Rivers as Dispersal Corridors for Primarily Expansion of Invasive Plant Species in

Renata Gagić Serdar*1, Tomislav Stefanović1, Svetlana Bilibajkić 1, Zoran Poduška 1, Ilija Đorđević1, Ljubinko Rakonjac1,2, Goran Češljar 1, Radovan Nevenić,1,3

1 Department for Spatial Planning, GIS and Forest Policy, Institute of forestry, Belgrade, Serbia 1, 2 Institute of forestry, Belgrade, Serbia 1,3 Department for Spatial Planning, GIS and Forest Policy, Institute of forestry, Belgrade, Serbia

[email protected]

Abstract Invasive plant species are often more successful within introduced areas when compared to their natural ranges. These species represented a suite of successful invaders within a young forest as pioneer vegetation on surfaces made by various transformations of native habitats and land use (river flood surfaces). River valleys represent biologically rich corridors, characterized by natural disturbances that create moist and barren sites suitable for colonization by native riparian plants, but also by woody invasive weeds. The most common, high aggressive and hydrochoric invasive plant in riparian forests in Serbia is false indigo bush (Amorpha fruticosa L.). Pattern of water regimes fluctuations and its influences on populations and stands of A. fruticosa was observed for this introduced riparian woody species along law land as well as few other rivers in Serbia. It was registered that spontaneous spreading of this plant is at the same time followed by certain level of infested seed by bruchid seed-beetle (Coleoptera, Bruchidae, Bruchinae). This evidence indicate that the seed parasites are endophagous herbivores. The variable effects of theirs abundance were recognized due to persistence of seed banks, asexual reproduction of plants, seasonal reproduction of plants, low or incomplete damage on seeds by the seed predators and competition between bruchid beetle and parasite whose larvae also detected feeding in seeds of A. fruticosa; genera Oedaule (Hymenoptera: Chalcidoidea: Pteromalidae). Therefore, if these biological conditions of weeds and seed-predators are overcome, the probability of biological control of weedy plants by seed-predators will increase. In the case of A. fruticosa control by bruchid beetle and parasitic wasp, as a potential condition has not been fully investigated and must be subjected to close examination in the future. Significant level of infested seeds (approximately 30% of all examined pods from localities where water regimes range are stabile) leads to conclusion that these seed parasites could be important reducing factor in successful suppression of A. fruticosa in non-flooded forest habitats. There is a hypothesis which needs to be proven by investigations that until now have labeled the hydrologic regime as the most important extrinsic factor of water-dispersed seeds along riparian corridors. Directly, fluctuations affect mortality of seed predators in significant level and that in a goal of biological measures of weed control utility host plant pods could be i.e. collected during extreme floods. Our results also suggest that planting native tree species along river corridors together with monitoring with prevailing of ecological and entomological research would help prevent invasion risks and contribute implementing natural diversity protection principles with invasive tree species aggressive expansion in floodplains in different invasion stages to prevent further water-mediated dispersal.

Keywords: Indigo bush, riparian corridors, expansion, native forests, seed predation, Serbia

BALWOIS 2012 - Ohrid, Republic of Macedonia - 28 May, 2 June 2012 1 Introduction False indigo bush, Amorfa fruticosa L. Fabaceae: Papilionaceae: Astragalae is the woody shrub plant species which had been introduced in Europe from North American continent in 1724 (1) when human also unintentionally starts the transportation and introduction of alien species that eventually colonized compatible ecosystems across the globe. It was introduced in the Balkan Peninsula, especially within flooding ranges of major rivers (Sava and Danube), at the beginning of the twentieth century, precisely in 1900 (1). Fast-growing bushes with purple with orange pollen illuminative flowers at first had been purposed for enriching urban greenery as one of the exotic woody plant, where still grows in urban areas as a cultivated decorative shrub expressed in rich, abundant and an attractive flowering (2). It had been proved to be useful in a way of preventing landslides, supportive for railroads stabilization or in other common ameliorative usage (3). One of its useful characteristics is that A. fruticosa belong to group of woody species of honey plants. These were also one of its initial using purposes, but very soon Indigo bush had reached out of planting controlled cultivation and spontaneously started spreading all over territory of Serbia in different forest habitats (4). Primarily in aquatic ecosystems in Serbia formed a numerous stands with outstanding abundant populations. False Indigo bush do not form such abundant populations in all law land forests communities in Serbia (4). It achieves self- reproductive forests, such as riparian forests of Pedunkulate oak, Ash, Ash- White Alder forests, Oak- Elm- Ash and Willow-Poplar forests. Localities where process of aggressively spreading of A. fruicosa were observed and where collecting of pods and other plant material was performed during period 2006-2011 were selected according to regarding self-reproductive key criteria. Degradation level of endangered phytocenological communities in great river planes (river Sava and river Danube), considering their natural regeneration attributes are in synecology and dinamics of abundance of A. fruicosa populations (5).Threatened influence of invasive species to native ecosystems and environment as global planetary disturbing phenomenon blocks the continuity of natural processes, even though the legislation in this area is very precise (6, 7). Permanent disruption of diversity and existing native food chains resulting with huge number of extinct species due to changed competitive species interrelationships as just some of the consequences is still actual and more progressive (6, 7). Surfaces of different land use were abundantly colonized with A. fruticosa during last few decades, on the territory of whole Republic of Serbia, especially in the valleys of large lowland rivers, with the broad indicative flood zones (8). For centuries forests had not been managed in a goal of intruder’s evident rising problem solving, and surfaces of occupied stands are constantly increasing. Changes happened in habitats crossed the critical values after alien population quantities. River flood surfaces and river valleys represent biologically rich corridors characterized by natural disturbances that create moist and barren sites suitable for colonization by native riparian plants, and also by woody invasive weeds. This paper are dealing with only sustainable development addressed method, use bio potential of bruchid beetle – indigo bush weevil (11, 12), recorded feeding in A. fruticosa seeds (pods) in Serbia in 2006 (12). Spontaneous spreading of this plant is, at the same time followed by certain level of infested seed by mentioned bruchid seed-beetle, Acanthoscelides pallidipennis (Motschulsky) Coleoptera: Bruchidae: Bruchinae. Native genera of parasite wasp Oedaule (Hymenoptera: Chalcidoidea: Pteromalidae) larvae had been found in significantly lower share of infested pods also determinated as seed predators of A. fruticosa. Previous hypothesis (9), that wasp was ektoparasitoid and predator of bruchid beetle larvae has been dismissed. Final conclusions is establishing of parasitoid complex of host plant – parasites system (A. fruicosa – A. pallidipennis; Oedaule sp.).This evidence may indicate that the seed parasites are endophagous herbivores, first registered in Serbia as indigo bush weevil, with the same origin as its host. It is assumed that it was introduced in the Balkan Peninsula at the beginning of the twentieth century, together with host plant, and its significant seed percentage predation were also established in previous studies. (9, 10). Pods yield natural insect toxic chemical (13), which indicate that this insects are resistant. Research findings are ecological performances of this positive useful insect in correlation with different environment parameters. The most effective on A. fruticosa growth and development are riparian microclimate conditions (specially moderate water level fluctuations in bank corridor formation stands and riparian cultures where Indigo bush is dominant in middle forest floor. Presence of light (if it reaches the ground conditions are ideal), and soil type are closely linked with the most common forest communities where A. fruticosa present serious harmful factor. It affects unsecured survival or much more fully unpossible growth and natural

BALWOIS 2012 - Ohrid, Republic of Macedonia - 28 May, 2 June 2012 2 regeneration of law land forests, especially with Quercus robur L. as dominant young tree in competition to (8). The research of the above representatives of entomofauna with the life cycle of this insect in Serbia started in 2006 in the aim of finding candidates for biological control and strategy of integral suppression of host plant. From late summer 2006, to October 2011 the pods had been collected from more than 20 localities in Vojvodina, Mačva, the vicinity of Belgrade, in Đerdap Gorge, Central Serbia (near river Morava valley) and South Serbia (river Vlasina), with minimal number of three different plant individuals per locality. Most of sample sites are located at indicative floded sites near rivers as dispersal corridors for primarily expansion (Danube, Sava, Tisa, Morava, Vlasina…) and near or in Special Nature Reserves such Obedska bara, Koviljski rit, Carska Bara which are old river meanders transformed in wide flooded wet lands. A. fruticosa, is also wide spread in Central and South East Europe and there is also treated as dangerous forest weed as woody plant (15). With consideration to natural attribute of pods being yield natural insecticide (13), and with powerful vegetative multiplication characteristic, integral control measures demanding serious and systematically planed combination of chemical, mechanical and biological measures trailed.

Experimental Purpose of answering to first research question was finding percentage of pods, or seeds infested by detected legume seed predators and its determination. The best explanation of methodology used for collecting data, could be simply described as using usually entomological research approach in field and laboratory methods. After survey of very limited literature sources, reachable study areas were chosen regarding several key criteria. Localities detecting was based from forests practice where host plant population were developed healthy habitus forms with obvious thick canopies, resulting the successfullnes of A. fruticosa adaptation to the intensive technological management treatment (combination of herbicides and mechanical measures of supression) i.e. all over PA Vojvodinašume forest units. Then, specially where environmental conditions like water accesability and enough Sun lights allows its evident aggresive spreading several sample fields were selected. Since it grows in the form of the shrub, amorpha is dangerous forest weed and significant hindering factor by shading endangered renewal of the native or cultivated trees. So material has been collected from plants in obvious competitive relations with native forest trees where nature regeneration was in starting process. In a goal of possibility of comparing studies and the most important, the seed matherial was collected in river flooded areas, in Forlands, from branches and from the ground during autmn, winther and spring, so that seed predators possibly could be drowned in floods. Pods were colected and host plants were monitored during summer in a goal of establishing insect bionomic, precise number of generations. Pods was collected from ruderal comunities, suburban areas near roads, from extreme dry places, from lake shores and it was preserved in the paper bages, and glass dishes closed by the marquis cloth and rubber, at exterior temperature. For the first season In March and April in spring 2007, 100 seeds from each locality were singled out and the flown openings on pods were counted. From October 2007 to February 2008 the material was collected in 15 localities in Vojvodina, Mačva, and the vicinity of Belgrade. The seed was collected in the paper bages at exterior temperature. In February 2008 the seed was placed in the cardboard light eclectors with flacons. After that, since February to March in 2008, 100 pods from each locality were dissected and 100 pods from each locality were places in the test tubes for growing (Figure 1 and 2). In order to obtain the parasitoids more easily, 50-100 g of the seed from each localities were transported in the plastic transparent bages so that it they could be recorded more easily. All the material is prepared and is being preserved in the Entomological Laboratory of the Faculty of Forestry of the University of Belgrade.

BALWOIS 2012 - Ohrid, Republic of Macedonia - 28 May, 2 June 2012 3

Figure 1 One flacon-one pod (daily checking for Figure 2 Same method-one season material eclosion to start in March, April..) Female A. Pallidipennis were dessicated, in order to record the beginning of oogenesis. The research and collection of material, in the season 2008/09 there was the aspiration to adapt and enhance the metodology, and first and foremost o avoid the damages in the grown insects caused by laboratory insects predator mites. The ripe false indigo seed was collected in the summer and autumn in 2007, 2008, 2009, 2010 and 2011 to determinate number of generations and make final confirmation of (Figure 3). Biological control (biocontrol for short) is the use of animals, fungi, or other microbes to feed upon, parasitize or otherwise interfere with a targeted pest species. Successful biocontrol programs usually significantly reduce the abundance of the pest, but in some cases, they simply prevent the damage caused by the pest (i.e. by preventing it from feeding on valued crops) without reducing pest abundance (18). Biocontrol is often viewed as a progressive and environmentally friendly way to control pest organisms because it leaves behind no chemical residues that might have harmful impacts on humans or other organisms, and when successful, it can provide essentially permanent, widespread control with a very favorable cost-benefit ratio. Therefore, before releasing a biocontrol agent (or using other methods), it is important to balance its potential to benefit conservation targets and management goals against its potential to cause harm.

Figure 3. Pods of A.fruticosa, still on branches Figure 4. Strong Power of Vegetative Reproduction- In April, spreads by spring floods bush cut by machete in previous Autumn

Results Regarding its cycle of growth in the condtions of our country, it mainly refers to the phase of adult. It is assumed that it has the double generation. The majority of population survive through the winter in the last larval stage (generation which survives through the winter) in the dry (completely ripe) seed.

BALWOIS 2012 - Ohrid, Republic of Macedonia - 28 May, 2 June 2012 4 By the eclosion, according to the results from the first year of investigation, an adult insect feeds in the ripe seed, in order to achieve sexual maturity. It was proved by the dissection of female bruchid beetles immediately after eclosion. It was determined that the females which did no take food were with undeveloped ovaries. The females of the first generation lay eggs on semi-ripe seed of the full size, and the eggs develop when the seed ripe (13).

Figure 5 Bruchid larvae in legume pod Figure 6 Adults after eclosion and The small number of the representatives of the second generation occur in October/ November and they survive through the winter in the stage of adults. The majority og generation fly out the next spring. The variable effects of A. pallidipennis abundance were recognized due to persistence of seed banks, asexual reproduction of plants (Figure 4), seasonal reproduction of plants, low or incomplete damage on seeds by the seed predators and competition between bruchid beetle and parasite wasp larvae also detected feeding in seeds of A. fruticosa; genera Oedaule (Hymenoptera: Chalcidoidea: Pteromalidae) (9). Therefore, if these biological conditions of weeds and seed-predators are overcome, the probability of biological control of weedy plants by seed-predators will increase. In the case of A. fruticosa control by bruchid beetle and parasitic wasp, as a potential condition has not been fully investigated and must be subjected to close examination in the future. Significant level of infested seed, about one third of all examined seeds from localities where water regimes range are relatively stable or even more from localities far from water (10), leads to conclusion that this seed parasites could be important reducing factor in successful suppression of A. fruticosa in forest habitats. Average number by infested, and by that seed with disabled viability is showed in Table 1. Percentage is result of combined assessment of all methods, but the dissection and one flacon-one pod were the most statistically proven. Weevil is native to North America, where it is widely distributed (from California in the north, to Arizona in the south and Texas in the east). In Europe it was first recorded in Bulgaria by Borowiec in 1980. As the spermatophaga of false indigo it was recorded in all places where the host plant was studied: Borowiec published research in 1988; Scenteesi published first record in 1999; and Šefrová & Laštůvka in Chesh Republic (14). The biology of false indigo bush spermatophaga in Serbia is completely related to one host plant, and in infected pods it reaches all larvae stages. According to the previous data, it does not have preference for the other related plant species, which is also confirmed by the studies of the biology of this bruchid species conducted in Hungary (10, 11). During winther seasons 2009/10 and 2010/11 we confirmed this important fact, in laboratory by exposing pods and seeds of others host plants widely spread in Serbia : Phaseolus sp. Gleditcia triacanthos L., Robinia pseudoacacia L. and Glycine max (L.) Merr. to false indigo bush weewils adults, groups of males and females right after eclosion. Closely related generas of Bruchinae subfamily members (Megabruchidius, Bruchidius and Sulchobruchus) are also related to one host plant from Fabaceae family but polyphagia performances of this insects are still under investigation, especially when they are potentially biocontrol candidates(16). Native parasite wasp genera Oedaule (Hymenoptera: Chalcidoidea: Pteromalidae) larvae had been found in significantly lower share of infested pods also determinate as seed predators of A. fruticosa. Previous hypothesis (9), that wasp was ektoparasitoid and predator of bruchid beetle larvae have

BALWOIS 2012 - Ohrid, Republic of Macedonia - 28 May, 2 June 2012 5 been dismissed. Final conclusions is establishing of parasitoid complex of host plant – parasites system (A. fruicosa – A. pallidipennis; Oedaule sp.). Parasitoid complex are parasitic wasps from next genera: Eupelmus and Anastatus (Hymenoptera: Chalcidoidea: Eupelmidae) – ectoparasitoids of weevil larvae Syntomaspis sp. and Torymus sp. (Hymenoptera: Chalcidoidea: Torymidae) –possibility of seed predation and superparasitism, both need to be proven Tetrastichus sp. (Hymenoptera: Chalcidoidea: Eulophidae) is known to encompass parasitoids of the first and second order, so it is needed to proceed the research in order to determine their status. For now, superparasitism phenomena demands experimental “tricks” (Hymenoptera: Chalcidoidea: Scelionidae) fresh bruchid beetle egg parasite, investigation in a goal of geting more specimens, data, status confirmation and at least genera determination (Hymenoptera: Chalcidoidea: Diapriidae) superparasitoid of Eupelmus and Torymus genera, more than until now cached and prepared (Hymenoptera: Chalcidoidea: Eupelmidae), Xanthellum transilvanicum Erdos&Novicky1951 Wingless ♀

On Figures 7-10 are some parasitoid complex representatives. There is a hypothesis needs to be proven by investigations which until now have labeled the hydrologic regime as the most important extrinsic factor of water-dispersed seeds along riparian corridors. Parameters that affect mortality of seed predators in significant level and that in a goal of biological measures of weed control utility could be e.g. collecting pods before extreme floods, collecting pods from other localities with significant seed infestation and preparing it for use in a field. That preparing could increase number of seed predators by storage material in proper condition. This hypothesis is proven in laboratory. After the floods, instead of drowned useful insects, they could be turned back and released to localities where they normally continuous to infest rest of the pods or move to near also by invasive host plant possessed forests. This process repeated by several years, precisely winters, could effects on disenabling generative reproduction of Indigo bush by higher number of seed predators than it supposed to survive in flooded forests.

Figure 7. Seed predator Oedaule sp ♂ , Figure 8. Ten ♀ and ten ♂ with food, on prepared observation

BALWOIS 2012 - Ohrid, Republic of Macedonia - 28 May, 2 June 2012 4

Figure 9. Parasitoid Eupelmus, the Figure 10 Xanthellum transilvanicum Erdos&Novicky most common one, ♀ 1951 Wingless ♀

Without finding solution that would exclude combined application of too expensive mechanical suppressing measures and environmentally eligible suspected pesticides it is possible to predict unstoppable expansion of this plant and facing with serious major problem in the near future. Often its crowns completely closed source of sunlight for ground vegetation and young trees in dense forests. The middle floor of trees is filled to such an extent that the domestic, native shrubs and trees in floristic way are unrecognizable. Dominant native shrubs and trees with primary generative reproduction hardly have or have no offspring at all, or it is obviously rather stunted to failure because of no competitive possibility with Indigo bush offspring or luck of sunlight, space and soil nutrition taken by alien plant adults (8). The subject of experimental use and results of natural enemies of Indigo bush recorded in previous research will be shown in the paper. Amorpha fruticosa L. is native to North America, where its population has a normal abundance. In our region for the past few decades, many years were characterized by low level of of precipitation, luck of rainfall during spring season, frequent drought occurrences in the vegetation period and it is evident that climate change is a slight but measurable progressive increase in average air temperature (20). Extremely high river water levels are also explained in terms of climate imbalances and thereby large areas of forests, agricultural land and wetland meadows are flooded, usually just at a time when maturely A. fruticosa pods falling and dispersed in a large scale. A. fruticosa pods dispersed over

BALWOIS 2012 - Ohrid, Republic of Macedonia - 28 May, 2 June 2012 7 huge distances by water, especially rivers and during floods (Figure 5). False Indigo Bush is very frequent and aggressive, so, this bush could be called or consider as conflict species.

Figure 5. Map of Serbia with fresh water resources presented, localities of collecting plant material and A. fruticosa populations monitoring within flooded areas with fresh water resources presented

Table 1. Research Sampling Periods, Localities, Plant communities, Water Regimes, Infested seed (in %) and Presents of Parasitoids

Flooded Infested No Locality 2006/07 2007/08 2008/09 2009/10 2010/11 2011 Area pods in %

a 1 Ada Ciganlija, Oct-07 Oct-08 dec.09 mar.10 mar.11 34 Oedaule sp. Taložnik,Oak-Elm-Ash forest Eupelmus 2 Kovilj, Swamp Bank, dec.08 dec.09 jun.10 maj.11 22 Willow-Poplar forest P sp Obedska bara A (Matijevica- Kadionica) 3 Kupinovo, nov.07 nov.09 26 Pedunkulate Oak Young Forest (20 years old) Obedska bara B (Obreške širine) Anastatus 4 Kupinovo Mixture feb.08 feb.09 apr.11 25 Pedunkulate Oak-Ash sp Forest (50 years old) Obedska bara C (Kupinske grede 39, 5 40) Kupinovo, dec.07 dec.08 May-10 16 Pedunkulate Mature P Virgin Oak Forest (100 years old)

BALWOIS 2012 - Ohrid, Republic of Macedonia - 28 May, 2 June 2012 8 Čortanovacka Forest, Xanthellum 6 Danube Bank, Willow- nov.07 dec.08 dec.09 17 transilvanicum, Poplar forest P Wingless ♀ 7 Obrenovac, Road jan.08 dec.09 44 Buffer greenery Šabac, River Sava 8 Bank, Willow-Poplar- dec.06 nov.07 nov.08 jan.10 10 Torymus sp. Ash Forest P Bačko Gradište, DTD Anastatus 9 Channel Bank, Willow nov.07 jan.09 jan.10 2 forest P sp Ostružnica, Bridge b 10 Sava River Bank feb.08 Aug-08 31 Greenery Tetrastichus 11 Vračev Gaj, Lake nov.07 dec.08 May-10 4 Shore Greenery P sp. Ašanja, Lošinjci, Tetrastichus 12 Coppice Pedunkulate nov.07 nov.08 May-10 apr.11 20 Oak Stand sp. Makiš, Devastated 13 Pedunkulate Ash Elm jan.07 nov.07 dec.09 32 Stand Bački Monoštor,Siga Eupelmus 14 Pedunkulate Ash feb.08 mar.09 feb.10 19 Virgin forest P sp 15 Carska bara, Perlez, nov.07 28 Swamp bank, P 16 Borkovac, Lake Shore mar.08 feb.09 feb.10 feb.11 2 Greenery PP Tetrastichus 17 Ada Ciganlija Lake nov.06 nov.07 nov.08 mar.10 36 Shore Greenery. sp. Tetrastichus sp.; c Jan 18 mar.08 Jan-09 jan.10 apr.11 20 Scelionidae, Zasavica I - Mačv. Mar-11 P Mitrovica, Swamp Anastatus Bank Vegetation sp Ada Ciganlija, Willow- Eupelmus 19 Poplar Bank Forests, nov.07 nov.08 mar.10 24 Sava River sp Senajske Bare, 20 Klenak, EA Poplar nov.06 nov.07 11 Culture P d Eupelmus 21 Šimanovci, Ruderal feb.07 nov.08 Nov-09 nov.10 41 Vegetation sp Opovo, Banat; EA 22 Poplar Clone 214 dec.08 17 Culture Đerdap Gorge, Dobra, 23 Road Buffer sep.08 Aug-10 15 greenary(Black Locust) Grabovačko- Vitojevačko Eupelmus 24 ostrvo,Klenak jan.07 feb.09 dec.09 May-10 May-11 4 Pedunkulate Oak Ash P sp Young Culture Stand Torymus sp 25 Futog Adica, River Oct-08 dec.09 dec.10 15 Anastatus Danube Bank- Willow- P Poplar forests sp(5. ♀) Zoljevo, Jelašnica and Korbevačka rivers, 26 Bank-Willow-Elm jul.08 Oct-08 Yield 16 Beech, latitude ≈≈1000m 27 Vlasina river bank, jul.08 Oct-08 Yield 21 Vlasotince,Willow

28 jul.08 Oct-08 Yield 25 , , Road Buffer greenary (with Black Locust) Sremska Rača, 29 Višnjićevo, Mature maj.08 jan.09 Aug-10 18 Pedunkulate Oak-Ash P Forest Barunovac, 0,5 ha 30 Homogenous Stand of nov.10 I-XII 11 32 A.fruticosa 31 Topčiderska reka, nov.10 27 Ruderal Vegetation Diapriidae, 32 Kamenjar, Danube mar.11 12 Anastatus river bank, Oak-Elm- e Ash forest sp 33 Klenak, Dobreč, sep.11 37 Juglans nigra Mature

BALWOIS 2012 - Ohrid, Republic of Macedonia - 28 May, 2 June 2012 9 (50 years old) Culture Stand

Notes related to Table 1. Superscripts:

a.) By seed dissection first time confirmed hypothesis that Eupelmus sp. is solitary ectoparasite of Acanthoscelides pallidipennis larvae. Remains of bruchid beetle pupae were found, change of pupae colorization were detected (It was partially eaten). This findings suggest observation of all “soft” pods (with larvae inside, but not dissected). Confirmed hypothesis – after eclosion of wasp adults which leave pods and bruchid larvae leftovers (mandibles cuticle...), finding of this as predicted dissection result, first time is reported from samples: Taložnik,Oak-Elm-Ash forest, from October 2008. Comparative, confirmative laboratory test have been repeated with pods from other locality sample in season 2010 /2011.

b.) Pod dissection first time confirmed hypothesis that parasite wasp Oedaule sp.- is endophagous herbivores, after emergence there were no any larvae remains, just eaten seed and diameter of emerging hole is smaller, diameter of emerging hole is smaller, and visible more ruff dentate rim-edges

c.) Identical result seed dissection fully confirmed hypothesis that Oedaule sp.- is endophagous herbivores, after emergence there were no bruchid larvae remains, just eaten seed remains and diameter of emerging hole is smaller, and visible more ruff dentate rim

d) By seed dissection verify hypothesis that Eupelmus sp. is solitary ectoparasite of Acanthoscelides pallidipennis larvae. Eupelmus adult remains bruchid beetle pupae and change of pupae colour were detected (It was also partially eaten). This findings suggest observation of all that “soft” pods (with larvae inside, but not dissected) will eventually resulted with wasp adult emergence and death of weevil beetle. Confirmed hypothesis – eclosion of adults which leave pods and bruchid leftovers (mandibles, cuticle...). Finding of this as predicted dissection result, first time is reported from samples from GVO, collected in May 2011 e) Parasitoid complex specimens, catching and preparing material taking part parallel with eclosions and emerging of adults of Acanthoscelides pallidipennis, Oedaule sp. Eupelmus sp. which three species are the most common. In several weeks of April and May, in one day could be collected about 20 specimens and more. In table 1 “data” are put in that colonel one by field, but indeed, data evidence about parasite wasp emergence looks like in Table 2., for all 5 autumn and spring seasons.

Table 2. The number of daily emerged parasites and parasitoids from A.fruticosa pods

The number of Locality Date Sci. Name flown parasitoids Dunav, Čortanovci 1-Nov-07 Oedaule sp 1♂ Šabac, Sava’s bank 7-Nov-07 Eupelmus sp 1♀ Kovilj, bank of 10-Nov-07 Eupelmus sp 1♂ Swamp ……………. ………………. ……………..

……………………….. ..

BALWOIS 2012 - Ohrid, Republic of Macedonia - 28 May, 2 June 2012 10 Ada Ciganlija 15-Mar-08 Oedaule sp 4♀+2♂ Ada Ciganlija 21-Mar-08 Oedaule sp 3♀+2♂ Ada Ciganlija 25-Mar-08 Oedaule sp 1♀ Ada Ciganlija 25-Mar-08 Eupelmus sp 3♀+3♂ Carska Bog, Perlez 25-Mar-08 Oedaule sp 1♀+3♂ Obrenovački road 25-Mar-08 Oedaule sp 1♂ …………….. ………… …………………. ……………………

Senajske bogs, 25-Mar-09 Oedaule sp 1♀+1♂ Klenak Zasavica 8-Apr-09 Eupelmus sp 1♀+2♂ Ada Ciganlija 10-Apr-09 Tetrastichus sp. 2 Ostružnica bridge 10-Apr-09 Eupelmus sp 1♀ Ostružnica bridge 10-Apr-09 Tetrastichus sp. 1

……………………. …………….. ………………. …………………

Zasavica 10-Apr-10 Eupelmus sp 1♀+1♂

……………. ………………… …………………… ……………………..

Zasavica 10-Apr-11 Tetrastichus sp. 1 Zasavica 15-Apr-011 Oedaule sp 1♀+1♂ Zasavica 25-Apr-11 Oedaule sp 3♀+2♂ Zasavica 28-Apr-11 Oedaule sp 1♂ Zasavica 28-Apr-11 Eupelmus sp 5♀+3♂ Zasavica 9-May-11 Oedaule sp 2♂

……………………….. ………. …………………. …………………

In example, there’s 36 rows for Eupelmidae wasps in table for season 2010/2011 per locality…

Discussion The human factor caused the destruction and degradation in many nature habitats. With for granted taking right to consider nature as the resource, man should caring also a responsibility to respect native communities that maintains life. Here ethic should be above the laws. However, laws are often either incomplete or too late. There’s just a hope that people will make the correct environmental decision in such cases. Cause - effect time discontinuous, in terms of self-reparation of nature, specifically in spreading weedy trees on Balkan Peninsula is over too much long. The wider scope of the good governance framework will be efficient policy making decisions in the organization of actions that would first present and introduce many with indigo bush repressive damaging attributes with the problem solving proposals, giving the concrete measures of placing under the control using integrated approach in forests protection. The logical sequence of events would be suggested by educated experts in topic and leads by purposely development. Whole process should be prepared on formulas as a study of predictive models for specific area, developed by one of the fastest model of risk assessment to increase the number of observed weed populations.

BALWOIS 2012 - Ohrid, Republic of Macedonia - 28 May, 2 June 2012 11 The mass production of the useful organisms is nowadays carried out in so-called bio factories in many countries. In the special laboratories the beneficial insects are massively breed for being are released out, since it is expected that their activity will be beneficial to the control of Indigo bush dispersed seed, ready to germination. In some parts of the world, very good results were achieved in the suppression of some very dangerous weed plants, such as our paper subject case. These studies in the world usually follow the positive examples of habitat rehabilitation after the intervention. Except on harmful host plant, ecosystem may react positively to the integration of useful species, here False indigo Bush weevil and wasps Pteromalidae. Another way would be integration of massive artificial indigenous Chalcid wasps also spermatophagous, and by including new food chain link on flooding river sites. Because of ecological and maybe much more population adaptive characteristics the parasite wasp indirectly discovered that they by having stabile abundance could prevent total lost and delaying chance of area not to be protected from dispersed seeds during flood waves in our riparian forests and river forelands. A further investigation of seed predation leads to possible use these kinds of strategies of in the future. Any conclusion from this research contributes topic it was very practicing unsatisfactory but wanting and something you should take genuine hopes. Reducing this alien plant particular will be professional marc and personally enthusiastic impress in finalizing PhDs thesis of this subject, on Forest Protection Cathedra, on Faculty of Forestry, Belgrade, started 2006 with “conflict legume tracking”. Our results also suggest that planting native tree species along river corridors together with monitoring with prevailing of ecological and entomological research would help prevent invasion risks and contribute implementing natural diversity protection principles with invasive tree species aggressive expansion different invasion stages to prevent further dispersal and water-mediated influence to biological control candidates. In a way of preserving autochthonous (indigenous) vegetation forms and native ecological indexes, regeneration should be led by certain role model (gradually cultivation of species grouped by eco index). In case of other agro-technical measures application to fully prepared land when it is assumed, that once densely populated stand with Indigo bush, (chemical control according to the type of pesticide is not permitted except on agricultural land and mechanical measures are overpriced expensive). Real situation actually getting more and more serious with our problematic alien plant in our forests, National Parks, protected wet lands as Bio Reservates with history and tradition in nature protection as famous Park Yellow Stone, on banks of our blue gorges rivers…It is not just a hypothesis that one non-native plants became invasive, superabundant and damaging, at least in part because it had escaped the control of their ‘natural enemies’, the herbivores and pathogens that checked its abundance in its native ranges. That all happened one century ago… Classical biocontrol addresses this would be by hand book example locating one or more herbivore species from the weed’s native range and introducing them so they can control the pest in its new range. That process also already happening for a long time, that two species are on our side, getting their enemies from our entomofauna which funded something for itself to their larvae. It is our Chalcidoid wasp parasitic wasp parasitoid complex with more and more species evolving and developing adaptation mechanisms. These herbivores are carefully selected during six years of studying and observation, and screened one test successfully in order to determine if they will attack crops or other non-target plant species. Successful classical biocontrol programs result would be in permanent establishment of the control agent(s) - False indigo Bush weevil and Oedaule parasite wasps and consequent permanent reduction in the abundance or at least the damaging impacts of the conflict weed- Amorpha fruticosa over all or in part of its introduced range. Especially in riparian forest and flooded areas where massive number of its every year produced seeds just is dispersed on more distances and in new habitats, closing that way magic circle of contagion. Classical biocontrol would not expected to eliminate the aggressive species completely and it probably should be take years or even decades after the initial release of control agents before their effects are become obvious. Classical biocontrol programs may fail for a variety of reasons. Some biocontrol agents never establish, or it may take repeated releases to establish viable populations. Some biocontrol agents may become established, but then have little or no detectable impact on the targeted weed. In our case water regimes fluctuation help A. fruticoca pods to “escape” from branches and drown all good insects in Amorpha’s seed predation phenomena. Some of ‘classical’ biocontrols greatest strengths are that once an agent is established, it will persist ‘forever’ and it may spread on its own to cover most or all of the area where the weed is present,

BALWOIS 2012 - Ohrid, Republic of Macedonia - 28 May, 2 June 2012 12 generally with little or no additional cost. That would be for a start just collecting and kipping dry all much more precious plant material dry until period of floods ends every year, only if it is necessary. Because of this, weed biocontrol researchers and successful control “belivers” take pains to locate and use agents that are specific to the targeted weed. This screening process contributes to the high cost and long time required for the discovery, testing, and approval of new biological control agents. But, for me, future vision of our planet is not one gigantic blue-green, but poisoned and lifeless blue- green “rock” in Universe.

References

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