1. Shortened Soybean Germ
Short Presentation of Survey Results
Good Morning/Afternoon – How is everyone doing? I am here to give a brief overview of my referee project. I do not have any data available yet (because seed hasn’t been sent out). But I do have a summary of my questionnaire results that I thought I would give a presentation on.

2. Purpose
The purpose of this referee is to obtain enough data to propose a rules change from an 8 day germination test to a 7 day germination test.
Some labs use a 7-day test period and other labs use an 8-day test period as specified by AOSA Rules. This referee will be used to determine whether we can standardize to a 7-day test and potentially create better uniformity in testing.
1. SCST and AOSA work together to conduct referees that are used to support changes to AOSA Rules. These rule changes are a key component to maintaining quality and uniformity in seed testing. This year BDI is responsible for coordinating a referee to determine whether it would be appropriate to change the number of days for germination in soybeans.
2. Ultimately, this referee would help harmonize with Canada.

3. Brief Overview of Planting Methods - AOSA
Substrata:
B (Between Blotters); T (Paper Toweling); S (Sand); TC (On Top of Creped Cellulose Paper Without a Blotter); TCS (On Top of Creped Cellulose Paper Without a Blotter and Covered with ½” to ¾” of Sand.
Temperature:
25C; 20-30C
First Count (Days): 5
Final Count (Days): 8
Specific Requirements:
Hard seeds: See section 6.2d and 6.9m(6).
6.2d: Hard Seeds – Seeds that remain hard at the end of the prescribed test period because they have not absorbed water due to an impermeable seed coat. Seeds known and recognized to contain hard seed are indicated in either the “Specific Requirements and Notes” column or “Fresh and Dormant Seed” column of Table 6A. The percentage hard seed is to be reported in addition to the percentage germination.
6.9m(6): Hard Seeds – The percentage of hard seeds it to be reported in addition to the percentage germination. If swollen seeds or seeds that have started to germinate are present at the end of the prescribed germination period, remove all hard seeds (record their number)…….continue the test for 5 additional days. The additional normal seedlings shall be included in the percentage of germination.
I’m going to start by giving a quick summary of each of the planting methods for Soybeans including AOSA, ISTA, and Canada M&P Rules.

4. Brief Overview of Planting Methods – Canada M & P
Substrata:
Sand: Seeds are placed on a level layer of moist sand and covered with a layer of uncompressed sand to a depth of 10 to 20 mm, depending on the size of the seed. The quantity of water added will depend on the particle size of the sand and the characteristics of the seeds to be planted. As a guideline, for 24-mesh sand with no vermiculite added, add 125 ml water per litre of sand or add water until sand can be formed into a ball when squeezed in the palm of the hand, the ball breaking freely when pressed between two fingers.
RT (Rolled Paper (Towels)): Seeds are evenly spaced on two sheets of standard weight (38 lb.) or a single sheet of heavy weight (76 lb.) germination paper, then covered with a single sheet. The paper is rolled and placed in an upright position. The paper should be moistened until its wet weight is about three times that of its dry weight.
Temperature
25C
First Count (Days)
N/A
Final Count (Days) 7
Hard Seeds:
If hard seeds are found at a low level they are treated as dead.
If hard seeds are found at a higher level (5- 10%), the seeds are extended, then at the end of the extension any remaining hard seeds are called dead.

5. Brief Overview of Planting Methods - ISTA
Substrata:
BP (Between Paper): The seeds are germinated between two layers of paper. This may be achieved by:
Loosely covering the seeds with an additional layer of filter paper.
Placing the seeds into folded envelopes which may be placed in a flat or upright position.
Placing the seeds in rolled paper towels (the rolls must be placed in an upright position).
The substrates are kept in closed boxes, wrapped in plastic bags or placed directly on trays in a cabinet germinator provided the relative humidity in the germinator can maintained very near saturation.
TPS (Top of Paper Covered with Sand): The seeds are germinated on top of a moistened sheet of crepe cellulose paper which is covered with a 2 cm layer of dry sand. Crepe cellulose paper is a multi-layered paper pad (e.g. Versa- Pak).
S (Sand): The seeds are planted on a level layer of moist sand and covered with mm of uncompressed substrate, depending on the size of the seed. To ensure good aeration, it is recommended that the bottom layer be loosened by raking before sowing.

6. Brief Overview of Planting Methods - ISTA
Temperature:
20-30C; 25C
First Count (Days) 5
Final Count (Days) 8
No additional special instructions.
Notes:
The prescribed test period may be extended 7 days or up to half the prescribed period.
Hard seeds are counted and reported as such on the ISTA Certificate.

7. Comparisons of the Three Rules:
AOSA and ISTA are the most similar:
5 Day First Count
8 Day Final Count
20-30C or 25C
Hard Seeds are reported as such.
Canada M&P is slightly different:
No 1st Count
7 Day Final Count
25C
Hard Seeds are reported as dead seeds. (Unless there is a large percentage, then the test is extended).
1. The one difference between AOSA and ISTA is the extension period. AOSA can be extended for an additional 5 days, whereas ISTA has the option of extending up to 7 days or half the prescribed germination period (which leaves room for interpretation for as to how many days you may extend the sample.)
The other difference between AOSA and ISTA are the substrata options. ISTA goes into extensive detail on the types of materials allotted to use for substrata.
We are trying to harmonize with ISTA, however, I think a 7 day final count for Soybeans is sufficient. It is a general practice amongst most labs.

8. Types of Rules/Methods That Are Followed:
1. Here is an idea of what people are testing their samples. There were 49 responses to this question. Out of the 49 responses, all 49 labs/analysts use the AOSA Rules, 15 of those 49 also use Canada M&P, and 12 of those also use ISTA Rules.

9. Amount of Soybeans Tested
There is quite the variety of the level of Soybean testing amongst the analysts that answered the questionnaire. These made the responses interesting and informational.

10. AOSA Warm Test Methods
Because all of the analysts that participated use the AOSA Rules to test their Soybeans, I’m going to only cover the responses to the AOSA questions about Soybean Testing for right now. Once I have all of my data put into place, I will cover the Canada M&P responses along with the ISTA responses.
Trivia Question: What year were Soybeans introduced to the United States and from what country: and China.

11. Substrata Used in Testing
Out of the 39 responses to what substrata was being used in their day to day testing, 28 of those analysts are using Rolled Towels or Paper Toweling. The other responses were: “If mold is present, we add peat to the rolled towel test.” and “We do not test Soybeans.” - 2 people answered that response.
There were several different means of planting samples (meaning there were specifics to the way that samples were planted using those specific substrata), but opted to not share those (in case some were proprietary).

12. Sand – Preferred Method or Retest Method
The next question asked was if Sand was used, was it used as a preferred method or only as a retest method. It’s mainly used as retest method, however, some of the other responses were: “Used only in high disease years, otherwise rolled towel is the preferred method”; “At customer request” – 2 answered this; and 2 response of N/A.
When sand is being used, the majority of the analysts (20 out of the 23 who answered this question) will evaluate all of the seedlings in the tray. The remaining analysts will only evaluate the seedlings that have emerged through the soil/sand surface.

13. Temperatures and Light
Trivia Question: How many crayons can one acre of Soybeans produce? 82,368 crayons….That’s a lot of coloring! 
When asked what temperature do analysts use predominately with their Soybeans, 25C was used most often (25 out of 40 analysts). 15 out 40 analysts use the 20-30C, while 2 out of the 40 analysts use both temperatures (one noting that the 25C is only used for retest or comparative tests.)
The majority of the analysts state that they use light in their test, however, it is not a requirement for Soybeans. The “other” was noted that they used it for Sand Tests.

14. Duration of Test Period
1. Since my referee is about shortening the test duration of Soybeans, I thought it would be important to ask about what each analyst tested at. The majority of the analysts (23 out of 39) who answered this, test at 7 days. 14 out of 39 answered 8 days and 2 out of 39 answered 5 days.

15. Duration of Test – Early Termination
1. The question was asked if the analysts terminate the tests early. If the seeds are only getting tested to day 7, technically, the test is being terminated early. 20 out of the 39 analysts say that yes, they do terminate their tests early. 14 out of the 39 analysts say that no, they do not. The assumption is that they are going the full 8 days (or 7 days based on Canada M&P). The other 5 analysts chose other. Those comments were as follows: 1. Non-applicable- 2 answered this; 2. Only as a client request; 3. They check tests early for mold or infection. If the mold/infection is minimal, they let the test go the full 8 days. 4. For regulatory samples only. 2.

16. Duration of Test - First Counts
In the AOSA Rules, it notes that a first count at day 5 could be done. When asked if first counts were done, 10 analysts said yes, while 27 analysts said no.
Because the majority of the analysts stated that they did not do a first count, the next question had quite a few non-applicable answers (20 out of 31). 8 out of the 31 said that they did the first count on day 5, while the remaining 3 stated other. Those answers included: “First Count is performed only if seedlings look necrotic when observed;”, “5-6 Days”; and “4 days”.

17. Duration of Tests - Test Extensions
Trivia Question: What country leads the production in Soybeans? United States. What country is second in production? Brazil.
The next set of questions was about test extensions in Soybeans. 19 out of 41 analysts said that they do extend, while 17 out of 41 analysts say they do not. The remaining 5 analysts noted “Other”. Those responses were as follows: 1. Non-applicable; 2. If there are questionable seedlings, we may extend 2 days; 3. If there are late starters or they are swollen; 4. Sometimes, if it needs it; 5. If the lot shows small sprouts which have the potential to be a normal seedling.
Analysts were split on the idea of extending. Personally, in our lab, we would only extend seedlings that were small or firm/ungerminated that appeared to have the potential to be a normal seedling.
So if the analysts extended the tests, I had asked how long their extensions were for. 4 out of the 24 answered 4 days; 5 out of the 24 answered 5 days; The remaining 9 answered “Other”. Those comments included: Days - 6 answered this; to 5 Days; Days; 4. Non-applicable - 2 answered this; 5. Typically 2 days, but up to 4 days; 6. Usually 2 days, but no more than 5 days; Days; or 3 Days.
There appears to be a random assortment of extension days, but it was noted that most analysts either never extended or rarely extended Soybean seedlings.

18. Post Germ TZ
1. The majority of the analysts answered that they do not do a Post Germ TZ on Soybeans at the end of the test. 3 out of the 40 analysts stated “Other”. The comments included: 1. Not required. Seeds are not fresh. They are either dead or show abnormal growth; 2. If a TZ Test is done, it is often on fresh seed; 3. Yes, if there are swollen seeds.
2. The majority of the seeds that we encounter do not have any firm/ungerminated seeds at the end of the test, however, seeds that come from the organic industry, especially black soybeans do exhibit slow germination along with many firm/ungerminated seeds at the end of the test.

19. Conclusion
There are various nuances in how each laboratory is testing Soybean samples.
The goal of this referee is to obtain enough data to support an AOSA rules proposal.
THANK YOU!
Which says to me, that there is always room for more uniformity in testing.
Out of the 49 people who answered the survey, I only had 5 analysts/labs who contacted me about participating in the referee. It would great if you participated in the survey, if you would participate in the referee.
I will be shipping out seed as soon as we are back in the lab on Monday, June 8th.