青枯菌无致病力菌株FJAT-1458与强致病力株FJAT-91的竞争生长作用
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摘要
为了明确青枯菌无致病力菌株FJAT-1458的生防竞争机制,本研究采用体外共培养方法研究了碳源、氮源和培养时间对菌株FJAT-1458和青枯菌强致病力菌株FJAT-91竞争生长的影响;同时,研究了菌株FJAT-1458和FJAT-91在番茄植株体内的竞争生长。结果表明,碳源含量低于20%时,菌株FJAT-1458不能生长,而菌株FJAT-91可生长;无氮源条件下,FJAT-1458和FJAT-91均不能生长;碳源和氮源含量达100%时,FJAT-1458的菌体浓度(2.16×109 cfu/mL)显著低于FJAT-91的菌体浓度(3.24×109 cfu/mL)。混合培养24h前,FJAT-1458生长量大于FJAT-91,24h后则相反。在单独接种或混合接种5d后,FJAT-1458和FJAT-91均在番茄植株体内出现最大定殖量,随后迅速减少;FJAT-91在单独接种15 d时,定殖数量为0;两种菌株单独接种的定殖数量均大于混合接种(接种15 d除外)。先接种FJAT-1458,3 d后接种FJAT-91对番茄青枯病的防效(100%)显著高于同时接种FJAT-1458和FJAT-91(74.67%)。本研究表明,在体外无致病力菌株FJAT-1458对碳源和氮源的营养竞争能力弱于强致病力菌株FJAT-91;在体内无致病力菌株FJAT-1458会抑制强致病力菌株FJAT-91的生长;因此,FJAT-1458的生防竞争机制中,营养竞争起非主导作用,而位点竞争可能是主导因素。
The aim of this study is to determine the biocontrol competition mechanism of Ralstonia solanacearum avirulent strain FJAT-1458. Effects of carbon source, nitrogen source and culture time on the competitive growth between avirulent strain FJAT-1458 and virulent strain FJAT-91 were analyzed by co-culture in vitro. Furthermore, the competitive growth between FJAT-1458 and FJAT-91 was also in the tomato plant. The results showed that the virulent strain FJAT-91 could grow but the avirulent strain FJAT-1458 could not, when they were co-cultured in the media with content of carbon source less than 20%. Both strains could not reproduce when there was no nitrogen source. The cell concentration of FJAT-1458(2.16×109 cfu/mL) was significantly less than FJAT-91(3.24×109 cfu/mL) when the contents of carbon source and nitrogen source content were 100%. The growth of avirulent strain FJAT-1458 was faster than the virulent strain FJAT-91 before 24 h, but became slower after 24 h. The colonization number of FJAT-1458 and FJAT-91 reached the highest 5 d after sole inoculation or co-inoculation, and then quickly decreased. Moreover, the colonization number of FJAT-91 was 0 15 d after sole inoculation. The colonization number of both FJAT-1458 and FJAT-91 by sole inoculation was higher than that by co-inoculation(except for 15 d). The pre-inoculation of FJAT-1458 for 3 d had higher control efficiency(100%) than co-inoculation of FJAT-1458 and FJAT-91 at the same time(74.67%). It was concluded that the utilization abilities of carbon and nitrogen source for FJAT-1458 were poorer than those for FJAT-91 when co-cultured in vitro, but FJAT-1458 could restrained by the growth of FJAT-91 when co-inoculation in tomato plants. In the biocontrol competition mechanism of strain FJAT-1458, site competition may be the dominant factor, but not nutrient competition.
The aim of this study is to determine the biocontrol competition mechanism of Ralstonia solanacearum avirulent strain FJAT-1458. Effects of carbon source, nitrogen source and culture time on the competitive growth between avirulent strain FJAT-1458 and virulent strain FJAT-91 were analyzed by co-culture in vitro. Furthermore, the competitive growth between FJAT-1458 and FJAT-91 was also in the tomato plant. The results showed that the virulent strain FJAT-91 could grow but the avirulent strain FJAT-1458 could not, when they were co-cultured in the media with content of carbon source less than 20%. Both strains could not reproduce when there was no nitrogen source. The cell concentration of FJAT-1458(2.16×109 cfu/mL) was significantly less than FJAT-91(3.24×109 cfu/mL) when the contents of carbon source and nitrogen source content were 100%. The growth of avirulent strain FJAT-1458 was faster than the virulent strain FJAT-91 before 24 h, but became slower after 24 h. The colonization number of FJAT-1458 and FJAT-91 reached the highest 5 d after sole inoculation or co-inoculation, and then quickly decreased. Moreover, the colonization number of FJAT-91 was 0 15 d after sole inoculation. The colonization number of both FJAT-1458 and FJAT-91 by sole inoculation was higher than that by co-inoculation(except for 15 d). The pre-inoculation of FJAT-1458 for 3 d had higher control efficiency(100%) than co-inoculation of FJAT-1458 and FJAT-91 at the same time(74.67%). It was concluded that the utilization abilities of carbon and nitrogen source for FJAT-1458 were poorer than those for FJAT-91 when co-cultured in vitro, but FJAT-1458 could restrained by the growth of FJAT-91 when co-inoculation in tomato plants. In the biocontrol competition mechanism of strain FJAT-1458, site competition may be the dominant factor, but not nutrient competition.
引文
[1]Hayward A C.Biology and epidemiology of bacterial wilt caused by Pseudomonas solanacearum[J].Annual Review of Phytopathology,1991,29(1):65-87.
[2]Wicker E,Grassart L,Coranson-Beaudu R,et al.Ralstonia solanacearum strains from Martinique(French West Indies)exhibiting a new pathogenic potential[J].Applied and Environmental Microbiology,2007,71(21):6790-6801.
[3]Norman D J,Zapata M,Gabriel D W,et al.Genetic diversity and host range variation of Ralstonia solancearum strains entering North America[J].Phytopathology,2009,99(9):1070-1077.
[4]Cellier G,Prior P.Deciphering phenotypic diversity of Ralstonia solanacearum strains pathogenic to potato[J].Phytopatholy,2010,100(11):1250-1261.
[5]Suga Y,Horita M,Umekita M,et al.Pathogenic characters of Japanese potato strains of Ralstonia solanacearum[J].Journal of General Plant Pathology,2013,79(2):110-114.
[6]Balabel N M,Ewed W E,Mostapha M I,et al.Some epidemiological aspects of Ralstonia solanacearum[J].Egypt Journal of Agriculture Research,2005,83(4):1547-1563.
[7]Frey P,Prior P,Marie C,et al.Hrp-mutants of Pseudomonas solanacearum as potentiall biocontrol agents of tomato bacterial wilt[J].Applied and Environmental Microbiology,1994,60(9):3175-3181.
[8]Trigalet A,Trigalet-Demery D.Use of avirulent mutants of Pseudomonas solanacearum for the biological control of bacterial wilt of tomato plants[J].Physiological and Molecular Plant Pathology,1990,36(1):27-38.
[9]肖田,肖崇刚,邹阳,等.青枯菌无致病力菌株对烟草青枯病的控病作用初步研究[J].植物保护,2008,34(2):79-82.
[10]程本亮,车建美,刘波.青枯菌Tn5转座子无致病力突变株构建及其生物学特性[J].农业生物技术学报,2011,19(1):26-38.
[11]刘波,朱育菁,蓝江林,等.植物免疫系统的研究与应用[J].中国农学通报,2007,23(S):163-172.
[12]郑雪芳,朱育菁,刘波,等.番茄青枯病植物疫苗胶悬剂的制备及其对病害的防治效果[J].植物保护,2017,43(2):208-211.
[13]Kelman A.The relationship of pathogenicity in P.solanacearum to colony appearance on a tetrazolium medium[J].Phytopathology,1954,44(12):693-695.
[14]郑雪芳,刘波,朱育菁.福建省青枯雷尔氏菌无毒基因组成及与致病力关系分析[J].农业生物技术学报,2016,24(1):98-106.
[15]刘波,朱育菁,林抗美,等.青枯雷尔氏菌在植株体内分布及其致病力的异质性研究[J].中国农业科学,2007,40(7):1559-1566.
[16]Feng D X,Tasset C,Hanemian M,et al.Biological control of bacterial wilt in Arabidopsis thaliana involves abscissic acid signalling[J].New Phytologist,2012,194(4):1035-1045.
[17]Chen W Y,Echandi E.Effects of avirulent bacteriocin producing strains of Pseudomonas solanacearum on the control of bacterial wilt of tobacco[J].Plant Pathology,1984,33(2):245-253.
[18]Tanaka H,Negishi H,Maeda H.Control of tobacco bacterial wilt by an avirulent strain of Pseudomonas solanacearum M4S and its bactriophage[J].Annals of the Phytopathological Society of Japan,1990,56(2):243-246.
[19]杨宇红,刘俊平,杨翠荣,等.无致病力hrp-突变体防治茄科蔬菜青枯病[J].植物保护学报,2008,35(5):433-437.
[20]葛慈斌,刘波,林营志,等.生防菌对青枯菌强致病力和无致病力菌株生长竞争的影响[J].植物保护学报,2006,33(2):151-156.
[21]朱育菁,周涵韬,刘波,等.番茄青枯菌的强致病力与无致病力菌株生长竞争关系的研究[J].厦门大学学报,2004,43(S):97-101.
[22]Zargaraan A,Kamaliroosta L,Yaghoubi A S,et al.Effect of substitution of sugar by high fructose corn syrup on the physicochemical properties of bakery and dairy products:a review[J].Nutrition and Food Sciences Research,2016,3(4):3-11.
[23]Shahina Z,Hossain M T,Hakim M A.Variation of protease production by the bacteria(Bacillus fastidiosus)and the fungus(Aspergillus funiculosus)[J].Journal of Microbiology Research,2013,3(4):135-142.
[24]周帮菊,肖崇刚,肖田,等.青枯无致病力菌株对烟草青枯病控病研究及机理初探[J].南方农业,2014,8(27):166-170.
[25]陈庆河,翁启勇,胡方平.无致病力青枯菌株对番茄青枯病的防治效果[J].中国生物防治,2004,20(1):42-44.
[2]Wicker E,Grassart L,Coranson-Beaudu R,et al.Ralstonia solanacearum strains from Martinique(French West Indies)exhibiting a new pathogenic potential[J].Applied and Environmental Microbiology,2007,71(21):6790-6801.
[3]Norman D J,Zapata M,Gabriel D W,et al.Genetic diversity and host range variation of Ralstonia solancearum strains entering North America[J].Phytopathology,2009,99(9):1070-1077.
[4]Cellier G,Prior P.Deciphering phenotypic diversity of Ralstonia solanacearum strains pathogenic to potato[J].Phytopatholy,2010,100(11):1250-1261.
[5]Suga Y,Horita M,Umekita M,et al.Pathogenic characters of Japanese potato strains of Ralstonia solanacearum[J].Journal of General Plant Pathology,2013,79(2):110-114.
[6]Balabel N M,Ewed W E,Mostapha M I,et al.Some epidemiological aspects of Ralstonia solanacearum[J].Egypt Journal of Agriculture Research,2005,83(4):1547-1563.
[7]Frey P,Prior P,Marie C,et al.Hrp-mutants of Pseudomonas solanacearum as potentiall biocontrol agents of tomato bacterial wilt[J].Applied and Environmental Microbiology,1994,60(9):3175-3181.
[8]Trigalet A,Trigalet-Demery D.Use of avirulent mutants of Pseudomonas solanacearum for the biological control of bacterial wilt of tomato plants[J].Physiological and Molecular Plant Pathology,1990,36(1):27-38.
[9]肖田,肖崇刚,邹阳,等.青枯菌无致病力菌株对烟草青枯病的控病作用初步研究[J].植物保护,2008,34(2):79-82.
[10]程本亮,车建美,刘波.青枯菌Tn5转座子无致病力突变株构建及其生物学特性[J].农业生物技术学报,2011,19(1):26-38.
[11]刘波,朱育菁,蓝江林,等.植物免疫系统的研究与应用[J].中国农学通报,2007,23(S):163-172.
[12]郑雪芳,朱育菁,刘波,等.番茄青枯病植物疫苗胶悬剂的制备及其对病害的防治效果[J].植物保护,2017,43(2):208-211.
[13]Kelman A.The relationship of pathogenicity in P.solanacearum to colony appearance on a tetrazolium medium[J].Phytopathology,1954,44(12):693-695.
[14]郑雪芳,刘波,朱育菁.福建省青枯雷尔氏菌无毒基因组成及与致病力关系分析[J].农业生物技术学报,2016,24(1):98-106.
[15]刘波,朱育菁,林抗美,等.青枯雷尔氏菌在植株体内分布及其致病力的异质性研究[J].中国农业科学,2007,40(7):1559-1566.
[16]Feng D X,Tasset C,Hanemian M,et al.Biological control of bacterial wilt in Arabidopsis thaliana involves abscissic acid signalling[J].New Phytologist,2012,194(4):1035-1045.
[17]Chen W Y,Echandi E.Effects of avirulent bacteriocin producing strains of Pseudomonas solanacearum on the control of bacterial wilt of tobacco[J].Plant Pathology,1984,33(2):245-253.
[18]Tanaka H,Negishi H,Maeda H.Control of tobacco bacterial wilt by an avirulent strain of Pseudomonas solanacearum M4S and its bactriophage[J].Annals of the Phytopathological Society of Japan,1990,56(2):243-246.
[19]杨宇红,刘俊平,杨翠荣,等.无致病力hrp-突变体防治茄科蔬菜青枯病[J].植物保护学报,2008,35(5):433-437.
[20]葛慈斌,刘波,林营志,等.生防菌对青枯菌强致病力和无致病力菌株生长竞争的影响[J].植物保护学报,2006,33(2):151-156.
[21]朱育菁,周涵韬,刘波,等.番茄青枯菌的强致病力与无致病力菌株生长竞争关系的研究[J].厦门大学学报,2004,43(S):97-101.
[22]Zargaraan A,Kamaliroosta L,Yaghoubi A S,et al.Effect of substitution of sugar by high fructose corn syrup on the physicochemical properties of bakery and dairy products:a review[J].Nutrition and Food Sciences Research,2016,3(4):3-11.
[23]Shahina Z,Hossain M T,Hakim M A.Variation of protease production by the bacteria(Bacillus fastidiosus)and the fungus(Aspergillus funiculosus)[J].Journal of Microbiology Research,2013,3(4):135-142.
[24]周帮菊,肖崇刚,肖田,等.青枯无致病力菌株对烟草青枯病控病研究及机理初探[J].南方农业,2014,8(27):166-170.
[25]陈庆河,翁启勇,胡方平.无致病力青枯菌株对番茄青枯病的防治效果[J].中国生物防治,2004,20(1):42-44.
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