A rapid technique for screening banana cultivars for resistance to Xanthomonas wilt

The banana Xanthomonas wilt disease (BXW) has threatened the livelihood of millions of farmers in East Africa. Use of resistant varieties is the most cost-effective method of managing this bacterial disease. A reliable and rapid screening method is needed to select resistant banana varieties. An in vitro screening method was developed for early evaluation of Xanthomonas wilt resistance using small tissue culture-grown plantlets. Eight cultivars of banana were screened with sixteen isolates of Xanthomonas campestris pv. musacearum using this method. There were significant differences (P < 0.0001) in susceptibility among the various banana cultivars tested, whereas no significant difference (P = 0.92) in pathogenicity was observed between the pathogen isolates. The cv. Pisang Awak (Kayinja) was found to be highly susceptible and Musa balbisiana resistant. Nakitembe was found to be moderately resistant while cvs Mpologoma, Mbwazirume, Sukali Ndiizi, FHIA-17 and FHIA-25 were susceptible. The susceptibility of these cultivars was further tested in vivo by artificial inoculation of potted plants with similar results. This study shows that an in vitro screening test can serve as a convenient, cheap and rapid screening technique to discriminate BXW-resistant from BXW-susceptible banana cultivars.     

Effect of leaf scald (<Emphasis Type="Italic">Xanthomonas albilineans</Emphasis>) on polyamine and phenolic acid metabolism of two sugarcane cultivars

Polyamine and phenolic acid levels as well as activities of some enzymes of their biosynthetic metabolism were examined in two sugarcane (Saccharum officinarum) cultivars differing in susceptibility to leaf scald, a disease caused by the bacterium, Xanthomonas albilineans. Juice obtained from both infected cultivars showed significantly increased levels of free putrescine and ornithine decarboxylase activity. However, the pathogen induced different changes in the two cultivars in subsequent metabolic steps. Whereas acid insoluble conjugated spermidine completely disappeared from the highly susceptible cv. C 439-52, an increase in acid insoluble conjugated polyamines was observed in the moderately susceptible cv. L 55-5. Phenolic acid metabolism also differed in the two cultivars. Since total phenolic acid content and phenylalanine ammonium lyase activity was greater in both cultivars after infection, distribution of phenolic acids between free or conjugated forms diverted into different pathways. The level of susceptibility of the two cultivars is discussed in terms of changes in these compounds.     

瓜类果斑病菌致病基因及靶标基因的预测

哈密瓜果斑病是严重危害瓜类的种传细菌性病害,其病原菌为嗜酸菌属西瓜种(Acidovorax citrulli)。本文首先用Bioedit软件建立瓜类果斑病菌全氨基酸序列本地资源库,运用比较基因组学的方法,通过大量查找嗜酸菌属、欧文氏菌属、假单胞菌属、黄单胞菌属、劳尔氏菌属、土壤杆菌属、木质部小菌属中的致病基因和参考文献中已报道的致病基因,下载相关致病基因的氨基酸序列,与瓜类果斑病菌的全氨基酸序列进行localblast,取E<10-5的为候选基因,得到果斑病菌中与致病性相关的基因,并对未知蛋白进行产物预测。同时,根据已经公布的靶标基因,用同样的比对方法,得到果斑病菌中潜在的靶标基因。本研究通过保守估计,得到77个致病性相关蛋白,预测了其中7个未知蛋白的产物,得到46个靶标蛋白。致病基因和靶标基因的分析预测,对以后研究此类基因提供了指导作用,为更好地防治该病害提供了理论基础。     

Disease levels in winter wheat,rye and triticale grown on soil artificially inoculated withCephalosporium gramineum

Field experiments with winter cereals grown on soil inoculated withC. gramineum showed that wheat and rye cultivars possess some resistance to the pathogen, while the triticale cultivars were the most susceptible. Higher tolerance of the tested wheat cultivars was connected mainly with slow development of disease symptoms; rye cultivars had, on average, lower percentages of plants infected byC. gramineum. The greatest variation in susceptibility toC. gramineum occurred among the selected cultivars of triticale.     

Symptom development, pathogen isolation and Real-Time QPCR quantification as factors for evaluating the resistance of olive cultivars to Verticillium pathotypes

Verticillium wilt is the most serious olive disease in the Mediterranean countries and worldwide. The most effective control strategy is the use of resistant cultivars. However, limited information is available about the level and source of resistance in most of the olive cultivars and there are no published data using microsclerotia, the resting structures of Verticillium dahliae, as the infective inoculum. In the present study, we correlated symptomatology and the presence of the fungus along with the DNA relative amount (molecules μl⁻¹) of a defoliating (D) and a non-defoliating (ND) V. dahliae strain in the susceptible cv. Amfissis and the tolerant cvs Kalamon and Koroneiki, as quantified by the Real-Time QPCR technology. The viability of the pathogen in the plant tissues was confirmed by isolating the fungus on PDA plates, while symptom assessment proved the correlation between the DNA relative amount of V. dahliae in plant tissues and cultivar susceptibility. It was further demonstrated that the D and ND strains were present at a significantly higher level in cv. Amfissis than in cvs Kalamon and Koroneiki. It was finally observed that the relative amount of the pathogen in roots was lower than in stems and shoots and declined in plant tissues over time. These data constitute a valuable contribution in evaluating resistance of olive cultivars or olive root-stocks to V. dahliae pathotypes.     

Development of Stem-base Pathogens on Different Cultivars of Winter Wheat Determined by Quantitative PCR

The progress of development of stem-base pathogens in crops of second winter wheat was plotted in nine experiments in three years. The amount of each pathogen present was determined by quantitative PCR. Where Tapesia yallundae was present in quantifiable amounts, it usually developed earlier than the other eyespot pathogen, T. acuformis. Both species were usually present in greater amounts on cultivars which are more susceptible to eyespot. The sharp eyespot pathogen, Rhizoctonia cerealis, developed more erratically than either of the Tapesia spp. and there were no consistent effects on different cultivars. Fusarium spp., the cause of brown foot rot, were rarely present in quantifiable amounts, but Microdochium nivale was usually present as one or both of the varieties nivale and majus. Late-season (after anthesis) decreases in M. nivale suggest that any brown foot rot symptoms attributable to this fungus would have fully developed earlier. Cultivar differences in amounts of M. nivale were most clear in stems during internode extension and when relatively large amounts of DNA were present. Such differences approximately reflected eyespot susceptibility, cv. Soissons containing most and cv. Lynx containing least DNA. The results emphasise the difficulty in relating diagnoses, by quantitative PCR or other means, at early growth stages when decisions to apply fungicides against stem-base disease are made, to later disease severity.     

Relative tolerances of wild and cultivated barley to infection by Blumeria graminis f.sp. hordei (Syn. Erysiphe graminis f.sp. hordei). II—the effects of infection on photosynthesis and respiration

The relative levels of tolerance of two wild barley lines (Hordeum spontaneum), B19909 and I-17-40, and one cultivated barley (Hordeum vulgare), cv. Prisma, to Blumeria graminis f.sp. hordei were determined by comparing the effects of different levels of infection on the photosynthesis and respiration rates of the third leaf. Infection caused the early onset of senescence in all three lines, and in particular in cv. Prisma, and was accompanied by decreases in gross and net photosynthesis rates, increases in respiration rates, and loss of chlorophyll. The onset of senescence occurred at approximately the same time in infected leaves of the two wild lines, but once triggered, photosynthesis rates and chlorophyll levels declined more rapidly in I-17-40 than in B19909. A burst of respiratory activity accompanied the onset of senescence, and this was greatest in cv. Prisma. Conidial production was higher in B19909, indicating a higher level of tolerance in this line.     

Differences in resistance in onion cultivars to pink root rot disease in Iran

Forty-five onion (Allium cepa) cultivars were evaluated for disease resistance to the fungal pathogen Pyrenochaeta terrestris (pink root disease). Forty-three Iranian cultivars and two commercially exotic resistant cultivars (Texas Early Grano and Yellow Sweet Spanish) were grown in a glasshouse for two successive years (2003 and 2004) in Isfahan, Iran. Susceptibility of the cultivars was determined using a scale of 0–3. Six cultivars had significantly lower susceptibility of pink root compared to the exotic ones. High susceptibility tended to be associated with high mean scores (2–3), and the highly resistant cultivars had the lowest scores (0–1). In addition, the 45 examined cultivars were ranked from 1 to 45 according to their markedly differing reactions to P. terrestris, which differed markedly. Of 32 cultivars that were less resistant, 30 with an intermediate status or tolerance (1–2) differed considerably in their rank order. No pink root symptoms were seen in the early stages, but on blotter paper or wheat-straw agar some roots turned pink with no pycnidia formation, whereas on potato dextrose agar medium (PDA) with a few fresh healthy and sterile onion roots on the surface, mycelia and pycnidia were recovered. Variance and cluster analysis showed similar results among the cultivars with various levels of resistance, tolerance and susceptibility as determined by scoring scales.     

Screening of cultivated and wild adzuki bean for resistance to race 3 of <Emphasis Type="Italic">Cadophora gregata</Emphasis> f. sp. <Emphasis Type="Italic">adzukicola</Emphasis>, cause of brown stem rot

Two diseases of adzuki bean, brown stem rot (BSR, caused by Cadophora gregata f. sp. adzukicola) and adzuki bean Fusarium wilt (AFW, caused by Fusarium oxysporum f. sp. adzukicola), are serious problems in Hokkaido and have been controlled using cultivars with multiple resistance. However, because a new race of BSR, designated race 3, was identified, sources of parental adzuki bean for resistance to race 3 were needed. Therefore, we examined 67 cultivars and lines of cultivated and wild adzuki bean maintained at the Tokachi Agricultural Experiment Station using a root-dip inoculation method. Consequently, nine adzuki bean cultivars, one wild adzuki bean accession and 30 lines (including two lines resistant to all the three races of BSR and AFW) were confirmed to be resistant or tolerant to race 3 of BSR, and we found a cultivar Akamame as well as a wild adzuki bean Acc2515 to be a new source for a resistance gene to the race 3. This cultivar also holds promise as a source of resistance against other races of BSR and AFW.     

Elucidation of defence responses and signalling pathways induced in Arabidopsis thaliana following challenge with Phytophthora cinnamomi

Arabidopsis thaliana (Arabidopsis) Col-0 was inoculated with Phytophthora cinnamomi to assess the interaction and defence responses involved. Pathogen ingress and asexual reproduction occurred on root tissue but not leaf tissue. The colonisation of root tissue did not cause disease symptoms or plant death, indicating that Arabidopsis Col-0 was tolerant of the infection. The induction of several plant defence responses including the expression of defence-related genes were found, with differences displayed between inoculated root and leaf tissue. Arabidopsis defence-related gene mutant/over-expressing lines were also inoculated with P. cinnamomi but none of the lines tested exhibited a marked increase in susceptibility to the pathogen.     

Assessment of real-time PCR as a method for determining the presence of <Emphasis Type="Italic">Verticillium dahliae</Emphasis> in different Solanaceae cultivars

Real-time PCR was used to detect and quantify Verticillium dahliae and to assess the susceptibility of four Capsicum annuum cultivars (Luesia, Padrón, SCM331 and PI201234) and the Capsicum chinense cv. C118 to this pathogen. The symptoms which developed after infection included stunting and yellowing, and were more acute in the cv. SCM331, which also suffered defoliation in later stages of the disease and in C118, which suffered severe stunting. Quantification of the pathogen DNA in roots 23 and 34 days post-inoculation (dpi) revealed that there were significantly higher amounts of Verticillium dahliae DNA in C118 than in the other cultivars, followed by SCM331, Padrón and PI201234. The lowest amounts of fungal DNA in roots were found in Luesia. In hypocotyls, the highest amounts of fungal DNA were found in SCM331, while Luesia, Padrón and PI201234 had much lower amounts, and C118 had intermediate levels. When a compatible versus an incompatible system was studied, using the near-isogenic tomato lines LA3030 (susceptible) and LA3038 (resistant to V. dahliae), we were able to detect fungal DNA in both lines. As expected, the fungus/plant DNA ratio was lower in LA3038 than in LA3030 and it decreased with time in LA3038. The amount of Verticillium dahliae DNA in the roots of LA3030 remained constant between days 23 and 34 post-inoculation, but increased 10-fold in collars. Finally, when real-time PCR was applied as a diagnostic method to samples from pepper plants, soil and water collected from farms in northwest Spain, we were able to detect V. dahliae DNA in these samples even when symptoms of the disease were not evident.     

Novel methodologies in screening and selecting olive varieties and root-stocks for resistance to <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

An innovative inoculation process, involving the drilling of a trunk hole in 3 year-old olive trees and injecting a dense conidial suspension of Verticillium dahliae, was developed to study differentiation in foliar symptom expression between olive cultivars tolerant or susceptible to the pathogen. It was demonstrated that V. dahliae conidia could be translocated and colonize the xylem at the same distance above and below the point of trunk injection in both cultivars. However, the pathogen could be subsequently isolated at statistically significant percentages in susceptible cv. Amphissis compared to the tolerant cv. Kalamon, indicating operation of resistance mechanisms in the vascular phase of the disease. Consequently symptom development in the susceptible cultivar was at least sixfold more intensive compared to the tolerant cultivar, 6–11 months after trunk inoculation. Perennial olive orchard experiments, aimed at selecting Verticillium-resistant root-stocks, were conducted by applying the novel method in 2–3 year-old root-stock suckers of Amphissis olive trees and in the tolerant cvs Lianolia of Corfu and Koroneiki. It was indicated that potentially resistant root-stocks could be obtained following the trunk drilling technique. Resistance differentiation between cvs Amphissis and Kalamon was further verified through root inoculation by various V. dahliae microsclerotial concentrations and demonstrated that the trunk drilling inoculation procedure is equally efficient in resistance evaluation of olives to Verticillium wilt. The trunk inoculation procedure could be useful in selecting and screening root-stocks for resistance to V. dahliae and other vascular pathogens and could elucidate resistance mechanisms in woody plants against vascular wilt diseases.     

西瓜品种资源抗细菌性果斑病苗期鉴定

为筛选高抗瓜类细菌性果斑病的西瓜资源,以24份西瓜品种资源为试材,采用苗期喷雾接种法分别接种分离自甜瓜上的瓜类嗜酸菌Acidovorax citrulli菌株pslb96和ZZ-1,鉴定各品种资源对瓜类细菌性果斑病的苗期抗性。结果表明,24份西瓜品种资源中未发现有对2株菌株表现免疫的材料,有7份资源对菌株pslb96表现高抗,12份资源对菌株ZZ-1表现高抗;9份资源对菌株pslb96表现中感或感病,7份资源对菌株ZZ-1表现中感和感病;对2株菌株均表现高抗的品种资源有野生型种质资源A9及商品种华欣、申蜜968、申选958和申抗988,占总品种资源的20.83%。部分品种资源A4、A13和申蜜7号对菌株pslb96和ZZ-1的抗性表现出明显的差异,表明相同寄主来源的2株不同菌株致病力存在差异。     

Analysis of head and leaf reaction towards <Emphasis Type="Italic">Microdochium nivale</Emphasis>

This research examined the variation in the response of eight commercial wheat cultivars to Microdochium nivale isolates using both in vivo FHB tests (AUDPC and RHW measurements) and in vitro detached leaf assays (LGR). Irrespective of fungal variety, the two Italian cvs Fortore and Norba exhibited the greatest amount of visual disease symptoms (mean AUDPC=2.2 and 2.3, respectively), being significantly more susceptible than the other six cultivars (AUDPC 1.24) (P < 0.05). Irrespective of fungal variety, the Italian cv. Norba and the Irish cv. Falstaff were more susceptible than the other cultivars (except Fatima 2) in terms of RHW (P < 0.05), while the cvs Fortore, GK Othalom and Consort were more resistant than the other five cultivars (P < 0.05). In the detached leaf assay, the Hungarian cv. GK Othalom and the Italian cv. Norba were more susceptible (mean LGR=0.79 and 0.81 mm day^–1, respectively) to M. nivalethan the other six cultivars (mean LGR=0.51–0.72) (P < 0.05). Analysis of the relationship between head and leaf reaction to M. nivaleinfection revealed no significant correlation.     

Emmer wheat, a potential new host of Tilletia indica

Two representative Italian emmer wheat (Triticum dicoccum) landraces, two selected lines and three improved emmer wheat cultivars, derived from crosses with durum wheat (Molisano landrace × ‘Simeto’), were tested for their susceptibility to Tilletia indica, the cause of Karnal bunt of wheat. Plants of emmer wheat were inoculated by injecting allantoid sporidial suspensions into the boot cavity of plants, just prior to ear emergence. A highly susceptible Indian spring wheat cultivar (Triticum aestivum) was used as a comparative control. At maturity of the plants, the seeds were harvested and assessed for incidence and severity of disease. All emmer wheat genotypes tested were infected but showed differing levels of susceptibility. The percentage of infected seeds for individual genotypes ranged from 5.4 to 75.0% compared with 99.1% for WL-711. The severity of infection was less in the old landraces, but it was higher in all the improved emmer wheat cultivars. In conclusion, Italian cultivars of emmer wheat were found to be highly susceptible to T. indica, and are potentially able to support the establishment of the pathogen.Authors L. Riccioni and M. Valvassori contributed equally to this work and should both be considered as first author.     

Occurrence of Fusarium oxysporum f. sp. melonis Race 1 in Japan

In 1994, Fusarium wilt of melon cultivars which are resistant to races 0 and 2 of Fusarium oxysporum f. sp. melonis was observed in southern area of the Lake Biwa region, Shiga prefecture. In commercial fields, mature plants of cv. Amus which were grafted onto cv. Enken Daigi 2, and of cv. FR Amus showed yellowing, wilting and finally death before harvesting of fruits. Diseased plants had vascular and root discolorations, and their stem sections yielded typical colonies of F. oxysporum. When the Shiga strains were tested for their pathogenicity to 12 species of cucurbits, they caused wilts only on melon. Using race differential cultivars of melon, the Shiga strains were classified as race 1 of F. oxysporum f. sp. melonis, which has not been reported in Japan. To further characterize their pathogenicity, the strains were used to inoculate 46 additional cultivars of melon, oriental melon and oriental pickling melon. All the race 1 strains were pathogenic to the cultivars tested, and their host range was apparently different from those of strains belonging to other races (races 0, 2 and 1,2y). DNA fingerprinting with a repetitive DNA sequence, FOLR3, differentiated race 1 strains from strains of races 0 and 2, but not from race 1,2y strains. Received 2 July 1999/ Accepted in revised form 30 September 1999     

Characterization ofRhizoctonia solani isolates from potatoes in turkey and screening potato cultivars for resistance to AG-3 isolates

A total of 304Rhizoctonia solani isolates and 60 binucleateRhizoctonia-like fungi were recovered from stems and tubers of infected potato plants over a 2-yr period in northeast Turkey.R. solani isolates were identified to 11 anastomosis groups (AGs): AG-1 (0.66%), AG-2-1 (5.6%), AG-2-2 (0.99%), AG-3 (83.9%), AG-5 (4.6%), AG-6 (0.66%), AG-8 (1.32%), AG-9 (0.33%), AG-10 (1.32%), AG-12 (0.33%), and AG-13 (0.33%). In the greenhouse tests, most of the AG-3 isolates were significantly more virulent than isolates belonging to other AGs on potato cv. Batum. Isolates of other anastomosis groups differed in their virulence. Results indicated that AG-3 is an important pathogen on potatoes grown in the study area. Five of 22 commercial and local potato cultivars evaluated for their reaction toR. solani AG-3 isolates (TP-2) under greenhouse conditions were highly resistant; the remaining cultivars exhibited different levels of susceptibility to the pathogen isolate. http://www.phytoparasitica.org posting July 14, 2005.     

Identification of Resistance to Common Bacterial Blight Disease on Bean Genotypes Grown in Turkey

Common bacterial blight (CBB) in edible beans (Phaseolus vulgaris), incited Xanthomonas campestris pv. phaseoli, reduces bean yields and seed quality. The main objective of this study was to determine resistance to common bacterial blight in bean genotypes. Twenty-two bean genotypes grown in Turkey including common and snap bean cultivars/lines were collected from different parts of Turkey and tested for resistance against to Xanthomonas campestris pv. phaseoli strain MFD-11. All the common and snap bean lines/cultivars tested were moderately susceptible, susceptible or highly susceptible, except AG-7117 which was found resistant to Xanthomonas campestris pv. phaseoli. This is the first report of a resistance source in a common bean line (AG-7117) against Xanthomonas campestris pv. phaseoli.     

Susceptibility of Broccoli Cultivars to Bacterial Head Rot: In Vitro Screening and the Role of Head Morphology in Resistance

Head rot is a major disease of broccoli caused by the soft rot pathogens Pseudomonas fluorescens and Erwinia carotovora. Two in vitro pathogenicity tests were evaluated as a methods to identify broccoli cultivars susceptible or resistant to bacterial head rot. One test used mature heads excised from the plant and inoculated with squares of cotton lint which had been soaked in a bacterial suspension. The other test involved stab-inoculating axenically grown seedlings. With the excised head test, susceptible cultivars showed a black soft rot, whilst less susceptible or moderately resistant cultivars showed only watersoaking, or browning and slight softening of the tissue. No cultivar was completely resistant. Ten cultivars were tested, and their susceptibility ratings corresponded with previously recorded field data, with one exception. This laboratory test could be used to screen for susceptibility to head rot in broccoli breeding programmes. The seedling test distinguished differences in aggressiveness among bacterial isolates but not cultivar susceptibility. Increasing head size correlated negatively with disease resistance. Head shape, i.e. cultivars which showed a domed shape rather than a flat shape, was positively correlated with disease resistance. Thus small domed heads are more resistant to head rot than large flat heads. Other morphological characteristics, viz. floret prominence and number, and sepal stomatal number were not correlated with host resistance.     

Management of melon bacterial blotch by plant beneficial bacteria

Plant beneficial bacteria (PBB) have shown potential for disease control and are particularly important in the management of bacterial diseases, which are poorly controlled by conventional methods. In melon, bacterial fruit blotch caused by Acidovorax citrulli is a seedborne disease that is particularly destructive under certain conditions. PBB strains were screened for their ability to protect seeds and leaves from bacterial fruit blotch, and their antibiosis activity and plant colonization were studied. When Bacillus sp. RAB9 was applied to infected seeds, it reduced the area under the disease progress curve (AUDPC) by 47% and increased the incubation period (the time between inoculation and the first visible symptoms) by 35%. Three of the selected strains (JM339, MEN2 and PEP91) displayed antibiosis against A. citrulli. The RAB9^Rif-Nal mutant colonized seeds epiphytically and roots and stems endophytically. Paenibacillus lentimorbus MEN2 sprayed on melon seedlings protected leaves, and when challenged with A. citrulli, it reduced the AUDPC (by 88%), disease index (by 81%) and incidence (by 77%). Given that the production of both melon seedlings and commercially grown greenhouse melons is increasing, biocontrol strategies may well be integrated into bacterial blotch management programs.