This area is characterized by a mountainous climate with a dry and windy spring, rainy summer, cool and foggy autumn, PD98059 and cold and long winter. The mean annual temperature varies between 3.3°C and 7.3°C,

with a mean summer temperature ranging from 8.7°C to 19.3°C and a mean winter temperature ranging from −23.3°C to −16.1°C. The annual solar radiation is 124 MJ m−2. The annual mean precipitation is over 1,400 mm, which is the highest in North-Eastern China [12] and [13]. A mixed hardwood forest was located in this area prior to ginseng cultivation. Albic luvisols were developed from the parent material of loess. After deforestation, a binary mixture of the humus and albic horizons (generally 1:1) was used to create an elevated bed for growing ginseng. Prior to seed sowing and/or seedling transplantation in the spring, the soils were fertilized with composted manure. The bed width was approximately 170 m and was separated by 40-cm walkways. Local SCH 900776 cost farmers constructed artificial plastic shades approximately 80 cm above the ginseng bed. The plastic covers were used from May through to September. Ginseng is a tender perennial. The first frost kills the leafy top, but a new top emerges the following spring from an underground bud on the perennial root. It takes 5 yrs or 6 yrs of ginseng cultivation

to grow into a mature product. Ginseng was planted on the same land for 3 yrs, then the root tissues were replanted into the newly-mixed bed soils for another 2 yrs or 3 yrs prior to harvest. Soil samples were collected from beds with different-aged ginseng plants in April (spring) of 2009 before the plastic shades were put into place. A 0.01 m2 area was plotted, and the ginseng was carefully removed. The soil was sampled at 0–5 cm (upper roots), 5–10 cm (root zone), and 10–15 cm (down root) using an auger in three Metalloexopeptidase replicates. We logged the

location using a global positioning system (garmin eTrex Venture HC; Garmin International Inc., Olathe, KS, USA) and re-sampled the soils in July (summer) of 2009, September (autumn) of 2009, and April of 2010 (the next spring). The re-sample location was just 1 m from the original plot. Parts of the soil samples were stored at 4°C to determine nitrate content. The remainder were air-dried and sieved through a 2-mm screen for laboratory analysis. Winter sampling was not conducted because of the difficulty of sampling frozen soils. The bulk density and moisture content of the soil was determined using general methods in the laboratory. The pH in water (w:v, 1:2.5) was measured with a pH meter (PHS-3C; Shanghai Precision Scientific Instrument Co., Ltd., Shanghai, China). The total organic carbon (TOC) was determined using a dry-combustion method. The soil nitrate was extracted using a 1M KCl solution and was analyzed using dual-wavelength UV spectrophotometry (Shimadzu UV-2450; Shimadzu Corporation, Kyoto, Japan) according to Norman et al [14].