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Urease producing microorganisms under dairy pasture management in soils across New Zealand

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Urea, the most commonly used nitrogen fertiliser in New Zealand, can be quickly lost from the system via ammonia volatilisation or nitrate leaching following hydrolysis of urea by urease producing soil microorganisms (UPSMs). Hossein Alizadeh investigated UPSMs involved in urea degradation for upcoming research to reduce soil urease activity.

Image courtesy of interviewee. April 24, 2024

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This video video discusses the significance and implications of urea, the most commonly used nitrogen fertilizer worldwide, and its impact on agriculture and the environment. Urea was first synthesized in 1828, challenging earlier theories about the origins of organic compounds. Urease, the enzyme that breaks down urea into ammonia and carbon dioxide, is produced by certain soil microorganisms and can lead to substantial nitrogen loss (80-90%) shortly after application due to volatilization and leaching. This video video loss reduces nitrogen availability for plants and contributes to greenhouse gas emissions and environmental degradation.

This video video also highlights the importance of identifying urease-producing microbes in soil, as they may aid in nitrogen utilization or contribute to its degradation. The study involved isolating microbes from diverse pasture soils in New Zealand, revealing a variety of urease-producing bacteria and fungi. A significant number of these microbes were identified to belong to different taxonomic groups, with some demonstrating particularly high urease activity.

The researchers noted that various environmental factors, such as soil moisture and temperature, influence urease activity. They proposed future work to manipulate these soil microbes to improve the persistence of urea in the soil, thereby enhancing nitrogen use efficiency in plants. Additionally, the research indicated a shift in understanding regarding the culturable versus non-culturable nature of soil microorganisms, emphasizing that more focus should be placed not only on enzyme activity but also on the microbial populations that produce these enzymes. Overall, the findings could inform better agricultural practices to maximize the effectiveness of urea while minimizing environmental impacts.

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X-TIMESTAMP-MAP:LOCAL=00:00:00.000,MPEGTS=900000

00:00.235 --> 00:03.495
Uh, you might know that the uria is the most common,

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commonly used nitrogen fertilizer in the world.

00:07.835 --> 00:10.335
And uria was the first organic

00:11.295 --> 00:15.295
molecular ever synthesized in the lab, in 1828,

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challenging the Vitalist theory on the origin

00:19.035 --> 00:20.535
of organic compounds.

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Urease, which can degrade uria,

00:24.435 --> 00:28.815
is a niche dependent enzyme, which can catalyze

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uria to ammonia and carbon dioxide.

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And it is interesting. Urease also was the first enzyme

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that crystallized.

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Yeah. And urias can be found in plants, fungi, bacteria,

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and some invertebrates, but not animals.

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The problem is when, farmers apply uria in the field,

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uria can be quickly lost from the system wire

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Nia Volatilization

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or nitrate leaching, following hydrolysis by,

01:05.555 --> 01:09.775
urease enzyme, which, this enzyme is produced by

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urease producing soil microorganisms.

01:13.995 --> 01:18.895
So, that's interesting that, I tell you,

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between 80 to 90% uria can be lost

01:23.655 --> 01:26.935
because of this enzyme within one or two days

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after, application.

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And therefore, nitrogen avail availability

01:33.075 --> 01:35.615
to the plant is, reduced.

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And, we'll have subsequent production of nitrous oxide,

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which is a greenhouse gas,

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and also leaching of soil nitrate to waterways,

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which are contributors, to environmental damage.

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H

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but why microbes, why some microbes should have urease?

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Because, some references, mentioned

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that it helps them to use, nitrogen,

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uria as a nitrogen source, for example.

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Uh, in general, UES en enables some micro,

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some bacteria like, they belong to klei,

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genus to use, uria as a sole source

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of nitrogen.

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And also, ues in some human pathogens,

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can act as a viral dense factor, like,

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helico is the famous bacteria, bacteria that could cause,

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human, stomach ulcer produces,

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the ues.

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So, we have these microbes in soil as well.

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And as I said, they could, make problem

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and waste uria.

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When we apply to soil, farmers are using,

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the uase inhibitors, some uase inhibitors, which,

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they can be applied in field, then they can block es,

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enzyme, enzyme, and

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consequently, uria can last in soil for longer time,

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and plant can uptake them.

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So we'll have less wastage, especially ammonia,

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volatilization.

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Also, farmers sometimes use, coated uria,

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which is like slow release fertilizer

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and will release, uria little by little,

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so doesn't have, much chance to be degraded quickly,

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like in between one or two days, up to 80 to 90%.

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And, soil URI activity could be affected,

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by some factors, including moisture, soil moisture, I mean,

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temperature, soil, organic matter, pH,

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and also rate of fertilizer,

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applied to soil.

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So, we, what, we were doing,

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for this study was to identify soil microbes,

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which produce, urease,

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the time we started working.

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Uh, not many report about that,

04:20.585 --> 04:23.565
but recently scientists started working on

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lytic microorganisms distribution,

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because they are also very important for remediating

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of soil, heavy metal based on carbonate

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precipitation in soil.

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And also, I've seen some of these microbes are being used

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for self-healing, concrete production,

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and it's very interesting.

04:43.865 --> 04:47.925
But, because most of, papers, had worked on,

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s enzyme

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and, ES inhibitors,

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we were thinking if we could isolate some microbes

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and identify them,

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maybe we can interfere their population in soil

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and try to reduce es activity

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with different ways in soil

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and see if we can do something that u last in soil

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for a longer period, then plant will have, some chance

05:17.425 --> 05:18.565
to use them.

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And we started, isolating culture

05:22.625 --> 05:24.485
microorganisms that time.

05:25.305 --> 05:29.405
Uh, scientists believe that 99% of

05:30.195 --> 05:33.085
soil microorganisms are not culturable.

05:33.685 --> 05:36.485
Actually, we are working on 1% of microbes

05:36.485 --> 05:41.005
that are culturable, which might not be a good indication,

05:41.345 --> 05:44.445
but even 1% doesn't mean

05:44.445 --> 05:47.685
that they produce less urease that 99%.

05:47.785 --> 05:50.005
But recently, a paper published

05:50.345 --> 05:53.685
and which, corrected that paradigm

05:53.715 --> 05:58.255
that 99% of, microorganisms

05:58.255 --> 05:59.815
of soil are non culture,

05:59.915 --> 06:02.095
and now they believe that majority

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of soil microorganisms are culturable.

06:05.595 --> 06:08.415
So that's the work we started, to do.

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And, we decided to work on soil s producing microbes.

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Uh, we started, collecting soil from,

06:17.075 --> 06:18.295
dairy pasture.

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Uh, soils are across New Zealand,

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with a pasture species component of rye grass

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and white clover.

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'cause in New Zealand, mostly pastures are,

06:31.655 --> 06:36.055
composed these two, plant species and pastures.

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Uh, we choose, age between nine months even.

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We had pasture, with, 60 years old,

06:44.685 --> 06:48.255
yeah, being used as a pasture to, use

06:48.315 --> 06:49.735
for animal grazing.

06:50.635 --> 06:55.255
And we started collecting soil from,

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those, areas.

06:57.035 --> 07:01.095
Uh, they look at located at different, provinces,

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oline Canterbury, Manawa to Malboro Nelson, Otago Taranaki,

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Waikato Ra and West Coast.

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And, to collect soil samples, mostly,

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we choose a diagonal from corner to corner, a straight

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sampling line, and took six samples from

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each and mixed together.

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And then we took, one kg soil as representative of each

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location, transferred them to lab,

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and made the some, serial dilutions.

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We culture them on URIs detection media

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and started isolating different microbes.

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Of course, as I said, they are cultural microbes

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because, they are growing.

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If they are not cultural, they are not going

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to grow on the medium.

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Then also, we, when we, purified them

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and got single colony, then,

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we started the protein extraction,

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and then we check again, ues activity of

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each microorganism.

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Then we needed to identify them.

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So what we did, we extracted total genomic

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DNA from either fungi or bacteria.

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And we used different primers for fungi.

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We used the, the internal transcript regimen

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after ribosomal, we amplified that,

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area.

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And also for bacteria, we use 16 SR RNA,

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which, using two primers, we opl amplified.

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Then, the amons we sent, for sequencing

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and identified those microbes.

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And totally, we identified 54 different species

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from different soil around the con country.

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Uh, fungal general belong to diverse taxonomical groups,

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like, phyla, ascota, pio, myta,

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and zemy Cota.

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And also bacteria we identified belong to

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gammaproteobacteria and also beta operator bacteria.

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Some of them, like, RIA

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and Mucor showed very, very strong RAs activity.

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And there was, the very fast, amazing

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how the s activity strong.

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And also, I should add that, we, for first time,

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we report, s activity from three different,

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general polonia, Mariana, and also gliomas.

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Uh, in previous reports,

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these microorganisms hadn't been reported for ES production

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and also, in different climates.

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It was, interesting.

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Like in Nelson, we found that soil dominant

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Es producers are bacteria compared with export, which,

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were fgi.

10:07.705 --> 10:11.445
So, depends on the area as well.

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We got, different, microbes, either bacteria

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or fungi.

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And also the least includes a wide range of microorganisms,

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including spro, for example.

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Ki is Ide

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and, like human pathogen.

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Also, we isolated, from soil trichosporon, which,

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is a human pathogen could produce, es

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and also insect and nematode pathogens, like s

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or Polonia, where in the lease.

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And also some plant pathogens like fusarium soi

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interestingly produce, is, producing

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urease as well.

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So some, microbes were very widespread,

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could find it all, all almost everywhere, like fpor.

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But, some of them just very dependent on like,

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soil type, which was very important.

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Or like geos, we only found it, at Lincoln.

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No, not any other area.

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And we did some, experiment on there,

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later on pH.

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And, you know, some of the factors which are very important

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to use it on future for, some,

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experiments to, find a way to reduce soil

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activity in, New Zealand pastures.

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Yeah, those, fungi, as I said, they, they belong

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with, very different,

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Uh, like a diverse,

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Taxonomical group, like Oscar mic,

11:58.115 --> 11:59.115
COTA. We had

11:59.115 --> 12:01.805
the, like, do

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Doms Or

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Euros, lows and mce

12:09.425 --> 12:12.485
and lumio, the myta.

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We had Treme Loyce. And also

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For Zy myta, we

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Had Mucor,

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Which, all of them should,

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es activity for bacteria. So,

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Gammaproteobacteria, Uh, beta bacteria, which,

12:31.165 --> 12:32.325
as I

12:32.325 --> 12:33.605
Said before, some of them were

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Very, very Strong

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Ess activity.

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And, They could show

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In, Uh,

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Em, in, in emmic Cota, I can't,

12:47.885 --> 12:50.005
mention some different,

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Uh, Genea like,

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Uh, Cladosporium Livia showed

12:55.685 --> 12:58.005
Very good activity. And,

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From, class Two

13:00.795 --> 13:04.445
CETs and Pomy penicillium, also from

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CETs and ce from Tio, mice showed

13:10.285 --> 13:12.045
A very, very good activity.

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And from Sidio myta,

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Uh, crypto Cocus Trichosporon,

13:19.375 --> 13:20.805
which I, as I said, is

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A human ion.

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And Zi, micro Cota, we found ob,

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Uh, sidia, which is, Again, another,

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Uh, fungus And,

13:31.425 --> 13:34.045
For, bacteria.

13:34.435 --> 13:39.045
Also, as I said, cop, cop reduce,

13:39.095 --> 13:41.005
which is very, very,

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Strong, Activity has shown.

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And It's,

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It belongs to, Uh, Beto proto, bacteria,

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for, the, like, the,

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Depends on the Soil type. Like,

13:57.505 --> 14:00.885
Uh, some of them only could be found in ent,

14:01.235 --> 14:03.085
Area, Some microbes

14:03.245 --> 14:04.245
We had. And,

14:04.245 --> 14:07.565
some of them could be only in the

14:08.015 --> 14:10.445
Canterbury area. So,

14:10.905 --> 14:12.365
All depends on the location.

14:12.705 --> 14:15.045
And maybe some of,

14:16.185 --> 14:17.565
Uh, Them depends on the

14:17.835 --> 14:20.485
Also soil type as well, Depends on really

14:21.505 --> 14:23.925
To, soil type. So

14:24.425 --> 14:27.925
We couldn't find, also, I should add no gram-positive

14:28.325 --> 14:29.325
Es Producing

14:30.045 --> 14:33.005
Bacteria Or actinomyces from New Zealand. So,

14:33.705 --> 14:36.005
Uh, That's very interesting,

14:36.005 --> 14:38.365
Because, Uh, some, gram-positive

14:39.445 --> 14:41.405
Bacteria Have been reported

14:41.475 --> 14:43.085
That they produce UREs.

14:43.095 --> 14:44.245
Maybe they are not

14:44.835 --> 14:46.565
Pasture solar Residents.

14:46.565 --> 14:49.365
That's the reason we couldn't find them,

14:49.785 --> 14:50.925
Uh, In New Zealand.

14:51.745 --> 14:55.565
So some, gram-negative bacteria, including

14:56.455 --> 14:59.685
opium bor, Borella

15:00.065 --> 15:02.685
or Bo Bohor,

15:03.385 --> 15:06.525
and, were also not isolated from pasture.

15:06.865 --> 15:10.965
So, so means that they are likely, they are not,

15:11.385 --> 15:14.565
the soil in ha in habitat.

15:14.705 --> 15:17.165
So they are not, living in soil.

15:18.145 --> 15:20.965
Uh, other soil u is producing bacteria,

15:21.315 --> 15:23.005
including two capsule

15:23.465 --> 15:26.405
and Euro plasma, Euro co,

15:26.745 --> 15:30.125
or fungus, co co

15:31.315 --> 15:34.005
have not been previously reported in New Zealand.

15:34.305 --> 15:38.845
So it means that, they are absent in New Zealand soil.

15:38.845 --> 15:40.325
That's the reason we couldn't find,

15:40.435 --> 15:42.325
they have been reported in other papers

15:42.355 --> 15:43.845
that they produce Es,

15:43.865 --> 15:48.005
but they are not in New Zealand soil, so we couldn't I,

15:48.025 --> 15:49.325
identify them.

15:50.305 --> 15:53.205
So, also,

15:53.765 --> 15:56.925
emer law was not found in New Zealand,

15:57.065 --> 16:01.125
but it has been, found in sewage sludge in,

16:01.125 --> 16:02.285
previous study.

16:02.345 --> 16:06.245
And as we collected soil samples from pasture,

16:06.705 --> 16:11.285
no wonder we couldn't, detect them in soil.

16:11.785 --> 16:16.545
Uh, and I would

16:16.685 --> 16:21.345
say, again, as Es is an,

16:21.485 --> 16:24.745
as an enzyme that, is produced

16:24.805 --> 16:27.905
by soil es producing microbes.

16:28.445 --> 16:31.945
So, reduction of population

16:31.945 --> 16:34.705
of these microbes could be very, very important.

16:34.965 --> 16:38.465
But, most of research, as I said, has been focused on

16:39.265 --> 16:43.545
ES activity, which is enzyme,

16:43.555 --> 16:45.905
which is released by these microbes,

16:45.925 --> 16:49.385
but not much work has been done on those microbes,

16:49.385 --> 16:51.065
and we try to identify them.

16:51.985 --> 16:54.345
Cultural s producing microorganisms

16:54.705 --> 16:55.905
isolated from New Zealand.

16:55.905 --> 16:59.145
Pasture soils belong to various

16:59.915 --> 17:01.425
systematic categories

17:01.565 --> 17:05.905
and fungal species were predominant, where we didn't get,

17:05.925 --> 17:08.825
many bacterial isolates.

17:08.845 --> 17:12.385
So these microorganisms, which were found in New Zealand

17:12.385 --> 17:16.705
through throat New Zealand, are responsible for

17:17.325 --> 17:21.225
ure degradation, and then resulting in nitrogen loses

17:21.295 --> 17:22.505
from soil.

17:23.205 --> 17:25.665
Uh, so future work, what we are going to do

17:25.665 --> 17:27.065
to work on these microbes

17:27.065 --> 17:31.345
and see how we can somehow do something that,

17:31.925 --> 17:36.105
uria can last in soil for longer

17:36.205 --> 17:39.625
to improve plant nitrogen use efficiency

17:40.175 --> 17:41.785
following uria application.

17:43.025 --> 17:46.985
A number of genea, including por,

17:48.125 --> 17:53.045
pseudo and saraia were isolated from both

17:53.535 --> 17:56.005
north and south island in New Zealand,

17:57.385 --> 18:02.205
but some, including cords, hu Geomi, crypto Cocus

18:02.225 --> 18:04.725
and Mariana were also

18:05.795 --> 18:08.125
were only found in South island.

18:08.385 --> 18:11.365
We couldn't found these microbes in north island.

18:12.505 --> 18:16.685
And, soil, collected, as I said, were the,

18:16.745 --> 18:19.725
in a range of age, between nine months to

18:20.255 --> 18:22.405
60, years old.

18:22.505 --> 18:26.045
And most of, those pastures had a seal.

18:26.465 --> 18:29.925
Ilt Lo texture, fomo,

18:30.025 --> 18:32.045
was isolated from the youngest

18:32.185 --> 18:35.125
and old pasture in this study, suggesting

18:35.125 --> 18:38.445
that this fungus present in both, islands

18:38.445 --> 18:42.685
and campes in soil also, for even long time.

18:44.075 --> 18:48.245
Some, macroorganisms, for example, fusarium, p**n, col,

18:48.245 --> 18:50.125
cladosporium, PASIs F

18:50.265 --> 18:53.525
and pseudomonas were found in different soil types,

18:54.375 --> 18:56.845
while some were in only one soil type.

18:57.555 --> 19:00.725
Keto, for example, was only in one kind of soil,

19:00.825 --> 19:05.045
or trichosporon, also, which is a human pathogen.

19:05.505 --> 19:09.365
We only found it in one kind of soil, not everywhere.

19:09.465 --> 19:13.005
And that's good because it's human, pathogen.

19:14.675 --> 19:19.285
Also, the, proportion of soil bacteria, as I said,

19:19.785 --> 19:23.245
that time were thinking that culture, well is a small,

19:23.245 --> 19:24.805
maybe one to 10%.

19:25.385 --> 19:29.645
But recently, a paper reported that,

19:29.775 --> 19:33.405
culturable microorganisms are, majority

19:33.465 --> 19:34.685
of soil microbes.

19:34.685 --> 19:37.565
And this indicates that those microbes we have isolated

19:38.225 --> 19:43.205
are very important in, degradation of, uria.

19:44.265 --> 19:48.045
And, we believe that, can give us an indication

19:48.155 --> 19:53.045
that which one are mostly involved, to,

19:53.375 --> 19:56.445
deree, to urea degradation on their pasture.

19:56.705 --> 20:00.085
So, so these isolates, can be used

20:00.085 --> 20:01.765
for further studies, as I said.

20:01.825 --> 20:05.805
And, the two studies, related to optimization

20:05.865 --> 20:09.485
of uria application based on microbial.

20:09.535 --> 20:13.485
While fa favored environmental conditions to increase

20:14.245 --> 20:18.085
nitrogen use efficiency in plants and.

00:00 - you might know that the uria is the most common, commonly used nitrogen fertilizer in the world. And uria was the first organic molecular ever synthesized in the lab, in 1828, challenging the Vitalist theory on the origin of organic compounds.

00:21 - Urease, which can degrade uria, is a niche dependent enzyme, which can catalyze uria to ammonia and carbon dioxide. And it is interesting. Urease also was the first enzyme that crystallized. Yeah. And urias can be found in plants, fungi, bacteria, and some invertebrates, but not animals. The problem is when, farmers apply uria in the field, uria can be quickly lost from the system wire Nia Volatilization

01:00 - or nitrate leaching, following hydrolysis by, urease enzyme, which, this enzyme is produced by urease producing soil microorganisms. So, that's interesting that, I tell you, between 80 to 90% uria can be lost because of this enzyme within one or two days after, application. And therefore, nitrogen avail availability to the plant is, reduced. And, we'll have subsequent production of nitrous oxide,

01:39 - which is a greenhouse gas, and also leaching of soil nitrate to waterways, which are contributors, to environmental damage. H but why microbes, why some microbes should have urease? Because, some references, mentioned that it helps them to use, nitrogen, uria as a nitrogen source, for example. in general, UES en enables some micro, some bacteria like, they belong to klei,

02:20 - genus to use, uria as a sole source of nitrogen. And also, ues in some human pathogens, can act as a viral dense factor, like, helico is the famous bacteria, bacteria that could cause, human, stomach ulcer produces, the ues. So, we have these microbes in soil as well. And as I said, they could, make problem and waste uria.

02:55 - When we apply to soil, farmers are using, the uase inhibitors, some uase inhibitors, which, they can be applied in field, then they can block es, enzyme, enzyme, and consequently, uria can last in soil for longer time, and plant can uptake them. So we'll have less wastage, especially ammonia, volatilization. Also, farmers sometimes use, coated uria, which is like slow release fertilizer

03:30 - and will release, uria little by little, so doesn't have, much chance to be degraded quickly, like in between one or two days, up to 80 to 90%. And, soil URI activity could be affected, by some factors, including moisture, soil moisture, I mean, temperature, soil, organic matter, pH, and also rate of fertilizer, applied to soil. So, we, what, we were doing, for this study was to identify soil microbes,

04:11 - which produce, urease, the time we started working. not many report about that, but recently scientists started working on lytic microorganisms distribution, because they are also very important for remediating of soil, heavy metal based on carbonate precipitation in soil. And also, I've seen some of these microbes are being used for self-healing, concrete production,

04:42 - and it's very interesting. But, because most of, papers, had worked on, s enzyme and, ES inhibitors, we were thinking if we could isolate some microbes and identify them, maybe we can interfere their population in soil and try to reduce es activity with different ways in soil and see if we can do something that u last in soil

05:12 - for a longer period, then plant will have, some chance to use them. And we started, isolating culture microorganisms that time. scientists believe that 99% of soil microorganisms are not culturable. Actually, we are working on 1% of microbes that are culturable, which might not be a good indication, but even 1% doesn't mean that they produce less urease that 99%.

05:47 - But recently, a paper published and which, corrected that paradigm that 99% of, microorganisms of soil are non culture, and now they believe that majority of soil microorganisms are culturable. So that's the work we started, to do. And, we decided to work on soil s producing microbes. we started, collecting soil from, dairy pasture.

06:19 - soils are across New Zealand, with a pasture species component of rye grass and white clover. 'cause in New Zealand, mostly pastures are, composed these two, plant species and pastures. we choose, age between nine months even. We had pasture, with, 60 years old, yeah, being used as a pasture to, use for animal grazing. And we started collecting soil from,

06:55 - those, areas. they look at located at different, provinces, oline Canterbury, Manawa to Malboro Nelson, Otago Taranaki, Waikato Ra and West Coast. And, to collect soil samples, mostly, we choose a diagonal from corner to corner, a straight sampling line, and took six samples from each and mixed together. And then we took, one kg soil as representative of each location, transferred them to lab,

07:31 - and made the some, serial dilutions. We culture them on URIs detection media and started isolating different microbes. Of course, as I said, they are cultural microbes because, they are growing. If they are not cultural, they are not going to grow on the medium. Then also, we, when we, purified them and got single colony, then, we started the protein extraction,

08:01 - and then we check again, ues activity of each microorganism. Then we needed to identify them. So what we did, we extracted total genomic DNA from either fungi or bacteria. And we used different primers for fungi. We used the, the internal transcript regimen after ribosomal, we amplified that, area. And also for bacteria, we use 16 SR RNA,

08:37 - which, using two primers, we opl amplified. Then, the amons we sent, for sequencing and identified those microbes. And totally, we identified 54 different species from different soil around the con country. fungal general belong to diverse taxonomical groups, like, phyla, ascota, pio, myta, and zemy Cota. And also bacteria we identified belong to gammaproteobacteria and also beta operator bacteria.

09:16 - Some of them, like, RIA and Mucor showed very, very strong RAs activity. And there was, the very fast, amazing how the s activity strong. And also, I should add that, we, for first time, we report, s activity from three different, general polonia, Mariana, and also gliomas. in previous reports, these microorganisms hadn't been reported for ES production and also, in different climates.

09:56 - It was, interesting. Like in Nelson, we found that soil dominant Es producers are bacteria compared with export, which, were fgi. So, depends on the area as well. We got, different, microbes, either bacteria or fungi. And also the least includes a wide range of microorganisms, including spro, for example. Ki is Ide

10:29 - and, like human pathogen. Also, we isolated, from soil trichosporon, which, is a human pathogen could produce, es and also insect and nematode pathogens, like s or Polonia, where in the lease. And also some plant pathogens like fusarium soi interestingly produce, is, producing urease as well. So some, microbes were very widespread, could find it all, all almost everywhere, like fpor.

11:08 - But, some of them just very dependent on like, soil type, which was very important. Or like geos, we only found it, at Lincoln. No, not any other area. And we did some, experiment on there, later on pH. And, you know, some of the factors which are very important to use it on future for, some, experiments to, find a way to reduce soil activity in, New Zealand pastures.

11:46 - Yeah, those, fungi, as I said, they, they belong with, very different, like a diverse, Taxonomical group, like Oscar mic, COTA. We had the, like, do Doms Or Euros, lows and mce and lumio, the myta. We had Treme Loyce. And also

12:16 - For Zy myta, we Had Mucor, Which, all of them should, es activity for bacteria. So, Gammaproteobacteria, beta bacteria, which, as I Said before, some of them were Very, very Strong Ess activity. And, They could show

12:40 - In, Em, in, in emmic Cota, I can't, mention some different, Genea like, Cladosporium Livia showed Very good activity. And, From, class Two CETs and Pomy penicillium, also from CETs and ce from Tio, mice showed A very, very good activity.

13:12 - And from Sidio myta, crypto Cocus Trichosporon, which I, as I said, is A human ion. And Zi, micro Cota, we found ob, sidia, which is, Again, another, fungus And, For, bacteria. Also, as I said, cop, cop reduce, which is very, very,

13:41 - Strong, Activity has shown. And It's, It belongs to, Beto proto, bacteria, for, the, like, the, Depends on the Soil type. Like, some of them only could be found in ent, Area, Some microbes We had. And, some of them could be only in the Canterbury area. So,

14:10 - All depends on the location. And maybe some of, Them depends on the Also soil type as well, Depends on really To, soil type. So We couldn't find, also, I should add no gram-positive Es Producing Bacteria Or actinomyces from New Zealand. So, That's very interesting, Because, some, gram-positive

14:39 - Bacteria Have been reported That they produce UREs. Maybe they are not Pasture solar Residents. That's the reason we couldn't find them, In New Zealand. So some, gram-negative bacteria, including opium bor, Borella or Bo Bohor, and, were also not isolated from pasture.

15:06 - So, so means that they are likely, they are not, the soil in ha in habitat. So they are not, living in soil. other soil u is producing bacteria, including two capsule and Euro plasma, Euro co, or fungus, co co have not been previously reported in New Zealand. So it means that, they are absent in New Zealand soil. That's the reason we couldn't find,

15:40 - they have been reported in other papers that they produce Es, but they are not in New Zealand soil, so we couldn't I, identify them. So, also, emer law was not found in New Zealand, but it has been, found in sewage sludge in, previous study. And as we collected soil samples from pasture, no wonder we couldn't, detect them in soil.

16:11 - and I would say, again, as Es is an, as an enzyme that, is produced by soil es producing microbes. So, reduction of population of these microbes could be very, very important. But, most of research, as I said, has been focused on ES activity, which is enzyme, which is released by these microbes, but not much work has been done on those microbes,

16:49 - and we try to identify them. Cultural s producing microorganisms isolated from New Zealand. Pasture soils belong to various systematic categories and fungal species were predominant, where we didn't get, many bacterial isolates. So these microorganisms, which were found in New Zealand through throat New Zealand, are responsible for ure degradation, and then resulting in nitrogen loses

17:21 - from soil. so future work, what we are going to do to work on these microbes and see how we can somehow do something that, uria can last in soil for longer to improve plant nitrogen use efficiency following uria application. A number of genea, including por, pseudo and saraia were isolated from both north and south island in New Zealand,

17:57 - but some, including cords, hu Geomi, crypto Cocus and Mariana were also were only found in South island. We couldn't found these microbes in north island. And, soil, collected, as I said, were the, in a range of age, between nine months to 60, years old. And most of, those pastures had a seal. Ilt Lo texture, fomo, was isolated from the youngest

18:32 - and old pasture in this study, suggesting that this fungus present in both, islands and campes in soil also, for even long time. Some, macroorganisms, for example, fusarium, p**n, col, cladosporium, PASIs F and pseudomonas were found in different soil types, while some were in only one soil type. Keto, for example, was only in one kind of soil, or trichosporon, also, which is a human pathogen. We only found it in one kind of soil, not everywhere.

19:09 - And that's good because it's human, pathogen. Also, the, proportion of soil bacteria, as I said, that time were thinking that culture, well is a small, maybe one to 10%. But recently, a paper reported that, culturable microorganisms are, majority of soil microbes. And this indicates that those microbes we have isolated are very important in, degradation of, uria. And, we believe that, can give us an indication

19:48 - that which one are mostly involved, to, deree, to urea degradation on their pasture. So, so these isolates, can be used for further studies, as I said. And, the two studies, related to optimization of uria application based on microbial. While fa favored environmental conditions to increase nitrogen use efficiency in plants and.

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