10.6092/issn.2531-7342/8613
application/pdf
2531-7342
8613
en
Maria Agnese Sabatini
Italian Journal of Mycology
Collembola, soil fungi, wheat plants: images of their interactions
Text.Serial.Journal
https://italianmycology.unibo.it/
47
Editorial
2018-10-09
2018-10-09
2018-01-29
2020-01-20
Ital J Mycol
Maria Agnese Sabatini
2018/10/09
10.6092/issn.2531-7342/8613
University of Modena and Reggio Emilia
2531-7342
en
Collembola, soil fungi, wheat plants: images of their interactions
https://italianmycology.unibo.it/article/download/8613/8372
47
G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613                      Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




                     Collembola, soil fungi, wheat plants:
                          images of their interactions


                         Gloria Innocenti
                 Department of Agricultural and Food Sciences Alma Mater Studiorum University of
                 Bologna, Italy; e-mail: gloria.innocenti@unibo.it

                       Maria Agnese Sabatini
                Department of Life Sciences University of Modena and Reggio Emilia, Italy;
                e-mail: mariaagnese.sabatini@unimore.it



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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613            Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




                                     Why do we publish these images?

    Trophic interactions of Collembola with some soil fungi and their effects on
    wheat plants health and animal life have been our field study for about twenty
    years. The collaboration between a zoologist and a plant pathologist, both
    passionate about their organisms, has allowed to explore this field with different
    eyes.
    The results have been published along the way, and we hope that they have been
    useful for those working in this field of study. We also hope that they will be
    useful in the future for all the possible applications the interactions Collembola–
    fungi might generate.
    We have documented our works with images that have been only partially used
    for oral presentations or posters.
    We want to share and provide free access to these images. The Italian Journal of
    Mycology makes it possible. The images are introduced by a brief text.



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DOI: https://doi.org/10.6092/issn.2531-7342/8613        Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




       The soil is a reservoir of organisms with the same aim: feeding !
       Collembola, commonly known as springtails, are among the most
       important groups of soil mesofauna as they live in all climatic
       environments from Arctic and Antarctic to tropical areas, and are largely
       dependent upon fungal food sources.

        Collembola interact with propagules of soil fungi, and in particular with
       those of the following fungi that influence plant growth and health:

       - pathogenic fungi which are responsible of diseases

       - antagonistic fungi which act against pathogenic fungi by different
       mechanisms reducing their inoculum and activity

       - arbuscular mycorrhizal (AM) fungi which promote plant growth through their
       role in nutrition and protection against pathogens.

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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613                      Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




    Since the disease is the result of the competiton between pathogenic,
    antagonistic and AM fungi        for rhizoplane/rhizosphere colonisation
    (competence) the question is :




                                                           can trophic activity of
                                                           Collembola modulate
                                                        plant-fungi interactions by
                                                   reinforcing or switching the existing
                                                        relationship between fungi?




               We are going to exam these interactions separately

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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613   Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




              Interactions between Collembola and soil borne
               plant pathogenic fungi: in vivo and in vitro tests




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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613                 Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




                                    Collembola used in in vitro tests




                                         Folsomia candida Willem
                                         Mesaphorura krausbaueri Börner
                                         Kalaphorura tuberculata (Moniez)
                                         Onychiuroides pseudogranulosus (Gisin)
                                         Protaphorura armata (Tullberg)



              In the image specimens of P. armata feeding on F. culmorum hyphae and sporodochia

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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613                           Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




               Fungal pathogens and symptoms they cause on wheat plants




            Gaeumannomyces
                                                   Fusarium culmorum
            graminis var. tritici
                                                   Bipolaris sorokiniana
                                                                                            Rhizoctonia cerealis )

                                                   These fungi are responsible for important
                                                   diseases of winter cereals indicated with the
                                                   generic name of foot and root rot. They usually
                                                   coexist in the same soil portion, even though one
                                                   is the prevalent depending on edaphic conditions.
                                                   Severe infections cause white-headed plants as
                                                   shown in the image.
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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613          Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




     In in vitro multiple choice tests, Collembola were able to interact
     contemporaneously with R. cerealis (Rc), G. graminis tritici (Ggt), B. sorokiniana
     (Bs) and F. culmorum (Fc) inoculated at different times to have colonies with
     similar size when animals were introduced into the Petri dishes.




                                                   Ggt           Rc                         Ggt
                 Rc
                                                                 Bs                           Fc
                     Bs                            Fc



        In the images distribution of P. armata specimens 3 h (left) and 50 h (right)
        after their introduction into the Petri dish with the fungal colonies on agarised
        medium. No animals are feeding on Bs colony in both images.

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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613                Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




         Distribution of Collembola at different times. Total animas = 120




                                                                     Fc
                                                                                        M. krausbaueri




                                                                                    O. pseudogranulosus
                                                                     Ggt
                                                                      Rc
                                                                      Bs


                                                   P. armata.
                                                                                        K. tuberculata

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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613         Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




               Fc                                        Specimens of P. armata
                                                         feeding on F. culmorum (Fc)
                                                         and G. graminis tritici (Ggt)
                                                         colonies. Note the absence of
                                                         hyphae in the animal feeding
                                                         area.




                                                   Ggt
       Note also that the gut color
       depends on       the hyphal color,
       carmine red for Fc, and black for Ggt.
       Faecal pellets of the same color of
       hyphae are also present as indicated
       by arrows.



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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613       Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




      No animals are on young non- sporulating colony area of B. sorokiniana (a) ,
      whereas collembolans feed on mature colonies of the fungus, characterised
                   by an abundant presence of melanised conidia (b).

              a

                Rc
                                                   b




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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613         Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




     Subsequent long term (150 d) experiments with the parthenogenetic
     collembolan M. krausbaueri showed that:

     . mycelia of F. culmorum, G. graminis tritici and R. cerealis were appropriate
     food for reproduction, all eggs were viable, all eggs developed up to hatching,
     and all newborns reproduced by thelitoky

     . a diet of G. graminis tritici determined early maturation, oviposition of more
     eggs over shorter time, early loss in fertility and early death

     . a diet of F. culmorum determined maximum eggs production later

     . a diet of R. cerealis determined the production of the highest total number of
     eggs, but late maturation, oviposition of more eggs very late, and very late death

     . a diet of B. sorokiniana conidia determined the lowest fertility.


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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613   Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




             Interactions     between      Collembola      and
             antagonistic fungi belonging to Trichoderma spp.




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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613             Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




      In in vitro multiple choice tests, in the presence of G. graminis tritici (Ggt), F.
      culmorum (Fc), R. cerealis (Rc) and the non sporulating young Trichoderma
      harzianum (Th) colonies, P. armata specimens avoided the Th colony, even if
      more developed than others.




                                               Ggt
                                                         Fc




                                                   Rc
                                                        Th


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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613                                    Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




         Distribution of Protaphorura armata specimens at different times on the
         mycelia of pathogenic fungi and of antagonistic fungus Trichoderma
         atroviride present contemporaneously in the same Petri dish.



                                               Hours after animals
                                                       Tempo        introduction
                                                             di osservazione

                                 1         2          6         12        24        36          48

             F. culmorum       8.7 a     10.0 a     11.2 a    18.2 a    17.7 a     18.7 a     17.7 a

             G. graminis       8.0 a      6.0 a     6.2 a      6.2 b    6.2 b      8.0 b      7.5 b

              R. cerealis      9.2 a     10.2 a     10.0 a    3.5 bc    4.2 bc     1.0 c       2.2 c

            T. atroviride      1.2 b      0.5 b     0.2 b      0.2 c     0 c        0 c        0 c




                   In the same column, values followed by the same letter are
                   not statistically different ( DMS, P < 0.05)




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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613               Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




                                              Can the Collembola spread fungal
                                                         propagules?

                                            Are conidia and hyphae viable after
                                                         gut transit?




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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613                 Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




     Conidia of F. culmorum (Fc) were palatable and no toxic for
     Collembola. They were damaged during the gut transit, indeed
     very few faecal pellets produced by animals fed by the fungus, gave
     rise to colonies of the fungus when transferred on agar medium.




        Conidia of Fc produced in pure
        colture.
                                                   Gut of a P. armata specimen with Fc conidia.

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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613                      Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




         Melanized macrohyphae of G. graminis tritici (Ggt) were fed by
         Collembola. No fungal colonies raised from faecal pellets produced
         by animals fed with the fungus when these were inoculated on
         agarised medium.
                                         Colony of Ggt in Petri dish on agarised medium with the
                                         typical melanised macrohyphae.




                                            Gut containing portions of Ggt macrohyphae.
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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613               Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




        Conidia of Trichoderma were palatable and no toxic for Collembola

                                                          b



         Colony of Trichoderma sp. in Petri dish on
            agarised medium, with masses of                    Mesaphorura krausbaueri
             mature conidia green in colour.
                   a                                  c




                                                               Protaphorura armata



          Collembola feeding on highly sprulating Trichoderma colony. Note the green
          colour of gut and faecal pellets (a). Particular of gut with masses of conidia (b, c).
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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613                    Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




                                                   Colonies of Trichoderma sp. at different
                                                   times of growth, arising from faecal
                                                   pellets produced on selective medium.




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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613      Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




                       Protaphorura armata                 Folsomia candida


          Back     Petri     dishes   with
          Trichoderma colonies from faecal
          pellets produced by different
          species of Collembola fed with
          conidia of the fungus.

                                                   Onychiuroides pseudogranulosus
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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613   Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




        From Petri dish to microcosm: effect of interactions
       between fungi and Collembola on wheat plant disease




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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613                    Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




                                         Petri dish




                                                      Microcosms: glass tubes (35
                                                      mm in diameter by 300 mm high)




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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613                    Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




                                                   Each microcosm


           - contains 150 g sterile sand mixed with 1.5 g sterile millet kernels
           colonised by F. culmorum or G. graminis tritici + 3 seeds of wheat cv
           Creso + 50 specimens of P. armata

           - was maintained for 4-5 weeks under controlled conditions, then
           seedlings were removed and animals collected by floating and counted

                The density of springtails corresponds to about 40,000 individuals
                per square meter, comparable to the density of Collembola in the Po
                Valley cultivated soils.

                The presence of Collembola did not damage plants, on the contrary
                increased plant dry weight, and reduced the disease severity caused
                by both pathogens.

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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613             Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




          Effect of F. culmorum (Fc), P. armata (Pa) and their interactions (Fc+ Pa) on
          wheat seedlings compared with untreated plants.




                       Untreated                   Pa   Fc     Fc+Pa

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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613                          Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




                  c           c
                                                                                Plant total dry
                                                                                weight (mg)
                                                      bb
                                         a
            Untreated        Pa




                             Disease index (%)
                                                                 b
                                                                                                a




                       Bars with the same letter are not statistically different ( DMS, P < 0.05)

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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613          Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




      Effect of G. graminis tritici (Ggt), P. armata (Pa) and their interactions (Ggt+Pa)
      on wheat seedlings compared with untreated plants.




                                 Ggt + Pa          Ggt    Untreated



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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613                        Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




          70
                                                                    Plant total dry weight (mg)
                  c
          60

          50
                                                       b
          40

          30

          20
                                       a
          10

           0
                untreated            Ggt           Ggt + Pa
                                        a



                                  Disease index (%)                     a

                                                                                                            a




                 Bars with the same letter are not statistically different ( DMS, P < 0.05)

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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613               Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




          What is the effect on plant health when F. culmorum (Fc) , G. graminis tritici
          (Ggt), and Collembola (C) were in the same substrate?


                                                   √ Collembolans reduced disease
                                                   severity.

                                                   √ Ggt was the prevalent pathogen.

                                                   √ The number of animals containing
                                                   only or mainly Fc conidia in their gut
                                                   was significantly higher than that of
                                                   specimens containing only or mainly
                                                   portions of Ggt macrohyphae.

                                                   √ The preference of collembolans for
                                                   Fc showed in Petri dish was
                                                   confirmed.
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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613                Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




                                    NO
                                 DISEASE !                    Do Collembola spread
                                                                  the disease?




                from Fc            untreated       from Ggt

      When springtails collected from microcosms containing propagules
      of F.culmorum (Fc) or G. graminis tritici (Ggt) were released into
      microcosms containing sterile sand and wheat seeds, no disease
      symptoms were observed on wheat seedlings.
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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613       Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




          Effect of Collembola-Trichoderma interaction on plant health

        Trichoderma harzianum (Th) inoculum,
        consisting of power formulation of conidia
        + ground rice kernels, was coated to seeds
        (c), or mixed with substrate (s) as Ggt
        inoculum just before wheat sowing and
        animals addition.




            Gut with Th conidial masses (blu arrow)
            and Ggt macrohyhae portions (red arrow).
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G. Innocenti, M. A. Sabatini
DOI: https://doi.org/10.6092/issn.2531-7342/8613                        Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




                                              Collembola: yes biocontrol

                   Collembola + Trichoderma in soil: no biocontrol

               Collembola + Trichoderma coated to seeds: yes biocontrol

                                               ..…probably when Trichoderma conidia were mixed
                                               with substrate, animals that fed on them were
                                               attracted not only by conidia, but also by ground rice
                      Why?                     kernels used to produce the inoculum. Their feeding
                                               activity on Ggt was not enough to control the disease.
                                               When Trichoderma was coated to seeds, it was
                                               present in a much smaller amount than Ggt and more
                                               localized, therefore collembolans turned mainly to
                                               Ggt as food source, reducing disease.


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DOI: https://doi.org/10.6092/issn.2531-7342/8613   Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




                     What is the effect of Collembola, arbuscular
                    mycorrhizal and pathogenic fungus interaction
                                   on plant health?




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DOI: https://doi.org/10.6092/issn.2531-7342/8613        Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




     Wheat plants in plastic bottles containing
     sterile sand with P. armata, G. graminis tritici
     (Ggt), and the AM fungus Glomus intraradices
     (Gi) inocula in growth chamber.




                                                              Disease index (%)


                                                         b

              Studies indicated compatibility                               a                 a
              between P. armata and Glomus
            intraradices to reduce the disease          Ggt             Gi+Ggt        Gi+ C +Ggt
                         index (%).

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DOI: https://doi.org/10.6092/issn.2531-7342/8613   Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




                           From microcosm to mesocosm




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DOI: https://doi.org/10.6092/issn.2531-7342/8613     Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




                Microcosms




       Mesocosms: polypropilene boxes (200 x 400 x 300 mm) with containers,
       each consisting in metallic frame enveloped by a wrap to allow passage of
       hyphae but not of animals (100 x 100 x 400 mm) in growth chamber.

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DOI: https://doi.org/10.6092/issn.2531-7342/8613           Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




     Each container was filled with 500 g of soil inoculated with G. graminis tritici
     (Ggt) propagules, then wheat seeds were sown and 350 specimens of P. armata
     were introduced to the surface. Three containers were placed in each microcosm.
     After three weeks plants and animals were collected, disease index and dry
     biomass were determined.




                                                                   Also under these
                                                                conditions, Collembola
                                                                        showed
                                                                biocontrol effect vs the
                                    2
                                                                disease caused by Ggt.
       1
                                                   3



     Containers with wheat plants grown in the substrate inoculated with Ggt in
     presence (fig. 1) or in absence of Collembola (fig. 2), untreated plants (fig. 3).
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DOI: https://doi.org/10.6092/issn.2531-7342/8613             Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




                                             Concluding remarks
        It has been found that Collembola feed preferably on
        pathogenic rather than on antagonistic or AM fungal
        propagules, and they don’t spread pathogenic fungal inoculum,
        thus springtails may reduce the inoculum of pathogens without
        counteracting the activity of beneficial fungi.



                                     Agronomists could study the effect of
                                     cultural practices to these animals,
                                     particularly for glasshouse crops.


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DOI: https://doi.org/10.6092/issn.2531-7342/8613                  Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




                                                   Main works
       Sabatini MA, Innocenti G (2000). Functional relationships between Collembola and plant
      pathogenic fungi of agricultural soils. Pedobiologia 44, 467–475 .
       Sabatini MA, Innocenti G (2001). Effects of Collembola on plant-pathogenic fungus interactions
      in simple experimental systems. Biology and Fertililty of Soils 33, 62–66.
       Sabatini MA, Grazioso P, Altomare C, Innocenti G (2002). Interactions between Onychiurus
      armatus and Trichoderma harzianum in take-all disease suppression in simple experimental
      system. European Journal of Soil Biology 38, 71-74.
       Sabatini MA, Ventura M, Innocenti G (2004). Do Collembola affect the competitive relationships
      among soil-borne plant pathogenic fungi? Pedobiologia 48, 603—608.
       Sabatini MA, Innocenti G, Montanari M, Ganassi S (2006). Survival and feeding activity of
      Protaphorura armata in different composts. Pedobiologia 50, 185-190.
       Innocenti G, Ganassi S, Montanari M, Branzanti MB, Sabatini MA (2009). Response of plant
      growth to Collembola, arbuscular mycorrhizal and plant pathogenic fungi interactions. Bulletin of
      Insectology 62, 191-195.
       Innocenti G, Montanari M, Ganassi S, Sabatini MA (2011). Does substrate water content influence
      the effect of Collembola-pathogenic fungus interaction on plant health? A mesocosm study.
      Bulletin of Insectology 64, 73-76.
       Innocenti G, Sabatini MA (2018). Collembola and plant pathogenic, antagonistic and arbuscular
      mycorrhizal fungi: a review. Bulletin of Insectology 71, 71-76.


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DOI: https://doi.org/10.6092/issn.2531-7342/8613                  Italian Journal of Mycology vol. 47 (2018) ISSN 2531-7342




        Could Collembola modulate the plant disease by their
               feeding activity on fungal propagules?


                                                   WE THINK SO!




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