Pal K K, Tilak K V, Saxena A K, Dey R, Singh C S
Division of Microbiology, Indian Agricultural Research Institute, New Delhi.
Microbiol Res. 2001;156(3):209-23. doi: 10.1078/0944-5013-00103.
A plant growth-promoting isolate of a fluorescent Pseudomonas sp. EM85 and two bacilli isolates MR-11(2) and MRF, isolated from maize rhizosphere, were found strongly antagonistic to Fusarium moniliforme, Fusarium graminearum and Macrophomina phaseolina, causal agents of foot rots and wilting, collar rots/stalk rots and root rots and wilting, and charcoal rots of maize, respectively. Pseudomonas sp. EM85 produced antifungal antibiotics (Afa+), siderophore (Sid+), HCN (HCN+) and fluorescent pigments (Flu+) besides exhibiting plant growth promoting traits like nitrogen fixation, phosphate solubilization, and production of organic acids and IAA. While MR-11(2) produced siderophore (Sid+), antibiotics (Afa+) and antifungal volatiles (Afv+), MRF exhibited the production of antifungal antibiotics (Afa+) and siderophores (Sid+). Bacillus spp. MRF was also found to produce organic acids and IAA, solubilized tri-calcium phosphate and fixed nitrogen from the atmosphere. All three isolates suppressed the diseases caused by Fusarium moniliforme, Fusarium graminearum and Macrophomina phaseolina in vitro. A Tn5:: lacZ induced isogenic mutant of the fluorescent Pseudomonas EM85, M23, along with the two bacilli were evaluated for in situ disease suppression of maize. Results indicated that combined application of the two bacilli significantly (P = 0.05) reduced the Macrophomina-induced charcoal rots of maize by 56.04%. Treatments with the MRF isolate of Bacillus spp. and Tn5:: lacZ mutant (M23) of fluorescent Pseudomonas sp. EM85 significantly reduced collar rots, root and foot rots, and wilting of maize caused by Fusarium moniliforme and F. graminearum (P = 0.05) compared to all other treatments. All these isolates were found very efficient in colonizing the rhizotic zones of maize after inoculation. Evaluation of the population dynamics of the fluorescent Pseudomonas sp. EM85 using the Tn5:: lacZ marker and of the Bacillus spp. MRF and MR-11(2) using an antibiotic resistance marker revealed that all the three isolates could proliferate successfully in the rhizosphere, rhizoplane and endorhizosphere of maize, both at 30 and 60 days after seeding. Four antifungal compounds from fluorescent Pseudomonas sp. EM85, one from Bacillus sp. MR-11(2) and three from Bacillus sp. MRF were isolated, purified and tested in vitro and in thin layer chromatography bioassays. All these compounds inhibited R. solani, M. phaseolina, F. moniliforme, F. graminearum and F. solani strongly. Results indicated that antifungal antibiotics and/or fluorescent pigment of fluorescent Pseudomonas sp. EM85, and antifungal antibiotics of the bacilli along with the successful colonization of all the isolates might be involved in the biological suppression of the maize root diseases.
从玉米根际分离出的荧光假单胞菌属的一种促进植物生长的菌株EM85以及两种芽孢杆菌菌株MR-11(2)和MRF,被发现对串珠镰刀菌、禾谷镰刀菌和菜豆壳球孢菌具有强烈的拮抗作用,它们分别是玉米根腐病和枯萎病、颈腐病/茎腐病以及根腐病和枯萎病、炭腐病的病原菌。假单胞菌属菌株EM85除了具有促进植物生长的特性,如固氮、溶解磷、产生有机酸和吲哚-3-乙酸外,还产生抗真菌抗生素(Afa+)、铁载体(Sid+)、HCN(HCN+)和荧光色素(Flu+)。而MR-11(2)产生铁载体(Sid+)、抗生素(Afa+)和抗真菌挥发物(Afv+),MRF表现出产生抗真菌抗生素(Afa+)和铁载体(Sid+)。芽孢杆菌属MRF还被发现能产生有机酸和吲哚-3-乙酸,溶解磷酸三钙并从大气中固氮。所有这三种菌株在体外都能抑制由串珠镰刀菌、禾谷镰刀菌和菜豆壳球孢菌引起的病害。对荧光假单胞菌EM85的一个Tn5::lacZ诱导的同基因突变体M23以及这两种芽孢杆菌进行了玉米原位病害抑制评估。结果表明,两种芽孢杆菌的联合应用显著(P = 0.05)降低了菜豆壳球孢菌引起的玉米炭腐病,降幅达56.04%。与所有其他处理相比,芽孢杆菌属MRF菌株和荧光假单胞菌属EM85的Tn5::lacZ突变体(M23)处理显著降低了由串珠镰刀菌和禾谷镰刀菌引起的玉米颈腐病、根腐病和根脚腐病以及枯萎病(P = 0.05)。接种后发现所有这些菌株在定殖玉米根际区域方面都非常有效。利用Tn5::lacZ标记评估荧光假单胞菌属EM85的种群动态,利用抗生素抗性标记评估芽孢杆菌属MRF和MR-11(2)的种群动态,结果显示所有这三种菌株在播种后30天和60天均能在玉米的根际、根表和根内成功增殖。从荧光假单胞菌属EM85中分离出四种抗真菌化合物,从芽孢杆菌属MR-11(2)中分离出一种,从芽孢杆菌属MRF中分离出三种,并进行了体外和薄层色谱生物测定。所有这些化合物都强烈抑制茄丝核菌、菜豆壳球孢菌、串珠镰刀菌、禾谷镰刀菌和茄镰刀菌。结果表明,荧光假单胞菌属EM85的抗真菌抗生素和/或荧光色素,以及芽孢杆菌的抗真菌抗生素,再加上所有菌株的成功定殖,可能参与了对玉米根病的生物抑制作用。