные лишайники, листоватые лишайники
