scholarly journals Isolation of Nonpathogenic Mutants of Fusarium oxysporum f. sp. melonis for Biological Control of Fusarium Wilt in Cucurbits

2002 ◽  
Vol 92 (2) ◽  
pp. 164-168 ◽  
Author(s):  
Stanley Freeman ◽  
Aida Zveibil ◽  
Haim Vintal ◽  
Marcel Maymon

Two nonpathogenic mutant strains 4/4 and 15/15 of Fusarium oxysporum f. sp. melonis (race 1,2) were isolated by a continuous dipinoculation technique following UV mutagenesis of the virulent wild-type isolate FOM1.2. No disease symptoms or detrimental effects were observed following inoculation of muskmelon seedlings by strain 4/4. In contrast, strain 15/15 caused mortality of susceptible cultivars although to a lesser extent than the wild-type isolate. Strain 4/4 colonized a variety of muskmelon and watermelon cultivars. In muskmelon cv. Ein Dor, seedlings were dipped in a conidial suspension of strain 4/4 and planted in medium amended with the mutant to achieve 100% colonization of roots and between 30 to 70% of the lower stem tissues 7 days after planting. Similar percent colonization of watermelon seedlings by strain 4/4 was recorded. In cross-protection experiments with muskmelon cultivars, significant reduction in seedling mortality was observed between 4/4-colonized FOM1.2. challenged plants compared with that of wild-type challenged plants alone. Similarly, strain 4/4 was able to significantly reduce mortality of watermelon seedlings caused by F. oxysporum f. sp. niveum race 2. This novel approach of generating nonpathogenic mutants for biological control in Fusarium spp. and other fungal pathogens from virulent wild-type isolates may be beneficial for control, because the mutant strains, lacking only in pathogenicity, may compete more efficiently than other biocontrol organisms against the pathogen of origin.

1963 ◽  
Vol 16 (1) ◽  
pp. 55 ◽  
Author(s):  
A Kerr

At least four fungal pathogens are involved in the root rot-Fusarium wilt complex of peas which is a serious problem following intensive cropping of peas in South Australia. The pathogens are Fusarium oxysporum f. pisi race 2 Snyder & Hansen, F. solani f. pisi Snyder & Hansen, Pythium ultimum Trow, and Ascochyta pinodella L. K. Jones. In susceptible pea cultivars there is a marked interaction between F. oxysporum and P. ultimum. P. ultimum alone causes initial stunting from which plants gradually recover; F. OX1Jsporum alone probably CRuses little damage; both fungi together CRuse initial stunting followed by severe wilt symptom about 6 weeks after sowing and death 2 weeks later. The importance ofF. solani and A. pinodella has not been fully determined, but they probably cause only minor damage.


2005 ◽  
Vol 30 (4) ◽  
pp. 409-412 ◽  
Author(s):  
Juliano C. da Silva ◽  
Wagner Bettiol

This study was done to evaluate the efficiency of non-pathogenic Fusarium oxysporum isolates (141/3, 233, 233/1, 245, 245/1, 251, 251/2, 251/5, and 257) in controlling vascular wilt caused by F. oxysporum f. sp. lycopersici, race 2 (isolates C-21A, TO11, and TO245) in tomato (Lycopersicon esculentum) cv. Viradoro seedlings. In order to determine the effect of non-pathogenic F. oxysporum isolates in tomato plants, the root system of 30-day-old seedlings was immersed in conidial suspensions (10(6) ml-1) of each isolate and the seedlings were transplanted to a cultivation substrate. Thirty-five days after transplanting it was observed that the non-pathogenic F. oxysporum isolates were not pathogenic to the cv. Viradoro nor did they affect seedling development. The efficiency of the non-pathogenic F. oxysporum isolates in controlling Fusarium wilt was determined by immersing the tomato seedling roots in the conidial suspension (10(6) ml-1) of each isolate and then transplanting them into substrates previously infested with isolates of F. oxysporum f.sp. lycopersici, race 2 (10(5) conidia ml-1 of substrate). Evaluations were performed 35 days after transplanting, for severity in scale with 1=healthy plant to 6=dead plant or plant showing vessel browning and wilted leaves up to the leader shoot and seedling height. The non-pathogenic F. oxysporum isolates were efficient in reducing the severity of the disease and maintaining normal plant development. These results provide evidence of the antagonistic activity of non-pathogenic F. oxysporum isolates in controlling vascular wilt caused by F. oxysporum f. sp. lycopersici race 2 in tomato.


2008 ◽  
Vol 98 (6) ◽  
pp. 695-701 ◽  
Author(s):  
S. Li ◽  
C. C. Jochum ◽  
F. Yu ◽  
K. Zaleta-Rivera ◽  
L. Du ◽  
...  

Lysobacter enzymogenes C3 is a bacterial biological control agent that exhibits antagonism against multiple fungal pathogens. Its antifungal activity was attributed in part to lytic enzymes. In this study, a heat-stable antifungal factor (HSAF), an antibiotic complex consisting of dihydromaltophilin and structurally related macrocyclic lactams, was found to be responsible for antagonism by C3 against fungi and oomycetes in culture. HSAF in purified form exhibited inhibitory activity against a wide range of fungal and oomycetes species in vitro, inhibiting spore germination, and disrupting hyphal polarity in sensitive fungi. When applied to tall fescue leaves as a partially-purified extract, HSAF at 25 μg/ml and higher inhibited germination of conidia of Bipolaris sorokiniana compared with the control. Although application of HSAF at 12.5 μg/ml did not reduce the incidence of conidial germination, it inhibited appressorium formation and suppressed Bipolaris leaf spot development. Two mutant strains of C3 (K19 and ΔNRPS) that were disrupted in different domains in the hybrid polyketide synthase-nonribosomal peptide synthetase gene for HSAF biosynthesis and had lost the ability to produce HSAF were compared with the wild-type strain for biological control efficacy against Bipolaris leaf spot on tall fescue and Fusarium head blight, caused by Fusarium graminearum, on wheat. Both mutant strains exhibited decreased capacity to reduce the incidence and severity of Bipolaris leaf spot compared with C3. In contrast, the mutant strains were as efficacious as the wild-type strain in reducing the severity of Fusarium head blight. Thus, HSAF appears to be a mechanism for biological control by strain C3 against some, but not all, plant pathogenic fungi.


Plant Disease ◽  
1997 ◽  
Vol 81 (4) ◽  
pp. 404-409 ◽  
Author(s):  
Y. Luo ◽  
D. O. TeBeest

Colletotrichum gloeosporioides f. sp. aeschynomene causes an anthracnose of northern jointvetch, Aeschynomene virginica. Infection components, including lesion number, latent period, lesion expansion rate, and sporulation, were measured in experiments conducted in controlled environments. Two wild-type strains (3-1-3 and CLA 5A), four benomyl-resistant strains (B13, B15, B18 and B21), and four nitrate nonutilizing mutant strains (Nit A, Nit R, Nit L, and Nit T) of the pathogen were tested. Nitrate nonutilizing strains caused significantly fewer lesions on northern jointvetch than did wild-type and benomyl-resistant strains. Latent periods were significantly shorter for the wild-type strain CLA 5A than for most other strains. Lesion expansion rates of all benomyl-resistant strains were significantly slower than those of the wild- type strains. Large variations in sporulation were observed for most strains, and no differences in sporulation were found between wild-type and mutant strains. The usefulness of infection component analysis for the identification of competitiveness of strains of fungal pathogens for biological control of weeds is discussed.


2008 ◽  
Vol 98 (10) ◽  
pp. 1099-1106 ◽  
Author(s):  
Y. Iida ◽  
T. Kurata ◽  
Y. Harimoto ◽  
T. Tsuge

Fusarium oxysporum produces three kinds of asexual spores, microconidia, macroconidia, and chlamydospores. We previously found that the transcript level of the nitrite reductase gene of F. oxysporum, named FoNIIA, was markedly upregulated during conidiation compared with during vegetative growth. FoNIIA was also found to be positively regulated by Ren1 that is a transcription regulator controlling development of microconidia and macroconidia. In this study, we analyzed the function of FoNIIA in conidiation of F. oxysporum. Conidiation cultures showed markedly higher level of accumulation of FoNiiA protein as well as FoNIIA mRNA than vegetative growth cultures. FoNIIA protein was significantly decreased in cultures of the REN1 disruption mutant compared with that of the wild type. These results confirmed that FoNIIA expression is upregulated during conidiation and is positively regulated by REN1. The FoNIIA disruption mutants produced microconidia, macroconidia, and chlamydospores, which were morphologically indistinguishable from those of the wild type. The mutants, however, produced significantly fewer macroconidia than the wild type, although the wild type and mutant strains produced similar numbers of microconidia and chlamydospores. These results demonstrate that nitrite reductase is involved in quantitative control of macroconidium formation as well as nitrate utilization in F. oxysporum.


2003 ◽  
Vol 51 (3) ◽  
pp. 257 ◽  
Author(s):  
Khaled A. El-Tarabily

Twenty-one streptomycete and 15 non-streptomycete actinomycetes were isolated from surface-disinfested lupin roots and evaluated for their potential to produce chitinase and to inhibit the growth of Plectosporium tabacinum, the causal agent of lupin root rot in Egypt. The most inhibitory isolate was identified as Actinoplanes missouriensis which produced relatively high levels of chitinase and degraded the hyphae of P.�tabacinum in vitro, causing extensive plasmolysis and cell-wall lysis. A crude culture filtrate of A. missouriensis exhibited antifungal activity and significantly (P < 0.05) reduced spore germination and germ-tube growth of the pathogen. The antagonist was recovered from inside the root at all samplings up to 8 weeks after inoculation, indicating that the roots of healthy lupin may be a habitat for the endophyte. A. missouriensis significantly (P < 0.05) reduced the severity of root rot under glasshouse conditions. An endophytic isolate of Actinoplanes italicus incapable of producing chitinase and a mutant strain of A. missouriensis that did not produce detectable levels of chitinase, did not lyse hyphae of P. tabacinum or reduce root rot in the glasshouse experiments, although colonisation of the lupin root by both these isolates was similar to that of the chitinase-producing wild-type isolate of A. missouriensis. This study is the first record of control of a soil-borne plant pathogen by a chitinolytic actinomycete, endophytic in plant roots.


1996 ◽  
Vol 74 (11) ◽  
pp. 1841-1851 ◽  
Author(s):  
Joeke Postma ◽  
Anita J. G. Luttikholt

A nonpathogenic isolate of Fusarium oxysporum, 618-12, added to soil prior to the pathogen, suppressed fusarium wilt (F. o. f.sp. dianthi race 2) in a susceptible cultivar of carnation by 80% compared with the treatment with the pathogen only. The possibility of systemically induced resistance by the nonpathogenic isolate was assessed by inoculating antagonist and pathogen at different locations (stem versus soil, soil versus stem, and in a split-root system). No significant disease suppression was found with any of these spatially separated inoculations. However, inoculation of antagonist and pathogen at the same location within the stem (i.e., mixed stem inoculation) resulted in significant and reproducible disease reductions compared with stem inoculation with the pathogen alone. This reduction was found for different inoculum densities and different cultivars. Several other nonpathogenic Fusarium isolates could also reduce wilt symptoms in the susceptible carnation cultivar after mixed stem inoculation with the pathogen. This disease-suppressive effect after mixed stem inoculations may be caused by locally induced resistance or competition between isolates within the stem. Plants showed vascular browning around the inoculation point following inoculation with nonpathogenic isolates. Disease suppression, as well as vascular browning, were absent when dead conidia of the isolate 618-12 were used. After its addition to soil, the isolate was recovered from 44–78% of carnation stems. Spread of the nonpathogenic isolate within the stem occurred only in the first 4 days after stem inoculation, and it remained confined to limited distances from the inoculation point between 4 and 59 days after inoculation. The pathogenic isolate could be isolated at increasingly greater distances from the inoculation point during this period. These data suggest that the nonpathogenic isolate is not actively spread through the plant by growth of the fungus. Keywords: biological control, competition, induced resistance, wilt.


HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 519c-519 ◽  
Author(s):  
Brent K. Harbaugh ◽  
Jeffrey B. Jones ◽  
Lee E. Jackson ◽  
Gail Somodi ◽  
Joseph E. Flaherty

Bacterial blight, caused by Xanthomonas campestris pv. pelargonii (XCP), is considered the most serious disease of geraniums (Pelargonium × hortorum). A novel approach that uses bacteriophages (phages, viruses that kill bacteria) for the biological control of geranium blight will be presented. Phages were once abandoned as biological control agents due to the emergence of bacterial mutants resistant to the phages employed. However, our approach utilizes a mixture of three to eight different phages including host-range mutants (H-mutants). H-mutants are spontaneously derived from their wild-type parent phages and lyse not only parent wild-type bacteria, but also phage-resistant mutants originating from parent bacteria. Two phages specific for XCP initially were isolated from soil samples from Florida and California. These phages produced virulent reactions in six of 30 XCP strains, and lysogenic reactions in 22 strains. After selection of these phages for increased virulence and additional phages were isolated from MN and UT, 17 phages were evaluated for sensitivity to 21 XCP strains from around the world. Four to 14 phages produced virulent reactions in the 21 XCP strains. Five phages produced virulent reactions in at least 17 XCP strains. A mixture of five phages tested against the 21 XCP strains produced virulent reactions for all 21 XCP strains. Geraniums in 10-cm pots were inoculated with XCP and placed on a greenhouse bench in the middle of 5 non-inoculated plants. After 2 weeks of daily spraying plants with a phage solution (109 pfu phage/ml) or water, there was a 71% reduction in the number of bacterial lesions on phage-treated plants.


1998 ◽  
Vol 11 (2) ◽  
pp. 91-98 ◽  
Author(s):  
Antonio Di Pietro ◽  
M. Isabel G. Roncero

pg1 encoding the major in vitro extracellular endopolygalacturonase of the tomato vascular wilt pathogen Fusarium oxysporum f. sp. lycopersici was cloned and sequenced. The deduced mature protein had a calculated molecular mass of 35.5 kDa and a pI of 6.2, and showed significant similarity with other fungal endoPGs. pg1 mRNA was induced in vitro by citrus pectin, tomato vascular tissue, 0.1% D-galacturonic acid, and polygalacturonic acid, and repressed by 1% D-galacturonic acid and 1% glucose. Reverse transcription-polymerase chain reaction revealed pg1 expression in roots and lower stems of tomato plants infected by F. oxysporum f. sp. lycopersici. Three naturally occurring F. oxysporum f. sp. melonis isolates deficient in PG1 were transformed with the cloned gene. The PG1 enzyme secreted by the transformants had the same molecular mass, pI, and glycosylation pattern as those of the donor isolate. Polygalacturonase activity in cultures of transformants grown in vitro on citrus pectin and on melon plants, but not on glucose, increased 10- to 20-fold, compared with the PG1-deficient wild-type isolate, whereas mycelial dry weight increased two- to threefold. Transformants exhibited the same degree of virulence toward susceptible muskmelon cultivars as the wild-type isolate and were avirulent on a resistant cultivar.


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