all the delegates from the different parts of the country. I will be speaking on soil biodiversity theme, which is a soil biodiversity saving the future food system, theme three. And my topic is on bio-fertilizer education in India for future prospects. I'm speaking from Indian Institute of Soil Science, which is located in Central India. The place is Bhupal. And with me, Dr. Esar Mohanty and Dr. JK Thakur, they are my colleagues working on the different aspects of bio-fertilizers as well as soil biodiversity. I just, since the time is very short, 10 minutes, just I'll briefly, you know, bio-fertilizer and agriculture throughout the world having a lot of importance in the present time because of the increasing cost of fertilizers. And also there are many other problems related to fertilizers. So we are talking more on the integrated nutrient management where bio-fertilizers is one components along with chemical fertilizers, organic manuals, different kinds. And when we talk about the Indian context, we have got, you know, more than 140 million hectares of lands, but majority of the farmers are small and marginal farmers. And the balance fertilizers, when we talk the balance fertilizers based on soil and the costly inputs fertilizers. So much fertilizers often could not be used for that. So in that, the bio-fertilizers is a very promising and having a lot of advantages. India is one of the 12 mega biodiversity countries of the world with only 2.5% of the land area, which is the biodiversity is 7 to 8% of the recorded species of the world. But the problem is with the increasing food productions and the intensive agriculture, the soil count is, you know, decreasing, which is now less than about 0.5%. As a result, the soil fertility, soil health is declining, you know, and that is a matter of concerns. So in this, the role of bio-fertilizers is enormous and the small and marginal farmers as I was telling. And bio-fertilizers are known to not only improve yields and productive quality of the product, but also improve the nutrient use efficiency.  So there's a lot of problem in Indian context. And these are some of the bio-fertilizers and we have studied our institutes having the all India coordinated research on the bio-fertilizers and biodiversity. At 18 locations, we are working on the different bio-fertilizers and these are the list of bio-fertilizers, rhizobium, azodobacter, azospirilum, brookalagi. So, and you see the ranges of benefits, 50 to 300 kilogram nitrogen per hectare per year when we apply the rhizobium along with the different crops like ground nuts, these are the different crops. And the benefits we have observed, which is highly significant. And this is the most important thing. And you see that is why the use of bio-fertilizers in Indian agriculture is very significant. But if you compare the zones of the country where, you know, how the bio-fertilizers are being used and you will find that some of the states are doing very good, some of the areas. See from here, the south zone of India, the increasing, there is increase of bio-fertilizers. That is because of the benefits, is the response of the bio-fertilizers, the different scenarios and also the awareness and the benefits which have been observed. So, there is increasing in some places, but some places they're almost constant because of different kinds of problems. And then recently, number of bio-fertilizers has been introduced in the country like, you know, acid factor, diatrophic gas, which is endophyte dig, nitrogen fixers in sugar cane. Similarly, the potassium, potassium mobilizers, which is also very much promising as just for your information, that 100% of the potassium fertilizers is imported to India. So, the role of potassium bio-fertilizers is very, very important. So, a lot of work has been done on this space and beneficial the organisms which having the potassium mobilizing capacities have been identified and being used. Similarly, the micronutrients, gene solubilizers. And then the effective organisms, sometimes the consortia, a mixture of different kinds of yeast, electro-bacillus, and rhodo-pseudomonas, they're all excellent for the residue degradation and for the fixation of the fixed nutrients solubilizations. And the, for the promotions, for the promotions for increasing the area more of using bio-fertilizers, a lot of schemes. Government of India is being undertaken. Some of these examples are that, you know, that one, we developed the national centers for organic farming, having different regional centers where the focus is also for increasing the use of bio-fertilizers on the nitrogen bio-fertilizers the phosphorus, potassium, the micronutrients and different kinds. And, you know, in India, just the demand is very high but still the capacity, the capacity and the production per year is not matching that much. So there is a lot of scope of the production of bio-fertilizers, increasing production as well as the use almost only the half of the requirements of the bio-fertilizers is being produced and which could be supplied for the use along with the fertilizers. And these are the, you know, other things is the financial support as well as and the financial support is also provided for, you know, the incurring, enter things, the, you know, at the demonstrations these are being demonstrated at the farmer's fields and also the different kind of, you know, incentives given, the financial supports given so that the bio-fertilizers, the bio-fertilizers units production of bio-fertilizers can be made and only their production as well as their applications at the, along with the, you know, the different packages of bio-fertilizer applications. And so these are some of the setting up of state of the art bio-fertilizer units where 100% assistance is given. Similarly, setting up of bio-fertilizer testing and quality control or strengthening of existing laboratory units under fertilizer control order. So here also the majority, the maximum amount of the assistance given the promotional contractors on farmer's fields, you know, 50% or more costs subject to limit of 5,000 per hectare each beneficiaries are given so that they understand the benefits and subsequently they can use. So as I was telling you that we have all India-coordinated network project on soil biodiversity and bio-fertilizers, these are the different products which have been developed by this institute and these are being manufactured at the different locations of the country given to the industries and thereby contributing for the use of bio-fertilizers in India. And there also the recent developments on the front of India. So there are many different approaches, the mechanized seed coating on the different crops and these are the different mechanisms. Also a lot of awareness about the use of bio-fertilizers working with the farmers in the farmer's field. I was telling these small and marginal farmers more about 85% where the land holding is about two hectares of my average land holdings of these small and marginal farmers. So the balanced fertilizers indicated nutrient management where the bio-fertilizers should be. You have one minute left, Ashok, please. Please. This is about the, it's an urgent conclusion so we heavily depends on the steel chemical fertilizers and pesticides but the overuse and misuse of those are very, very unfavorable for the quality of the environment where the importance of bio-fertilizers in the present time is tremendous. As I was telling the role and benefits is well known and that is being encouraged and promoted in Indian context. And but there are a lot of works to be done like the selection and effective screens which are the multifunctional qualities and different crops, different soils, different agricultural zones. And also the quality control system is very, very important. Often, you know, often the spurious materials may be and where these not if it is stored properly the availability of the organisms. So that is why of many times the farmers, they apply bio-fertilizers but they do not get the response. So there are a lot of interventions and control mechanisms, those things also require that. Establishment of our fertilizer act also in India, we are working. So with these few, thank you very much. Thank you very much each of you for your kind patience. Thank you very much Ashok for this very nice presentation with this country, country vision of the state of the use of bio-fertilizers which I think that it's very enlightening. We go to the second presentation of today by Mrs. Makipa Riza from Natural Resources Institute, Finland with the presentation, what the good microbiota contributes to soil by your diversity. Riza, the floor is yours. So you can unmute yourself and share your screen please. Riza, you have to unmute yourself. Yes. Can you start please? Yes, we can see your presentation. Please put it in presentation mode and you can start. Thank you. And can you see the full screen now? Perfectly. Yeah, thank you. So my name is Riza Makipa and I will talk about what redwood microbiota contributes to soil by diversity. So this is not related to agriculture but it's soil by diversity in forest ecosystems. And we have been studying this kind of stand where we have all-growth semi-natural forest with lots of dead wood which is typical to Norway's bruce stancing in Finland and in Northern Europe. And from this particular site, we have identified 2,000 species of fungi in this one single stand. And we have analyzed that two-third of them live in decaying wood. So without this long-lasting substrate dead wood, they won't occur in forest. And especially those one-third, almost 600 species they exclusively inhabit wood. So they don't occur in forest soil. And this is what we have analyzed there who occur in soil and who occur in dead wood. And these are just 15 most abundant species and those who inhabit both substrates are here in the middle and those who are unique to wood are on right-hand side and on the left-hand side, we have species which are specific to soil. I don't go into the detail to the species. Some of you know these when you operate in similar ecosystems but maybe these are not typical species for tropical agricultural soils. But this is what we find in boreal forest soil and dead wood. And what happens in decaying wood, this is as I told long-lasting substrate and we classify these decay bases in decay glass first. It's recently fallen very solid dead wood and during the decay density decrease and in decay class five, this is very soft wood almost transforming to soil organic matter. And what happens to species richness in this substrate in wood, it's increasing all the time. So the highest biodiversity we see or observe in decay class five. And if we compare that to the soil biodiversity, soil fungal richness, we see that from soil samples which are taken very close from decaying wood, there is no change during this decay base of wood. Sorry, I'm talking about soil but this is soil sample taken just close to wood. And from area where we don't have decaying wood, species richness is at similar level. And more detail about species and functionally cruelty what we have. Again, I'm using these decay glasses and first is just recently dead wood fallen one and then five is the late decay base. And we see that white rotors dominate in early phase of decay and brown rotors come soon after them. And then if we have micro-asal species which are light brown in this figure and they start to invade decaying wood in decay class two and in late decay phase, decomposing wood is inhabited by micro-asal species and they are dominating group. And what happens to species? Here I have 30 most abundant species. You won't see these names but these are four different species and we see that in early phases of decay there is not dramatic changes in dominant species but then we go to later decay phase there is a transition to more diverse species. Community and total different species and further on in decay class five which is here again, different species and they are very close to common destruction which we observed from soil. This last column here is the abundance of 30 most common species in soil. After saying that it's not the surprise that when we analyze stable isotopes patterns we see that decaying wood in early phases from one to three they are rather similar but then something happens in late decay phase and isotope pattern seems to be closer to that what we observe from soil which is this code here different locations of soil and interpretation of this isotope pattern is that there is the active transfer of nitrogen carbon between soil and host plants. Further on we have analyzed that in decaying wood which is originally very solid hard for microbes during the decay there is the asymptotic nitrogen fixation taking place in decaying wood and that activity is really strongly dependent on temperature in all decay phases and optimum is somewhere in 25 degrees Celsius and it's also highly dependent on decay class. So in early decay nitrogen fixation activity is low and it's highest in relatively late decay phase and then drop when you go to decay wood which is dominated by mycorrhizal species but anyhow nitrogen fixation activities is there and because of this activity we also see that when decay process proceed from early decay to late decay this is the density which decrease during the decay then we observe increasing nitrogen concentration and also amount of nitrogen in decay wood increase so it's partly because of transfer from soil to decaying wood and partly because biological nitrogen fixation and early decay wood is also optimal substrate for seedlings to grow because there are mycorrhizal community available and higher nitrogen concentration or increasing nitrogen concentration so they have also nutrients in there. I will skip that quickly just saying that we identified also what species are active in nitrogen fixation and how number of niph copies increase with decay phase and then decrease in latest decay phase and observation is that these species who has capacity to fix nitrogen, they are many of them are assigned to older ritsubials and most common are metane oxidizing so we have a undirect evidence that there are species who has capacity to fix nitrogen and they might use methane as the energy source for that but this is undirect evidence for that. And then some conclusion from these studies we conclude that decaying wood has an important contribution to overall fungal diversity in forest ecosystems and many species, large proportional species are really dependent on this substrate and dead wood specialist species, so fungal species specialized on dead wood they might be lost from managed forest where we have very marginal amount of dead wood left because of the harvesting and interaction between soil and wood inhabiting punchy is tight and we show evidence that there is active transfer of carbon and nitrogen by punchy and microbiota inhabiting dead wood has importance to forest nitrogen cycling because of this asymptotic nitrogen fixation which is taking place in the dead wood and here are some of the references and I thank you for your attention. Thank you so much Faisa for this very interesting talk because well at least for me it's amazing to know new sources of asymptotic nitrogen fixation and how the transfers between wood, soil and plant proceed so thank you very much. Without any more delay we are going to the third presentation and by Mrs Esperanza Huerta, Wanger she's talking about us about earthworms Esperanza, cuando quieras Gracias Hello, good afternoon Good evening, good morning to everybody so I will try to share my screen Faisa, can you Ah, okay Yeah, okay So now with a mode of presentation Yes Then I do this to the side So yes I will go to the title Yes indeed I will talk about earthworms Microbial diversity on the conventional and organic farms and what is the interaction of them with pesticides in the presence of actual and inherited pesticides This is a team work done at Waggingen University and research but also with Universidad Politécnica de Cartagena among other institutions Okay, so earthworms as indicators of soil quality we know we have heard in this congress how important are earthworms on their soil invertebrates and earthworms are named also soil ecosystem engineers Okay, they promote several soil ecosystem services for instance organic matter decomposition, infiltration, migration and they can impeccable reflect what occurs above ground So if there is an affectation above ground, we can see it with a city of soil invertebrates and earthworms can inform a lot of that Okay, soil microorganism we have heard also during the congress how important are they just a previous presentation we will hear how they participate in the decomposition of material so they are responsible of different biochemical processes they interact as the earthworms with the actual and inherited soil conditions So but what happened what pesticides are present so we already know what we have heard that when there is a concentration of pesticides in soils there is microbial degradation where it is leaching sulphur runoff effect on soil micro and macro found including not target species percolation of pesticides in water table, deterioration of soil properties So this is some of the affectations mentioned by Migliani and Beach 2019 Also just to inform that in soils, unfortunately we don't have only one type of pesticides we have a cocktail of pesticides so this is important to mention because when the studies are done to test a pesticide they normally do with one only one single pesticides and not with a combination of them so we don't know really what is happening with the combination with these cocktails over soil invertebrates or over soil I invite you when you have time to read this work done by Hazen et al 2021 and then there are a cocktail of pesticides in soils some of them are actual and inherited when I'm talking about inherited for instance, DDTs we still can find DDTs in the soils so of course this is more abundant in the conventional or in the Australian farms and in the organic, unfortunately yes, we can find few concentrations few, few, few but also from the inherited pesticides well ok, so what was the aim of this study? The aim of this study was to inside of age 2020 project International Project Iber Farming this project enhances soil quality was searching the enhancing of soil quality through plant diversification and rotation we also heard this morning in the plenary how important is the diversification and rotation and diversity in soil so this project was also promoting this activities but also assessing earthworms and soil microbial diversity was also an objective of this study and of course to assess pesticides, residues and in those areas where they have done long-term studies long-term farms, so long-term rotation diversification ok, so well the study was done in the Netherlands in Groningen so we can see here the map at the north of the Netherlands and we took the samples in 2019 so here just to have a vision of the different farms so we have conventional farms and also organic farms and we assess them so the crops that were present are mainly potatoes you know that the Netherlands is famous also for the production of potatoes so we had farms with potatoes and also with fodder for animals so we took samples for microorganisms samples for earthworms samples for pesticides and we determined also the characteristics of the soils and the pesticide determination was done by this map ok so results yeah in relation to the earthworms diversity we found the highest diversity in the organic farms so this is something that we normally were expecting the highest diversity in organic farms and also in those farms where they have more practice in this activity so this is also very important to say because sometimes we want to find results in few years but we need to wait to have a significant results more than 10 years 20 years with this activity we can have a rich abundance and diversity of earthworms so this where the species found I'm not going to go to mention all the names because of the time but we can see that was a nice diversity in contrast the farms, the conventional farms they had only one color so only one species you can see it also yeah ok in relation to the micro organisms so what we can see here those organic farms are those here in the middle but the highest abundance for instance relative abundance of those micro organisms that are nitrogen fixation who fix nitrogen those are organisms that are related with good practices and with organic matter of the position indeed the moment is reflected below ground so those organisms that are more intervening in nitrogen fixation in the organic farms also here at the right you can see the groups for instance the proto bacteria yeah but because of the time I'm not going to go to mention all the names so what about pesticides so we found 17 pesticide residues DDT, proscihol carve AMPA AMPA is the first metabolic of life you have heard this on every site so we found that also and in this graph you are seeing the concentration of pesticides and the density of the earthworms so what is the relationship here Grosso modo is that when you find highest concentrations pesticides is lower the density of earthworms and here it is very clear how the highest concentrations and cocktails more diversity of pesticides you find less earthworms so it is important to mention it is not only the concentration but the diversity of these pesticides the cocktails is what is damaging the abundance and diversity of earthworms okay so also here clearly the correlation now here we are seeing concentration and here different types of families micro-organized families so we can see the highest abundance relative abundance is at the left side of the graph then how the concentration of the pesticides is increasing increasing you see less yeah it is like this Esperanza you have one minute left yes thank you almost finished yeah so yeah the right side you can see the groups of the micro-organized thank you and here in discussion earthworms and micro-organized diversity are clearly influenced by actual and inherited pesticides when stress factors are present that biomass abundance and diversity may decrease so yes just to mention first those are the practices in green that promotes diversity below ground so rotation perennial crops no tillage no chemicals and pesticides and thank you very much thank you for your attention thank you very much Esperanza for your complete and comprehensive report on this project and the results we are going to the next speaker who is Mr. Otanilo Augusto Silvestre and yeah the floor is yours you should unmute yourself and put your presentation in a presentation mode please hello yeah we can listen to you yeah it's all clear yes you can start okay are you are you starting okay you can if you can and share your screen that's good Danilo you can start whenever you want can you manage one minute please otherwise we can upload it from the GSP site from the meeting site if you have problems do you want us to upload it I can't find yeah do you prefer that we upload this presentation and then you indicate when to change the slide yes it will be better yeah Isabel can you manage this please that's good so Danilo you can start whenever you want and then you indicate when to change the slide the floor is yours please okay good afternoon everybody let me introduce myself my name is Danilo I'm from Brazil MPP student agronomy in the department of soil at the University of Rio de Janeiro it's a pleasure to be here today to talking about my research I complete my research in 2019 it was done in the molecular microbiology lab at the University of New Brazil in Canada with the collaboration of Professor Shadow Padding in today's presentation I'd like to show you the construction of Newton strained from the cyclotolerant soil back there to evaluate the contribution of the biology nitrogen fixation propellants the next slide is okay thank you so nitrogen nitrogen is the module limit nutrient for crop growth crops needs high inputs of nitrogen fertilizers and I know nitrogen fertilization impacts the environment because if you introduce a lot of nitrogen fertilizer on the crops production you can contaminate the soil and agro systems and biology nitrogen fixation is safe for environment it's well new in legumes such as soybean however it is necessary to understand better nitrogen fix bacteria in non-legumes crops such as maize, sugarcane, rice, wheat okay the next please so nitrogen fix bacteria is bacteria I use there is Sedomonas cng was isolated from the roots of an arctic grass in Canada Sedomonas is a strong candidate for development as soil in loculant for temperature and colder regions some studies with bacteria in loculation of canola tomatoes and lettuce has resulted in substantial promotion of satellite satellite roots grow okay this this is slide and nitrogenase complex it's very it's very very important for biology nitrogen fixation because nitrogen fix bacteria are able to convert nitrogen from atmosphere to ammonia okay please next slide here you can see nitrogenase structural genes nipid genes ADK okay here I show nipid genes locals in Sedomonas cng I use the deletion of nipid genes ADK in this seat so the objective this study to construct a mutant of Sedomonas with a precise marco less deletion of nipid genes including the nitrogenase complex to evaluate the contribution of biology nitrogen fixation crops of agriculture okay the methodology this work was I use Sedomonas cng 30 degrees for growth E. coli 37 degrees for growth media LB vector with our genes and biology 10 percent here here was steps for the construction this mutants first step one five sequences AB funky nipid genes by PCR after that join A and B fragments by overlap extension using PCR after that digest and insert into suicide vector and finally introduce this vector into Sedomonas by eletroporation okay here after after eletroporation I had transformation of Sedomonas with vector I I straight purify with LB eager with antibiotic after that I took single colony I put into media LB no antibiotic after that I I streak on the plate with LB with sucrose 10 percent for first the recombination after that I select and step five replicate plate on well be antibiotic and LB with sucrose 10 percent then identify columns that are sense to antibiotic after that streak purify on well be no antibiotic and confirm loss of genes by PCR okay result at PCR product confirms the deletion of nipid genes in the BNA of Sedomonas and we can see in the letter freezes properly a difference between wheel type and new of 4.3 cup a using any we can see in the bottle that nipid mutants growth white type was able to growth in this condition it can be seen by the change of color from yellow to blue so it show that nipid mutants lost its capacity of nitrogen okay discussion with the construction of nipid mutants it's possible to study the contribution of biology nitrogen fixation to crop growth the transfer of fixed nitrogen from sedomonas to crop plants would be a significant mechanism for plant growth promotion in this association the effect of transfer fixed nitrogen on nitrogen metabolism in the host plant can be evaluated okay you have one minute left okay in conclusion deletion of sedomonas by all models recombination was confirmed by presence of 1.2 pcr 2 columns of the deletion mutants were verified by sequence the amplification of the construction of the plant is an important strategy to understand the function of genes on the association between plants and bacteria okay would you like to thank you Prof. Shadow Palatin Prof. Leandro Zevedo Santos and Government Brazil Canada okay thank you thank you very much Daniela for this very interesting presentation showing us the detailed methods of how these genetic engineers procedures apply to nitrogen fixation genes work because normally we stay outside looking at the product but we don't see how it really works so thank you very much okay thank you well thank you for the discussion session now and I have seen that several questions have been already answered through the chat so please have a look but I have have chosen some of them in particular one question to Mr. Ashok Patra and this question is the following I wonder if the trend in use of bio fertilizers in northeast India is an increasing because maybe they already use other sustainable practices such as only certified organic practices in Sikkim also are traditional practices also being considered to make the bio fertilizer use successful in areas where success has not been observed so the combined use of bio fertilizers with traditional practices yeah if you can give us some information about this Ashok yeah you know in the of course in the northeastern states you know and the organic more of the organic agriculture is followed there and is also of lot of emphasis and focus of the government that this area use minimum fertilizers and maximum the organic practices and the use of bio fertilizers not really picked up in those because in the organic farming the bio fertilizers is also a very important component but and that is also cheaper inexpensive and sustainable but maybe one thing is that the easily availability in the area and proper demonstrations another aspect is the viability and these area is you know the acidic soil most of these areas acidic soil high acidic soils in many parts so they are survival of these different bio fertilizers in the regions also need to be looked into so different factors are there the dominant factors to be identified why the bio fertilizer is not really picking up that is what I can tell at this moment thank you very much I have chosen another question for Mrs Makipa and well she already answered this question with the references of two papers but I would like her to give more insight into the first question that she got the question is did you identify the specific species with asymbiotic nitrogen in the game wood and I think that the second question is what I would be interested in for you to answer sounds like the game would be used like a source of nitrogen for so what do you think about this possibility of using wood as fertilizer or as an amendment thank you for the questions and thank you for all the questions in the chat we identify species who have capacity to fix nitrogen but maybe some of you who work with the crop land species and crop species already recognize whether they are common species what you have seen I don't know because I have not been working with agricultural soils but what is interesting for me is that this soil microbiota is so intelligent complex that there seem to have bacteria who are using methane and fungi who are producing methane and others are using methane as a source of energy and so it's complex and inter-linkages between species are really great and in our system we have this decaying wood where nitrogen fixation is taking place we know that it's a favorable substrate for tree seedlings and and so tree seedlings with they might they might benefit from these nitrogen fixing bacteria already in our system but that's linkage to the crop species trees so they might be useful but I think that by studying our ecosystem in your region you might find directly those species what you need there. Thank you very much so there's still some research to do to see if it can be used in agriculture thank you well I have seen that most of the other questions have been being answered on the chat but I would like our last speaker to answer one question before passing to the second hour of our session our session well the first one was what target plants were covered in the aim of this study and if you are planning to do breeding house experiments comparing nitrogen fertilizer and be sitting at the plants so Danilo if you can I did not use any plants I did have no time but I would like to test in other crops for example canola it's a better association with bacteria and I would like I would like to test and you there you are okay so can you please mute yourself thank you I would like to pass on the crops using nitrogen fertilizer and it's bacteria from our grasses and it was 30 years ago but there's very little study about these bacteria in genetics is manipulated is to compare nitrogen fertilizer together bacteria yeah okay that's all okay well you can look at the chat for the answers to the rest of the questions since we are a little bit short of time well we have recovered a little bit the time before that we lost at the beginning but we will pass to the second to the second hour of our parallel session 5 so for those that are coming in for the second hour that's which is from other sessions it's a pleasure to welcome you to the second hour and my name is Ros Maria I am the chair of the ITPS of the Global School Partnership and I will moderate this last hour session um well I will kind of remind the presenters to give to the 10 minutes so that we have time for the question and answer session which is very important and then I will uh I will tell you at the minute nine of your intervention that you have only one minute left um before starting the second hour I kindly ask you to check the zoom chat you should click the icon at the bottom of your screen and then there will be some rules and information coming up and for the question and answer question please when you have a question to any of the presenters write the number of the presenter and then post your question on the chat they will be collected and then at the end of the four presentations we will be choosing some of them for the discussion for the like discussion the rest of the questions the presenters should answer them directly on the chat the host of the meeting is Yulita is here to help you for technical issues on the video on the sound whatever so write to her directly using the chat with a direct message to her and then without further delay I would like now to give the floor Mr. Manuel Languita Maeso from Spain so Manuel yes thank you very much everyone going to share the presentation can you see properly it's okay yes it's very clear so you can start okay so hi everyone my name is Manuel Languita Maeso I came from the Institute of Sustainable Agriculture in Spain and I'm going to present this a word that is called soil and physiological properties seasonality, plant niche and plant genotype affect bacterial fungi communities in olive orchard soil but let's start wondering why olive tree well I'll give you several reasons the first one is because it keeps the landscape conservation it's a millenary crop it can be cultivated in less accessible areas promoting the soil fissure it serves as an ecological niche for other species and because olive oil and table oil are the basic food for the Mediterranean diet talking about olive oil production we observe that the European Union is the main producer of olive oil in the world followed by Tunisia and Turkey and Spain is the main producer in Europe taking into account the Mediterranean Basin countries followed by Italy and Greece however this productivity and the phytosanitary status of olive trees is being affected currently by several pathogens among which we can find verticillium dahlia that is a soil borne basquare food that cause verticillium will there are several factors that can influence the development of this disease the plant age it has higher incidence in plants between 2 and 10 years soil humidity it's increased in an irrigated soil the soil and earth temperature the property is at 25 degrees and the growth management that it's increased with tillage the symptoms of the disease start with the chlorosis and the chlorosis of the leaves followed by the death of the branches and finally the death of the whole tree because of the abstraction of silen vessel where the silencer flow on the other hand olive like other living organism is colonized by a series of microbial communities that we call microbiome this microorganism can be beneficial, neutral and helpful for the host plant can break into the host through the lithosphere and the phytosphere mobilizing within the silen vessel in addition they interact each other and with a biotic and a biotic factor to express a series of key responses that affect on plant growth and plant health so my hypothesis is that the olive silen microbiome can be used as a control strategy to mitigate or suppress the development of verticillin growth and the objective of this work is to characterize the effect of soil physical chemical properties seasonality, plant needs, plant genotype on the assembly and shrift of the bacterial fungal communities present in olive located in the south of Spain so the experimental and the methodology start in the south of Spain specifically in the province of Haim and we selected three olive genotype white-legged cultivating in Spain that are Piqual, Arvequina, and from Toyu each of the genotype came from the same mother plant so they share the same microbiome within each genotype we selected four trees for the genotype and from each tree we collect a sample of root with a sphere and soil into China in autumn and in spring the sample different examples follow different treatment the soil were drying then go underwent directly to the DNA extraction the park the root and we made small woody chips with a sterile scalpel doing a extract that was used for DNA extraction and with the resource first we insert in sterile water and after sonicating we create an extract that we use for DNA extraction too then we perform the 16S and ITS libraries, the sequencing and the bainformatian statistical analysis and the results so that the talking about the bacteria we observed there is three main fields and bacteria and bacteria and we observed that these three fields reduces its relative abundance between autumn to spring we also identified a 271 genera and we observed that this genera differ according to olive genotype and plant niche being in Arbequina the genotype with the most genera and the plant niche in the soil talking about now about fungal community we observed that basidiomicota mortylamicota and ascomicota were the three main fields and basidiomicota increased its relative abundance from autumn to spring while mortylamicota and ascomicota decreased its relative abundance here talking about the fungal communities we detected a total of 106 genera and that vary according to genotype and plant niche too in this case, Piqua Arbequina show a similar genera and the same occurred with plant niche in soil and the rithosphere it's were quite similar also we measured different soil physical chemical properties and we detected a significant increase of phd between autumn and spring while we observed a decrease in water, nitrate, magnesium and sodium in addition we observed a significant increase of calcium in soil under Piqua genotype and reduction of copper and the soil of Frontoio genotype the region and diversity according to the region and diversity we observed less region and diversity in fungal community in contrast to the bacteria and also the root were the plant niche less abundant according to the seasonality we observed a slightly reduction of the region and diversity in bacteria from autumn to spring but this region and diversity remain quite similar when talking about fungal communities the principle coordinate analysis of with the unit practice and the vertidiversity give us the olive plant niche so we observed that the olive plant niche were significant in both population while the seasonality resulted in only for bacteria community but not in fungal communities and the olive genotype had a effect in this structure of the population so my final conclusion is that bacteria community so higher diversity that fungal community in soil resource fair and root that microbial community very mainly according to plant niche showing the greatest diversity the soil so in the diversity followed by the resource fair and the roots that the fungal communities were less affected by two environmental changes due to seasonality while the bacteria population resulted significantly affected and that the host plant genotype had a minor effect of the microbial that different genotype presented different microbial community each one so as a final I would like to share with you this take home message is that indicating only that the plant niche and seasonality strongly affected the diversity and abundance of distribution of microbial communities while the olive genotype so a relative minor role as a driver of microbial composition thank you for my group all the institution that founded this project and this is my mail and my Twitter account if you want to keep updated with all my research thank you thank you very much Manuel you have been very fast you have finished even before 10 minutes or before the 9th minute and also thank you for your well illustrated presentation showing us the factors concerning soil microbioma and including time and season so this is not very frequent to find and also nice that they vary depending on the only variety which is also very interesting okay we are going to the second presentation of the second session so I will give the floor to Mrs. Linda Maria Dimitrova from the Swedish University of Agricultural Sciences so the floor is yours and you can start talking about perennial crops for sustainable soil management same beauty benefit from cultivation of perennial soil in Europe yeah so you can start so good afternoon thank you for this very nice and important conference and thank you for inviting me to speak about our research so my name is Linda Maria Dimitrova Mortensen and I'm affiliated at the Swedish University of Agricultural Science in the southern most of Sweden okay now it's moving so perennial crops are more and more often suggested to be then we cannot see your presentation we can only see your outlook currently okay that's not very helpful let me try again okay is it better now yes good thank you so I start from the beginning my name is Linda Maria from Sweden and so perennial crops are more and more often suggested to be introduced into our crop production to provide different benefits especially to the soil ecosystem and also the surrounding environment and synopyram into medium or as it's called with its common name intermediate wheatgrass is one of those candidates so this species has been taken in for domestication at the land institute in Kansas in the US and it was selected from a hundred of candidates because of its promising features so now it's enrolled in the breeding program of the land institute and it is aiming to be a perennial cereal crop with a quite nice yield but also with the benefits of a perennial crop so it is still under development we don't really know where the yields are landing in the end right now the yields are very low but the environmental and ecological benefits might be very large so and when the grain is harvested for human consumption which is a very nice thing with this grass it is trade-barked as currency my presentation don't want to skip change slide but here it is so in comparison to annual cereal crops the perennials they are expected to provide a range of benefits as I said and also to contribute less to the negative influence agriculture have on the surrounding environment so for example the perennial crop is of course expected to require a low frequency of soil tillage resulting in lower magnitude of soil perturbation and potentially because of this perennial feature these crops will be better at acquiring the nutrients that they need and that means that the requirement for additional inputs as fertilizers is also lower and taking these two features together lower tillage and lower fertilization will lead to a lot of benefits such as improved organic matter content in the soil and reduced respiration and losses of nutrients via the atmosphere via the water leaching from the systems and also in the burning of fossil fuels in the different production and transportation and distribution in the parts of the process so these perennials also are expected to protect the soil habitats and here we have already seen results showing that the soil microbial biomass and the diversity in general is higher under the perennial stands compared to the annual cereal stands and also the microisal colonization as the previous speaker talked about also have a better potential to colonize the soil and the roots contributing to a better nutrient acquisition by the crops and if we have a more living environment in the soil the capacity of the soil to provide ecosystem services will also be better such as nutrient delivery nutrient decomposition and possibly also increased carbon storage and possibly there is also if the soils are improving to the expected account then the production system in its soul might also provide drought resilience being better of taking on heavy rainfalls and also coping with dry periods so just to give you a glimpse from our research which is quite new because the crop is new and the production system in our fields are also new we recently started so for example we went out in our Swedish fields you can see the full scale experiment here it covers 14 hectares of land so it's quite large and we are cultivating this currency crop so we grow it alone but we grow it also with leucine for a better nitrogen household in the system but we also sampled a one year standard currency in France together with PhD student Olivier Duchan at ISARA well he's not a PhD student any longer but he was at the sampling time so and Anna Bareiro my postdoc she was the one doing all the hard work in the lab extracting biomarkers for example migraic alpha fungi in the field from the field so our results here we have a lot of bars so I have given the most important one circle so here we have the amount or concentration of the bio indicator the biomarker for migraic alpha fungi and it's two different so that's why I have both black and white bars and in the left in this first figure we have samples from the Swedish experiment and we can see we have the conventional annual co-op organic annual co-op then perennial system and the perennial inter crop the one of alpha as well so here we can see that we have a much higher amount of the biomarker for mycorrhiza when we have the perennial crop so possibly there is an interaction between the crop and this particular group of microorganisms the results from France to the right they were not as clear we can see some slightly higher value in the perennial system compared to an annual serial but the system is rather young so the soil might need some time for maturing and also the annual field was managed with reduced tillage which also resembles the perennial stand in many ways so why is this important yeah that's why all are gathered here today we want to protect our soils both because there was something for themselves and there were organisms inhabiting them but also because they deliver a lot of different services to us for our survival and this is what we are doing in our research and we'll do the coming three years in a new project so we will dig down deep into the soils under perennials to study these different aspects of ecosystem services so we hope that we can be back with even more results later and Olivier and his colleagues has also proposed the uses as I said from the beginning the yields are very low still in this crop and there is a risk that it will not be taken into the crop production land because it doesn't give the farmer an adequate income so we are also thinking about how can this crop be used it can give father for animals, it can be used for bioenergy conversion and then the grain can be consumed as a niche crop or sold as a niche crop but it can also be put in the landscape to protect the environment so we combine environmental protection with production so thank you very much thank you very much Linda you have also been very good in sticking to the time and thank you also for showing us the additional advantages of perennial crops in Europe and how additional ecosystem services can be studied and quantified and thanks also for the nice watercolors in your slides they were very nice, thanks so we go to the following presentation by Mrs. Miriam Mementza from the Universidad Nacional Agraria La Molina from Peru and she will speak to us about volatile organic compounds produced by selected antagonistic bacteria again solid one-fit phantogenic fungi so Miriam there has been a change in the presenter so the presenter will be Miriam Mementza it's okay she's around thank you good afternoon everybody I will present let me put in full screen please good morning everybody my name is Miriam Mementza today I will present the research about volatile organic compounds produced by selected antagonistic rhizobacteria again soil borne phytopathogenic fungi this research was developed in the Universidad Agraria La Molina Lima, Peru okay we know that fungal pathogens can cause most of the diseases that occur in agriculture fields in my country, Peru fungal pathogens distribution is very wide powered by climate conditions especially the temperature and humidity these microorganisms are capable of affecting different tissues of plants causing causing symptoms such as leaf spud, rust wheel, blight, canker and roots it's well known that agrochemical has been used to minimize the impact of pathogen fungi on plant health and gel however they have adverse effects environment effects and also cause pesticide resistance for that reason it's an urgent need to reduce the use of pesticides and identify more sustainable crop protection strategies in this sense the use of plant growth promoting rhizobacteria could improve the quality of crops the idea of this study was to elucidate the antagonistic potential of two strains previously isolated from rhizospheric common bean plants and we will look for the potential of this strain to produce volatical organic compounds in order to control the micellium growth of three different fungi sclerotinia, fusarium and rhizoctonia in the methodology we previously isolated from from fields located in Viru la Libertad this state are located at 76 meters above sea levels we we collected a resource for samples from the common bean from the resource for common bean plant variety biochimum okay from these two strains we will look for the production of volatine organic compounds by the seedling play method bacteria slayed culture in triptychsoid broth at 28 degrees celsius and shaken at 150 rpm overnight after that 100 microlitre of the bacteria fermentation broth where it spread onto the bottom dish containing triptychsoid agar after that 5 millimeters of agar look of freshly growing micellium was taken from the margin of the colony using a sterile corker border and placed in the center of the fresh PDA plate it was then immediately inverted of the bacteria plate and sealed with parafilm the plates were incubated at 25 degrees celsius for 3 to 5 days we also considered controls the controls were only the fungi okay the antagonist activity bacteria through the production of volatine organic compounds was evaluated every day in order to see the reduction of the micellium growth now in order to identify the perfil of these organic volatine compounds produced by these rhizobacteria strains able to inhibit these fungi we used the gas chromatography combined with mass spectrometer technique for that 2 millimeters of bacteria suspicion was grown in 25 millimeters of tryptic soy broth culture at 28 degrees celsius for 24 hours then the solidified microfibre was inserted in the health space deal just about the inoculate medium and allowed to deliberate for 30 minutes the solidified microfibre was inserted and the sore at 70 degrees celsius for 1 minute in the injection in the injection of the injector we used gilium for the carrier gas at constant flow of 1 millimeter per minute in the results the results for strain were able to inhibit micellium growth of these fungi between 31 to 97 percentage as you can see for example the micellium of the fusarium was very different it turns transparent as you can observe in the case of frisctonia and sclerotinia we observed that micellium was inhibited if you can compare with the contrast you can see that change in the fungi that was in contact with the bacterial colonies okay volatil organic compounds profile measured by chromatography in combination with the mass spectrometer analysis was different between these two strains in total we identify 42 volatil organic compounds in the case of the first strain called tbps2.4 this analysis revealed for compounds with major abundance in this case we detected the medial delimony diametal disulfide metal butanoid and nonanon with a major abundance between all the compounds that we detected by this technique in the second strain tbps1.6 this analysis revealed also for principal peaks with a major abundance in this case we detected diametal disulfide delimony etal metal butanoid we also made an observation a light microscope of the chain in order to see the chain of the miscellaneous in the miscellaneous of the sclerotinia in this case we saw a a a minute thank you I almost finished in this case we saw morphological abnormalities in the E5 compared to the contrast as you can see the E5 is more thin and you can see also a vacuolation of the Ctoplas conclusions these two strains produce 42 volatile organic compounds detected by the gas chromatography combined with a mass spectrometer from them there is very important compounds that you can see diametal, disulfide, delimony and two non-anone and they have a new reporter for being antagonistic for half antagonistic activity against different soil-bore pathogens the result for us is to represent a preliminary screen for the PGPR from the Result for Automobile Plans in Peru that may start point to optimize the production of bioactive extracellular compounds that inhibit the miscellaneous growth of these harmful phytopathogens thank you very much for this very nice presentation on showing us another way to manage biodiversity by isolating the specific products from rhizobacteria to fight against pathogens we hope that further research will be successful well we will go to the last speaker of today Mr. Majid Rostami from Malayad University in Iran the floor is yours he will speak about the effect of different species of mycorrhizal fungi also on growth and physiological characteristics of sorghum in cadmium contaminated soil so whenever you want you can start Majid are you around I think that there are some problems maybe we are listening your voice we are hearing you you have your presentation ready yes so if you can share your screen you can see not yet you have to go below to the green button then if you have problems we can upload it for you no I try to start again if you have problems we can upload it Isabel can you share your screen and then you can tell Isabel when you want to change the Majid Rostami has started now yes that's good thank you very much so it works can you put it in presentation mode and start hello everybody thank you for coming here and participating in global symposium of soil biodiversity my name is Majid Rostami from Marajir University and the title of my presentation is effect of different species of mycorrhizal fungi in cadmium contaminated soil okay let's start with introduction and definition literally mycorrhizal means fungus root so mycorrhizal refer to the fungus and rhizal refer to the plant roots therefore we can define mycorrhizal as a useful relationship between the plant root and fungus this symbiotic relationship could establish in the root so in the root surface or inside the plant root so we have two main group of mycorrhizal as you can find here the first one is extra mycorrhizal in this group the hyphae don't penetrate the root cell and this bilateral association observed in about 10% of plant family the second type is endomycorrhizal which in this group more in this group you can find that hypha penetrate into the root and this type are more common and about 90% of plant have this type of association with fungus here you can find mycorrhizal extra mycorrhizal and as you see here in extra mycorrhizal there is visible structure that you can see without microscope but considering the endomycorrhizal you have to use microscope to see different structure of fungus and this is another picture related to endomycorrhizal and you can see that in endomycorrhizal there is different structures such as physical and harvest school harvest school structure have a care and increase the contact surface area between the hyphae and the cytoplasm of the cell the role of harvest school mycorrhizal is to improve uptake and translocation of water and different plant nutrients such as phosphorus and nitrogen to the plant but this relationship is not free for the plant in return about 20% of photosynthetic product such as sugar and lipid are transferred to the fungus here I summarize the K benefit of mycorrhizal fungi they are two mechanism for increasing absorption ability by mycorrhizal in physical mechanism mycorrhizal provide a large surface area for absorption of water and nutrient but chemical mechanism is related to different chemistry of cell membranes in mycorrhizal I have to mention also that our vascular mycorrhizal have a good ability to enhance plant tolerance for different environmental stress such as osmotic stress in some case however this relationship is not beneficial for example when the fungus is harmful for the plant it act like a phatogen and when the plant feed from the fungus this relationship is not positive also not all of the plant in the world have ability to start a mycorrhizal plant association for example aquatic plant and many of hypofit are not able to start this type of association for the terrestrial plant also we can say that about 90% have this type of ability so you can find some major important crop among the less 10% that are non-mycorrhizal plant for example sugar beet and canola are among the plant that couldn't establish this type of relationship this is the result of different study that I used for literature review and you can see in all of them mycorrhizal have a positive effect on the plant growth especially in the root growth let's start with material in order to study the effect of to our bascular mycorrhizal species on sorghum plant under cadmium stress I do the current experiment as a factor here and here we have to concentration of cadmium 0 and 40 mg per kilogram of soil and for level of mycorrhizal application in the F0 F0 or control we didn't use any fungi in the F1 treatment we use galomous massae and in F2 treatment we use galomous intradices in F3 treatment we use simultaneous we use simultaneous lead galomous massae and galomous intradices so it was corrected from the field and dried and sterilized in order to delete the native microorganism and all of the physiological measurement was down based on the scientific method I know that we have short time so it has to result here you can find summarized result as you see in this slide in cadmium contaminated soil relative water content of leaf significantly was lower than soil without cadmium different mycorrhiza treatment increased relative water content in cadmium contaminated soil the highest amount of relative water content this one was major after co-inuclation with both of mycorrhiza species in the absence of cadmium mycorrhiza treatment here significant effect on chlorophyll content but under the cadmium stress condition as you find here as a result of application of mycorrhiza the leaf chlorophyll content increased and this is you can find the catalyst activity in normal condition and without cadmium stress this one the catalyst activity significantly was lower and in this condition mycorrhiza application didn't have any effect on enzyme activity but under the cadmium stress condition please the catalyst activity increased decreased significantly after application of mycorrhiza you know when we mycorrhiza the activity of catalyst significantly decreased and it means that plant tolerant and plant facing with lower stress condition cadmium stress significantly decreased the root weight in the absence of cadmium mycorrhiza treatment didn't have significant difference in contamination with both of mycorrhiza species in this one significantly increased the root dry weight leaf phosphorus content under cadmium stress condition was lower than cadmium treatment in the absence and presence of cadmium application of mycorrhiza significantly increased phosphorus content here you can see that even when we didn't have cadmium stress this one application of mycorrhiza increased phosphorus content here you can find the relationship between the root biomass and phosphorus uptake or phosphorus content higher root biomass resulted as you find here one minute left the higher root biomass resulted in higher phosphorus in leaf of and finally see the change in different treatment in contaminated soil significantly was lower than on contaminated soil in both soil application of mycorrhiza increased the root dry weight but only in contaminated soil simultaneous application of two mycorrhiza species significantly increased the root biomass in conclusion I can say that application of our vascular mycorrhiza improved the studied characteristics of sorghum the positive effect of mycorrhiza treatment in contaminated soil was higher under cadmium stress condition mono-inuculation with Columbus-Massie or Columbus-Intradices had less positive effect on plant growth so I can suggest that simultaneous application of different species of our vascular mycorrhiza to enhance plant growth in contaminated soil is recommended thank you for your attention and I am at your service for any question that's very good thank you very much well at the beginning for your comprehensive review on the role of the mycorrhiza and also how they can be managed in a synergistic way to improve the productivity of sorghum on contaminated soil okay well I have here some questions on the chat some of them have already been been answered I will start with the last one that I have now on the screen is in Spanish but I'm going to translate it for Majid the mycorrhiza through what mechanism it could reduce the content of cadmium in these plants I don't know if the effect is the process is the removal of cadmium or that they can grow with cadmium in the environment so could you please answer this question the role of mycorrhiza is that mycorrhiza absorb cadmium and in the root surface but it didn't translocated this absorbed cadmium in the material part of the plant and in this case plant can't tolerate more okay I hope it has been understood I will shortly do it in Spanish mycorrhiza absorb cadmium and I transfer it to the areas and it doesn't generate any adverse effect okay then I will many questions as I said at least what I noted from the first speaker to Manuel Anguita Maeso he already answered very in a very complete way the two questions that were on the chat then the questions for directed to Linda Maria de Mitrova she already answered it then there are a couple of questions to Mrs. Miriam Memenza and well the first one is can you describe the benefit of using SPME instead of using a gas sample for GCMS I don't know exactly what this I mean it's not my field of expertise so could you please answer that Miriam okay no problem the the principal difference between those techniques is that the SPM allow us to collect the the major of the compounds present in the medium broth is the principal difference because this fiber can absorb a lot of compounds including the compounds that present in a little quantities so this is the principal difference that we found thank you there is a second question for you the question is I wonder if you are considering extracting these compounds and testing them as effective by your controls in greenhouse experiments as I know that currently some legume plants struggle from fusarium infections which is a major reason for the local division by some farmers this will be a breakthrough which is important from the future of legume crops as a protein source as an nitrogen source and great work yeah okay thank you for that question yeah some trades some trades in the greenhouse conditions we tested a lot of trades with different abilities antagonistic abilities including that production of a lot of organic compounds and also another activity like production of antifungal compounds production of the iron collating syrup for example and production of the hydrolyte enzymes that we saw in the laboratory condition that control the mycelium growth so we already made some trills in the greenhouse using legumes and other crops also and we saw very interesting results that these strains demonstrate that can control different diseases caused by these fungus risottonia, these solar fungus so now we started that other compounds that could be involved in this antagonistic activity so especially we try to identify the name of these compounds so also we made some fields, field trails also okay, thank you okay there is another question in Spanish for the Majid I'm going to translate it the measurement of glomalin is an indicator that maybe there is a decrease of cannium content in the soil the question is in your study have you considered the measurement of this protein in the treatments and the content of cannium in the plant? no we didn't measure this type of protein we just measured the traits related to the plant most physiological traits of sorghum we didn't measure anything related to the soil you know and then if you look at the chat there is another question for you from Maria Lisa I already played this you have played it the last one I don't see the answer the AMF fungi results are significantly higher in the cannium can it maybe yes it is possible because when we delete the other micro-organisms from the soil one type of micro-organisms especially in our study will be dominant and in this case we will see the more positive effect comparing the the soil because we delete the other micro-organisms from the soil yeah okay good so I don't see thank you very much for all the answers and for the interactive discussions that has been very active in this chat I also see that there is an open I have from Erika Bauer there is an open 24 hours free to join a café since we don't have any coffee break there is a place to to have informal chats and well if there are no more questions if there are no more questions we would close this session and first I would thank only all the presenters I think that well I will try to summarize a little bit the different aspects we had very interesting presentations showing the benefits of soil biotomy different aspects of food production mainly no gene availability, pest control and growth of crops in polluted soils and also how we can manage them some of the presentations today have shown us the inverse relationship between conservation of the soil and biodiversity I think that this is a general trend in most of the presentations and also the specificity of the soil of micro biomas depending even on the species of the cultivar use also in the case of AMF how they increase productivity and how they can be managed in relation to the control of local pathogens we have seen how some volatile compounds from rhizobacteria can be isolated that used to act against them we have been convinced also on the potential of nitrogen fixing bacteria to be used as inoculants to non-leguminous plants and thus reducing nitrogen fertilizer inputs and also on the existence of sufficient nitrogen fixing fungi living in that wood and I want to finish with the results of the first presentation that we have seen that it is possible to convince farmers and governments on the advantages of using biofertilizers as substitutes of agrochemicals for fertilization and pest control so this technology transfer is possible and can be successful to achieve a more sustainable agriculture and a higher soil biodiversity at the same time so thank you very much to all it has been a very fruitful session and I hope to meet you tomorrow in the last day thank you so thank you very much and see you tomorrow thank you much to all, see you tomorrow thank you


24th. It's Wednesday. It's 11 o'clock. That can mean only one thing. It's Trump week. I'm Tim Apachele, your host. And this week, like last week and the week before that, we have so many things to discuss and so little time in which to discuss it. I want to jump right in and talk about the title of this show is testing needs to slow down. At the Trump rally in Tulsa, Oklahoma, we heard Donald Trump say the following. When you do testing to that extent, you're going to find more people. You're going to find more cases. So I said to my people, slow the testing down, please. The next day, the media was all over that one saying, are you kidding? There's 120,000 American deaths. And it's through testing. Do we try to eliminate or minimize COVID-19 cases and deaths? And the White House, to their challenge, had to come up with some answer. And that some answer was, well, come on, you guys, Donald Trump was kidding. It was tongue in cheek. Well, for those who've been watching Donald Trump for the last three and a half years, we know it's not tongue in cheek. And I'll tell you why. Because on May the 15th, he said the following, testing was overrated. When you test, you have a case. And when you test, you find something's wrong with people. If we didn't do any testing, we would have very few cases. And then on 15th of June on Twitter, he says, if we stop testing, we would have fewer cases. So if it's a joke, it's an ongoing running joke. But we all know that Donald Trump doesn't have a sense of humor to begin with. So to throw his staff under the bus on the 23rd in an interview with Scripps TV, with one of the station reporters, he said, if you did slow down, frankly, I think we're way ahead of ourselves to tell you the truth.  COVID-19 is in his rearview mirror, and it's on to the election. And 120,000 Americans' deaths be damned. Who cares? I need to get re-elected. And with that, I'm going to jump in and say hello to our guests. Special today, we have Jay Fidel. He's made a return trip. We have Stephanie Dalton and Cynthia Sinclair. Welcome, everyone. Hello. Thanks for having us, Tim. Oh, it's a pleasure. And I'm glad to see you all back. And Jay, I'm just going to jump right into you regarding the comment from Donald Trump at the Tulsa rally. Your thoughts? What was your initial reaction when you heard this yet again from Donald Trump? No, this is his way of dealing with the problem, lie. And the number of lies go on and on. He lied to begin with to say that we don't need testing. And he lied about what he said. And he lied about the joke. So what we have is an ongoing pattern. Pattern is hardly the word for it. It's a full tilt boogie on lies from the beginning till now. And just to add one thought to that is that we cannot have a president in office who lies to us. This is what happens. 120,000 lies. What do they think about him? Well, and what about those who are in an age category where their immune system is compromised or they're older? And do you think that they're thinking that Donald Trump has their best interest in mind as he moves forward? Nobody who's rational thinks Donald Trump has their best interest in mind. But I'm sorry to say there are a lot of people in this country who nevertheless, irrationally think that believe him, which is a problem that we have to deal with. I think we should examine that all of us to see why this country, anybody in this country believes him when he's a congenital liar. We can all have our opinions about whether or not Donald's fringe support, which is beyond the 42% or whatever number you want to sign to that, whether or not they're shifting their opinions. But the polls seem to be suggesting just that, that white males are starting to shift away from Donald Trump and as well as college-educated white women. The suburban crowd is shifting away from Donald Trump in the polls that are coming out. Do you think that continues to widen? No, I did. I do think it continues to widen. I think it largely has to do with recent events. Possibly the Tulsa rally is an important event for that. And every single poll that's been reported shows that he's lost ground. He's in 35% as against Biden's way over 50% all across the board. And that gives you two thoughts.  And the photographs of him getting off the helicopter after Tulsa, he looked pretty bedraggled and unhappy. A broken man, if you will, because of his under attendance at that rally. So yes, I think it's clear that he's desperate. And the other point that comes out of it is desperate people do desperate things. You're going to see him in the next 120 days do the most remarkable things to, as you said, Tim, to play to the base, to play to the people who would still support and believe him and so forth. You're going to see him do the most remarkable things about election day too. God knows what kind of emergencies we're going to have.  So it goes both ways. One is he doesn't look so good and people see that. The other is he's going to do some crazy stuff. All right. Thank you, Jay. Hey, Stephanie, what was your opinion about the Tulsa rally, specifically him trying to slow down testing, but also other things that we saw there. And that was quite frankly the sparse crowds, the crowds that he had thought were going to be in excess of 40,000 people. He even prepared an outside seating stage and seating arrangement for overflow guests. Yet we all saw that he packed them in like sardines wherever he could just to try to get some density images. But if you pan the cameras up a little bit, the whole upper tier was vacant, empty. The source of my spot of good feeling for the week, which I nurtured my little flame. Anyway, yes. And because it missed expectations and by two thirds, at least two thirds, because they said the arena was only about third full, plus he had the outdoor overflow. So it missed by a long shot. And that sounds like a little bit of gaming out there in the country. It sounded like it was something maybe from youngsters, teens, people on the internet playing the game of let's sign up for the rally, even though we're not going to go. So a little taste of his own medicine perhaps there was another a little bit of a bump to the good feeling that it's about time tables turn some. So I also found that I didn't watch the whole thing. I saw whatever clips I saw on the other programs, but it didn't look like he had practiced anything or put together anything new. There really wasn't a coach. Well, I think he did put something in there new and he talked about for about 15 minutes, the fact that he had to explain his his his shuffle, you know, down the ramp at West Point, he went into great lengths to try to convince the audience that there's nothing wrong with me. And it was just the fact that it was a metal ramp and there was no railings and it was wet and boy went the liberal media loved to watch me fall down and make hay of that. So he spent 15 minutes just discussing that. Yeah, he's getting old. So yeah, and that that was kind of a little turnaround fair play too, given what they whoever did to Hillary during the last round. I'm not speaking as Hillary saw, you know, as like my camp, my favorite, but I'm just mentioning that a lot of that went on with her. Yeah, so I've seen some Republican ads that are touting that the Democrats want to do all of these things while they stay off in their mansions, and they themselves keep themselves safe. And they of course keep their incomes, but yet they spend everybody else's money on all of these communistic and socialistic policies.  And for instance, here is Donald Trump standing up on the stage when he did invite participants to the stage. It was extraordinarily distance. I mean, like off kilter for the TV camera, maybe 40 feet. Protecting himself from, you know, any possibility. And of course, he never mentions, and I don't know why Fauci and these other people don't mention that he has protection just around the clock and also people checking on him continuously. Well, that's a good point.  I find it really amazing that because you had so many empty seats that someone right then and there didn't say, Hey, you know what, we can actually space some people out here and protect them. I did, why didn't that, but they are not in that headset. So they're not following directions and they're not buying into any of the scientific evidence. So this is another version of seeing their belief systems and their devotion. Yeah, yeah. Okay. And he's not doing them well as that kind of a foe leader. Yeah. Okay. Thank you very much, Stephanie. Hey, Cynthia. Were there any jaw dropping moments for you by watching some of the excerpts from the Tulsa rally? You're melt on. I actually watched the whole thing and the whole thing made my jaw drop. Oh, okay. So no one, no one particular moment, the entire, the entire rally. Okay. The entire rally, but before they even started, they all had to sign a release saying that they wouldn't sue him if they got sick, which was the jaw dropping moment to start with for me.  And then he thinks they, they started to applaud. So of course he decides he wants to do it again, drinks a little more water and then shows how tough he is by throwing it down on the stage. And I, that was a special moment. I, I, I agree with that.  Do you have to be to, to, to do something like that?  I don't know. I was just blown away by all of that. It was a lot to take in. And I will tell you that the Lincoln project did make great, great use of his slowing down testing in the form of a political ad that was, in my opinion, quite, it was a home run. And we'll see how that plays not only in, in, in, in Oklahoma, but we'll see how it plays in Arizona and all these other states that he insists on cramming people together and exposing them to COVID-19, all for his political pleasure. Right. And two days later, not even two or two days later, we've got all of the scientists. We've got Fauci. We've got Redfield.  Yes, we need to socially distance. The numbers are going up and up and up. Texas and, and Arizona, both just today have the highest number of cases they've ever had before. They're breaking records. And I guarantee you in two weeks, you'll see those numbers accelerate from where they are even today. Yes, we will. I agree with you. But I'm sure on Corona, Coronaville, Jay, you'll have that down and you'll be all over that topic. Yes, we will. Yeah. All right. Well, let's, one last, one last thing. Let's touch on at the Tulsa Rally. He used the term for the second time now, kung flu. And that seems to have made a mark for his loyal base, but it's not selling well to the rest of the public. And I'm just kind of curious, Jay, let's go to you on this. What was, what was, what's his strategy about bringing up kung flu and No, it's racist. It's racist. It's no other way to put it. It's been characterized as racist when he started doing it in the first place. Everybody knows the signal he's sending and the signal is, I don't like the Chinese and you shouldn't like the Chinese either. They're bad people.  What's interesting is the remarkable thing that happened at the same time. Bolton wrote a book and published the book this week. I think it came out yesterday, but it was parts of it were out for a while in which he said that Trump made a deal with Xi Jinping to give him a break on agricultural product pricing and other things in return for Xi Jinping's supporting Trump in the election. God knows what nefarious things that means. It could be, you know, the same kind of social media gambit that Putin did last time around. Maybe we'll have it from both sides this time. But, you know, Trump was, he said he was joking at the time. He said, oh, I was only joking when he said that he was going to try to get Xi Jinping to do that. Now it's clear it's documented. He is trying to get Xi Jinping to do that at the same time. He's been quoted on TV saying, if George Stephanopoulos is saying they called me up, I'd listen to what they had to say about my opponent. Why not? But now we have hard copy. It wasn't him waiting for a call. It was him making a call. And by the way, it was only three days ago that Pompeo was here in Hawaii, furthering those discussions about the pricing on agricultural products. And that has to be part of the discussions, this unholy deal. You never heard more about that. We never even really heard much press that Pompeo was here. And we certainly didn't hear what they talked about. And we didn't hear exactly what, you know, the nature of this deal was. The only person that comes from right now is Bolton. I believe Bolton. Well, I, you know, I think you said it yourself in the next 120 days, we're going to see all sorts of outrageous things. Jerry Nadler was interviewed saying, you know, this discussion about firing the Manhattan District U.S. Attorney Jeffrey Berman. And he goes, yeah, bar should be impeached, but we got 120 days. I doubt that's going to happen politically. So to what degree do we see travesty after travesty in the next 120 days? And what can you do about it? And Bolton's book, whether I think it's probably factually accurate, and, you know, getting China involved with Donald Trump and the reelection here, to what degree do we stop it? Can we do anything about it? No. And as you said, and we both said it's going to get worse. I mean, the Trump thing with Michael Flynn, who pleaded guilty on a couple of occasions, the Department of Justice, you know, tried to withdraw that prosecution. Judge Sullivan in District Columbia found that he wasn't so quick, he's not going to be so quick to allow the withdrawal of the charges. Because it's obvious that everything that Barr does is a pass through from Trump. They are one and the same. Okay. And Judge Sullivan put it out for some amicus briefs. And Judge Sullivan, you know, hired a master in order to help him make the ruling. And then the government, that is, Barr appealed, the government appealed that ruling by Sullivan to the D.C. Circuit. And this morning, if you haven't noticed, the D.C. Circuit, through a judge who was appointed by Trump, okay, ruled that the case had to be withdrawn and that Barr rules. So to go to your point, Tim, that's what's going to happen from year to election day. The most outrageous things. One commentator said, this is the end of the rule of law. I agree. Well, that was also in testimony of the DOJ, former DOJ senior official. I believe his name is Donald Ayer. He said exactly that. This is the end of the rule of law. And then we had Aaron Zelinski, a former prosecutor in the Department of Justice, basically said, hey, I know we were told that Roger Stone had preferential treatment from Barr and Donald Trump. So we kind of hands off. We put hands off on it. So these are coming to light. They're being known at what can one do about it. And I think your answer is good, not a whole lot. So Cynthia, let me ask you something. Right now, Barr is showing his hand explicitly that he is the lap dog and the lackey for Donald Trump. Whatever Donald Trump wants, Barr is more than happy and willing to do it. Here's what Barr said this week. Right now, a foreign country could print up tens of thousands of counterfeit ballots and it would be harder for us to detect them, which was right and which ones were wrong. That was for a Fox News interview. So William Barr seems to be playing hand in hand with the idea that Donald Trump thinks that all mail-in ballots are fraudulent. And if you watched a little bit of his Arizona rally yesterday, it got a lot scarier. He basically said the Democrats are rigging this election. They're rigging it now, and we can't allow that to happen. So like 2016, Donald Trump is throwing that up in the air saying, if I don't win, this whole thing was taken away from me and from you, most importantly, voters, that my election has been stolen. Right. And you're right. That's exactly what he's doing. He's setting the stage for that exact thing. We also have to remember the element of projection because everything that he does and says basically is a projection, which means he's doing it. So when he says the Democrats are going to cheat, that means he's going to cheat. When he says the Democrats are doing such and such, it means that he is doing that thing. So I think it's always important to remember, but he does all this gaslighting stuff, and that's what it's all about, so that by the time we do get, and we talk a lot about that here on the show, right? He was talking about last week. Yeah. And so by the time we do get to the election, if he does lose, he's going to say, oh, it was rigged. Everybody cheated. And he's already laid the groundwork for that. And that's what gaslighting is. So it's the perfect example of outright gaslighting. Well, when you have your top cop of the country, the Attorney General, basically on the same song sheet as that all mail-in ballot should be perceived as fraudulent or ripe with fraud. And we have several states that do it. What's the end result here if he doesn't win the election? Are we going to have the Boogaloo factions, the high military extreme right take to the streets, or are we going to peacefully get through all this? No, I don't think we will peacefully get through it. And I think a really good sort of microcosm example of it is just this weekend, there was a rally or not a rally, I'm sorry, a protest set up for Black Lives Matter to come forward in a small town in Ohio.  These people that were there to represent counter-protesting were armed with rifles.   And instead of getting in between the protesters and the counter protesters, it seemed like the police were almost pushing the counter protesters forward to be closer to the protesters. So you're thinking they're trying to agitate? I'm sorry, what now? You're thinking the police are actually trying to agitate the crowd versus to keep it separate? You can totally tell. And I think the thing that struck me the most was that they're calling all the people, the protesters, not the counter protesters, but the protesters, are being called traitors. Traitors, go back where you came from.  And I thought, now where did we hear that from? Where did we get this from? Oh yeah, that's what Trump has been saying. Obama was a traitor. The Democrats are traitors. And so now his base has picked up that moniker and off they run with it. And they're attaching it to everyone. All right, Stephanie, what are your thoughts about William Barr and the fact that he's joined, fully joined Donald Trump with the fraud of mail-in ballots and of course all the influence with either Roger Stone or Michael Flynn cases? Well, George Washington University Law School has written him a letter or made a statement about his malfeasance and the need for him to resign. So there's professional pressure coming up. That was his alma mater, was it not? This is alma mater, yes. We all know people from there, but not that we all know, but that's a big school and right there in D.C. But I think that he is, we've got to put on our seat belts, strap on the seat belts because he is going to probably maybe be taken to the Supreme Court. This case, that is one of Sullivan's options is he can choose to move it on up to the Supreme Court or there's another option to do something within the district circuit court or whatever it is. Jay, maybe you can talk about that later. But I think that we need to remember with our strap on seat belts, we got to keep our eyes on the prize because this is just going to continue because there's no holds barred on him. And the bigger picture is if we get a Mr. Biden in, then the questions are going to be, what are we going to do to protect ourselves from this in the future? Is that much of a piece of the agenda? Is that going to be? And of course he can't talk about that ahead of time, but he's got a lot of cleanup to do. And then we've got to decide, this is going to be a very interesting issue for the president to take on. And I'm sure with Obama's influence and support and his constitutional background, maybe we can figure out how to get protected. Does the House Oversight Committee, do they get any bang for their buck by having him testify in July? William Barr? Yes, I'm very pleased. I don't know that he'll go. He's king of the hill. He may or may not go. We knew nothing about anything until it leaks out one way or another. And here again is the media issue. Are they doing enough of their job?  And now here we're in the mess of, it's always about the latest piece that's upsetting. What about the research and the background stuff? Okay, like you were saying earlier in the chess game, what's going on on the back bench, Tim? And who, what's coming next? And let's hear more. I mean, we're going to be overblown by all of these judges who are inexperienced at least, if not incompetent, and considered so by the ABA and other certifying organizations. What about all this? What is the rest? Well, as Jay said, expect more in the next 120 days. I don't have an answer for that. I don't know if any of us do. I'm not sure. Half of Congress has an answer on how to apply the brakes to this runway train of a president. But we're almost out of time. Let's switch gears. JTU, Donald Trump just extended the ban on visas, entry visas for foreign workers. You think that's going to have an impact on farm produce and employment? What are your thoughts about that? Well, he does, he makes these moves without having a plan, aside from the bad temper of them and the corruption of them. I mean, he just got finished negotiating the sale of additional agricultural products to China. So now he's cutting off a labor supply that is necessary to grow those agricultural products. Go figure. We're in a reopening, aren't we? We need labor. We need to engage labor in the field. We need to get back to productivity. So you terminate a source of an important source of labor. I can't figure that out. Don't you think a lot of Republicans are all in favor of illegal immigration in order to pull the crops out of the fields? It's just as you said before, Tim. This is his way of playing to the anti-immigration crowd in the country. He thinks they're going to solidify behind him and vote for him.  It's really back more of the same. It's the wall. He played that card yesterday in Arizona. He talked a lot about the wall again. So in fact, he visited the wall and he wrote his name on the wall. So I don't know what more he could do. He's certainly not building a wall. I think he can do a lot more and I think he will do a lot more.  He wants to be reelected. He will put it another way. He wants to stay in power and we're going to see the most incredible machinations from him. He has no boundaries, no limits, moral, legal or otherwise and he is destroying. In many regards, he's already destroyed the balance of power in the government. In many ways, he's divided the country beyond redemption. We're in a terrible crisis right now. Agreed and it doesn't seem like anyone can do anything to stop it. Only the election, but he is trying to undermine the election. Imagine getting up and saying, I want to stop the post office because people who vote by mail will vote against me. Unbelievable statement. He wants to suppress voting. He has Republican parties in so many states working to do that. He is trying really hard and then he says, I do not necessarily agree with the election. If I win, I agree with it. If I lose, it was all rigged. He said the same thing in 2016. And that's dangerous. That's promoting insurrection and quite frankly, he may succeed partially in this. Okay, we have run out of time. Real quick. Cynthia, predictions for next week? Real quick. Well, I think we're going to hear more about Barr because obviously they're in the process of looking at him. And hopefully, Stephanie, you'll have another bit of good news to hold on to coming up this week. All of us will have a little bit of good news. The one thing that I say is good news from all of this. Nobody died in Tulsa and it could have easily been that. Nobody died yet. Because of that. Yeah, thank you for that. Okay. Hey, we're out of time. Jay Fidel, thank you for joining us. Stephanie Dalton, thank you. Cynthia Sinclair as always. Thank you for joining us. I'm Tim Appachella. This is Trump Week. We'll see you next Wednesday, 11 o'clock. Aloha.