ISO/TR 23016-3:2021

TECHNICAL ISO/TR
REPORT 23016-3
First edition
2021-10
Fine bubble technology — Agricultural
applications —
Part 3:
Guidelines for the minimum
viable number concentration of
ultrafine bubbles for promoting the
germination of barley seeds
Technologie des fines bulles — Applications agricoles —
Partie 3: Lignes directrices relatives à la concentration minimale en
nombre viable de bulles ultrafines pour favoriser la germination des
graines d'orge
Reference number
ISO/TR 23016-3:2021(E)
© ISO 2021

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ISO/TR 23016-3:2021(E)
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ISO/TR 23016-3:2021(E)
Contents Page
Foreword .iv
Introduction .v
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Test subjects . 2
5 Judgement of significant difference in T . 2
50
6 Seeds and measurement device . .2
6.1 Seeds for germination test . 2
6.2 Measurement device for UFB size and concentration . 2
6.3 Examination range of UFB number concentration . 2
7 Measurement data . 2
7.1 Necessary storage period of barley seeds longer than that of dormancy breaking . 2
7.2 Germination promotion effects in association with each number concentration of
UFB and the storage period of seeds . 3
7.3 Results of the data analyses . 4
7.3.1 Typical example . 4
7.3.2 Analyses results of the remaining data (No. 2, 3, 4, 6, 7, 9, 10 and 11 in
Table 1) . 8
8 Suggestion .20
Annex A (informative) Example of measured data of size distribution and number
concentration of UFB .21
Bibliography .23
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ISO/TR 23016-3:2021(E)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards
bodies (ISO member bodies). The work of preparing International Standards is normally carried out
through ISO technical committees. Each member body interested in a subject for which a technical
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organizations, governmental and non-governmental, in liaison with ISO, also take part in the work.
ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of
electrotechnical standardization.
The procedures used to develop this document and those intended for its further maintenance are
described in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the
different types of ISO documents should be noted. This document was drafted in accordance with the
editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/directives).
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of
any patent rights identified during the development of the document will be in the Introduction and/or
on the ISO list of patent declarations received (see www.iso.org/patents).
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constitute an endorsement.
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www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 281 Fine bubble technology.
A list of all parts in the ISO 23016 series can be found on the ISO website.
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html.
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ISO/TR 23016-3:2021(E)
Introduction
The market for technologies using fine bubbles have been rapidly growing in many applications
throughout the industrial, domestic and academic sectors. Most notably, the applications of ultrafine
bubble (UFB) technology to the agricultural area are drawing very high interest as they are thought to
be part of the key technologies improving the productivity and efficiency of agriculture. Consequently,
this contributes to the promotion of the United Nations Sustainable Development Goals (SDGs) in terms
of providing sufficient food and maintaining water resources on land, for example.
As part of the strategy for standardizing the agricultural applications of fine bubbles, ISO 23016-2 was
published, describing the test method for promoting barley seed germination by application of a UFB
water generation system. Although UFB number concentration is closely related to the promotion of
barley seed germination, the effects of concentration were not specified in it.
For the practical application of UFB technology to seed germination, it is important to provide data
specifying the minimum number concentration of UFB necessary for promoting seed germination. This
data provides useful information to users in their selection of an appropriate UFB generation system.
This document describes the collected data indicating the experimental observations between various
number concentrations of UFB and their promotion effects on barley seed germination.
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TECHNICAL REPORT ISO/TR 23016-3:2021(E)
Fine bubble technology — Agricultural applications —
Part 3:
Guidelines for the minimum viable number concentration
of ultrafine bubbles for promoting the germination of
barley seeds
1 Scope
This document demonstrates guidelines for promoting the germination of barley seeds with a lower
number concentration of ultrafine bubbles (UFB). This is achieved by taking the data concerning the
germination ratio of barley seeds conforming to ISO 23016-2 as a starting point and then evaluating the
minimum number concentration range of ultrafine bubble water necessary for promoting the effect on
germination of barely seeds by changing germination time.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content
constitutes requirements of this document. For dated references, only the edition cited applies. For
undated references, the latest edition of the referenced document (including any amendments) applies.
ISO 23016-2, Fine bubble technology — Agricultural applications — Part 2: Test method for evaluating the
promotion of the germination of barley seeds
3 Terms and definitions
For the purposes of this document, the terms and definitions given in ISO 23016-2 and the following
apply.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at http:// www .electropedia .org/
3.1
median germination period
T
50
time where the inferred germination ratio marks 50 % of the maximum germination ratio
Note 1 to entry: See Reference [1].
3.2
storage period
time length when barley seeds are stored under 20 °C
3.3
dormancy
failure of an intact viable seed to complete germination under favourable conditions
Note 1 to entry: The related term 'dormancy breaking' describes the breaking of dormancy. See Reference [2].
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ISO/TR 23016-3:2021(E)
4 Test subjects
The items subject to the test are air containing UFB water stored in bottles or other containers for
preservation and transport, and the UFB generating system used to generate the UFB water. UFB water
is generated by supplying raw water to the UFB generating system. Distilled water with a quality of
1)
A2 or greater is used as raw water according to ISO 23016-2.
The size, quantity and concentration of UFB in UFB water are measured and an example of measured
data is given in Annex A.
5 Judgement of significant difference in T
50
After determining the correlation curve shown in ISO 23016-2:2019, Formula (2), the sum squared of
residual (SSR) and standard error (SE) around the time T are calculated. From this curve, the 95 %
50
confidence interval of T is determined.
50
6 Seeds and measurement device
6.1 Seeds for germination test
Barley seeds stored longer than the period of dormancy breaking were used as the seeds for the
germination test. Barley seeds are not obtained from a specific location, but can be obtained anywhere.
It was not necessary for the seeds to be rinsed in water for use; the test was initiated using the dry
seeds as supplied. The germination test was conducted at the constant temperature of 25 °C.
6.2 Measurement device for UFB size and concentration
The air containing UFB water was generated using the pressure dissolution system with the pressure
just after the pressurizing pump at around 700 kPa and around 300 kPa at the saturator. These
samples had their size and concentration measured using a particle tracking analysis instrument (see
2)
ISO 19430). The instrument used was a Nano Sight (Malvern Panalytical, UK) , allowing a measuring
range from 50 to 1 000 nm. The wavelength of laser light source was 635 nm, the power was 40 mW,
and a camera with black and white charge coupled device (CCD) was used in conjunction with analysis
software of NTA 3.1 Build 3.1.46. Preset parameters are given in Annex A. The measuring temperature
was at a room temperature of around 22 °C.
6.3 Examination range of UFB number concentration
As the UFB generation system given in ISO 23016-2 stably generates UFB in the number concentration
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ranges from 10 /ml, 10 /ml and greater, the promotion effect in the lower region of number
7 8
concentration of UFB was examined from between10 /ml to 10 /ml. The air UFB number concentration
in this range is known to be within the range able to be measured by the commercially available device
mentioned in 6.2.
7 Measurement data
7.1 Necessary storage period of barley seeds longer than that of dormancy breaking
As barley plants grown under cool temperatures produce highly dormant seeds, extended periods of
after-ripening are required to break dormancy. For example, storage for 3 to 4 months is required to
[3]
allow dormancy to decay . It is hoped that the barley seeds to be used are stored longer than the
1) Electrical conductivity 0,1 mS/S (25 °C), total organic carbon (TOC) 0,5 mg-C/l or less, zinc 0,5 µg-Zn/l or less,
- 2-
silica 50 µg-SiO /l or less, chloride ions µg-Cl /l, sulphide ions µg-SO /l.
2 4
2) "Nano Sight" is an example of a suitable product available commercially. This information is given for the
convenience of users of this document and does not constitute an endorsement by ISO of this product.
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ISO/TR 23016-3:2021(E)
period of dormancy breaking. The seeds used in this test described in this document were stored for 11
months or more, which means dormancy breaking seeds were used.
7.2 Germination promotion effects in association with each number concentration of
UFB and the storage period of seeds
Every measurement result is shown in Table 1. Judging from evaluation of the median germination time
(T ), it is shown that there is a clear turning point for the deterioration of seeds’ quality for a storage
50
period between 34 and 38 months.
In terms of germination promotion effects, in all cases, the median germination time was T < T ,
50,UFB 50,C
(where " " represents the control section). This shows the improvement in germination speed when
C
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UFB is used. However, in sample No. 1 (1,1×10 /ml) and sample No. 3 (3,5×10 /ml), statistically
significant differences were not seen. This is considered to have been caused by an excessively low
number concentration being below the threshold required for the promotion of germination.
Meanwhile, the results with a number concentration of UFB about three times as high that of sample
7
No. 1 under a similar storage period for the seeds are shown as sample No. 2 (3,1×10 /ml) and sample
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No. 3 (3,5×10 /ml). On the contrary to the case of sample No. 2, where there was a significant difference
between T and T a significant difference was not recognized in the case of sample No. 3.
50,C 50,UFB,
Therefore, it was judged that a statistically stable effect on the promotion of germination was not
expected at this number concentration level.
Furthermore, when the number concentration increased, i.e., in the region of sample No. 4 to sample
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No. 11 (from 7,2×10 /ml to 1,4×10 /ml), the germination promotion effect with statistically significant
[4]
difference was seen regardless of the storage period (seed quality) . Seeds used from samples No. 1
to 4, and samples No. 8, 9 and 11 were shown to have high qualities, supported by G (where " "
max,C C
represents the control section) being higher than 90 %. On the other hand, seeds from samples No. 5 to
7 and No. 10 were shown to have low quality, with G being less than 65 %.
max,C
Table 1 — Germination promotion effect on barley seeds with UFB number concentration and
storage period
Number Storage T T G G
50,C 50,UFB max,C max,UFB
Significant
b
Exponent No. concentration duration P ± ΔP
a a
difference
(%) (%)
(/ml) (month) (h) (h)
The UFB water with the bubble number concentration of the
Not appli-
6
6 0 order of 10 /ml did not show any germination promotion effect NO
cable
...