[Objective] For preparing the biopesticide of Bacillus subtilis B579 with high spore concentration,the sporulation conditions were optimized.[Method] Two-step fermentation control strategy was used,in which,the first ...[Objective] For preparing the biopesticide of Bacillus subtilis B579 with high spore concentration,the sporulation conditions were optimized.[Method] Two-step fermentation control strategy was used,in which,the first phase（0-10 h）was to improve cell growth,and the second phase（10-30 h）was to promote spore formation.Four factors including initial glucose concentration,fermentation pH,temperature（in the second phase）and shaking speed（in the second phase）were optimized using the methods of single factor test and orthogonal experiment.[Result] The initial glucose concentration showed a significant effect on sporulation.The optimal conditions for the spore formation of B.subtilis B579 were as follows：initial glucose concentration 5 g/L,fermentation pH 7.0,the temperature for the first phase 37 ℃,and the shaking speed for the first phase 180 r/min,the temperature for the second phase 40 ℃,and the shaking speed for the second phase 200 r/min;in addition,the first phase was 10 h and the second phase of fermentation was conducted for 30 h.Under such conditions,the spore concentration and spore formation rate could reach 9.43×108 CFU/ml and 90.99%,respectively,which represented 6.70-fold and 2.43-fold increase compared with those before optimization.[Conclusion] The spores concentration of biocontrol agent was improved using two-step control strategy,which provided the basis for biopesticide production in large scale.展开更多
[Objective] The aim was to understand defense mechanism of pear after inoculated Botryosphaeria berengeriana f. sp. piricolan and mechanism of antioxidant enzymes of biocontrol bacteria. [Method] Pears were treated by...[Objective] The aim was to understand defense mechanism of pear after inoculated Botryosphaeria berengeriana f. sp. piricolan and mechanism of antioxidant enzymes of biocontrol bacteria. [Method] Pears were treated by Botryosphaeria berengeriana f. sp. piricolan and bio control bacteria, and the change of antioxidant enzymes were determined. [Result] The biocontrol bacteria had little effect on MDA;the content of MDA treated by B. berengeriana reached high peak in 48 h, was 10.22nmol/g which was 1.86 times of CK; the content of MDA treated by B. berengeriana and biocontrol bacteria reached high peak in 24 h, was 8.92 nmol/g which was1.62 times of CK. The content of SOD treated by biocontrol bacteria reached high peak in 48 h, was 126.69 U/[g（FW）·min] which was 1.54 times of CK; the contents of SOD treated by B. berengeriana as well as B. berengeriana and biocontrol bacteria reached high peak in 24 h, were 122.10 and 135.32 U/[g（FW）·min] which were 1.48 and 1.65 times of CK respectively; the contents of POD on biocontrol bacteria treatment, B. ana treatment as well as B. berengeriana and biocontrol bacteria treatment reached high peak in 24 h, were 385.34, 342.50 and 290.00 U/[g（FW）·min] which were 1.83, 1.62 and 1.38 times of CK respectively. The contents of CAT on biocontrol bacteria treatment, B. rengeriana treatment as well as B. berengeriana and biocontrol bacteria treatment reached high peak in 6 h, were 133.33,114.17 and 113.35 U/[g（FW）·min] which were 1.33, 1.14 and 1.13 times of CK respectively. The biocontrol bacteria had little difference in CK; the content of PPO of B. berengeriana treatment reached high peak in 12 h, was 81.86 U/[g（FW）·min]which was 1.76 times of CK; B. berengeriana and biocontrol bacteria treatment reached high peak in 24 h, was 70.00 U/[g（FW）·min] which was 1.50 times of CK.[Conclusion] B. berengeriana and biocontrol bacteria had more effect on MDA; both B. berengeriana and biocontrol bacteria could increase the excitation of SOD enzyme activity; both B. berengeriana and biocontrol bacteria could increase the excitation of POD enzyme activity; both B. berengeriana and biocontrol bacteria could increase the excitation of CAT enzyme activity; using biocontrol bacteria alone had not obvious effect on PPO, B. berengeriana could increase the excitation of PPO enzyme activity.展开更多
The study was taken up with the objective of testing whether the endophytic organisms isolated from crops that are normally non-hosts to the bacterial wilt pathogen Ralstonia solanacearum possessed pathogen-antagonist...The study was taken up with the objective of testing whether the endophytic organisms isolated from crops that are normally non-hosts to the bacterial wilt pathogen Ralstonia solanacearum possessed pathogen-antagonistic activity and to evaluate the selected isolates for the alleviation of wilt disease in the target tomato crop through horizontal movement of promising organisms. Sixteen endophytic bacteria (EB) isolated from the micropropagated cultures of grape, watermelon and papaya were tested for potential antagonistic effects against R. solanacearum tomato isolate “NH-01” through agar-well diffusion assay. Enterobacter cloacae from papaya (EB-11) displayed the maximum antagonistic effect followed by Bacillus subtilis (EB-06) and B. flexus (EB-07) from watermelon and B. pumilus (EB-02) from grape. Testing the above organisms for crop protection through seed fortification of susceptible tomato cv. Arka Vikas at sowing in R. solanacearum inoculated (Ral+) organic cocopeat showed EB-02 and EB-11 promising (33% and 32% survival, respectively, four weeks after sowing against 15% in Ral+ control). A second trial showed 37%, 28%, 21% and 55% seedling survival 6 weeks after sowing for EB-02, EB-06, EB-07 and EB-11 respectively, compared to 2.5% in non-treated control. Assessing the four endophytes for crop protection in Ral+ sick-soil through seedling fortification at transplanting indicated less disease incidence in treated sets (40%, 40%, 20% and 20% survival, respectively, six weeks after transplanting) over non-fortified control (5%). Endophytic fortification of seedlings through hypocotyl inoculation showed some systemic resistance induction upon seedling transplanting to sick soil but not with petiole fortification. Seedling growth was enhanced by the isolates EB-06 and EB-07. The study thus identifies four endophytic organisms from crops unrelated to tomato possessing potential antagonistic activity against the wilt pathogen and prospects for exploitation as biocontrol agents coupled with seedling growth promotion effects.展开更多
Silicate solubilizing bacteria (SSB) can play an efficient role in soil by solubilizing insoluble forms of silicates. In addition to this some SSB can also solubilize potassium and phosphates, hence increasing soil fe...Silicate solubilizing bacteria (SSB) can play an efficient role in soil by solubilizing insoluble forms of silicates. In addition to this some SSB can also solubilize potassium and phosphates, hence increasing soil fertility and enhancing plant defense mechanisms. A total of 111 bacterial strains were isolated from various habitats of Pakistan and screened for solubilization of silicate, phosphate and potassium on respective media. Out of these, 35 bacterial isolates were capable of solubilizing either silicate, phosphate or potassium. Amongst these 7 bacterial isolates were capable of solubilizing all three minerals tested. The highest silicate (zone diameter 54 mm) and phosphate solubilization (zone diameter 55 mm) was observed for bacterial isolate NR-2 while the highest potassium solubilization was observed for NE-4b (zone diameter 11 mm). Dual culture antagonistic assays were carried out by using these bacterial isolates against four plant pathogenic fungi Magnaporthae grisae, Rhizoctonia solani, Altarnaria alternata and Macrophomina pheasolina. Mean zone of inhibition of these bacterial isolates against the four pathogenic fungi ranged between 4 mm to 39 mm. The largest zone of inhibition against all four bacterial strains was recorded for bacterial isolate NR-2 followed by NE-4b. These strains will be further investigated for their plant growth promoting activities in the future.展开更多
The present study evaluates the possible modes of action of antagonistic bacteria and their tolerance to UV radiation, temperature and osmotic stress. The partial 16S-23S rRNA gene sequencing of eight antagonistic bac...The present study evaluates the possible modes of action of antagonistic bacteria and their tolerance to UV radiation, temperature and osmotic stress. The partial 16S-23S rRNA gene sequencing of eight antagonistic bacteria had a high match with three bacterial genera: <em>Curtobacterium</em>, <em>Pantoea</em> and <em>Bacillus</em>. In this study, the three <em>Bacillus</em> isolates showed the most relevant production of enzymes, volatile organic compounds and antibiosis against <em>Exserohilum turcicum</em>. Respect to UV radiation and temperature <em>Pantoea</em> and <em>Bacillus</em> isolates were more tolerant;whereas the eight isolates were tolerant to osmotic stress in varying degree. The three <em>Bacillus</em> isolates have the greatest potential as biocontrol agents for foliar diseases in maize. The antagonistic action could be explained through different modes of action such as enzymes, volatile organic compounds and/or direct antibiosis by other secondary metabolites. <em>Bacillus</em> isolates tolerance to environmental stresses including UV radiation, temperature and osmotic stress is relevant for survival and persistence on the leaf surface. This work provides new information about the mode of action of antagonistic bacteria with proven efficacy against maize leaf pathogens. In addition, it provides information about the tolerance of antagonistic bacteria against different stress conditions. The data of the present study could contribute to the development of a successful foliar biofungicide.展开更多
产铁载体细菌通过与病原菌竞争铁元素而具有良好的生防效果。本研究从人参根组织及根际土壤中分离细菌,对其抑菌作用和产铁载体能力进行筛选,并对所筛选的1株菌株进行发酵条件的单因素和响应面优化。结果表明,共分离得到细菌菌株207株,...产铁载体细菌通过与病原菌竞争铁元素而具有良好的生防效果。本研究从人参根组织及根际土壤中分离细菌,对其抑菌作用和产铁载体能力进行筛选,并对所筛选的1株菌株进行发酵条件的单因素和响应面优化。结果表明,共分离得到细菌菌株207株,其中8株菌株既具有产铁载体的能力又对人参黑斑、根腐和锈腐病菌具有良好抑制作用;经筛选发现菌株GR-39在限铁、富铁培养基上对黑斑病菌抑菌作用差异显著,并鉴定该菌株为普城沙雷氏菌Serratiaplymuthica。通过单因素试验确定菌株GR-39在蔗糖、蛋白胨5 g/L、温度34℃、p H 8时产铁载体相对含量为74.6%;通过响应面法建立了铁载体量与自变量的回归模型,得出最佳发酵条件为蔗糖6 g/L、蛋白胨5 g/L、p H 8时铁载体产量最大达77.13%。研究表明,菌株GR-39具有较强产铁载体能力并且对人参黑斑病菌防效显著,可为多功能微生物菌剂研制提供良好菌种资源。展开更多
基金Supported by the Natural Science Foundation of Tianjin(09JCZDJC19100)Scientific Research Fund of Tianjin Scienceand Technology University(20090403)~~
文摘[Objective] For preparing the biopesticide of Bacillus subtilis B579 with high spore concentration,the sporulation conditions were optimized.[Method] Two-step fermentation control strategy was used,in which,the first phase（0-10 h）was to improve cell growth,and the second phase（10-30 h）was to promote spore formation.Four factors including initial glucose concentration,fermentation pH,temperature（in the second phase）and shaking speed（in the second phase）were optimized using the methods of single factor test and orthogonal experiment.[Result] The initial glucose concentration showed a significant effect on sporulation.The optimal conditions for the spore formation of B.subtilis B579 were as follows：initial glucose concentration 5 g/L,fermentation pH 7.0,the temperature for the first phase 37 ℃,and the shaking speed for the first phase 180 r/min,the temperature for the second phase 40 ℃,and the shaking speed for the second phase 200 r/min;in addition,the first phase was 10 h and the second phase of fermentation was conducted for 30 h.Under such conditions,the spore concentration and spore formation rate could reach 9.43×108 CFU/ml and 90.99%,respectively,which represented 6.70-fold and 2.43-fold increase compared with those before optimization.[Conclusion] The spores concentration of biocontrol agent was improved using two-step control strategy,which provided the basis for biopesticide production in large scale.
文摘[Objective] The aim was to understand defense mechanism of pear after inoculated Botryosphaeria berengeriana f. sp. piricolan and mechanism of antioxidant enzymes of biocontrol bacteria. [Method] Pears were treated by Botryosphaeria berengeriana f. sp. piricolan and bio control bacteria, and the change of antioxidant enzymes were determined. [Result] The biocontrol bacteria had little effect on MDA;the content of MDA treated by B. berengeriana reached high peak in 48 h, was 10.22nmol/g which was 1.86 times of CK; the content of MDA treated by B. berengeriana and biocontrol bacteria reached high peak in 24 h, was 8.92 nmol/g which was1.62 times of CK. The content of SOD treated by biocontrol bacteria reached high peak in 48 h, was 126.69 U/[g（FW）·min] which was 1.54 times of CK; the contents of SOD treated by B. berengeriana as well as B. berengeriana and biocontrol bacteria reached high peak in 24 h, were 122.10 and 135.32 U/[g（FW）·min] which were 1.48 and 1.65 times of CK respectively; the contents of POD on biocontrol bacteria treatment, B. ana treatment as well as B. berengeriana and biocontrol bacteria treatment reached high peak in 24 h, were 385.34, 342.50 and 290.00 U/[g（FW）·min] which were 1.83, 1.62 and 1.38 times of CK respectively. The contents of CAT on biocontrol bacteria treatment, B. rengeriana treatment as well as B. berengeriana and biocontrol bacteria treatment reached high peak in 6 h, were 133.33,114.17 and 113.35 U/[g（FW）·min] which were 1.33, 1.14 and 1.13 times of CK respectively. The biocontrol bacteria had little difference in CK; the content of PPO of B. berengeriana treatment reached high peak in 12 h, was 81.86 U/[g（FW）·min]which was 1.76 times of CK; B. berengeriana and biocontrol bacteria treatment reached high peak in 24 h, was 70.00 U/[g（FW）·min] which was 1.50 times of CK.[Conclusion] B. berengeriana and biocontrol bacteria had more effect on MDA; both B. berengeriana and biocontrol bacteria could increase the excitation of SOD enzyme activity; both B. berengeriana and biocontrol bacteria could increase the excitation of POD enzyme activity; both B. berengeriana and biocontrol bacteria could increase the excitation of CAT enzyme activity; using biocontrol bacteria alone had not obvious effect on PPO, B. berengeriana could increase the excitation of PPO enzyme activity.
文摘The study was taken up with the objective of testing whether the endophytic organisms isolated from crops that are normally non-hosts to the bacterial wilt pathogen Ralstonia solanacearum possessed pathogen-antagonistic activity and to evaluate the selected isolates for the alleviation of wilt disease in the target tomato crop through horizontal movement of promising organisms. Sixteen endophytic bacteria (EB) isolated from the micropropagated cultures of grape, watermelon and papaya were tested for potential antagonistic effects against R. solanacearum tomato isolate “NH-01” through agar-well diffusion assay. Enterobacter cloacae from papaya (EB-11) displayed the maximum antagonistic effect followed by Bacillus subtilis (EB-06) and B. flexus (EB-07) from watermelon and B. pumilus (EB-02) from grape. Testing the above organisms for crop protection through seed fortification of susceptible tomato cv. Arka Vikas at sowing in R. solanacearum inoculated (Ral+) organic cocopeat showed EB-02 and EB-11 promising (33% and 32% survival, respectively, four weeks after sowing against 15% in Ral+ control). A second trial showed 37%, 28%, 21% and 55% seedling survival 6 weeks after sowing for EB-02, EB-06, EB-07 and EB-11 respectively, compared to 2.5% in non-treated control. Assessing the four endophytes for crop protection in Ral+ sick-soil through seedling fortification at transplanting indicated less disease incidence in treated sets (40%, 40%, 20% and 20% survival, respectively, six weeks after transplanting) over non-fortified control (5%). Endophytic fortification of seedlings through hypocotyl inoculation showed some systemic resistance induction upon seedling transplanting to sick soil but not with petiole fortification. Seedling growth was enhanced by the isolates EB-06 and EB-07. The study thus identifies four endophytic organisms from crops unrelated to tomato possessing potential antagonistic activity against the wilt pathogen and prospects for exploitation as biocontrol agents coupled with seedling growth promotion effects.
文摘Silicate solubilizing bacteria (SSB) can play an efficient role in soil by solubilizing insoluble forms of silicates. In addition to this some SSB can also solubilize potassium and phosphates, hence increasing soil fertility and enhancing plant defense mechanisms. A total of 111 bacterial strains were isolated from various habitats of Pakistan and screened for solubilization of silicate, phosphate and potassium on respective media. Out of these, 35 bacterial isolates were capable of solubilizing either silicate, phosphate or potassium. Amongst these 7 bacterial isolates were capable of solubilizing all three minerals tested. The highest silicate (zone diameter 54 mm) and phosphate solubilization (zone diameter 55 mm) was observed for bacterial isolate NR-2 while the highest potassium solubilization was observed for NE-4b (zone diameter 11 mm). Dual culture antagonistic assays were carried out by using these bacterial isolates against four plant pathogenic fungi Magnaporthae grisae, Rhizoctonia solani, Altarnaria alternata and Macrophomina pheasolina. Mean zone of inhibition of these bacterial isolates against the four pathogenic fungi ranged between 4 mm to 39 mm. The largest zone of inhibition against all four bacterial strains was recorded for bacterial isolate NR-2 followed by NE-4b. These strains will be further investigated for their plant growth promoting activities in the future.
文摘The present study evaluates the possible modes of action of antagonistic bacteria and their tolerance to UV radiation, temperature and osmotic stress. The partial 16S-23S rRNA gene sequencing of eight antagonistic bacteria had a high match with three bacterial genera: <em>Curtobacterium</em>, <em>Pantoea</em> and <em>Bacillus</em>. In this study, the three <em>Bacillus</em> isolates showed the most relevant production of enzymes, volatile organic compounds and antibiosis against <em>Exserohilum turcicum</em>. Respect to UV radiation and temperature <em>Pantoea</em> and <em>Bacillus</em> isolates were more tolerant;whereas the eight isolates were tolerant to osmotic stress in varying degree. The three <em>Bacillus</em> isolates have the greatest potential as biocontrol agents for foliar diseases in maize. The antagonistic action could be explained through different modes of action such as enzymes, volatile organic compounds and/or direct antibiosis by other secondary metabolites. <em>Bacillus</em> isolates tolerance to environmental stresses including UV radiation, temperature and osmotic stress is relevant for survival and persistence on the leaf surface. This work provides new information about the mode of action of antagonistic bacteria with proven efficacy against maize leaf pathogens. In addition, it provides information about the tolerance of antagonistic bacteria against different stress conditions. The data of the present study could contribute to the development of a successful foliar biofungicide.
基金云南省万人计划“云岭学者”项目(YNWR-YLXZ-2018-018)国际原子能机构(International Atomic Energy Agency)项目“An IntegrativeApproach to Enhance Disease Resistance Against Fusarium Wilt(Foc TR4)in Banana–Phase II”(D23033-26673)。
文摘产铁载体细菌通过与病原菌竞争铁元素而具有良好的生防效果。本研究从人参根组织及根际土壤中分离细菌,对其抑菌作用和产铁载体能力进行筛选,并对所筛选的1株菌株进行发酵条件的单因素和响应面优化。结果表明,共分离得到细菌菌株207株,其中8株菌株既具有产铁载体的能力又对人参黑斑、根腐和锈腐病菌具有良好抑制作用;经筛选发现菌株GR-39在限铁、富铁培养基上对黑斑病菌抑菌作用差异显著,并鉴定该菌株为普城沙雷氏菌Serratiaplymuthica。通过单因素试验确定菌株GR-39在蔗糖、蛋白胨5 g/L、温度34℃、p H 8时产铁载体相对含量为74.6%;通过响应面法建立了铁载体量与自变量的回归模型,得出最佳发酵条件为蔗糖6 g/L、蛋白胨5 g/L、p H 8时铁载体产量最大达77.13%。研究表明,菌株GR-39具有较强产铁载体能力并且对人参黑斑病菌防效显著,可为多功能微生物菌剂研制提供良好菌种资源。
