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 cold-adapted 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 +35ºCK-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 germination, % / Conidia germination,% to control / Difference with 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 germination, % / Conidia germination,% to control / Difference with 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 germination, % / Conidia germination,% to control / Difference with 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
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