We plan to deploy samples for an experiment using the litter collected in our previous visit. Here are the details of the protocol: Experimental outline of transplant bags in UC Reserves Items needed: Nylon membrane (0.2 um) ordered from Tisch Scientific Alleen’s fabric glue ordered through amazon.com Paper clips Duct tape or gorilla tape stakes aluminum labels chicken wire Cut the nylon membrane 10cm by 20 cm / window screen Fold the membrane along the longer side to form a square Add glue on the 2 open sides and fix them with paper clips After glue is hardened cover the three open sides with duct /gorilla tape Fill the bags / gamma irradiate (not for window screen bags) Inoculate with specific microbial community (not for window screen bags) Glue the final open side of the bag and reinforce closure with duct /gorilla tape add aluminum identifier encage the bag in chicken wire stake bags down flat on the ground, stake ibutton flag area After filling the bags with leaf litter (as a substrate), we gamma irradiate the bags to eliminate (knock down) the original community. We will inoculate the irradiated site specific substrate leaf litter with a community from the site specific leaf litter (4 bags) and with a community from the Irvine Grassland (4 bags). We will also inoculate the irradiated Irvine Grassland leaf litter with a community from the Irvine Grassland leaf litter (4 bags) and with the community from the site specific leaf litter (4 bags). See figure below. In total there are 8 bags that will contain the site specific communities and 8 bags that will contain the Irvine Grassland communities. In addition we will have 4 bags of each litter type (site specific and Irvine Grassland) non-irradiated in window screen bags. A total of 28 bags per site (20 nylon bags and 8 window screen bags) will be staked down flat onto the ground. The bags will stay in their place until the end of the rainy season (between April and June). The area of the bag location will be flagged with yellow flags labeled AB-UCI-# In addition we will place an ibutton to measure temperature and humidity in the area of the bags. Nylon bags encased in chicken wire to prevent rodent/ mammal destruction: The shown bag is bigger than what we will be using. Window screen bag enclosed in chicken wire:

Approved