The research town includes c. dos km dos regarding types-rich deceased calcareous grassland within the western Estonia (°Letter, °E). The brand new grasslands set-up over the years due to a lot of time-term detailed management, somewhat grazing because of the residential animals, get together hay and you can cutting shrubbery and you may trees to own electricity. , 1999 ). The latest grasslands traditionally got a maximum juniper (Juniperus communis) defense from 29%, but are now increasingly overgrown that have Scots pine (Pinus sylvestris; as much as 80% cover).
Testing is actually presented when you look at the on websites where consistently treated (mown) open grassland really stands (GR) was basically enclosed by stands away from varying quantities of overgrowth: transitional grasslands (TR) and you may younger pine forest (FO). The three successional levels (GR, TR, FO) means a temporal series away from regeneration series covering c. 60 year, and therefore are characterised because of the a steady overgrowth away from open grasslands by the bushes and you will woods, that is typical to own abandoned calcareous grasslands about data area (Neuenkamp et al., 2016 ; Supporting Advice Fig. S1). During the transformation grasslands administration stopped c. 30–thirty five year prior to sampling and you will grasslands have been partially overgrown by oak and juniper; young oak woods had been former grasslands in which administration ceased c. sixty year in advance of testing and you may which have been youngsterally overgrown of the pine and you may juniper. A site is actually identified as a place where the three successional grade was basically found contained in this 50 m each and every almost every other (GR–TR–FO). Completely, twenty five internet sites was indeed tested. An arbitrarily receive 1 ? step 1 m spot was created in for each and every successional stage (GR, TR, FO) of any website, leading to a maximum of 75 plots of land (twenty-five internet sites ? about three successional levels). Another services was indeed explained each spot: the field layer bush society, In the morning fungal teams (in plant origins, and rhizosphere ground) and you will ecological details.
Each step 1 ? 1 m patch, most of the vascular bush variety about floor-layer neighborhood was submitted as well as their wealth was estimated since commission coverage (Peet & Roberts, 2013 ). Likewise, fee safeguards prices off bushes and you will woods within this good ten ? ten meters town to for each and every spot were submitted. Hereafter, a floor-level plant society is referred to as ‘bush community’ and all of analyses is held using this type of area unless of course indicated if you don’t.
Good ten ? 10 ? ten cm surface center try built-up about middle of each and every area and you will bush roots were separated regarding surface, dried to have 24 h on fifty°C and you may held dry for further molecular investigation out-of options-colonising Have always been fungus. Are fungal diversity in plant sources ended up being analysed out-of mixed root examples. Origins in the for every single sample had been floor using liquids nitrogen and you may carefully combined as the explained because of the Garcia de- Leon ainsi que al. ( 2016 ). An effective subsample away from 70 milligrams of any combined means attempt was useful unit analyses.
Bush teams was in fact explained in the a bigger spatial scale weighed against In the morning yeast groups, since this scale will be just as much as mirror the new bush means people captured by http://www.datingranking.net/cs/fetlife-recenze an effective 10 ? 10 ? 10 cm soil key. Small-size plant assortment into the grasslands try high below ground than just more than soil (Hiiesalu et al., 2014 ), partly once the roomy root program and you may enough time rhizomes of many grassland herbs connect this new origins found in a localized soil key which have significantly more than-surface organs growing 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 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).