Antifungal potential of colloidal silver

alternative method in seeds

pathogens control

Coordinator Professor: Authors names:

Assoc. Prof. Adrian Peticila Oana Venat
Assoc. Prof. Beatrice Iacomi

International Student Symposium "Hortus Academicus March 31, 2017 - Bucharest, Romania
 We are not alone
Pathogens on humans
Variola viruses The new worl was infected by europeans over the ocean; in 1967 over 2 mil people
died from variola, even the cure is available.
Spanish influenza 1918 - between 50-100 mil people died over the world; consider the most lethal
pandemic disease from the beginning of human kind.
Black death - bubonic fever?/plague? consider by scientists the first form of Ebola; 1348 - killed
over half on European population; spread by a bacteria call Yersinia pestis; if is a plague, is still
here.
Malaria - cause by for Plasmodium microbes species and hosted by mosquitoes and humans as well;
still here.
Tuberculosis - The great white plague - discover in Egyptian mummies caused by a air speeded
bacteria called Mycobacterium tuberculosis; 8 mil people are infected yearly and over 2 mil are
dying.

And so on......Yellow fever, AIDS (spread by virus), cholera (spread by bacteria Vibrio cholerae)

International Student Symposium "Hortus Academicus March 31, 2017 - Bucharest, Romania
 Pathogens on plants/ Fungi
Crt. Fungal pathogen Romanian name Author of fungal description
number

1 Magnaporthe oryzae Arsura orezului Ralph Dean

2 Botrytis cinerea Putergaiul cenusiu Jan A. L. van Kan

3 Puccinia spp. Rugini (specii de Puccinia) Zacharias A. Pretorius

4 Fusarium graminearum Fuzarioza graului Kim Hammond-Kosack

5 Fusarium oxysporum Ofilirea fuzariana Antonio Di Pietro

6 Blumeria graminis Fainarea graului Pietro Spanu

7 Mycosphaerella graminicola Septarioza frunzelor de grau Jason J. Rudd

8 Colletotrichum spp. Antracnoze Marty Dickman

9 Ustilago maydis Taciunele comun al porumbului Regine Kahmann

10 Melampsora lini Rugina inului Jeff Ellis

Source: Journal Molecular Plant Pathology nominate which fungal pathogens could take a place in a Top 10 based on scientific/economic
importance.
The survey generated 495 votes from the international community, and resulted in the generation of a Top 10 fungal plant pathogen list
for Molecular Plant Pathology. (2012)

International Student Symposium "Hortus Academicus March 31, 2017 - Bucharest, Romania
 Pathogens on plants/Bacteria

Crt Bacterial pathogen Romanian name Author of bacterial description

Number
1 Pseudomonas syringae pathovars Arsura bacteriana John Mansfield

2 Ralstonia solanacearum Ofilirea bacteriana Stphane Genin

3 Agrobacterium tumefaciens Cancerul bacterian al plantelor Shimpei Magori, Vitaly Citovsky
4 Xanthomonas oryzae pv. oryzae Arsura bacteriana a orezului Malinee Sriariyanum, Pamela Ronald
5 Xanthomonas campestris pathovars Nervatiunea neagra a verzei, arsri Max Dow
bacteriene
6 Xanthomonas axonopodis pv. manihotis Arsura bacteriana a maniocului Valrie Verdier

7 Erwinia amylovora Focul bacterian al rozaceelor Steven V. Beer

8 Xylella fastidiosa Boala lui Pierce la vita de vie, Marcos A. Machado
declinului maslinului etc
9 Dickeya (dadantii and solani) Innegrirea bazei tulpinii la cartof Ian Toth
10 Pectobacterium Putregaiul umed al plantelor George Salmond
carotovorum (and P. atrosepticum)

Source: Journal Molecular Plant Pathology nominate which bacteria could take a place in a Top 10 of plant viruses based on their perceived
importance, scientifically or economically.
The survey generated 458 votes from the international community, and resulted in the generation of a Top 10 bacteria plant pathogen list
for Molecular Plant Pathology. (2012)

International Student Symposium "Hortus Academicus March 31, 2017 - Bucharest, Romania
 Pathogens on plants/Viruses
Crt Bacterial pathogen Romanian name Author of bacterial description
Number
1 Tobacco mosaic virus (TMV) Virusul mozaicului tutunului Karen-Beth G. Scholthof

2 Tomato spotted wilt virus (TSWV) Boala petelor de bronz la tomate Scott Adkins

3 Tomato yellow leaf curl Virusul rasucirii si ingalbenirii frunzelor de Henryk Czosnek
virus (TYLCV) tomate
4 Cucumber mosaic virus (CMV) Virusul mozaicului castravetilor Peter Palukaitis

5 Potato virus Y (PVY) Virusul y al cartofului Emmanuel Jacquot

6 Cauliflower mosaic virus (CaMV) Virusul mozaicului conopidei Thomas Hohn and Barbara Hohn

7 African cassava mosaic Virusul mozaicului african al maniocului Keith Saunders

virus (ACMV)
8 Plum pox virus (PPV) Virusul varsatului prunului Thierry Candresse

9 Brome mosaic virus (BMV) Virusul bromusului Paul Ahlquist

10 Potato virus X (PVX) Virusul mozaicului x al cartofului Cynthia Hemenway

Source: Journal Molecular Plant Pathology nominate which viruses could take a place in a Top 10 of plant viruses based on their perceived
importance, scientifically or economically.
The survey generated 250 votes from the international community, and resulted in the generation of a Top 10 viruses plant pathogen list
for Molecular Plant Pathology. (2012)

International Student Symposium "Hortus Academicus March 31, 2017 - Bucharest, Romania
About 80% of plant diseases can be traced to fungi, which have
a great capacity to reproduce themselves both sexually and Alternaria alternata
asexually.

More than 300 plant viruses have been identified, new strains
continually appear because these organisms are capable of
mutating.

Bacteria can be spread by water, insects, infected soil, or

contaminated tools.

Pesticide resistance is a natural ability of a biotype of an

organism to survive exposure to a pesticide that would normaly
kill an individual of that species.

Development of Resistance: Organisms can develop resistance

by:
1) developing a means of detoxifying the pesticide;
2) altering the target site (part of the bodys metabolism affected
by the pesticide) to reduce its sensitivity or the pesticides
ability to bind; Fusarium oxysporum
3) decreasing pesticide penetration.
International Student Symposium "Hortus Academicus March 31, 2017 - Bucharest, Romania
 Objectives

Integrated pest and disease control intend to limit the effects on plants that
chemicals can have, the impact on the environment and on the humans health.

The main objective of the research willing to finding solutions refers to non-
chemical, non-toxic and non-residual pesticides for the replacement even partial of
conventional chemicals that already producing drug resistant on diseases and pests.

Colloidal silver and SNP are a new direction in this area

 Materials and Methods
I. In vitro tests on mycelial growth
Pathogens Host CODE PROVENANCE

Fusarium nivale R.sativus seeds F065 FR* France

Aspergillus niger R.sativus seeds An1* Romania

Aspergillus flavus R.sativus seeds Av1* Romania

Alternaria brassicicola R.sativus seeds Ab43* France

*fungal collection

II. In vitro tests on seeds

Pathogens Host Isolate CODE Seed PROVENANCE

R.Sativus
Alternaria brassicicola Cherry Bell Ab43 Romania
(Starsem)
France
seeds harvested from plants inoculated
R. Sativus at the end of flowering and at silique
Alternaria brassicicola Ab43
Flamboyant 5 formation stage with Ab43;
experimental station FNAMS, Brain sur
L'Aution
International Student Symposium "Hortus Academicus March 31, 2017 - Bucharest, Romania
 Materials and Methods
I. In vitro test on mycelial growth

Methods: poisoned food technique

Tested compounds
CONCENTRATION
TRADE NAME PRODUCER CODE
(ppm)
Water with colloidal silver
(food supplement) Medicer M 3.75, 7.5, 15

Colloidal silver Crafts V 3.75, 7.5, 15

V15 ppm M15 ppm

Results are expressed as Inhibitory concentration IC50

and IC 90, based on inhibition of mycelial growth.

International Student Symposium "Hortus Academicus March 31, 2017 - Bucharest, Romania
 Materials and Methods
II. In vitro tests on seeds

Methods
II A: Colloidal Silver Seed Treatment (artificially contaminated seeds)

Tested variants
Variant CODE
Disinfected seeds
SD
Disinfected Contaminated Seeds
(seed artificially inoculated with A. brassicicola spores suspension
SDC
105 spores/ml)

Disinfected Contaminated Treated Seeds

with colloidal silver 19 ppm (crafts), 30 minutes SDCT

International Student Symposium "Hortus Academicus March 31, 2017 - Bucharest, Romania
 Materials and Methods

II. In v itro tests/seeds

Me thods
II B: Colloidal Silver Seed Treatment (on seeds harvested from inoculated plants)

Tested variants
Variant CODE

Non-Disinfected Seeds
(seeds harvested from inoculated plants) SND

Treated Non-Disinfected Seeds

SNDT
with colloidal silver T19 ppm (crafts), 30 minutes

International Student Symposium "Hortus Academicus March 31, 2017 - Bucharest, Romania
 Materials and Methods
II. In v itro tests on seeds
Results expressed as:

1) Developmental stages (0-3 grade scale)

0 1 3 5 7 9

2) Disease severity (0-9 grade scale)

3
3) Attack incidence on seedlings (%) = n/N x 100
( n = number of attack seedlings, N = total number of seedlings)

International Student Symposium "Hortus Academicus March 31, 2017 - Bucharest, Romania
 Results and discussions
I. In vitro test: Sensitivity of Fusarium nivale to colloidal silver

M edicer CS Crafts CS

Fusarium 065/Crafts colloidal Silver

100
y = 7.4621x - 25.32
80 R = 0.9933
CI50 CI90
60
10.09 15.45
40

20

0
0 5 10 15 20

Fusarium 065/Medicer colloidal Silver

35
y = 2.6472x - 6.87
30
CI50 CI90 25
R = 0.9336

21.48 36.59 20
15
10
5
0
0 5 10 15 20

I nternational Student Symposium "HortusAcademicus March 31, 2017 - Bucharest, Romania

 Results and discussions
I. In vitro test: Sensitivity of Aspergillusniger to colloidal silver
M edicer CS Crafts CS
Aspergillus niger 1/Crafts colloidal Silver
90
80
y = 5.2457x - 1.19
70 R = 0.9671
CI50 CI90 60
9.75 17.38 50
40
30
20
10
0
0 2 4 6 8 10 12 14 16

Aspergillus niger 1/Medicer colloidal

Silver
25
CI50 CI90 20 y = 1.8143x - 9.525
32.8 54.85 15 R = 0.8929
10
5
0
-5 0 2 4 6 8 10 12 14 16

I nternational Student Symposium "HortusAcademicus March 31, 2017 - Bucharest, Romania

 Results and discussions
I. In vitro test: Sensitivity of Aspergillusflavusto colloidal silver
M edicer CS Crafts CS

Aspergillus flavus/Crafts colloidal Silver

70
60 y = 6.1981x - 32.54
R = 0.8929
50

CI50 CI90 40

13.75 19.77 30
20
10
0
0 5 10 15 20
-10
-20

CI50 CI90
>15 >15

I nternational Student Symposium "HortusAcademicus March 31, 2017 - Bucharest, Romania

 Results and discussions
I. In vitro test: Sensitivity of Alternaria brassicicola to colloidal silver

M edicer CS Crafts CS 90.00 Alternaria brassicicola/Crafts Colloidal Silver

80.00 V
70.00
60.00
CI50 CI90 50.00

7.01 17.55 40.00 y = 3.7937x + 23.415

30.00 R = 0.912

20.00
10.00
0.00
0 5 10 15 20

35.00 Alternaria brassicicola/Medicer Colloidal

30.00
Silver

25.00

20.00

15.00 y = 1.0587x + 15.87

R = 1
CI50 CI90 10.00
32.25 70.06 5.00

0.00
0 5 10 15 20

I nternational Student Symposium "HortusAcademicus March 31, 2017 - Bucharest, Romania

 Results and discussions
II. A In v itro tests /artificially contaminated seeds

Effect of craft colloidal silver on seed development, necrosis index and incidence of attacked
seedlings

Treatment DI IN Attack frequency (%)

SD 3 0 0
SDC 1.4 7 100
SDCT 19 ppm 2.7 3.92 21.60

Disinfected seeds

International Student Symposium "Hortus Academicus March 31, 2017 - Bucharest, Romania
 Results and discussions
II. A In v itro tests /artificially contaminated seeds
Effect of craft colloidal silver on seed development, necrosis index and incidence of attacked
seedlings

Development index Necrosis index Attack frequency

3 100%
3 2.7 7
8 1
2.5
6 SD SD
2 SD 3.92
1.4 0.5 SDC
1.5 4 SDC 21.60%
SDC
1 SDCT 19 ppm 0 SDCT 19 ppm
SDCT 19 ppm 2
0
0.5 0
0
0 0
1
1 2

Necrosis index - Ab43 attack on seed

Mycelial growth of Ab43

International Student Symposium "Hortus Academicus March 31, 2017 - Bucharest, Romania
 Results and discussions
II. B In v itro tests on seeds harvested from contaminated plants

Effect of craft colloidal silver on seed development, necrosis index and incidence of attacked
seedlings

Treatment DI NI Attack frequency (%)

SND 2.9 4.62 36.60

SNDT 2.91 4.05 21.10

SND SNDT

International Student Symposium "Hortus Academicus March 31, 2017 - Bucharest, Romania
 Results and discussions
II. B In v itro tests/seeds harvested from contaminated plants

Effect of craft colloidal silver on seed development, necrosis index and incidence of attacked
seedlings

Development index Necrosis index

2.91
2.91
4.62
4.8
4.6
2.905
4.4 SND
2.9 4.05
4.2 SNDT
2.9
4
3.8
2.895
3.6
SND SNDT
1
Attack frequency

36.60%
40.00%
30.00% 21.10% SND
20.00% SNDT
10.00%
0.00%
1

International Student Symposium "Hortus Academicus March 31, 2017 - Bucharest, Romania
 Conclusions and perspectives

Efficiency of colloidal silver on mycelial growth inhibition of different

fungi isolated from radish seeds (starting at 15 ppm).

Seed treatment with colloidal silver at 19 ppm reduces Alternaria

brassicicola incidence on seedlings (about 80%) compared to control.

Ong oing research on foliar treatment on p lants with colloidal silv er, p athosystem A.
brassicicola - cruciferous p lants.

Preliminary results on:

Colloidal silver as preserv ative solution to improve cut flowers self life;

Potential disinfected age nt on fungal meristems culture contamination

International Student Symposium "Hortus Academicus March 31, 2017 - Bucharest, Romania