Mildew Of Lettuce Research Articles

Identification and mapping of new genes for resistance to downy mildew in lettuce

Key messageEleven new major resistance genes for lettuce downy mildew were introgressed from wild Lactuca species and mapped to small regions in the lettuce genome.Downy mildew, caused by the oomycete pathogen Bremia lactucae Regel, is the most important disease of lettuce (Lactuca sativa L.). The most effective method to control this disease is by using resistant cultivars expressing dominant resistance genes (Dm genes). In order to counter changes in pathogen virulence, multiple resistance genes have been introgressed from wild species by repeated backcrosses to cultivated lettuce, resulting in numerous near-isogenic lines (NILs) only differing for small chromosome regions that are associated with resistance. Low-pass, whole genome sequencing of 11 NILs was used to identify the chromosome segments introgressed from the wild donor species. This located the candidate chromosomal positions for resistance genes as well as additional segments. F2 segregating populations derived from these NILs were used to genetically map the resistance genes to one or two loci in the lettuce reference genome. Precise knowledge of the location of new Dm genes provides the foundation for marker-assisted selection to breed cultivars with multiple genes for resistance to downy mildew.

Essential Tea Tree Oil Activity against Bremia lactucae in Lettuce

Downy mildew caused by the oomycete Bremia lactucae Regel is a serious disease of lettuce in field and greenhouse production. Here we report on the antifungal activity of essential Australian tea tree oil (TTO) derived from Melaleuca alternifolia against downy mildew in lettuce. Preventive treatments with Timorex Gold (STK Ltd., Petach Tikva, Israel), fungicide containing TTO, suppressed the development of B. lactucae on cotyledons and young lettuce plants. Epifluorescence microscopy showed that TTO had a moderate inhibitory effect on spore germination of the pathogen but a strong inhibitory effect on sporophore emergence and hence on sporulation. Timorex Gold (22.3 EC W/V) was as effective as copper hydroxide in controlling the disease in organic field plots. It was as effective as the fungicides Bellis (Boscalid + Pyraclostrobin; BASF, Germany) and Commet (pyraclostrobin; BASF, Germany) in conventional production. The results demonstrate that Timorex Gold effectively suppresses the development of downy mildew in lettuce in growth chambers as well as in the field, and thus suitable biopesticide for its control.

RESIDUAL EFFECT OF CERTAIN COPPER FUNGICIDES USED IN CONTROLLING DOWNY MILDEW OF LETTUCE

the fungicidal residues of certain copper fungicides, i.e. copper oxychloride, copper hydroxide and copper salts of fatty and rosin acids used against downy mildew in lettuce were studied. Data showed the occurrence of Cu residues in the plant, at different values according to the forms and rates used. Copper compounds residues were digested from lettuce plants samples then the residues were determined by microwave plasma atomic emission spectrometry. Results indicated that the pre-harvest interval (PHI) for lettuce plants were 1, 3 and 4 days for copper salts, copper hydroxide and copper oxychloride, respectively.

Resistance to Downy Mildew in Lettuce 'La Brillante' is Conferred by Dm50 Gene and Multiple QTL.

Many cultivars of lettuce (Lactuca sativa L.) are susceptible to downy mildew, a nearly globally ubiquitous disease caused by Bremia lactucae. We previously determined that Batavia type cultivar 'La Brillante' has a high level of field resistance to the disease in California. Testing of a mapping population developed from a cross between 'Salinas 88' and La Brillante in multiple field and laboratory experiments revealed that at least five loci conferred resistance in La Brillante. The presence of a new dominant resistance gene (designated Dm50) that confers complete resistance to specific isolates was detected in laboratory tests of seedlings inoculated with multiple diverse isolates. Dm50 is located in the major resistance cluster on linkage group 2 that contains at least eight major, dominant Dm genes conferring resistance to downy mildew. However, this Dm gene is ineffective against the isolates of B. lactucae prevalent in the field in California and the Netherlands. A quantitative trait locus (QTL) located at the Dm50 chromosomal region (qDM2.2) was detected, though, when the amount of disease was evaluated a month before plants reached harvest maturity. Four additional QTL for resistance to B. lactucae were identified on linkage groups 4 (qDM4.1 and qDM4.2), 7 (qDM7.1), and 9 (qDM9.2). The largest effect was associated with qDM7.1 (up to 32.9% of the total phenotypic variance) that determined resistance in multiple field experiments. Markers identified in the present study will facilitate introduction of these resistance loci into commercial cultivars of lettuce.

Effects of stacked quantitative resistances to downy mildew in lettuce do not simply add up.

In a stacking study of eight resistance QTLs in lettuce against downy mildew, only three out of ten double combinations showed an increased resistance effect under field conditions. Complete race nonspecific resistance to lettuce downy mildew, as observed for the nonhost wild lettuce species Lactuca saligna, is desired in lettuce cultivation. Genetic dissection of L. saligna's complete resistance has revealed several quantitative loci (QTL) for resistance with field infection reductions of 30-50 %. To test the effect of stacking these QTL, we analyzed interactions between homozygous L. saligna CGN05271 chromosome segments introgressed into the genetic background of L. sativa cv. Olof. Eight different backcross inbred lines (BILs) with single introgressions of 30-70 cM and selected predominately for quantitative resistance in field situations were intercrossed. Ten developed homozygous lines with stacked introgression segments (double combinations) were evaluated for resistance in the field. Seven double combinations showed a similar infection as the individual most resistant parental BIL, revealing epistatic interactions with 'less-than-additive' effects. Three double combinations showed an increased resistance level compared to their parental BILs and their interactions were additive, 'less-than-additive' epistatic and 'more-than-additive' epistatic, respectively. The additive interaction reduced field infection by 73 %. The double combination with a 'more-than-additive' epistatic effect, derived from a combination between a susceptible and a resistant BIL with 0 and 30 % infection reduction, respectively, showed an average field infection reduction of 52 %. For the latter line, an attempt to genetically dissect its underlying epistatic loci by substitution mapping did not result in smaller mapping intervals as none of the 22 substitution lines reached a similar high resistance level. Implications for breeding and the inheritance of L. saligna's complete resistance are discussed.

Fine mapping quantitative resistances to downy mildew in lettuce revealed multiple sub-QTLs with plant stage dependent effects reducing or even promoting the infection.

Three regions with quantitative resistance to downy mildew of non-host and wild lettuce species, Lactuca saligna , disintegrate into seventeen sub-QTLs with plant-stage-dependent effects, reducing or even promoting the infection. Previous studies on the genetic dissection of the complete resistance of wild lettuce, Lactuca saligna, to downy mildew revealed 15 introgression regions that conferred plant stage dependent quantitative resistances (QTLs). Three backcross inbred lines (BILs), carrying an individual 30-50cM long introgression segment from L. saligna in a cultivated lettuce, L. sativa, background, reduced infection by 60-70% at young plant stage and by 30-50% at adult plant stage in field situations. We studied these three quantitative resistances in order to narrow down their mapping interval and determine their number of loci, either single or multiple. We performed recombinant screenings and developed near isogenic lines (NILs) with smaller overlapping L. saligna introgressions (substitution mapping). In segregating introgression line populations, recombination was suppressed up to 17-fold compared to the original L. saligna×L. sativa F 2 population. Recombination suppression depended on the chromosome region and was stronger suppressed at the smallest introgression lengths. Disease evaluation of the NILs revealed that the resistance of all three BILs was not explained by a single locus but by multiple sub-QTLs. The 17 L. saligna-derived sub-QTLs had a smaller and plant stage dependent resistance effect, some segments reducing; others even promoting downy mildew infection. Implications for lettuce breeding are outlined.

Evaluation and QTL mapping of resistance to powdery mildew in lettuce

Lettuce (Lactuca sativa) is the major leafy vegetable that is susceptible to powdery mildew disease under greenhouse and field conditions. Quantitative trait loci (QTLs) for resistance to powdery mildew under greenhouse conditions were mapped in an interspecific population derived from a cross between susceptible L. sativa cultivar Salinas and the highly susceptible L. serriola accession UC96US23. Four significant QTLs were detected on linkage groups LG 1 (pm‐1.1), LG 2 (pm‐2.1 and pm‐2.2) and LG 7 (pm‐7.1), each explaining between 35 to 42% of the phenotypic variation. The four QTLs are not located in the documented hotspots of lettuce resistance genes. Alleles for the disease resistance at the four QTLs originated from both parents (two from each), demonstrating that even highly susceptible accessions may provide alleles for resistance to powdery mildew. These QTLs appeared to operate during limited periods of time. Results of the field trials with F2:3 and F3:4 families derived from a Soraya (moderately resistant) × Salinas cross demonstrated effective transfer of resistance to powdery mildew in this material. An integrated rating approach was used to estimate relative levels of resistance in 80 cultivars and accessions tested in a total of 23 field and greenhouse experiments. Generally, very low resistance was observed in crisphead‐type lettuces, while the highest relative resistance was recorded in leaf and butterhead types. Comparison of two disease assessment methods (percentage rating and the area under the disease progress steps, AUDPS) for detection of QTLs shows that the two approaches complement each other.

Activity of carboxylic acid amide (CAA) fungicides against Bremia lactucae

Four carboxylic acid amide (CAA) fungicides, mandipropamid (MPD), dimethomorph (DMM), benthiavalicarb (BENT) and iprovalicarb (IPRO) were examined for their effects on various developmental stages of Bremia lactucae, the causal agent of downy mildew in lettuce, in vitro and in planta. Spore germination in vitro or on leaf surfaces was inhibited by all CAA fungicides (technical or formulated). MPD was more effective in suppressing germination than DMM or BENT, whereas IPRO was least effective. CAA induced no disruption of F-actin microfilament organisation in germinating spores of B. lactucae. CAA applied to germinating spores in vitro prevented further extension of the germ tubes. When applied to germinated spores on the leaf surface they prevented penetration. Preventive application of CAA to intact plants inhibited infection. MPD was more effective in suppressing infection than DMM or BENT, whereas IPRO was least effective. Curative application was effective at ≤3 h post-inoculation (hpi) but not at ≥18 hpi. CAA (except IPRO) applied to upper leaf surfaces inhibited spore germination on the lower surface and hence reduced infection. CAA suppressed sporulation of B. lactucae on floating leaf discs and when sprayed onto infected plants two days before onset of sporulation. BENT and DMM were more effective in suppressing sporulation than MPD or IPRO. Epidemics of downy mildew in shade-house grown lettuce were suppressed by CAA. A single spray applied to five-leaf plants before transplanting controlled the disease for 50 days. The results suggest that CAA are effective inhibitors of spore germination and therefore should be used as preventive agents against downy mildew of lettuce caused by B. lactucae.

Comparative genetic analysis of field resistance to downy mildew in the lettuce cultivars ‘Grand Rapids’ and ‘Iceberg’

Downy mildew on lettuce is currently controlled using host resistance genes (Dm genes) that confer race-specific resistance in seedlings. Field resistance (FR) that is active in adult plants but not seedlings was identified in the cvs. Grand Rapids and Iceberg. The goal of our study was to evaluate the utility of Grand Rapids as a source of novel Bremia resistance alleles, particularly in comparison with Iceberg. To measure FR, downy mildew symptoms were evaluated following natural infection in field experiments. The responses of Grand Rapids and Iceberg were similar in many respects. Although both cultivars had a small percentage of plants exhibiting disease symptoms, the average disease ratings were as low as for cultivars with effective Dm genes. We observed no evidence for race specificity. FR was effective over 3 years of our study, despite documented variation within pathogen populations. Both cultivars lacked all known seedling resistance genes except Dm13, which was not responsible for the resistance observed in field experiments. Similar segregation of FR was observed in F2 populations for both Grand Rapids and Iceberg. The presence of highly susceptible families within Grand Rapids × Iceberg populations suggested the presence of at least one unique resistance allele in each cultivar. Preliminary genetic analysis of FR from Grand Rapids revealed a high estimate of narrow-sense heritability that suggested simple inheritance, but single gene models did not fit the observed data. Our results suggest that Grand Rapids may represent an underutilized resource for controlling downy mildew in lettuce.

Lactuca saligna, a non-host for lettuce downy mildew ( Bremia lactucae), harbors a new race-specific Dm gene and three QTLs for resistance.

Lactuca sativa (lettuce) is susceptible to Bremia lactucae (downy mildew). In cultivated and wild Lactuca species, Dm genes have been identified that confer race-specific resistance. However, these genes were soon rendered ineffective by adaptation of the pathogen. Lactuca saligna (wild lettuce) is resistant to all downy mildew races and can be considered as a non-host. Therefore, L. saligna might be an alternative source for a more-durable resistance to downy mildew in lettuce. In order to analyze this resistance, we have developed an F(2) population based on a resistant L. saligna x susceptible L. sativa cross. This F(2) population was fingerprinted with AFLP markers and tested for resistance to two Bremia races NL14 and NL16. The F(2) population showed a wide and continuous range of resistance levels from completely resistant to completely susceptible. By comparison of disease tests, we observed a quantitative resistance against both Bremia races as well as a race-specific resistance to Bremia race NL16 and not to NL14. QTL mapping revealed a qualitative gene ( R39) involved in the race-specific resistance and three QTLs ( RBQ1, RBQ2 and RBQ3) involved in the quantitative resistance. The qualitative gene R39 is a dominant gene that gives nearly complete resistance to race NL16 in L. saligna CGN 5271 and therefore it showed features similar to Dm genes. The three QTLs explained 51% of the quantitative resistance against NL14, which indicated that probably only the major QTLs have been detected in this F(2) population. The perspectives for breeding for durable resistance are discussed.

Meteorological factors affecting periodicity and concentration of airborne spores of Bremia lactucae

Several systems for forecasting downy mildew of lettuce are based on the assumption that spores infect leaves in the early morning. However, there is no clear indication that spores are present in sufficient amounts at that time. Spores of Bremia lactucae were trapped using a 7-day volumetric spore trap for about 35 days in each of 1997 and 1998. Three periodicity patterns were observed. The most frequent pattern had a peak, representing more than 75% of the daily catch, near 10:00 a.m. The second pattern was similar except that the peak occurred later. The third pattern was observed on days with low numbers of spores and was characterized by a random distribution, though most spores were trapped during daylight hours. Spore release was associated with a drop in relative humidity (RH) and increases in temperature and wind speed. The fluctuations in numbers of airborne spores followed fluctuations in disease severity. Overall, daily spore concentrations and number of spores per lesion were not correlated with daily weather variables. However, when spore concentrations were grouped into three categories (low: <10 spores/L; intermediate: ≤10 to < 100 spores/L; high: ≥100 spores/L), a good correlation (r = 0.79) was obtained with night hours of high RH (>95%). For both years, the first significant spore catch was observed when about 200 degree-night hours of RH > 95% (base 0°C) (DNRH) had accumulated. The relative spore concentration (RSC) was modeled as a function of DNRH and the model that best explained (R 2= 0.97) this relationship was RSC = 1×(1 + e–0.0183×DNRH)1/1−1.005.

BREMCAST: Development of a system to forecast risk levels of downy mildew on lettuce (Bremia lactucae)

A forecasting system was developed to predict risk levels of downy mildew of lettuce ( Bremia lactucae ). The criteria for the forecasting system were derived from the relationships of weather variables to sporulation, dissemination and infection processes of B. lactucae . In addition the presence of inoculumsource, or the disease, in the field was taken into consideration. User-friendly software (BREMCAST) for use within the Windows (Microsoft Corp.) operating system was developed to calculate sporulation, infection, disease severity and cumulative disease severity values. The system predicted the occurrence of downy mildew on 84 and 80% of the days, under commercial and field plot conditions, respectively. The potential of the system for use as an intelligent decision tool in managing downy mildew is discussed.

Phytogard® and DL-β-amino Butyric Acid (BABA) Induce Resistance to Downy Mildew (Bremia Lactucae) in Lettuce (Lactuca sativa L)

Downy mildew of lettuce (Bremia lactucae) is a serious disease. An alternative to chemicals is the application of disease resistance inducers. The aim of this study was to test whether DL-β-amino butyric acid (BABA) and Phytogard® (K2HPO3) could induce resistance in downy mildew susceptible plants. Aqueous solutions of BABA (0, 10, 20, 30, 50, 80, 100 mM) and Phytogard® (0.0, 5.8, 29.0, 40.6, 58.0 and 87.0 ppm) were sprayed on seven-day-old seedlings 0, 3, 7 and 15 days before or 1–3 days after inoculation with B. lactucae. Results obtained showed that Phytogard®- and BABA-induced resistance was dose-dependent. At 40.6 ppm for Phytogard® and 10 mM for BABA, complete protection was obtained. Both compounds had a curative effect and the induced resistance lasted for at least 15 days. It was also shown that both compounds induced systemic resistance in lettuce against downy mildew. Phytogard® at 40.6 ppm completely inhibited spore germination while BABA at 20 mM did not. Pathogenesis related (PR) protein analysis showed that BABA induced weak accumulation of PR-2, but not PR-1, PR-5 and PR-9. Phytogard® induced none of these proteins. The use of these two compounds to protect lettuce from B. lactucae is discussed.

A demonstration of the inheritance of field resistance to lettuce downy mildew (Bremia lactucae Regel.) in progeny derived from cv. Grand Rapids

The inheritance of field resistance to downy mildew in lettuce derived from cv. Grand Rapids was studied in F2, F3 and F4 progeny from a cross with the more susceptible cv. Lobjoits Green Cos. The results for 32 randomly produced F3 families and the F2 population corresponded closely although the absolute level of resistance varied from season to season according to disease pressure and environmental conditions. The field resistance character showed a quantitative pattern of inheritance but the rapid response to selection at F3 implied that only a few genes may be involved. F4 families raised from families selected from the two extremes of the F3 distribution fell into distinct categories with resistance levels corresponding to that of their F3 parents. An association between resistance and morphological features of the resistant parent was present at F2 but not in subsequent generations when individuals and families which were as resistant as cv. Grand Rapids and yet bore little morphological resemblance to this cultivar were readily recovered.

Inheritance of Resistance to Downy Mildew (Bremia Lactucae Reg.) in Lettuce1

Abstract The inheritance of resistance to California race(s) of downy mildew in lettuce was studied in cvs. Bourguignonne, Meikoningin, Proeftuin’s Blackpool, Red Salad Bowl, Salad Trim, Ventura, and PI 164937. Resistance in cvs. Bourguignonne, Red Salad Bowl, and Salad Trim is conferred by a single dominant allele. This is the same allele found in resistant cvs. Calicel, Calmar, E-4, Imperial 410, Valrio, Valtemp, and Valverde, and is derived from resistant PI 91532. Resistance in ‘Meikoningin’ is controlled by a single dominant allele different from the one derived from PI 91532. Allelism tests indicate that the 2 alleles are independently inherited. ‘Proeftuin’s Blackpool’ and ‘Ventura’ each have 2 dominant alleles for resistance that are different from the single dominant allele derived from PI 91532. Allelism tests of ‘Proeftuin’s Blackpool’ and ‘Ventura’ indicate resistance is conferred by the same 2 dominant alleles. The 2 alleles in cvs. Proeftuin’s Blackpool or Ventura and the single allele derived from PI 91532 are independently inherited. Resistance in PI 164937 is conferred by 2 dominant alleles, one of which is the same as the single dominant allele derived from PI 91532.

Studies on two races of Bremia lactucae Regel

SUMMARYTwo races of Bremia lactucae are described; race W.1, which can attack various lettuce varieties now being grown commercially but cannot attack the resistant lettuces bred at Wellesbourne, and race W.2, which can infect not only the market varieties but also some of the W.1‐resistant lines. Race W.2 was shown to arise directly from race W.1, possibly by mutation.The length of incubation time from inoculation to sporulation of the mildew in lettuce leaves decreased as the number of spores applied in the inoculum increased. This fact was used to assess the concentration of race W.2 spores in naturally occurring spore populations, by comparing the effects on W.1‐resistant plants of graded dilutions of spores from the field with those of known concentrations of race W.2. These tests showed that race W.2 was most prevalent where W.1‐resistant lettuces had been grown for some years. Race W.2 spores were found, however, where no W.1‐resistant lettuces had been grown, and it appears that race W.2 may be a normal constituent of most Bremia populations. The importance of these findings in relation to future breeding for downy mildew resistance is discussed.