Analysis site
The research city include c. 2 km 2 away from kinds-rich lifeless calcareous grassland inside the western Estonia (°Letter, °E). The fresh new grasslands set up usually due to long-title comprehensive management, notably grazing from the residential dogs, gathering hay and you may reducing shrubbery and trees getting stamina. , 1999 ). The fresh grasslands generally got a maximum juniper (Juniperus communis) safeguards of 31%, but they are today much more overgrown having Scots oak (Pinus sylvestris; doing 80% cover).
Sampling build
Sampling was held when you look at the at web sites in which continuously treated (mown) unlock grassland really stands (GR) were enclosed by stands away from different quantities of over growing: transformation grasslands (TR) and you can younger oak woods (FO). The three successional grade (GR, TR, FO) function a temporal series out-of regeneration series level c. 60 year, as they are characterised by the a progressive over growing away from unlock grasslands because of the shrubbery and you will woods, that’s typical for given up calcareous grasslands from the analysis region (Neuenkamp ainsi que al., 2016 ; Supporting Advice Fig. S1). Within the transformation grasslands administration stopped c. 30–thirty five yr before sampling and you can grasslands was basically partially overgrown of the pine and you may juniper; younger pine forest have been previous grasslands where government stopped c. sixty yr prior to sampling and you can that have been completely overgrown by the pine and juniper. A webpage is recognized as a location in which most of the three successional amount was in fact discovered within this 50 yards of every almost every other (GR–TR–FO). Completely, twenty-five web sites had been tested. A randomly found step 1 ? step one meters area was created in for every successional phase (GR, TR, FO) of any webpages, leading to a total of 75 plots of land (twenty-five websites ? about three successional levels). Next attributes was basically discussed each spot: industry coating plant area, Was fungal organizations (in plant sources, and you may rhizosphere surface) and ecological details.
Data range
For every step one ? 1 meters spot, most of the vascular plant variety on the soil-covering people were filed as well as their wealth is actually projected while the fee cover (Peet & Roberts, 2013 ). Concurrently, payment coverage estimates regarding bushes and you may trees contained in this a great ten ? ten yards town doing for every single area was indeed filed. Hereafter, the floor-covering bush community is called ‘plant community’ as well as analyses was presented with this area until conveyed otherwise.
A good ten ? 10 ? 10 cm surface center are collected on centre of every spot and you can bush origins was in fact split regarding the crushed, dehydrated for twenty-four h on 50°C and you will stored dry for further unit data of root-colonising In the morning fungi. Have always been fungal diversity in-plant roots was then analysed off combined resources samples. Sources in the each test were floor using liquids nitrogen and you may thoroughly mixed because the explained by the Garcia de Leon et al. ( 2016 ). Good subsample of 70 mg of any blended means shot is actually utilized for molecular analyses.
Plant communities was demonstrated from the a larger spatial size weighed against In the morning fungal teams, since this level is to around echo brand new bush options area captured by the a great 10 ? 10 ? ten cm surface center. Small-size bush variety for the grasslands try highest underground than a lot more than soil (Hiiesalu et al., 2014 ), partly since roomy resources system and you may much time rhizomes of several grassland plants link the new sources contained in a localised crushed core with a lot more than-soil organs increasing several ).
To characterise AM fungal diversity in the soil, 10 g of soil was collected from each soil core, dried with silica gel and stored airtight at room temperature. A second subsample of 10 g of soil was collected from each soil core for measuring the concentration of an AM fungal neutral lipid fatty acid fungal abundance in the soil. This second soil sample was freeze-dried for 15 h at ?80°C and stored airtight citas introvertidas gratis at room temperature. The remainder of each soil core was air-dried for analysis of soil chemical properties (pHKCl, Ntot, P, K, Mg, Ca, Corg) following International Organization for Standardization (ISO) protocols (see Methods S1).