Biological control of Pythium aphanidermatum in cucumber with a combined application of Lysobacter enzymogenes strain 3.1T8 and chitosan

Pythium aphanidermatum (Edson) Fitzp., causing root and crown rot in cucumber, was successfully managed by Lysobacter enzymogenes strain 3.1T8. Greenhouse experiments were performed with cucumber plants grown in rockwool blocks up to 5 weeks with a recirculated nutrient solution. Application of L. enzymogenes 3.1T8 in combination with chitosan (the deacetylated derivative of chitin) reduced the number of diseased plants by 50–100% in four independent experiments relative to the Pythium control. Application of chitosan or the bacterial inoculant alone was not effective. Washed bacterial cells plus chitosan inhibited Pythium-induced disease, but the supernatant without bacterial cells combined with chitosan was not effective. The most effective and convenient type of commercially available chitosan was selected. Chitosan disappeared from the hydroponic system within 24 h after application, which we attribute to enzyme expression of L. enzymogenes 3.1T8 induced by the exposure to chitosan. Plate counts of the nutrient solution on a general bacterial medium showed the dominance of the inoculated strain, and an increased bacterial population growing on chitin and chitosan as single carbon source. The population density of L. enzymogenes 3.1T8 on the cucumber roots was investigated with a strain specific real-time TaqMan PCR. Highest chitosan concentrations applied (0.1 and 0.03 g/plant) resulted in the highest numbers of L. enzymogenes 3.1T8 present on roots; i.e. 10⁸–10⁹ cells/g root. Substantially higher numbers of bacterial cells were observed by scanning electron microscopy after application of chitosan; no morphological or other qualitative differences were found. The results indicate that addition of chitosan enhanced the biocontrol efficacy of L. enzymogenes 3.1T8; either chitosan serves as C- and N-source for the antagonist, induces antagonistic gene expression, or both.     

Biological control of rice bacterial blight by Lysobacter antibioticus strain 13-1

Bacterial leaf blight (BB) is a worldwide destructive rice disease caused by pathogen Xanthomonas oryzae pv. oryzae (Xoo). A novel strain of Lysobacter antibioticus, which was isolated from the rhizosphere of rice in Yunnan Province of China, can significantly inhibit the growth of various phytopathogenic bacteria and fungi, especially BB pathogen Xoo. In greenhouse experiments, whole bacterial broth culture (WBC) of strain 13-1 was more effective in reducing BB than other components of the culture, with disease suppression efficiency up to 69.7%. However, bacterial cells re-suspended in water, cell-free culture extracts, and heated cultures also significantly reduced BB severity. Suppression efficiencies ranged from 79.0% to 61.8% for undiluted to 100-fold dilution treatments and from 57.6% to 31.7% when the WBC of strain 13-1 (10⁸ CFU/mL) was applied at 3 days and 7 days prior to pathogen inoculation, respectively. In three field trials, strain 13-1 reduced BB incidence by 73.5%, 78.3%, and 59.1%, respectively. Disease suppression by strain 13-1 varied significantly among different rice cultivars, although efficacy was not directly related to the susceptibility level of the cultivars. Efficacy of biocontrol was also affected by different pathogen isolates, with some isolates of Xoo being more sensitive to 13-1 suppression than others. These results suggest that antibiotics and density of colonization on leaves may be involved for biological control of rice BB by strain 13-1. To our knowledge, this is the first report of L. antibioticus being a potential biocontrol agent for rice bacterial blight.     

Biological control of seedling blight of wheat caused by Fusarium graminearum with beneficial rhizosphere microorganisms

Fusarium graminearum is associated with the cereal damping-off complex which reduces germination, seedling stand and yield. Fifty-two bacterial strains and six Trichoderma spp. isolated from the wheat rhizosphere were evaluated for biocontrol of seedling blight of wheat caused by F. graminearum. Their potential as biocontrol agents was tested in vitro and in the greenhouse. Isolates varied in their ability to inhibit the mycelial growth of F. graminearum in agar plate bioassays by 0–79%. This parameter was not related with biocontrol efficacy of in vivo assays. In greenhouse trials, all isolates were initially evaluated for reducing disease on wheat cultivars Klein Centauro (moderately resistant to F. graminearum) and Pro INTA Oasis (susceptible) planted in sterilized soil artificially infested with the pathogen. Among the 25 bacteria and six fungal isolates that exhibited a pronounced suppressive effect, the most efficient 10 for both cultivars were further assayed on eight cultivars (Buck Candil, Buck Catriel, Buck Chambergo, Buck Poncho, Buck Topacio, Klein Cacique, Klein Centauro and Pro INTA Oasis) potted in cultivated–inoculated soil. Three weeks after sowing, plant stand, percentage of diseased emerging seedlings, plant height and dry weight were evaluated. Among the antagonists only Stenotrophomonas maltophilia was significantly better than the control for the average of the eight cultivars for plant stand, height and dry weight. Stenotrophomonas maltophilia also caused a non-significant decrease in the percentage of diseased plants. Three strains of Bacillus cereus and one isolate of Trichoderma harzianum gave also a good control in some cultivars. The ability of these isolates to affect the infection of wheat seedlings by F. graminearum may be of potential value in field trials.     

一种自溶性产酶溶杆菌新菌株LE16对温室番茄灰霉病的抑制作用

【目的】我国人多地少,设施农业日益普及,温室栽培保障了番茄的有效供给,在温室中种植的番茄容易发生灰霉病,造成减产。生物防治产品对人畜安全,环境友好,但亟待增加生防微生物种类,提高防效。【方法】以自主分离的自溶性产酶溶杆菌(Lysobacter enzymogenes)LE16为对象,利用纯培养试验明确其发酵液对灰霉病菌(Botrytis cinerea)的拮抗作用;生物化学、液相色谱-质谱联用和转录组技术从生防菌合成分泌的胞外水解酶、抗菌物质和病菌基因差异表达等方面,揭示生防菌拮抗病原真菌的机制;温室盆栽试验评估菌株LE16发酵液对番茄灰霉病的防治潜力。【结果】LE16能合成分泌铁载体、多种与抗病性相关的酶类(包括磷酸酶、蛋白酶、纤维素酶、β-1,3-葡聚糖酶和溶菌酶)以及9种抗真菌物质(毒菌素D、N-十一烷基苯磺酸、利福布汀、纳奈霉素、替加环素、米诺环素、间甲酚、肉桂酸和环戊酮)和激活植物系统获得性抗性的P-水杨酸。LE16发酵液粗提物显著抑制灰霉病菌生长繁殖,抑制率为34.99%-100%,显著高于产酶溶杆菌HYP18和撕裂蜡孔菌HG2011。此外,LE16发酵液显著诱导灰霉病菌基...     

Utilization of chemical inducers of resistance and Cryptococcus flavescens OH 182.9 to reduce Fusarium head blight under greenhouse conditions

Four chemicals [salicylic acid (SA), sodium salt of salicylic acid (NaSA), isonicotinic acid (INA), and DL-β-amino-n-butyric acid (BABA)] and the yeast antagonist Cryptococcus flavescens (=C. nodaensis nomen nudum) OH 182.9 were evaluated separately or together for the ability to reduce Fusarium head blight (FHB) of wheat in the greenhouse. When sprayed onto wheat heads at 3 days prior to pathogen challenge with Gibberella zeae, NaSA and INA at 10 mM significantly reduced FHB severity compared to the non-treated disease control. Applied at concentrations of 1 and 5 mM at 3 days before pathogen challenge, NaSA or INA in combination with OH 182.9 did not significantly reduce FHB severity compared to either treatment alone, though the lowest disease severity values frequently were associated with the combination treatments. When sprayed onto wheat heads just beginning to emerge from boot at 10 days prior to pathogen inoculation, NaSA, INA, and BABA at 1 mM significantly reduced FHB severity indicating that induced systemic resistance was at least partially responsible for the reduction of FHB disease. Induced FHB resistance was achieved by treating wheat with INA at concentrations as low as 0.1 mM. In only one instance was 100-kernel weight affected by any chemical or combination of chemicals with OH 182.9 treatment. Data from our studies in the greenhouse suggest that chemical inducers can induce resistance in wheat against FHB, and that further efforts are warranted to explore the potential of improved control of FHB disease by incorporating chemical inducers with the FHB biocontrol agent OH 182.9.     

Overexpression of defense response genes in transgenic wheat enhances resistance to Fusarium head blight

Fusarium head blight (FHB) of wheat, caused by Fusarium graminearum and other Fusarium species, is a major disease problem for wheat production worldwide. To combat this problem, large-scale breeding efforts have been established. Although progress has been made through standard breeding approaches, the level of resistance attained is insufficient to withstand epidemic conditions. Genetic engineering provides an alternative approach to enhance the level of resistance. Many defense response genes are induced in wheat during F. graminearum infection and may play a role in reducing FHB. The objectives of this study were (1) to develop transgenic wheat overexpressing the defense response genes α-1-purothionin, thaumatin-like protein 1 (tlp-1), and β-1,3-glucanase; and (2) to test the resultant transgenic wheat lines against F. graminearum infection under greenhouse and field conditions. Using the wheat cultivar Bobwhite, we developed one, two, and four lines carrying the α-1-purothionin, tlp-1, and β-1,3-glucanase transgenes, respectively, that had statistically significant reductions in FHB severity in greenhouse evaluations. We tested these seven transgenic lines under field conditions for percent FHB disease severity, deoxynivalenol (DON) mycotoxin accumulation, and percent visually scabby kernels (VSK). Six of the seven lines differed from the nontransgenic parental Bobwhite line for at least one of the disease traits. A β-1,3-glucanase transgenic line had enhanced resistance, showing lower FHB severity, DON concentration, and percent VSK compared to Bobwhite. Taken together, the results showed that overexpression of defense response genes in wheat could enhance the FHB resistance in both greenhouse and field conditions.     

小麦赤霉病拮抗菌JB7的拮抗活性及鉴定

由禾谷镰刀菌(Fusarium graminearum, Fg)引起的赤霉病是限制小麦生产的主要病害之一。生物防治是一种高效且可持续的防治方法。【目的】从小麦种子内筛选具有抑制禾谷镰刀菌的菌株并对其生防潜力进行评估,为小麦赤霉病生防制剂的开发与利用提供菌种资源及理论支撑。【方法】采用平板对峙、孢子萌发法和无菌上清液抑菌试验筛选小麦种子内对禾谷镰刀菌具有拮抗活性的内生菌株;利用扫描电镜(scanning electron microscope, SEM)和共聚焦扫描电镜(confocal laser scanning microscope, CLSM)观察并分析无菌上清液对Fg的分生孢子形态、膜完整性以及胞内活性氧的影响;通过盆栽试验验证内生菌对小麦赤霉病的生防效果;应用二代Illumina HiSeq测序平台进行全基因组测序。【结果】从小麦种子中分离出一株高效抑制Fg生长的内生菌株JB7,其衰亡期无菌上清液对Fg孢子萌发抑制率高达85.23%。菌株JB7的无菌上清液使Fg孢子表面凹陷,破坏其细胞膜,造成核酸和蛋白质的渗漏,诱导Fg菌丝活性氧的累积,引起Fg菌丝可溶性蛋白和丙二醛含量的显著升高。该菌株具有分泌蛋白酶、纤维素酶、葡聚糖酶和产铁载体的能力。盆栽试验表明菌株JB7能显著降低小麦赤霉病的病情指数(P<0.05)。经全基因组学鉴定为甲基营养型芽孢杆菌(Bacillus methylotrophicus) JB7,该菌株基因组中含有12个抑菌功能的次级代谢产物合成基因簇。【结论】菌株JB7能抑制禾谷镰刀菌的生长,对小麦赤霉病有较强的防效,可作为生物防治小麦赤霉病的候选菌株。     

Mapping of Fhb2 on chromosome 6BS: a gene controlling Fusarium head blight field resistance in bread wheat (Triticum aestivum L.)

Fusarium head blight (FHB) is one of the most important fungal wheat diseases worldwide. Understanding the genetics of FHB resistance is key to facilitate the introgression of different FHB resistance genes into adapted wheat. The objective of this project was to study the FHB resistance QTL on chromosome 6B, quantify the phenotypic variation, and qualitatively map the resistance gene as a Mendelian factor. The FHB resistant parent BW278 (AC Domain*2/Sumai 3) was used as the source of the resistance allele. A large recombinant inbred line (RIL) mapping population was developed from the cross BW278/AC Foremost. The population segregated for three known FHB resistance QTL located on chromosomes 3BSc, 5A, and 6B. Molecular markers on chromosome 6B (WMC104, WMC397, GWM219), 5A (GWM154, GWM304, WMC415), and 3BS (WMC78, GWM566, WMC527) were amplified on approximately 1,440 F_2:7 RILs. The marker information was used to select 89 RILs that were fixed homozygous susceptible for the 3BSc and 5A FHB QTLs and were recombinant in the 6B interval. Disease response was evaluated on 89 RILs and parental checks in the greenhouse and field nurseries. Dual floret injection (DFI) was used in greenhouse trials to evaluate disease severity (DS). Macroconidial spray inoculations were used in field nurseries conducted at two locations in southern Manitoba (Carman and Glenlea) over two years 2003 and 2004, to evaluate disease incidence, disease severity, visual rating index, and Fusarium-damaged kernels. The phenotypic distribution for all five-disease infection measurements was bimodal, with lines resembling either the resistant or susceptible checks and parents. All of the four field traits for FHB resistance mapped qualitatively to a coincident position on chromosome 6BS, flanked by GWM133 and GWM644, and is named Fhb2. The greenhouse-DS trait mapped 2 cM distal to Fhb2. Qualitative mapping of Fhb2 in wheat provides tightly linked markers that can reduce linkage drag associated with marker assisted selection of Fhb2 and aid the pyramiding of different resistance loci for wheat improvement.     

Osmotic stress adaptation, compatible solutes accumulation and biocontrol efficacy of two potential biocontrol agents on Fusarium head blight in wheat

Fusarium head blight (FHB) caused by Gibberella zeae (anamorph = Fusarium graminearum) is a devastating disease that causes extensive yield and quality losses to wheat in humid and semi-humid regions of the world. Biological control has been demonstrated to be effective under laboratory conditions but a few biocontrol products have been effective under field conditions. The improvement in the physiological quality of biocontrol agents may improve survival under field conditions, and therefore, enhance biocontrol activity. Bacillus subtilis RC 218 and Brevibacillus sp. RC 263 were isolated from wheat anthers and showed significant effect on control of FHB under greenhouse assays. This study showed the effect of water availability measured as water activity (a_W) using a growth medium modified with NaCl, glycerol and glucose on: (i) osmotic stress tolerance, (ii) viability in modified liquid medium, (iii) quantitative intracellular accumulation of betaine and ectoine and (iv) the biocontrol efficacy of the physiologically improved agents. Viability of B. subtilis RC 218 in NaCl modified media was similar to the control. Brevibacillus sp. RC 263 showed a limited adaptation to growth in osmotic stress. Betaine was detected in high levels in modified cells but ectoine accumulation was similar to the control cells. Biocontrol activity was studied in greenhouse assays on wheat inoculated at anthesis period with F. graminearum RC 276. Treatments with modified bacteria reduced disease severity from 60% for the control to below 20%. The physiological improvement of biocontrol agents could be an effective strategy to enhance stress tolerance and biocontrol activity under fluctuating environmental conditions.     

Genotype x strain interactions for resistance to Fusarium head blight caused by Fusarium culmorum in winter wheat

Summary In 3 consecutive years, a set of 17 winter wheat genotypes, representing a wide range of Fusarium head blight resistance, was inoculated with four strains of Fusarium culmorum. Fusarium head blight ratings were analyzed. The interaction between genotypes, strains, and years was described using a Finlay-Wilkinson model and an Additive Main effects and Multiplicative Interaction effects (AMMI) model. The interaction consisted primarily of a divergence of genotypical responses with increasing disease pressure, modified by genotype specific reactions in certain years. The divergence was mainly caused by one very pathogenic strain. The Fusarium head blight resistance in this study can be described as horizontal resistance in terms of Vanderplank, with the exception of three genotypes selected from one particular cross that showed a strain-year combination dependent resistance which was ineffective in 1 year.     

Genetic dissection of a major Fusarium head blight QTL in tetraploid wheat

The devastating effect of Fusarium head blight (FHB) caused by Fusarium graminearum has led to significant financial losses across the Upper Midwest of the USA. These losses have spurred the need for research in biological, chemical, and genetic control methods for this disease. To date, most of the research on FHB resistance has concentrated on hexaploid wheat (Triticum aestivum L.) lines originating from China. Other sources of resistance to FHB would be desirable. One other source of resistance for both hexaploid wheat and tetraploid durum wheat (T. turgidum L. var. durum) is the wild tetraploid, T. turgidum L. var. dicoccoides (T. dicoccoides). Previous analysis of the `Langdon'-T. dicoccoides chromosome substitution lines, LDN(Dic), indicated that the chromosome 3A substitution line expresses moderate levels of resistance to FHB. LDN(Dic-3A) recombinant inbred chromosome lines (RICL) were used to generate a linkage map of chromosome 3A with 19 molecular markers spanning a distance of 155.2 cM. The individual RICL and controls were screened for their FHB phenotype in two greenhouse seasons. Analysis of 83 RICL identified a single major quantitative trait locus, Qfhs.ndsu-3AS, that explains 37% of the phenotypic or 55% of the genetic variation for FHB resistance. A microsatellite locus, Xgwm2, is tightly linked to the highest point of the QTL peak. A region of the LDN (Dic-3A) chromosome associated with the QTL for FHB resistance encompasses a 29.3 cM region from Xmwg14 to Xbcd828.     

Purification and characterization of mannitol dehydrogenase and identification of the corresponding cDNA from the head blight fungus,Gibberella zeae (Fusarium graminearum)

The mannitol-2-dehydrogenase (MtDH) from Gibberella zeae was purified and the corresponding cDNA identified. Purification of MtDH was accomplished using a combination of ammonium sulfate fractionation, anion exchange and dye-ligand chromatography. Final purification was achieved following electroelution from a native gel. Molecular mass determination based on SDS-PAGE indicated that the denatured protein was 29 kDa. Native protein mass was determined to be 110 kDa using gel permeation chromatography, indicating a tetrameric form. The pH optima for mannitol oxidation and fructose reductase activities were 9.0, and 7.0, respectively. Activity with sorbitol as the substrate was 21% of activity with mannitol. Kinetic parameters were determined by direct-linear plots of enzyme activity vs. substrate concentrations. Fructose concentrations above 600 mM and NADPH concentrations above 0.3 mM caused substrate inhibition. Comparisons of predicted amino acid sequences of several fungal MtDHs indicated high conservation within the phyla. A possible role for MtDH in generation of turgor pressure for forcible ascospore discharge is discussed.     

DNA marker analysis for pyramided of Fusarium head blight (FHB) resistance QTLs from different germplasm

A pyramided FHB resistance line of wheat (WSY) was previously developed from three FHB resistant cultivars (Sumai 3, Wangshuibai, and Nobeokabouzu) in the Jiangsu Academy of Agricultural Sciences, China. In the present study, we analyzed the genetic relationship between WSY and the three parental cultivars using DNA markers in order to clarify how many and which resistance genes had accumulated in WSY. We analyzed 282 DNA markers from the 21 wheat chromosomes. WSY was found to include different chromosome regions that harbored putative FHB QTLs of the three parental germplasm. Haplotypes of DNA markers on these QTL regions revealed that the 1BL, 2BL, 5AS, and 7AL QTL regions were from Sumai 3, the 2AS, 2DS, 3AS, and 6BS QTL regions were from Wangshuibai, and the 3BS QTL region was from Nobeokabouzu. This study showed that different resistance genes from the different resistant germplasm had indeed accumulated in WSY. WSY is a potential resistant resource for FHB resistance in wheat breeding programs.     

Saturation and comparative mapping of a major Fusarium head blight resistance QTL in tetraploid wheat

Fusarium head blight (FHB) is a devastating disease of cultivated wheat worldwide. Partial resistance to FHB has been identified in common wheat (Triticum aestivum L.). However, sources of effective FHB resistance have not been found in durum wheat (T. turgidum L. var. durum). A major FHB resistance quantitative trait loci (QTL), Qfhs.ndsu-3AS, was identified on chromosome 3A of T. dicoccoides, a wild relative of durum wheat. Here, we saturated the genomic region containing the QTL using EST-derived target region amplified polymorphism (TRAP), sequence tagged site (STS), and simple sequence repeat (SSR) markers. A total of 45 new molecular marker loci were detected on chromosome 3A and the resulting linkage map consisted of 55 markers spanning a genetic distance of 277.2 cM. Qfhs.ndsu-3AS was positioned within a chromosomal interval of 11.5 cM and is flanked by the TRAP marker loci, Xfcp401 and Xfcp397.2. The average map distance between the marker loci within this QTL region was reduced from 4.9 cM in the previous study to 3.5 cM in the present study. Comparative mapping indicated that Qfhs.ndsu-3AS is not homoeologous to Qfhs.ndsu-3BS, a major FHB QTL derived from the common wheat cultivar Sumai 3. These results facilitate our efforts toward map-based cloning of Qfhs.ndsu-3AS and utilization of this QTL in durum wheat breeding via marker-assisted selection.     

Biological control of crown rot of bananas with Pichia anomala strain K and Candida oleophila strain O

The antagonistic activity of two yeast strains (Pichia anomala (E.C. Hansen) Kurtzman, strain K and Candida oleophila Montrocher, strain O) against the parasitic complex responsible for banana crown rot was evaluated. The strains were applied at three different concentrations (10⁶, 10⁷, 10⁸ cfu/ml) and their efficacy tested in vivo on three separate fungi (Colletotrichum musae (Berk. & Curt.) Arx, Fusarium moniliforme Sheldon, and Cephalosporium sp.) and on a parasitic complex formed by association of these three fungi. At the concentrations used C. musae appeared to be the most pathogenic. The complex showed intermediate aggressiveness between C. musae and both other fungi.Statistically significant antagonistic effects were observed on C. musae, F. moniliforme, and the fungal complex. The highest protection level (54.4%) was observed with strain O added at 10⁸ cfu/ml on crowns previously inoculated with the fungal complex. The level was lower when the fungi were inoculated separately.Furthermore, the antagonistic effect was strongly reinforced when strain O at 10⁸ cfu/ml was applied 24 h before fungal complex inoculation (59.9%), as compared to its application 15 min (24.3%) or 3 h (27.3%) after fungal complex inoculation. Bananas showed increased susceptibility to the fungal complex from March to June, and this influenced the level of protection by yeast, which decreased over the same period. A strict negative correlation (R² = 0.83) was highlighted between susceptibility of banana to crown rot and protection provided by yeast.     

Susceptibility to <Emphasis Type="Italic">Fusarium </Emphasis>head blight is associated with the <Emphasis Type="Italic">Rht-D1b</Emphasis> semi-dwarfing allele in wheat

Fusarium head blight (FHB) is an important disease of wheat worldwide. The cultivar Spark is more resistant than most other UK winter wheat varieties but the genetic basis for this is not known. A mapping population from a cross between Spark and the FHB susceptible variety Rialto was used to identify quantitative trait loci (QTL) associated with resistance. QTL analysis across environments revealed nine QTL for FHB resistance and four QTL for plant height (PH). One FHB QTL was coincident with the Rht-1D locus and accounted for up to 51% of the phenotypic variance. The enhanced FHB susceptibility associated with Rht-D1b is not an effect of PH per se as other QTL for height segregating in this population have no influence on susceptibility. Experiments with near-isogenic lines supported the association between susceptibility and the Rht-D1b allele conferring the semi-dwarf habit. Our results demonstrate that lines carrying the Rht-1Db semi-dwarfing allele are compromised in resistance to initial infection (type I resistance) while being unaffected in resistance to spread within the spike (type II resistance).     

The feruloyl esterase gene family of Fusarium graminearum is differentially regulated by aromatic compounds and hosts

Feruloyl esterases can liberate ferulic acid (FA) from plant cell wall polymers. They are expressed by plant pathogenic fungi and could play a role in pathogenicity, although this question has not been addressed yet. The fungus Fusarium graminearum is the principal causal agent of fusarium head blight (FHB) and gibberella ear rot (GER), major diseases of wheat, barley, and maize in all temperate regions of the world. The F. graminearum genome contains seven genes with strong homology to feruloyl esterase (FAE) sequences. Phylogenetic analysis showed that these included three type B, three type C, and one type D FAE genes. Expression profiling of the seven FAE genes showed complex regulation patterns unique to each gene. In F. graminearum-infected plant tissues, the FAE genes exhibited host-specific gene expression. On wheat, FAEB1 and FAED1 were strongly expressed while FAEB2, FAEB3, and FAEC1 were expressed at more modest levels. On maize, only FAEB3, FAEC1, and FAED1 were expressed and at low levels. When growing F. graminearum in liquid culture, only FAEB1 and FAEC1 were expressed. Both genes were induced by a small group of related aromatic compounds including FA, caffeic acid, and p-coumaric acid. FAEB1 was induced by xylose, while repressed by glucose and galactose. FAEC1 was constitutively expressed at low levels in the presence of those sugars. Expression of the other five FAE genes was not detected in the culture conditions used. To determine if FAE genes were important for pathogenicity of F. graminearum, mutant strains inactivated for faeB1?, faeD1? or both genes were constructed and tested on wheat plants. No statistically significant change in pathogenicity and no compensatory expression of the other FAE genes were observed in the fae gene mutants. Our results show that FAEB1 and FAED1 are not required for pathogenicity of F. graminearum on wheat.     

Correlation between screening procedures to select root endophytes for biological control of Fusarium verticillioides in Zea mays L

Biological control is an alternative to pesticides for protection against crop diseases. A key to progress in the field of biological control to protect maize against Fusarium verticillioides is to select in vitro the best agent to be applied in the field. This research was undertaken to evaluate the correlation between different screening methods and to suggest an adequate procedure that could be used to select bacterial agents with potential biocontrol against F. verticillioides in the maize rhizosphere. The results show an extensive Pearson correlation coefficient analysis between different screening procedures carried out for Arthrobacter spp., Azotobacter spp., Pseudomonas spp., and Bacillus spp. paired with 13 F. verticillioides strains isolated from maize endorhizosphere. The following screening methodologies were correlated: niche overlap index (NOI), indices of dominance, growth rate, lag phase, antibiosis, and fumonisin production. It was observed that NOI and antibiosis methodologies did not show any correlation. They were used to choose the best bacteria to apply under greenhouse conditions. Azotobacter armeniacus RC2 showed a NOI > 0.9 and inhibited all F. verticillioides strains assayed. Seed maize bacterization with A. armeniacus RC2 at 10⁶ and 10⁷ inoculum level resulted in significantly lower values of F. verticillioides counts compared to the control without bacteria at the root levels assayed. The analysis of variance indicated that there were significant interactions between two fungal repression assays. Antibiosis assays significantly correlated with greenhouse conditions (p < 0.01) at two inoculum levels tested. Therefore, the selection system adopted was adequate to choose the best bacterial biocontrol agent. The application of this methodology shows a way to outline the best biocontrol candidate.     

The potential of Ulocladium botrytis for biological control of Orobanche spp.

A strain of Ulocladium botrytis isolated from diseased Orobanche crenata shoots caused disease on the parasitic weed in pathogenicity tests. The potential of the fungus to be developed as a mycoherbicide for Orobanche spp. was further investigated. Although the fungus significantly decreased O. crenata germination in vitro by 80%, it did not generally lead to a decreased number of O. crenata shoots or tubercles in inoculated root chambers or pots. However, the number of diseased or dead tubercles and underground shoots was significantly increased compared to the noninoculated treatments. Postemergence inoculation of O. crenata shoots with a conidial suspension resulted in the death of almost all inoculated plants 14 days after application under greenhouse conditions. In preliminary host-range studies, the pathogen caused disease on Orobanche cumana on sunflower whereas on Orobanche aegyptiaca shoots parasitizing tomato only minimal disease symptoms could be detected after postemergence inoculation. Based on the results of our investigations, we conclude that Ulocladium botrytis has only a limited potential to be used as a biocontrol agent against Orobanche spp.     

Screening rhizobacteria for biological control of Ralstonia solanacearum in Ethiopia

Bacterial wilt caused by Ralstonia solanacearum (Smith) has become a severe problem mainly on potato and tomato in Ethiopia and no effective control measure is available yet. To explore possibilities for the development of biological control for the disease, 118 rhizobacteria, most of them collected from Ethiopia, were screened against an Ethiopian R. solanacearum strain. On the basis of in vitro screening, six strains (RP87, B2G, APF1, APF2, APF3, and APF4) with good inhibitory effect were selected for in planta testing in a greenhouse. In the greenhouse, soil and tomato seedlings were treated with the antagonists and their effects studied. The study showed that APF1 and B2G strains significantly reduced disease incidence and increased weight of tomato plants. Area under disease progress curves (AUDPC) was reduced by 60% and 56% in plants inoculated with APF1 and B2G strains, respectively. Plant dry weight increase in plants inoculated with APF1 and B2G strains was 96% and 75%, respectively. APF1 was found to be the most beneficial strain in disease suppression and also growth promotion resulting in 63% dry weight increase compared to untreated control. The study revealed that APF1 and B2G strains are promising strains whose effectiveness under field conditions and their mode of action should be investigated.