This trial examined the effects of various organic products on a mixed grass green. Myco-up, which contains an arbuscular mycorrhizal fungus, led to significantly greater root length than the control and other treatments. Several treatments improved turf color initially based on DGCI analysis, but effects were generally short-lived. Myco-up maintained the darkest green color longest. While most treatments improved NDVI readings initially, only Myco-up and the control maintained higher turf quality than Turf Bio over the trial duration. In summary, Myco-up promoted root growth and maintained darker green color and higher quality turf over a longer period compared to other treatments tested.

1. 1
Trial examining the impact of various organic/biological applications on a mixed Agrostis/Poa annua
green
Summary
This is by no means either a recommendation or otherwise of the products tested. All make claims
and in this particular situation and over the relatively short duration of the trial the results did
support these to some extent.
Possibly a longer run trial may well give more conclusive results but even over the duration of this
trial we did gain some interesting results with all the treatments.
In regard to the Mycoup colonization was evident throughout the root length as shown in the images
within this report.
Interestingly work by Gange et al (1999) looked at the relation between AM fungi and P. annua. In
greens where AM fungi were relatively common P. annua was rare, and vice versa. Furthermore,
when the fungi were common, abundance of the desirable turfgrass Agrostis stolonifera was greater.
Two explanations were suggested for these relations, a competitive one, in which AM fungi alter the
balance of competition between the two grasses, and an antagonistic one, in which the fungi may
directly reduce the growth of P. annua. Where mycorrhizal inoculum was added to a golf green, the
colonization level of A. stolonifera roots was enhanced, as was the abundance of this grass.
Furthermore, there was a suggestion that adding inoculum could decrease the abundance of P.
annua. This would be an interesting next step in any further work.

2. 2
Experimental protocol
A practice green comprising a mixed Agrostis/Poa annua sward at Bonnie Doon GC was marked out
into 64 x 1m2 plots with a 50mm buffer between each. The profile comprised approximately 50mm
of thatch overlying a sandy rootzone. A randomized block trial was set up comprising 16 treatments.
These were applied as shown opposite with 4 replicates of each including a control which was
untreated. This is the first summation of the trial data gathered relating to Glomus iranicum var.
tenuihypharum var. nov.,
The trial ran from 6th February to 16th April 2018.
Product was applied as per label rate in 400L of water (foliar application) apart from myco-up applied
in 800L/Ha (soil application). The water source used was rainwater. All treatments were monthly
apart from Chipco Signature applied every three weeks and myco-up and Turf-Bio applied on the 6th
February and 26th Feb. Initial application was made on 6th February with reapplication occurring on
23rd February, 12th March, 27th March, 4th April and 11th April,
Plots were monitored for turf health using an NDVI (Spectrum TCM 500 Turf Colour metre), and root
health before and at the cessation of the trial
A L J N O P P I
F I F M I K O B
B E L H C E J H
D G B J H G A E
M H E L D J L D
C A C A G M C N
P D G F B O F O
N K K I M N K P

3. 3
PRODUCTS: Active Ingredient Rate/Ha Rate/1m2
A Zinc sulphate
heptahydrate
ZnSO4H2O 200kg/Ha;
B Poacheck 175g/L endothal 2L/Ha 2.0ml/m2
C Control - -
D Poacheck +zinc
sulphate
175g/L endothall 1.5L/Ha + 25kg/Ha 1.5ml/m2 + 2.5g/m2
E
F K acetate + Ca
acetate + Mn
acetate + Mg
acetate + Silica SC
K acetate + Ca acetate + Mn acetate +
Mg acetate + Silica SC
30L + 15L + 15L + 15L
+ 7.5L
3ml + 1.5ml + 1.5ml + 1.5ml + 1.5ml
G Myco-up Glomus iranicum var. tenuihypharum
var. nov.
2.8Kg/Ha 0.28g
H Turf Bio Azospirillum, Herbaspirillum and
Bacillus bacteria
53g/Ha 0.0053g
I Phylgreen Electra Ascophyllum nodosum, calcium EDTA
7.8%; Boron 0.2%
20L/Ha 1ml
J Systacal 342g/kg Calcium phosphite 12.5Kg/Ha 1.25g
K Standard pigment Copper pthalimde 700ml/Ha 0.07ml
L Systacal plus
pigment
342g/kg Calcium phosphite + Copper
pthalimde
12.5kg/ha + 700ml/Ha 1.25g + 0.07ml
M Signature 800g/kg Fosetyl alumium plus Copper
pthalimde
12.5kg/Ha + 700ml/Ha 1.25g
N FeSO4 FeSO4.5H2O 20Kg/Ha 2g
O Encase - 20L/Ha 2ml
P K acetate + Ca
acetate + Mn
acetate + Mg
acetate + UAN
K acetate 39% w/v + Ca acetate 6% w/v
+ Mg aceate 6% w/v + Mn acetate 7%
w/v +UAN 28% w/v
30L + 15L + 15L +
15L+15L
3ml + 1.5ml + 1.5ml + 1.5ml + 1.5ml;

4. 5
Temperature
10
15
20
25
30
35
40
45
TempºC
Max/Min Temp ºC
Minimum temperature (░C) Maximum temperature (░C)

5. 6
Products trialled
Turf Bio (Thinkbio)
A patent protected plant growth promoting inoculant containing Azospirillum,
Herbaspirillum and Bacillus bacteria at 1 x 10⁹ CFU /gram.
Myco-up (Symbio)
This contains an arbuscular mycorrhizal fungus, Glomus iranicum var. tenuihypharum var.
nova, isolated from a sodium-saline soil and high production of external mycelium, intense
root colonization
Chipco Signature
Chipco Signature® (with Turf shield technology) is a true two-way systemic turf fungicide designed to
control tough turf diseases and improve overall turf quality. It protects turf from stresses during
summer and winter. The end result is turf with higher quality colour, vigour, uniformity and
playability.
Phylgreen Electra
A biostimulant solution based on GLT 100% extract of Ascophyllum nodosum coupled with
chelated calcium and boron.

6. 7
Results
Results were recorded for all trials and statistical analysis carried out (ANOVA Fisher LSD 95%
confidence)
Root length
Initial root length
Control Mycoup Phylgreen
Electra
Signature Acetates + UAN Turf Bio
Sample
1
5.5 9 5.5 7 6.7 6
Sample
2
9.1 8 6.5 7.5 11 7
Sample
3
7.3 9.3 7 5.5 11 7
Sample
4
10 9.8 6 10 7.5 5
Sample
5
8 10 11.2 7
Sample
6
8 7.3 12.5
Mean 7.98 9.03 7.00 8.08 9.05 7.42
ST DEV 1.545855 0.758837 1.767767 2.106577 2.275228926 2.61565798
Statistical analysis (ANOVA Fisher LSD 95% confidence) showed no significant difference
between the root lengths pretreatment)
4
5
6
7
8
9
10
11
12
Control Mycoup Phylgreen
Electra
Signature Acetates +
UAN
Turf Bio
Mean maximum root length cm pre
treatment

7. 8
Root length at cessation of trial
Control Mycoup Phylgreen
Electra
Signature Acetates + UAN Turf Bio
Sample
1
7.8 11.1 7.7 9.3 8.4 8.2000
Sample
2
8.2 8.9 7.4 7.5 8.4 6.00
Sample
3
8.8 8.8 8.4 9 9.3 8.3000
Sample
4
7.8 10.6 7.9 8.2 7.2 8.6
Mean 8.15 9.85 7.85 8.5 8.325 7.775
ST DEV 0.472582 1.173314 0.420317 0.8124038 0.861684397 1.19547759
Statistical analysis (ANOVA Fisher LSD 95% confidence) showed significant differences
between the root lengths post treatment) with Myco-up giving significantly greater root
length than the control and all other treatments.
There was no significant difference between the pre and post treatments.
5
6
7
8
9
10
11
12
Control Mycoup Phylgreen
Electra
Signature Acetates +
UAN
Turf Bio
Mean maximum root length cm post
treatment

8. 9
Surface Quality
DGCI Analysis
This is a method of quantifying turfgrass color using digital image analysis (DIA) using Image j
with the NCSU Turf DIA Turf Colour plugin (Zhang, C., et al 2017). NCSU Turf DIA is a plugin for
the open-source image processing software ImageJ. This tool offers rapid batch analysis of digital turf
images to report means and standard deviations of hue angle, saturation, and brightness of each
image.
DIA was carried out on 5 occasions namely April 1st
, April 2nd
, April 9th
, April 12th
and May 3rd
with
images being taken using an Olympus SZ-16,DZ-105.
Where on a scale of 0–1.0: 1.0 = darkest green
Statistical analysis (ANOVA Fisher LSD 95% confidence) showed on April 1st no significant
difference between the treatments. On April 2nd post treatment significant differences were
evident between the control and all treatments. On April 9th the only treatment giving
significantly darker colour than the control was Myco-up. On April 12th the only treatment
giving significantly darker colour was Turf Bio. There were no significant differences on May
3rd.
0.15
0.2
0.25
0.3
0.35
0.4
0.45
DGCI Mean April 1 DGCI Mean 2 April DGCI Mean 9 April DGCI Mean 12 April DGCI Mean 3 May
DGCI Mean
Control Myco-up Turf Bio Phylgreen Electra Systacal plus Pigment Signature Acetates + UAN

9. 10
In the case of DGCI SD the lower DGCI SD means more uniform turf colour and higher DCGI SD means
colour is less uniform
Statistical analysis (ANOVA Fisher LSD 95% confidence) showed on April 1st no significant
difference between the treatments. On April 2nd post treatment no significant differences
were evident between the control and all treatments. On April 9th, April 12th and May 3rd
there were no significant differences between any of the treatments in relation to colour
uniformity compared to the control.
0.03
0.035
0.04
0.045
0.05
0.055
0.06
0.065
0.07
0.075
DGCI SD Mean April 1 DGCI SD Mean 2 April DGCI SD Mean 9 April DGCI SD Mean 12 April DGCI SD Mean 3 May
DGCI SD Mean
Control Myco-up Turf Bio Phylgreen Electra Signature Acetates + UAN

10. 11
NDVI
Initial NDVI readings
Control Myco-up Turf Bio Phylgreen
Electra
Signature Acetates
0.6872 0.6727 0.6762 0.6818 0.6434 0.6779
0.6695 0.6757 0.6864 0.6688 0.6708 0.6827
0.6952 0.6822 0.6758 0.6592 0.7049 0.6978
0.691 0.67 0.6564 0.665091 0.6567 0.6718
Mean 0.685725 0.67515 0.6737 0.668723 0.66895 0.68255
SD 0.011299 0.005245 0.012533 0.009572 0.026449 0.011102
Statistical analysis (ANOVA Fisher LSD 95% confidence) showed that at the commencement
of the trial there was no significant difference between the treatments.
0.61
0.62
0.63
0.64
0.65
0.66
0.67
0.68
0.69
0.7
0.71
Control Myco-up Turf Bio Phylgreen
Electra
Signature Acetates
Initial pre treatment NDVI

11. 12
Post treatment NDVI readings
Statistical analysis (ANOVA Fisher LSD 95% confidence) showed that at the cessation of the
trial all the treatments including the control gave better turf quality than the Turf Bio
treatment.
Statistical analysis (ANOVA Fisher LSD 95% confidence) showed over the period of the trial
there was no significant difference between the treatments.
0.52
0.54
0.56
0.58
0.6
0.62
0.64
0.66
0.68
0.7
0.72
Post treatment NDVI
0.62
0.63
0.64
0.65
0.66
0.67
0.68
0.69
0.7
0.71
0.72
0.73
Control Myco-up Turf Bio Phylgreen
Electra
Signature Acetates
Mean NDVI Readings for duration of trial

12. 13
The following graphs show the impact of the individual treatments as indicated by the black
arrows. All the treatments gave a ‘’spike’’ in turf quality but this generally was short lasting.
0.6
0.62
0.64
0.66
0.68
0.7
0.72
0.74
12-Feb 19-Feb 26-Feb 5-Mar 12-Mar 19-Mar 26-Mar 2-Apr 9-Apr 16-Apr
NDVI Readings February 6 to April 16th
Control Myco-up Turf Bio
Phylgreen Electra Signature Acetates
0.6
0.62
0.64
0.66
0.68
0.7
0.72
0.74
12-Feb 19-Feb 26-Feb 5-Mar 12-Mar 19-Mar 26-Mar 2-Apr 9-Apr 16-Apr
Control

13. 14
0.6
0.62
0.64
0.66
0.68
0.7
0.72
0.74
12-Feb 19-Feb 26-Feb 5-Mar 12-Mar 19-Mar 26-Mar 2-Apr 9-Apr 16-Apr
Myco-up
0.56
0.58
0.6
0.62
0.64
0.66
0.68
0.7
0.72
0.74
12-Feb 19-Feb 26-Feb 5-Mar 12-Mar 19-Mar 26-Mar 2-Apr 9-Apr 16-Apr
Turf Bio

14. 15
0.6
0.62
0.64
0.66
0.68
0.7
0.72
0.74
12-Feb 19-Feb 26-Feb 5-Mar 12-Mar 19-Mar 26-Mar 2-Apr 9-Apr 16-Apr
Phylgreen Electra
0.6
0.62
0.64
0.66
0.68
0.7
0.72
0.74
12-Feb 19-Feb 26-Feb 5-Mar 12-Mar 19-Mar 26-Mar 2-Apr 9-Apr 16-Apr
Signature

15. 16
Statistical analysis (ANOVA Fisher LSD 95% confidence) showed that at the cessation of the
trial there was no significant difference between the treatments.
0.6
0.62
0.64
0.66
0.68
0.7
0.72
0.74
12-Feb 19-Feb 26-Feb 5-Mar 12-Mar 19-Mar 26-Mar 2-Apr 9-Apr 16-Apr
Acetates

16. 17
Root architecture
GIA Analysis
Characterizing root system architecture (RSA) is essential to understanding the development
and function of vascular plants. Identifying RSA-associated genes also represents an
underexplored opportunity for crop improvement. Software tools are needed to accelerate
the pace at which quantitative traits of RSA are estimated from images of root networks.
Testing was carried out using GiA Roots (General Image Analysis of Roots), a semi-automated
software tool designed specifically for the high-throughput analysis of root system images.
GiA Roots includes user-assisted algorithms to distinguish root from background and a fully
automated pipeline that extracts dozens of root system phenotypes. (Galkovskyi, T, et al
2012)
Control Phylgreen Acetates Myco-up Signature Turf Bio
Mean width (um) 23.21 24.25 30.60 27.23 29.27 30.03
SD 2.10 6.34 3.27 2.35 2.51 2.280
GIA
Max Number roots 39.75 51.89 39.9 44.33 52 44.3
Number of Connected
Components 40.16 52.89 45.6 55.75 59.8 51.33
Median Number of roots 27.91 40.77 28.8 34.58 31.7 29.83
Width
Microscopic analysis using a stereo microscope together with a Dino Eye digital microscope
running Dino Eye software revealed significant differences in root width with Signature, the
acetates and UAN and Turf Bio all giving significantly greater root widths than the control.
Signature and the acetates plus UAN gave significantly greater numbers of roots compared
to the control.
All the treatments gave a significantly greater number of connected components than the
control. With Myco-up giving the highest number recorded.

17. 18
Mycoup
This contains an arbuscular mycorrhizal fungus, Glomus iranicum var. tenuihypharum var.
nova, isolated from a sodium-saline soil and high production of external mycelium, intense
root colonization and resistance to high nutrient concentrations. This as discussed previously
was applied twice and the results below clearly show it had successfully colonized the root
network.
Images below are taken at a 2cm depth

18. 19

19. 20
Images below are taken at a 6cm depth.

20. 21

21. 22
References
Galkovskyi, T, Mileyko, Y., Bucksch, A., Moore, B., Symonova, O., Price, C.A., Topp, C.N., Iyer-
Pascuzzi, A.S., Zurek, P.R., Fang, S., Harer, J., Benfey, P.N. and Weitz, J.S. (2012) GiA Roots:
software for the high-throughput analysis of plant root system architecture. BMC Plant
Biology. 12:116.
Gange, AC., Lindsay, D.E., and Ellis, L.S., Can arbuscular mycorrhizal fungi be used to control
the undesirable grass Poa annua on golf courses? School of Biological Sciences, Royal
Holloway University of London, Egham, Surrey TW20 0EX. UK Journal of Applied Ecology
1999, 36, 909-919
Zhang, C., G.D. Pinnix, Z. Zhang, G.L. Miller, and T.W. Rufty. © 2017. Evaluation of Key
Methodology for Digital Image Analysis of Turfgrass Color Using Open-source Software. Crop
Science. https://www.turffiles.ncsu.edu/nc-state-turfgrass-dia-imagej-plugin/
Read more at: https://www.turffiles.ncsu.edu/nc-state-turfgrass-dia-imagej-plugin/