KARAULNAYA-2 CAVE AS A SOURCE OF COLD-ADAPTED STRAINS
FOR PLANTS PROTECTION IN THE BIOREGENERATIVE LIFE
SUPPORT SYSTEMS
Yu Ming Fu1, Vorobieva S. V. 2, Khizhnyak S. V. 2, Harlamova L. T. 2
1
Beihang University (Beijing University of Aeronautics and Astronautics), Beijing,
Peoples Republic of China
2
Krasnoyarsk State Agrarian University, Krasnoyarsk, Russia
Among 15 strains of cold-adapted bacteria isolated from the Karaulnaya-2
cave 9 strains suppressed conidia germination of Fusarium oxysporum (Table 2), 15
strains suppressed conidia germination of Bipolaris sorokiniana and 12 strains
suppressed conidia germination of Alternaria sp. Unability of these strains for
growth at human's body temperature allows to use them as a safety for people agents
for biological control of phytopathogens in Bioregenerative Life Support Systems.
Bioregenerative Life Support Systems (BLSS) are designed for a long-term
human life support by recycling of biogenic elements as it takes place in the natural
ecosystems. One of the key problems of BLSS based on the higher plants is plants
protection against diseases. The specificity of BLSS does not allow using chemical
pesticides for control of phytopathogens because of their toxicity for human. The
biological agents which are used for plants diseases control as an alternative to
chemical preparations are potentially dangerous for people if are applied in closed
environment. The use of cold-adapted microorganisms unable to grow at human's
body temperature for biological control of phytopathogens may solve the problem of
plants protection in BLSS.
Previous researches demonstrated that Middle Siberian limestone caves are
natural reservoir of cold-adapted bacteria and fungi [2, 3, 5, 6, 8]. These bacteria and
fungi are of big interest for low-temperature biotechnology and for biological plant
protection in boreal climate regions and in BLSS based on higher plants [1, 4, 7]. The
present study was devoted to estimation of the Karaulnaya-2 cave as a source of coldadapted strains for biological control of Bipolaris, Fusarium and Alternaria
phythopathogenic fungi species.
The Karaulnaya-2 cave (540 m length, 34 m depth) is located in limestone at
the distance 5 km from Krasnoyarsk in the Karaulensky karstic district, Eastern
Sayan Mountain area, Russia. Bacteria were isolated on the PD-cultural media (g/l:
peptone – 9,0; casein hydrolysate – 8,0; yeast extract – 3,0; NaCl – 5,0; Na2HPO4 –
2,0; agar – 20; pH = 7,0) from water samples at temperature +10ºC. Totally 21 strains
were isolated. Among them 17 strains are unable to grow at temperature +35ºC and
higher or demonstrate very slight growth with abnormal and unviable cells formation,
and only 4 strains are able to grow at +35ºC (Table 1).
Nine strains which are unable to grow at human's body temperature
demonstrated ability to suppress conidia germination of Fusarium oxysporum (Table
2), 15 strains suppressed conidia germination of Bipolaris sorokiniana (Table 3) and
12 strains suppressed conidia germination of Alternaria sp. (Table 4).
Strain
Growth at +10ºC
Growth at +35ºC
K-W1-PD1
Normal
No growth
K-W1-PD2
Normal
No growth
K-W1-PD3a
Normal
No growth
K-W1-PD3b
Normal
Normal with abnormal cells formation
K-W1-PD4
Normal
Normal
K-W1-PD15
Normal
No growth
K-W1-PD16
Normal
No growth
K-W2-PD8
Normal
No growth
K-W2-PD9
Normal
No growth
K-W2-PD11a
Normal
No growth
K-W2-PD11b
Normal
No growth
K-W2-PD12
Normal
No growth
K-W2-PD13a
Normal
Normal
K-W2-PD13b
Normal
No growth
K-W3-PD14
Normal
Slight with abnormal cells formation
K-W4-PD5
Normal
No growth
K-W4-PD6a
Normal
Normal
K-W4-PD6b
Normal
Slight with abnormal cells formation
K-W4-PD6v
Normal
Slight with abnormal cells formation
K-W4-PD6g
Normal
No growth
K-W4-PD7
Normal
No growth
Table 1 – An ability of strains to grow at different temperatures
Strain
Conidia
Conidia germination, Difference with control,
germination, %
% to control
significance level P
Control
84,9
100
K-W1-PD1
75,7
89,1
<0,05
K-W1-PD2
86,2
101,5
no
K-W1-PD15
70,6
83,1
<0,001
K-W1-PD16
90
106
no
K-W2-PD8
0,5
0,6
<0,001
K-W2-PD9
69,3
81,7
<0,001
K-W2-PD11a
79,8
94
no
K-W2-PD11b
75,2
88,5
<0,05
K-W2-PD12
21,9
25,8
<0,001
K-W4-PD5
34,9
41,1
<0,001
K-W4-PD6b
70,5
83
<0,001
K-W4-PD6v
91,2
107,4
0,05
K-W4-PD7
95,1
112
<0,001
K-W-PD10
85,5
100,7
no
Table 2 – Effect of cultural liquid on germination of Fusarium oxysporum conidia
Strain
Conidia
Conidia germination, Difference with control,
germination, %
% to control
significance level P
Control
100
100
K-W1-PD1
33,3
33,3
<0,001
K-W1-PD2
40,7
40,7
<0,001
K-W1-PD3a
14,3
14,3
<0,001
K-W1-PD15
30,6
30,6
<0,001
K-W1-PD16
41,9
41,9
<0,001
K-W2-PD8
17,2
17,2
<0,001
K-W2-PD9
40,4
40,4
<0,001
K-W2-PD11a
33,3
33,3
<0,001
K-W2-PD11b
40,5
40,5
<0,001
K-W2-PD12
44,3
44,3
<0,001
K-W4-PD5
20,8
20,8
<0,001
K-W4-PD6b
45,8
45,8
<0,001
K-W4-PD6v
46,9
46,9
<0,001
K-W4-PD7
81,5
81,5
<0,001
K-W-PD10
52,2
52,2
<0,001
Table 3 – Effect of cultural liquid on germination of Bipolaris sorokiniana conidia
Strain
Conidia
Conidia germination, Difference with control,
germination, %
% to control
significance level P
Control
92,5
100
K-W1-PD1
3,7
4
<0,001
K-W1-PD2
77,9
84,3
<0,01
K-W1-PD3a
3,6
3,9
<0,001
K-W1-PD15
67,3
72,8
<0,001
K-W1-PD16
78,6
84,9
<0,01
K-W2-PD8
3,7
4
<0,001
K-W2-PD9
63,8
68,9
<0,001
K-W2-PD11a
75
81,1
<0,01
K-W2-PD11b
85,4
92,3
no
K-W2-PD12
84,6
91,4
no
K-W4-PD5
1,8
2
<0,001
K-W4-PD6b
83,6
90,4
<0,05
K-W4-PD6v
83,2
89,9
0,05
K-W4-PD7
91,4
98,8
no
K-W-PD10
73,8
79,8
<0,001
Table 4 – Effect of cultural liquid on germination of Alternaria sp. conidia
Inability of these strains to grow at human's body temperature allows to use
them as a safety for people agents for biological control of phytopathogenic fungi in
BLSS.
References
1.
Ilients, I.R. Cave communities of micromycetes as a source of strains for
agricultural and environmental biotechnology: PhD thesis. Krasnoyarsk State
Agrarian University, Krasnoyarsk, 2011: 140 pages (in Russian)
2.
Khizhnuak, S., Tausheva, I., Berezikova, A. Supposed contribution of
cave microorganisms to a karstic process in the limestone caves of Easten Siberia.
Contributions to the IGCP448 meeting in Bowling Green, Ky, USA, June3-6 2003,
Part III-1 World comparison of karst ecosystem. Bowling Green, 2003: 99.
3.
Khizhnuak, S.V., Tausheva, I.V., Berezikova, A.A., Nesterenko, E.V.
Ecological and biological peculiarities of microorganisms from underground karst
cavities of the Middle Siberia. Homeostasis of forest ecosystems: Proc. X
International Symposium "Concept of Homeostasis: Theory, Experiments and
Application". Novosibirsk, 2001: 161-165.
4.
Khizhnyak, S.V., Lankina, E.P., Ilients, I.R. Estimation of effectiveness
of cold-adapted cave microorganisms in biological control of common root rot of
cereals. Vestnik Krasnoyarsk State Agrarian University, 2009, 6: 49-52. (in Russian)
5.
Khizhnyak, S.V., Nesterenko, E.V., Berezikova, A.A., Trusey, I.V. The
microbial communities of Middle Siberian cold caves. 3-rd International Workshop
on Ice Caves Kungur Ice Cave, Perm Region Russia, May 12-17: Volume of
Abstracts. Kungur, 2008: 56-57.
6.
Khizhnyak, S.V., Tausheva, I.V., Berezikova, A.A., Nesterenko, Y.V.,
Rogozin D.Y. Psychrophilic and Psychrotolerant Heterotrophic Microorganisms of
Middle Siberian Karst Cavities. Russian Journal of Ecology, 2003, 34 (4): 231-235.
7.
Lankina, E.P. Cave bacterial communities as a source of strains for
biological protection of plants against diseases: PhD thesis. Krasnoyarsk State
Agrarian University, Krasnoyarsk, 2011: 140 pages (in Russian)
8.
Nesterenko, E.V. Micromycetes of Middle Siberian Karst Cavities: PhD
thesis. Krasnoyarsk State Agrarian University, Krasnoyarsk, 2007: 134 pages (in
Russian)