Nitrogen fixing bacteria.

Simon Chiles

DD Moderator
With the recent escalation of Nitrogen prices it has occurred to me that growers are going to consider alternative sources of N. Having been involved with some initial work on N fixing bacteria I thought it might be useful to have a thread on here so that farmers can ask questions about some of the trial work that has been so that they can make some informed decisions. Having discussed this with @Chris F we have invited Natallia from Plantworks to join the forum and participate in the discussion. If you have any questions you’d like to ask please post them here and hopefully she’ll be able to answer them.
 

parker

Member
Arable Farmer
Location
south staffs
I have been using Biology for the last 10 years with very good results from fixing nitrogen releasing phosphorous and controlling disease.
@Natallia @PlantWorks I have been following the work that plantworks have been doing with great interest, all plants have different requirements from different biology to reach a fully balanced plant, I feel it would be interesting Natallia if you went into more detail how you have developed different combinations for different plants, because it is not one size fits all ime, Nature is far more complexed than that, and we still have so much to learn imo.
 
Location
Kent
I have been using Biology for the last 10 years with very good results from fixing nitrogen releasing phosphorous and controlling disease.
@Natallia @PlantWorks I have been following the work that plantworks have been doing with great interest, all plants have different requirements from different biology to reach a fully balanced plant, I feel it would be interesting Natallia if you went into more detail how you have developed different combinations for different plants, because it is not one size fits all ime, Nature is far more complexed than that, and we still have so much to learn imo.
This is a very good comment; thank you, @parker, and you are right about ‘one size doesn’t fit all'.

We initially commenced our research intending to design a universal consortium for all crops. However, early in our research, we quantified that crops responded more positively and consistently with tailored consortia. In any mono-crop system, the microbiome will adjust to support the crop; therefore, it is understandable that specific consortia are required to treat specific crops. When we identified this effect on the field, we returned to the greenhouse and laboratory for four years to profile crop responses against different bacterial isolates. This work led to decoding most of the rosetta stone- see a partial example below.

Furthermore, our research has shown that some isolates that have proven beneficial to one crop can be suppressive to some degree to another. Currently, PlantWorks has eight tailored consortia that have been tested and validated in field trials in the UK, with new consortia to be released in 2022, including one for maize.

It is noteworthy that it is not the case that a consortium needs many different bacteria; research has shown 3 or 4 dominant species have a significant effect.

1639433986721.png
 

N.Yorks.

Member
I have been using Biology for the last 10 years with very good results from fixing nitrogen releasing phosphorous and controlling disease.
@Natallia @PlantWorks I have been following the work that plantworks have been doing with great interest, all plants have different requirements from different biology to reach a fully balanced plant, I feel it would be interesting Natallia if you went into more detail how you have developed different combinations for different plants, because it is not one size fits all ime, Nature is far more complexed than that, and we still have so much to learn imo.

This is a very good comment; thank you, @parker, and you are right about ‘one size doesn’t fit all'.

We initially commenced our research intending to design a universal consortium for all crops. However, early in our research, we quantified that crops responded more positively and consistently with tailored consortia. In any mono-crop system, the microbiome will adjust to support the crop; therefore, it is understandable that specific consortia are required to treat specific crops. When we identified this effect on the field, we returned to the greenhouse and laboratory for four years to profile crop responses against different bacterial isolates. This work led to decoding most of the rosetta stone- see a partial example below.

Furthermore, our research has shown that some isolates that have proven beneficial to one crop can be suppressive to some degree to another. Currently, PlantWorks has eight tailored consortia that have been tested and validated in field trials in the UK, with new consortia to be released in 2022, including one for maize.

It is noteworthy that it is not the case that a consortium needs many different bacteria; research has shown 3 or 4 dominant species have a significant effect.

View attachment 1003043
Can you explain what your Figure 1 is actually telling us? What is crop 1,2,3 & 4?

Am I right in saying that bacteria isolate 'K' doesn't benefit Crop 1 but does benefit Crop 4? Isolate'O' doesn't benefit any of the crops 1 - 4 but 'E' benefits all Crops 1 - 4?
 
Location
Kent
Hi Natallia.

What are the bacteria called (binomial name) that form the typical product for wheat for example?

How do they fix N, is it atmospheric N gas that is fixed or N present already within the soil from fert/manure/organic matter?

If you were to slot your bacteria into a classic description of N cycles within a general soil, where do your bacteria fit in to that?

What is the number of bacteria that will be typically applied from a field application of your product?

Thanks.
What is the number of bacteria that will be typically applied from a field application of your product?

SR3 products contain monocultures of PGPR (10^10cfu). This number was carefully selected and validated in trials for the following reasons:
  • We seek not to dominate but modulate the population as we change crops annually.
  • The consortium that goes in the ground is in the optimum state – bacteria multiply in the presence of root exudates of the crop they are tuned to
  • CFU levels have been validated in the UK under standard farming practices. It is not always the case that higher CFU has higher benefits.
 
Location
Kent
Hi Natallia.

What are the bacteria called (binomial name) that form the typical product for wheat for example?

How do they fix N, is it atmospheric N gas that is fixed or N present already within the soil from fert/manure/organic matter?

If you were to slot your bacteria into a classic description of N cycles within a general soil, where do your bacteria fit in to that?

What is the number of bacteria that will be typically applied from a field application of your product?

Thanks.
How do they fix N, is it atmospheric N gas that is fixed or N present already within the soil from fert/manure/organic matter?


The answer to this questions is BOTH - bacteria uses soil sources of N and also fixes N from the air.

In terms of fixing N from the air – we grow these bacteria on N-free media and that ensures that bacteria we produce have to uptake N from the air.

Our experiments have shown with various levels of commercial N the bacteria have an increased effect on yield and if you go back to this blog you will see this. This will imply that bacteria are interacting with soil applied N and metabolising it. One of the factors we noticed that there is a correlation between sunlight, soil temperature and bacteria x N interaction. In poor photosynthetic conditions (cloudy dull summer) or in cold soil bacterial activity is slowed down.
 
Location
Kent
Hi Natallia.

What are the bacteria called (binomial name) that form the typical product for wheat for example?

How do they fix N, is it atmospheric N gas that is fixed or N present already within the soil from fert/manure/organic matter?

If you were to slot your bacteria into a classic description of N cycles within a general soil, where do your bacteria fit in to that?

What is the number of bacteria that will be typically applied from a field application of your product?

Thanks.
Hi @N.Yorks. - you are correct, a competent person using appropriate equipment can identify to the species level using the genetic techniques. As you might be aware, isolates within species vary in their performances and we spent a number of year refining the isolates. Bacterial isolates are proprietary of PlantWorks and won’t be disclosed to the public.

But in the spirit of full disclosure, therefore SR3 Wheat species are as follows (with some of their confirmed mode of actions):

Gluconoacetobactor diazotrophicus: (1) N-fixing, (2) phytohormone production, (3) P and Zn solubilising, (4) Induced systemic resistance, (5) Siderophore production.

Bacillus amyloliquefaciens: (1) N-fixing, (2) phytohormone production, (3) P and K solubilising, (4) Induced systemic resistance, (5) Siderophore production.

Derxia lacustris: (1) N-fixing, (2) phytohormone production, (3) Induced systemic resistance, (4) Siderophore production.

Agrobacterium strain: (1) phytohormone production, (2) Induced systemic resistance, (3) Siderophore production, (4) N-fixing.
 

N.Yorks.

Member
Hi @N.Yorks. - you are correct, a competent person using appropriate equipment can identify to the species level using the genetic techniques. As you might be aware, isolates within species vary in their performances and we spent a number of year refining the isolates. Bacterial isolates are proprietary of PlantWorks and won’t be disclosed to the public.

But in the spirit of full disclosure, therefore SR3 Wheat species are as follows (with some of their confirmed mode of actions):

Gluconoacetobactor diazotrophicus: (1) N-fixing, (2) phytohormone production, (3) P and Zn solubilising, (4) Induced systemic resistance, (5) Siderophore production.

Bacillus amyloliquefaciens: (1) N-fixing, (2) phytohormone production, (3) P and K solubilising, (4) Induced systemic resistance, (5) Siderophore production.

Derxia lacustris: (1) N-fixing, (2) phytohormone production, (3) Induced systemic resistance, (4) Siderophore production.

Agrobacterium strain: (1) phytohormone production, (2) Induced systemic resistance, (3) Siderophore production, (4) N-fixing.
Thanks for that Natalia, appreciate the full disclosure.
Would you mind giving a bit more detail on what 'Induced systemic resistance', 'Siderophore production' and 'Phytohormone production' are. I'm thinking these are additional benefits on top of the N fixing.
This is really interesting stuff!!
 
Location
Kent
Thanks for that Natalia, appreciate the full disclosure.
Would you mind giving a bit more detail on what 'Induced systemic resistance', 'Siderophore production' and 'Phytohormone production' are. I'm thinking these are additional benefits on top of the N fixing.
This is really interesting stuff!!
@N.Yorks. Chelation of divalent cations, including Cd2+, Cu2+, Ni2+, Pb2+ and Zn2+ ; trivalent cations, such as Mn3+, Co3+ and Al3+; and actinides, such as Th4+, U4+ and Pu4+ help plants acquire balanced nutrient needs. Microbes such as PGPR can release chelating agents that can aid with the plants' uptake of these minerals. Iron, for example, is an essential growth element for all living organisms. Siderophore production by PGPR under iron limiting conditions can promote plant growth by solubilising and directly supplying iron for plant utilisation. This removal of iron from the surrounding environment reduces availability for the growth of phytopathogens thereby reducing their competitiveness. Around 500 siderophores, a low molecular weight organic compound acting as chelators, have been identified to date.

Meanwhile, PGPR help plants to sequester these nutrients before they are lost or locked up again by encouraging them to grow more extensive root systems as well as shoots via phytohormone production. Indole-3-acetic acid (IAA), cytokinin (CK), gibberellin (GA) and abscisic acid (ABA) play an important role in cell division, root development, enlargement of the root surface area, transport of nutrients to the plant, seed germination, chlorophyll accumulation, leaf development and initiation of enzyme function.

Induction of systemic resistance can occur through biotic factors (e.g. infecting or feeding pathogens) or through chemical agents (e.g. salicylate). Puts plant's defence mechanisms under high alert. Various non-pathogenic, i.e. PGPR strains, have the ability to induce systemic disease resistance in plants against broad spectrum phytopathogens through the production of secondary metabolites, competition for resources or through the stimulation of the host’s immune system.
E.g. some PGPR can regulate plants' ethylene levels by producing 1-Aminoclopropane-1-carboxylate (ACC) deaminase to lower the levels of ethylene precursors. Although ethylene is essential for normal growth and development in plants the build-up of ethylene will cause plant growth inhibition and reduced ability to respond to different stressors, leading to reduced crop performance. ABA plays an important role in many physiological processes in plants and is crucial for the response to environmental stresses such as desiccation, salt and cold.
 

Simon C

Member
Arable Farmer
Location
Essex Coast
I think I am going to use the Plant Works PGPR over a fair few acres this year. I did some trials 2 or 3 years ago for them and have been looking over the results again recently. In almost every scenario, the bacteria produced a good yield increase, mostly around 1 tonne/Ha. That's when used with no N at all, and when applied as well as a moderate amount of N.

The only problem as I see it is that you do need to have the right conditions, they need to be washed in by rain within 24 or 48 hours and the soil needs to be warm enough for the bacteria to multiply and do their thing. I was going to try some again last year but with the long period of dry cold weather we had, I decided that it was never going to work.

Something that has been troubling me for some time is the actual quantities of bacteria that are being applied with these, or any type of biological inoculation. I have been reading "Soil Microorganisms and Higher Plants" by N A Krasilnikov on and off for a while, (very heavy going) and he says that there are about 7 to 9 tonnes of bacterial biomass in a hectare of highly fertile soil. Very poor soil may only have about a ton of bacteria per hectare, but after all these years of tying to improve my own soils health, I would like to think I am nearer the higher end of this range.

We have obtained the following data on the total microflora of the rhizosphere of vegetative plants. There are 2-2.5 kg of cells in a soil under lucerne in Central Asia, per 120 kg of soil; i.e., 6,000-7,000 kg of cells per hectare. Outside the root zone there are, according to our calculations, 1,500-2,000 kg bacterial cells per hectare of the upper (plow) layer. Consequently, there are about 7-9 tons of bacterial mass per hectare (Krasil'nikov, 1944).
In soils of medium fertility the total mass is considerably smaller. For example, in podsol soils under two-year clover and frequently fertilized we have found 1,000-3,000 millions of organisms per gram of soil in the rhizosphere and in the zone outside the roots, 300-800 million organisms per gram of soil. The total bacterial mass in the root zone amounted to 1,200-3,000 kg and outside the root zone about 350-1,000 kg. The total bacterial mass per hectare was 1,500-4,000 kg.
In the same soil under wheat, there were 800-1,200 million organisms per kg in the rhizosphere, and 100-200
million outside the roots. The total mass of bacteria was 1,100 kg per hectare.
In a poor. lightly cultivated soil (podsol) we have found under wheat, only 100-150 kg of bacterial mass per hectare in the upper (plow) layer. Eighty per cent of this mass was found in the rhizosphere.
Strugger (1948). on the basis of his investigations and those of Kendall, calculated that the total bacterial mass comprises 0.03-0.28% of the weight of the soil. Clark (1949) has shown that the bacteria constitutes 300-3,000 parts per million by weight of the soil. These data agree with our own.


As we are talking about "tonnes" of bacteria, whether 1 or 9, it would seem highly unlikely that a few grams of the inoculant could make any difference to the bulk of life already there. I don't think it would be possible to increase the total volume of life in a given soil so the introduced bacteria would have to elbow out a whole lot of others which are already living there in their perfect environment. I may be looking at this completely the wrong way, but the numbers don't add up to me.

To counteract this, @Natallia @PlantWorks , your ex colleague told me that there are as many bacteria in a sachet of your SR3 product as there are already living in 5 hectares of farmed soil. I may have misunderstood the comment, but it would be nice to understand how the inoculation and multiplication process works.

So two point of view, both of which seem pretty far fetched????


1643577374611.png1643577343389.png
 
With the recent escalation of Nitrogen prices it has occurred to me that growers are going to consider alternative sources of N. Having been involved with some initial work on N fixing bacteria I thought it might be useful to have a thread on here so that farmers can ask questions about some of the trial work that has been so that they can make some informed decisions. Having discussed this with @Chris F we have invited Natallia from Plantworks to join the forum and participate in the discussion. If you have any questions you’d like to ask please post them here and hopefully she’ll be able to answer them.
In the USA I like to use Twin N. It will replace about 25% of the N required. Would like to initiate some work in the UK
 
Location
Kent
I think I am going to use the Plant Works PGPR over a fair few acres this year. I did some trials 2 or 3 years ago for them and have been looking over the results again recently. In almost every scenario, the bacteria produced a good yield increase, mostly around 1 tonne/Ha. That's when used with no N at all, and when applied as well as a moderate amount of N.

The only problem as I see it is that you do need to have the right conditions, they need to be washed in by rain within 24 or 48 hours and the soil needs to be warm enough for the bacteria to multiply and do their thing. I was going to try some again last year but with the long period of dry cold weather we had, I decided that it was never going to work.

Something that has been troubling me for some time is the actual quantities of bacteria that are being applied with these, or any type of biological inoculation. I have been reading "Soil Microorganisms and Higher Plants" by N A Krasilnikov on and off for a while, (very heavy going) and he says that there are about 7 to 9 tonnes of bacterial biomass in a hectare of highly fertile soil. Very poor soil may only have about a ton of bacteria per hectare, but after all these years of tying to improve my own soils health, I would like to think I am nearer the higher end of this range.

We have obtained the following data on the total microflora of the rhizosphere of vegetative plants. There are 2-2.5 kg of cells in a soil under lucerne in Central Asia, per 120 kg of soil; i.e., 6,000-7,000 kg of cells per hectare. Outside the root zone there are, according to our calculations, 1,500-2,000 kg bacterial cells per hectare of the upper (plow) layer. Consequently, there are about 7-9 tons of bacterial mass per hectare (Krasil'nikov, 1944).
In soils of medium fertility the total mass is considerably smaller. For example, in podsol soils under two-year clover and frequently fertilized we have found 1,000-3,000 millions of organisms per gram of soil in the rhizosphere and in the zone outside the roots, 300-800 million organisms per gram of soil. The total bacterial mass in the root zone amounted to 1,200-3,000 kg and outside the root zone about 350-1,000 kg. The total bacterial mass per hectare was 1,500-4,000 kg.
In the same soil under wheat, there were 800-1,200 million organisms per kg in the rhizosphere, and 100-200
million outside the roots. The total mass of bacteria was 1,100 kg per hectare.
In a poor. lightly cultivated soil (podsol) we have found under wheat, only 100-150 kg of bacterial mass per hectare in the upper (plow) layer. Eighty per cent of this mass was found in the rhizosphere.
Strugger (1948). on the basis of his investigations and those of Kendall, calculated that the total bacterial mass comprises 0.03-0.28% of the weight of the soil. Clark (1949) has shown that the bacteria constitutes 300-3,000 parts per million by weight of the soil. These data agree with our own.


As we are talking about "tonnes" of bacteria, whether 1 or 9, it would seem highly unlikely that a few grams of the inoculant could make any difference to the bulk of life already there. I don't think it would be possible to increase the total volume of life in a given soil so the introduced bacteria would have to elbow out a whole lot of others which are already living there in their perfect environment. I may be looking at this completely the wrong way, but the numbers don't add up to me.

To counteract this, @Natallia @PlantWorks , your ex colleague told me that there are as many bacteria in a sachet of your SR3 product as there are already living in 5 hectares of farmed soil. I may have misunderstood the comment, but it would be nice to understand how the inoculation and multiplication process works.

So two point of view, both of which seem pretty far fetched????


View attachment 1013515View attachment 1013514
When people refer to bacteria in the soil it is important to understand that many of these do not interact directly with plants, the ones that do positively benefit plants are Plant Growth Promoting Rhizobacteria (PGPR) and it this group that inoculum are drawn from.

Our Bio-inoculant rhizobia seek to modulate the microbial community in the soil, not to dominate as the legacy of these bio-inoculant may pose other unwanted consequences on the subsequent crops. Our trials, and other academic work, have consistently shown that the CFU levels that we employ are sufficient for the bacteria consortia to initially become established and then multiple to positivity co exist within the background of other bacteria and benefit the host plants.

Factors that have positively effect the inoculum functions include soil temperature and photosynthetic activity (as this translates to increased root exudate).

The PGPR inoculants function to provide more nutrients such as nitrogen, phosphorus and potassium as well as other micro elements for their host plants, but also to support some of the other bacteria in the community. The inter relationship of soil bacteria ultimately is one of ‘balance’ and not of dominance.

The synergies between beneficial bacteria and other fungi and bacteria are well documented in research papers across different soils types, environments across the world. For example Arbuscular Mycorrhizal Fungi (AMF) work synergistically with beneficial rhizobacteria as the fungal hyphae help to feed the bacteria as well as taking up the nutrients produced by the rhizobacteria. Therefore, these bacterial inoculant seek to promote the populations of other beneficial microbes such as AMF that are already in the soil too.
 

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