Using Commercially Available Mycorrhizae Inoculant, Compost, or Mycorrhizae Inoculant and Compost when Transplanting Small Berry Bushes

PROJECT BACKGROUND
We have approximately 20 acres of land. It is a mixed wooded area and tillable acres. We are a family operation and we have the goal of seeking sustainable ways to make our property diverse and productive while earning income. For several years now we have had a vegetable garden that we market through the local farmers' markets. We have experimented with a few specialty items and find that our raspberries always do well at the market. We are trying to diversify our operation with other more unusual small specialty berries, such as elderberry, aronia, and haskap. When we eventually finish our berry planting project, we hope to have about 4 acres of berries.

Over the years, we have always sought to be stewards of the land by using sustainable practices. We try very hard to till the ground early to prevent the weeds before they become established, to use cover crops that we till under for organic matter, apply composted organic matter, and to use organic fertilizers and pesticides. I would like to do a better job learning how to use cover crops and using the concepts of permaculture to develop land that can be productive with less carbon footprint.

PROJECT DESCRIPTION
Goal: The focus of this experiment was a field experiment meant to help the small farmer to improve the survivability and plant development of transplants with commercially-available products.

Our goal was to see if adding commercially-available mycorrhizae and/ or commercially-available compost would affect the early plant growth of small berry bushes.

PROCESS
Preparing for the experiment:
Cuttings were made while the plants were dormant from aronia and elderberry bushes. We rooted our plants in a 2:1 perlite vermiculite mix. Dip n Gro rooting hormone was used to dip the cutting before putting them in the pots. The plants were monitored while the roots were developing. At this point no other fertilizer or amendment was added to avoid inoculating mycorrhiza. It is ideal to use a misting system to water the plants but this was not available to us at this time so we purchased an ebb-and-flood water table and a timer to provide regular watering to the plants. We found with the water table, the vermiculite was sucked to the bottom of the pot and kept the plants too wet. The aronia did not survive and the elderberry had an adequate but low survival rate. We continued to grow the elderberry and gathered softwood aronia cuttings in late June. At that time, the local horticulture school offered to let me plant the cuttings in their greenhouse and to use their mister. The instructor at the school suggested that we may be more successful with the same perlite: bark mix that the local growers use for rooting woody cuttings. Rooting of the aronia cuttings was much more successful with the bark mix and mister in the greenhouse.

Once the plants were rooted, they were compared for size and fullness of growth. The plants were arranged so the size of the plants were similar for each treatment. Each treatment had three plants.

Prior to planting, the ground was prepared by tilling and planting a clover/grass ground cover. Holes were dug about 10 inches deep and marked with flags labeling the holes with one of four treatments—1) control, 2) compost, 3) mycorrhiza and 4) compost and mycorrhiza mixed. Treatments were planted in groups of three with an untreated “guard” plant in between each treatment. The purpose of this was to prevent the possibility of a plant treatment affecting an adjacent differing plant treatment.

The experiment:
Large containers were put by each treatment area for the group of plants. (We used plastic kiddie pools.) We mixed one part Purple Cow Compost with two parts soil for the compost only holes, 1/3 package of PHC Treesaver mycorrhiza with two gallons of soil for the mycorrhiza only holes, and used a mix of both with the compost and mycorrhiza holes. The control plants had no amendments added to the soil when planted. The treated soil was used in the top 8 inches of the soil. The plants were watered consistently with a soaker hose.

Prior to planting, small pieces of root were harvested from each plant group and put into bags for processing in the lab. Roots were also harvested by digging up a small section of soil near the plant for processing at 2, 4, and 8 weeks. For consistency, we used the middle plant from each group of plants to harvest the roots. At four weeks the whole plant, including the root system, was removed to observe the differences in root system growth.

In the Lab:
One at a time, each root sample was washed and the finer roots were cut into small pieces about 1/4-1/2 inch long and placed into labeled 3 dram vials. The roots were covered with 10 percent solution of KOH to clarify the roots. The vials were then placed into a 90 degree C water bath for one hour. The vials were removed from the water bath, the KOH was drained using a metal strainer, and the roots were rinsed with water. The roots were covered with 2 percent HCl for 10 minutes to neutralize the KOH and drained without rinsing. The roots were then covered with a 1:1:1 solution of lactic acid:glycerol:water with 0.05 percent trypan blue stain. The vials were returned to the water bath for 20 minutes. The stained roots were drained and rinsed with water into a waste jar. The roots were then covered with a 1:1:1 solution of lactic acid:glycerol:water. The roots were left to stand for at least 8 hours in the refrigerator before further processing.

A slide is prepared and labeled with a root sample, 1:1:1 solution of lactic acid: glycerol: water, and a slide cover. A microscope is used to look for the presence of mycorrhizae. A picture is taken of a representative part of the slide for comparison with other data. Pictures will be compared with pictures of known mycorrhizae pictures and with other pictures within this experiment to determine if a plant is infected with mycorrhizae.

PEOPLE
Joan Benjamin
Joan is the NCR-SARE associate regional coordinator. She assisted me with my many questions. She helped to find me resources to complete my SARE application by inviting me to the National Small Farm Trade Show & Conference, finding me a ride to get there, finding me a person to share a room while at the conference, and introducing me to people who had the knowledge to help me. Without this initial support it would have been difficult to get started.

Addie Kinghorn
Some time ago, I had this vague notion that I wanted to do more with my property than I was doing. I spoke with Addie about her assisting me with a plan for my property. I expressed a desire to plant berry bushes and one of the people she recommended I talk to was Greg Berna at Paradigm Gardens. Since that time, Addie has been a gracious and knowledgable resource everytime I had a question on almost any horticulture subject.

Paradigm Gardens
Greg Berna, the owner of Paradigm Gardens, told me about his newly planted aronia berries and explained how he used commercially-available mycorrhizae when he planted bushes. At that time, I had never heard of mycorrhizae. Greg gave me just enough knowledge that I started to do more research, this time on both mycorrhizae and berries. Greg also introduced me to some local berry producers, including Charlie Caldwell. Charlie grows his own aronia plants and started the Midwest Aronia Association. Since then Greg's sons and the employees of Paradigm have used their knowledge to assist me many times. They have taken me through quite a varied educational journey. Also, much of the equipment and supplies for this experiment came from Paradigm.

Charles Shapiro
Charlie is a soil scientist at the University of Nebraska in Concord, NE. I was first introduced to him by Joan Benjamin when she suggested we carpool to the National Small Farm Trade Show & Conference in Missouri. Charlie had knowledge about soil and had grant writing experience so I spent the whole trip from Fremont, NE to Columbia, MO and back picking his brain on how I should set up my experiment and how to write a grant.

Patrick Byers
Patrick Byers is a berry expert that was speaking at the Small Farm Conference. He helped me to refine the design of my experiment.

Rhae Drijber
Part of my experiment included staining the roots of the plants to find mycorrhiza under the microscope. Dr. Drijber is a professor and soil scientist at UNL that studies mycorrhiza. Dr. Drijber invited me to her lab so her lab technician could assist me in doing a small experiment staining roots and looking for mycorrhiza.

Liz Jeske
Liz Jeske is the lab technician in Dr. Drijber's lab. She has been an invaluable resource. She not only walked me through the experimental process, she also assisted me in finding sources for obtaining the hazardous chemicals needed for this experiment and was very patient with my numerous questions about the many little details of this experiment.

Ellen Paparozzi
Dr. Paparozzi is a professor of horticulture at UNL. She spent the time with me to explain how to root woody cuttings.

Charlie Caldwell
Charlie Caldwell is an aronia grower from Iowa who was instrumental in starting the Midwest aronia association. He offered to give me aronia cuttings to use for my experiment. He has a successful grape and berry farm that he appears to market for wines. He also spent time educating me on aronia berries.

Terry Durham
I first met Terry at the National Small Farm Trade Show & Conference in Missouri. He is a grower of elderberry and works with the local University to grow and market elderberries. In both year one and two of my experiment I purchased elderberry plants from Terry.

Todd Morrissey
Todd is a horticulture instructor at Metropolitan Community College in Omaha, NE. His experience includes working for the Forestry service and propagating woody plants. When I had some difficulty rooting my aronia cuttings, Todd spent a significant amount of time helping me to analyze what had gone wrong, provided me with cuttings from his own aronia plants, and assisted me by providing me space in the college greenhouse under the mister to root the plants.
Lance Johnson
Dr. Johnson is a professor and head of the biology department at Midlands University in Fremont, NE. He has been extremely gracious in allowing us to use his lab and equipment to do our experiments, including space to store our chemicals and specimens. He has also been very receptive to allowing my children in his lab to assist in the experiments.

My husband and children
My husband spent many hours assisting me and bringing me places to accomplish the tasks of this experiment, including but not limited to preparing the ground, purchasing and planting seed, and assisting with the irrigation. My two children worked on the experiment and were invaluable. This experiment required a series of trials on a number of plants. There was always too much work for one person to complete the job. This was the first time for my eleven-year-old son to work in a lab. From what he expressed, this is one of the most exciting things he has done in his life. He became very proficient at performing the tasks required of this experiment, including learning about lab safety and clean up, pouring hazardous chemicals, preparing slides, and looking for mycorrhiza under the microscope. The children both presented at the Rural Advantage conference in Nebraska City, NE in February 2012 and did a wonderful job. They had many nice comments.

Many other people assisted me, sometimes by just referring me to other people, educating me about a small detail, or assisting me in finding a resource. Hopefully, I have given credit to all the people that have helped make this grant successful in a large way. We have shared or are in the process of sharing our data with all the people who have helped us or otherwise expressed an interest.

RESULTS
Aronia and elderberry rooted cuttings were planted in four groups; the control, commercially-available compost added, commercially-available mycorrhiza added, and both commercially-available compost and mycorrhiza added. Based on our earlier research, we expected the group with both the commercially-available compost and mycorrhiza would produce the most significant early growth. We were wrong. The compost only group appeared to develop mycorrhiza at two weeks with the aronia and at four weeks with the elderberry. The aronia both group also had mycorrhiza at two weeks. At four weeks sample plants were pulled from the ground and the compost only group in both plants had very bushy feeder like roots compared to the control group which was rather scrawny with both plants. The mycorrhiza only treatment was close behind the compost only treatment but still obviously less full. The both treatment had a completely different type of growth characterized by a very long root with little fullness. All the plants appeared to have mycorrhiza inoculation by eight weeks.

We will likely continue this experiment to look at the plants over the next year to see is the early start of the composted plants make a difference in plant growth over a longer period of time. If I were to do a variation of this experiment in the future, I would pay more attention to the appearance of the root systems. The elderberry had a heavier root system than the aronia. The aronia seemed to be more prone to damage from over or under watering and the hot water bath processing of the aronia roots seemed to sometimes damage the roots while being stained so I might cut down the hot water bath time on the aronia by a couple of minutes compared to the elderberry.

Since the aronia root system was somewhat sensitive to the type of media chosen for the experiment, the aronia used in this experiment were soft wood cuttings grown in a bark/perlite mix similar to what aronia growers use. This may have created an environment that promoted earlier development of mycorrhiza and may be a confounding part of this experiment when comparing the aronia to the elderberry. In the future, I would use a more porous soil with a mister for each treatment and see if the mycorrhiza formed at a similar time to the first experiment.

DISCUSSION
What I learned:
1. I learned about the balance of media, air, and water when growing plants.
2. I learned that this balance is tricky should you change one factor in the balance.
3. I learned that if you are going to propagate plants, a mister for keeping the cuttings consistently moist (not wet), is beneficial.
4. I learned that it is difficult to water plants consistently without automatic irrigation.
5. I learned that mycorrhiza is found in the ground and will eventually inoculate the plant. In our case it was about 8 weeks when the control showed consistent signs of mycorrhiza inoculation.
6. I learned that mycorrhiza in compost may actually inoculate plants faster than the dry mycorrhiza granules.
7. I learned that mycorrhiza does positively affect the early growth of plants.
8. I learned how to purchase hazardous chemicals and use them safely.
9. I learned how to identify at least some types of mycorrhiza.

Advantages to this project
1. Although I did not initially expect to have to propagate my own woody cuttings, I found it was very satisfying to learn how to propagate plants.
2. As a result of this project, I met many very gracious people who were willing to help with this project. I also hope to help others should they have similar projects. I already helped one person in Vermont.
3. This project allowed me to learn more about the plants I wanted to plant and how to plant them.
4. My son appears to really enjoy working with plants and in the lab. He became very proficient and it was wonderful to be able to give him this experience.
5. Although I have experience with a number of lab classes, prior to this experiment, I had never purchased and prepared chemicals for an experiment. Although I would probably still seek advice, I feel much more confident about doing future experiments, if not for SARE perhaps with my children as a learning experience for them.

Disadvantages to this project
1. Until I learned about the balance of air, media, and water when growing plants and how delicate the balance was, I lost a lot of plants.
2. I spent a tremendous amount of time and family resources to make this experiment work.
Recommendations to others who would like to do this experiment:
1. Seek out the resources to help you be successful early.
2. Some of the equipment used for this experiment is expensive. Look for a receptive person at the local schools who will allow you to borrow the equipment you need. Some might even allow you to use small amounts of their chemicals.
3. Write out your plan and seek knowledgeable people when you are uncertain about how to do something.

PROJECT IMPACTS
My project identifies a simple way to improve the early growth of a plant through the inoculation of mycorrhiza. Other studies have already shown that mycorrhiza inoculation helps plants in soils with heavy metals and that are low in nutrients. It also helps the plant uptake nutrients and moisture by increasing the surface area of the root system up to 700-1000 percent.

OUTREACH
Methods for sharing information about my project:
1. We worked with many people on this project. We listed 15 people above but have talked to many others. We have already contacted and shared results with many of these people and are continuing to do so.
2. Information on the SARE site led an individual desiring to work on a similar project to contact me to ask questions. I provided what assistance I could.
3. We spoke at the National Small Farm Trade Show & Conference in November 2011 and the Rural Advantage Conference in 2012.
4. If we can fund it, we were invited to speak at and will attend an Elderberry Festival in 2012.
5. I hope to continue gathering data on growth differences among the trial groups and intend to continue to share that information.

A video recording of a presentation from the 2012 NCR-SARE Farmers Forum at the Small Farm Trade Show & Conference can be found online on NCR-SARE's YouTube channel. Visit the following link to view it: https://youtu.be/VwhWDqL0CYw