Immunohistochemical changes in methyl-ester distribution of homogalacturonan and side chain composition of rhamnogalacturonan I as possible components of basal resistance in tomato inoculated with Ralstonia solanacearum

The composition and structure of pectic cell wall polysaccharides of stem sections were investigated in healthy and Ralstonia solanacearum-inoculated tomato genotypes L390 and Hawaii 7996, susceptible and resistant to bacterial wilt, respectively, by immunohistochemical analysis. Constitutive differences between genotypes manifested in methyl-ester distribution of homogalacturonan (HG), arabinan and galactan side chain composition of rhamnogalacturonan I (RG I) and arabinogalactan-protein (AGP) in the xylem parenchyma and in vessel cell walls. After inoculation increased labeling was observed with all the antibodies (JIM5, JIM7, LM2, LM5, LM6, LM7) specific for HG, RG I and AGP epitopes, in the xylem parenchyma and around xylem vessels of stem sections of L390, but not of Hawaii 7996. Also vessel cell walls were stronger stained after inoculation in L390, particularly for the non-blockwise de-esterification of HG, possibly indicating for the first time the non-blockwise action pattern of bacterial pectin methyl esterase. In genotype Hawaii 7996 a reaction to inoculation was observed only in vessel walls, with a significantly increased number of stained vessels—five- and nine-fold for arabinan and galactan epitopes of RG I, respectively. Differences in xylem cell wall structure may play a role as a constitutive resistance mechanism in the multigenic resistance of tomato against bacterial wilt, while changes after inoculation may contribute to induced basal resistance on cell wall level.     

Bacterial wilt-resistant tomato rootstock suppresses migration of <Emphasis Type="Italic">ralstonia solanacearum</Emphasis> into soil

Ralstonia solanacearum, the causal agent of bacterial wilt of tomato, grows in infected plants and migrates from the roots into the soil. We investigated the effectiveness of bacterial wilt-resistant tomato rootstock in reducing the migration of R. solanacearum from susceptible scions into the soil. Rootstock stems were either 3–5 cm tall (low-grafted, LG) or ≥?10 cm tall (high-grafted, HG). After inoculation of scions of the susceptible cultivar (SC) with R. solanacearum below the first flower, there was no difference in disease progression among LG, HG, and ungrafted SC plants, and plants had wilted by 2 weeks. However, the rate of detection of R. solanacearum in the soil of wilted plants was reduced by grafting. The size of the R. solanacearum population in the soil of fully wilted plants increased in the order of HG?<?LG?<?SC. These results show that grafting onto resistant rootstock strongly suppressed the migration of R. solanacearum into the soil by the time of full wilting, and the effect was stronger with a longer rootstock. Migration of R. solanacearum into soil increased with increasing disease severity in SC, LG and HG. These facts suggest that early uprooting of slightly infected plants could control the spread of the bacteria into the soil.     

Interaction between silicon amendment,bacterial wilt development and phenotype of Ralstonia solanacearum in tomato genotypes

Silicon amendment significantly reduced bacterial wilt incidence expressed as area under disease progress curve for tomato genotypes L390 (susceptible) by 26.8% and King Kong2 (moderately resistant) by 56.1% compared to non-treated plants grown in hydroponic culture. However, wilt incidence in silicon-treated plants of genotype L390 reached 100% at 13 days post-inoculation (dpi), while in genotype King Kong2, plant death was retarded by 6 days, with 20% reduction of final wilt incidence. Bacterial numbers were significantly lower in silicon-treated compared to non-treated plants in King Kong2 at 2 dpi in midstems and in all organs at 5 dpi, and in Hawaii 7998 (resistant) in all organs at 2 dpi. Differences between genotypes were obvious on midstem level (5 dpi), where bacterial populations were generally significantly lower compared to roots. Increased tolerance was observed in genotypes L390 and King Kong2 with silicon treatment.Silicon accumulated in roots and was low in stems and leaves. Inoculation with Ralstonia solanacearum did not significantly affect silicon uptake and distribution. Negative correlations between root silicon content and bacterial numbers of midstems in genotypes Hawaii 7998 and King Kong2 suggested an induced resistance. Indications for an influence of host genotype and silicon treatment on the phenotypic conversion of R. solanacearum strain To-udk2-sb from fluidal to non-fluidal colonies in planta were observed.This is the first report on the effect of silicon on a bacterial disease and in a silicon-non-accumulator plant.     

Biochemical and histochemical traits: a promising way to screen resistance against spot blotch (Bipolaris sorokiniana) of wheat

Spot blotch caused by Bipolaris sorokiniana is a serious disease of wheat in warmer and humid regions of the world. Three blighting components, area under disease progress curve (AUDPC), disease severity (DS) and lesion size along with four biochemical and histochemical factors viz., total phenol content (TPC), chlorophyll content (CHC), phenylalanine ammonia-lyase (PAL) activity and lignin deposition were studied in a recombinant inbred lines (RILs) population involving parents “Sonalika” (susceptible) and “Yangmai 6” (resistant). The objective was to identify one or more robust and reliable tools of resistance, physical, biochemical or histochemical, to facilitate selection against spot blotch. The DS, AUDPC and lesion size were higher in the susceptible parent and RILs compared to the resistant. The mean TPC (246 mg Gallic acid g^?1 fresh weight) of the most resistant RIL was significantly higher than the most susceptible (181.5 mg Gallic acid g^?1 fresh weight) at 48 h after inoculation (hai). The mean SPAD value for CHC varied between 48.8 in the most resistant RILs to 8.8 in the most susceptible, while the mean PAL varied between 928.4 and 96.0 μmoles Cinnamic acid mg^-1 fresh weight at 48 hai in resistant and susceptible RILs, respectively. Likewise, lignin deposition was significantly higher in resistant RILs compared to the susceptible. The biochemical and histochemical parameters were significantly correlated with resistance and appeared robust for facilitating screening of breeding material and for increased precision in phenotyping against spot blotch.     

Validation and use of a qPCR protocol to quantify the spread of Ralstonia solanacearum in susceptible and resistant eucalypt plants

Bacterial wilt caused by Ralstonia solanacearum is a serious disease of eucalypt in humid and high temperature areas worldwide. Spreading of the bacterium in the field or to other nurseries occurs mainly by symptomless infected plant material. The use of pathogen-free propagating material as well as planting of resistant genotypes are currently the only strategies used for disease control. Therefore, a reliable and sensitive method for detection of low titres of R. solanacearum in infected plant tissue is essential for the success of management programmes. In this work, we adapted an efficient intercalating dye-based real-time PCR protocol to detect the bacterium in symptomless eucalypt plants as well as to investigate its movement in eucalypt clones CLR172 and CLR371, which exhibit resistant and susceptible phenotypes, respectively. We found that the bacterium translocates acropetally and basipetally in inoculated but symptomless cuttings of the resistant clone, as in cuttings of the susceptible clone displaying symptoms. Nevertheless, a smaller concentration of bacterial DNA was detected in tissues of the resistant clone. Mature biofilms occluding the xylem vessels were present in the susceptible clone whereas only single cells or small aggregates were observed in the resistant clone. This work contributes to improve our knowledge of the colonization process of R. solanacearum in eucalypt clones with different levels of susceptibility and to understand how the defence mechanisms against bacterial wilt in Eucalyptus work. Our findings could aid in the selection of the most resistant eucalypt clones to be used in wilt disease management programmes.     

Diversity of Brazilian biovar 2 strains of Ralstonia solanacearum

Ralstonia solanacearum is responsible for bacterial wilt disease. Specific and accurate identification of this pathogen is essential for protection of susceptible crops as well as breeding resistant varieties. Historically, R. solanacearum has been classified into biovars based on the use of sugar and alcohol as carbon sources, into races based on its ability to infect different hosts, more recently into phylotypes based on the intergenic transcribed sequence of the ribosomal RNA genes 16S and 23S and into sequevars based on the endoglucanase gene (egl) sequence. Race 3 biovar 2 (R3Bv2) is widespread in South and Central America, and in Brazil it is present in all potato-producing regions as the most prevalent strain. In this study, we classified 53 Brazilian R. solanacearum biovar 2 (Bv2) strains by traditional and molecular methods. PCR with specific primers confirmed all 53 bacterial strains as belonging to the R. solanacearum species complex, and all were classified as biovar 2A or 2T based on acidification of sugars and alcohols. Multiplex phylotype PCR assigned all strains to phylotype II. Phylogenetic analysis of egl sequences showed that most Bv2 strains from Brazil analyzed in this study did not cluster with known sequevars and are less clonal than the R3Bv2 strains reported for other countries. This is the first study to address the diversity of a collection of Brazilian R. solanacearum strains using the phylotype and sequevar classification scheme.     

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.     

Resistance to Taiwanese race 1 strains of Ralstonia solanacearum in wild tomato germplasm

A total of 252 wild Solanum accessions and one population of 49 introgression lines of LA716 were screened for resistance to a race 1/biovar 4/phylotype I strain Pss186 of Ralstonia solanacearum. Most wild tomato accessions were highly susceptible. However, five accessions of S. pennellii, i.e. LA1943, LA716, LA1656, LA1732 and TL01845 were resistant to strain Pss186. These accessions were then challenged against two other race 1/phylotpye I strains Pss4 and Pss190, which were more aggressive. All the five S. pennellii accessions were susceptible to Pss4, but displayed high to moderate resistance to Pss190 with a percentage of wilted plants ranging from 0% to 60%. Pss190 is an aggressive strain that made a resistant tomato line Hawaii 7996 susceptible. Thus, the results found in this study provide evidence of the presence of strain-specific resistance. LA3501, which has an introgression segment on chromosome 6, was found to be resistant to Pss186 among the screened introgression lines. This confirms the importance of resistance trait loci on chromosome 6 that have been identified by other studies. This is the first report of S. pennellii being resistant to bacterial wilt. These new resistant sources will provide breeders with more resources to breed for stable resistance to bacterial wilt of tomato.     

<Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> upregulates marker genes of the salicylic acid and ethylene signaling pathways but not those of the jasmonic acid pathway in leaflets of <Emphasis Type="Italic">Solanum</Emphasis> lines during early stage of infection

Real-Time PCR assay was used to quantify the expression of marker genes of the salicylic acid, jasmonic acid and ethylene signaling pathways in seven Solanum lines after inoculation with a Ralstonia solanacearum phylotype I strain, R008. Four Solanum lycopersicum lines (CRA 66, Hawaii 7996, MST 32/1, Quatre carrées), one S. tuberosum line (Spunta), the wild Lycopersicon cerasiforme and Solanum commersonii were used for this investigation. Results revealed very little activation of the jasmonic acid pathway marker genes, lipoxygenase A (LoxA) and protease inhibitor II (Pin2), with no significant difference (p > 0.05) in fold change expression among the Solanum lines. In contrast the salicylic acid pathway marker genes, glucanase A (GluA) and PR-1a, and the ethylene pathway marker genes, osmotin-like (Osm) and PR-1b, were expressed at higher levels with a statistically significant difference (p < 0.05) in fold change expression among the Solanum lines. The resistant lines L. cerasiforme, CRA 66, Hawaii 7996 and S. commersonii showed stronger activation of the salicylic acid and ethylene marker genes than the moderately resistant cultivar (MST 32/1) and the susceptible lines (Quatre carrées and Spunta). The marker genes reached their highest expression levels earlier (4 h.p.i) in the resistant and moderately resistant lines than in the susceptible lines (48 h.p.i.). These results indicate that salicylic acid and ethylene signaling pathways have a significant role in defense against R. solanacearum. The timing and magnitude of the upregulation of gene expression may determine the plant ability to put up a defense response against the pathogen.     

Unique infection structures produced by Pseudocercosporella capsellae on oilseed crops Brassica carinata,B. juncea and B. napus in Western Australia

White leaf spot disease (Pseudocercosporella capsellae) is widespread across oilseed, vegetable and forage brassicas. Light (LM) and scanning electron (SEM) microscope studies were undertaken to investigate host–pathogen interactions on cotyledons of resistant and susceptible Brassica carinata, B. juncea and B. napus. Under LM, unique brown structures were present, particularly on susceptible genotypes, in two morphologically distinct forms: first, as thread‐like structures within cortical tissue by 24 h post‐inoculation (hpi) and secondly, as brown ropy strand structures either within cortical tissues (internal ropy strands), or extruded out through stomatal pores (ropy strand extrusions). Under LM, these brown structures were most prevalent in highly susceptible B. juncea ‘Vardan’ that had both a high incidence within cortical tissue (70%) and of ropy strand extrusions (73%), as did susceptible B. napus ‘Trilogy’ within cortical tissue (60%). Under SEM, both these genotypes showed thread‐like structures smaller than hyphae forming highly branched networks and ropy strand‐like structures. While there were fewer brown structures in susceptible B. carinata UWA #012 (35%), fine, thread‐like structures forming networks were again prominent (SEM). In contrast, for resistant genotypes, brown structures (LM) were of very low frequency or absent; only 5% in resistant B. juncea ‘Dune’ and none in resistant B. napus ‘Hyola 42’ or highly resistant B. carinata ATC94129P. Under SEM, fine, thread‐like structures were present in the resistant B. juncea ‘Dune’ and B. napus ‘Hyola 42’. Liquid chromatographic analyses of brown structures revealed that both internal ropy strands within cortical tissues and ropy strand extrusions contained the mycotoxin cercosporin.     

Identification of fusarium head blight resistance related metabolites specific to doubled-haploid lines in barley

Fusarium head blight (FHB) and deoxynivalenol (DON) mycotoxin produced by Fusarium graminearum reduce barley yield and quality worldwide. Hundreds of quantitative trait loci (QTLs) have been identified in wheat and barley but their functions are largely unknown. Metabolic profiling was applied to better understand the mechanisms of resistance and to identify potential FHB resistance biomarker metabolites in barley. Four FHB resistant (H15-2, H148-3, H203-2 and H379-2) and one susceptible (H97-2), two-row, purple, doubled-haploid (DH) lines of barley were inoculated with either the pathogen or mock-solution. The disease severity quantified as the area under the disease progress curve (AUDPC) significantly varied between the resistant and susceptible genotypes, but not among the resistant genotypes. Neither the amount of DON nor the detoxified product, proportion of total DON, was significant among lines. The resistance related (RR, higher in abundance in resistant than in susceptible) metabolites varied in numbers and fold changes among the DH resistant lines. A total of 144 RR constitutive (RRC) and 167 RR induced (RRI) metabolites were selected, of which 39 and 37, respectively, were putatively identified. These RR metabolites mainly belonged to six chemical groups: phenylpropanoids, hydroxycinnamic acid amides, flavonoids, fatty acids, terpenoids, and alkaloids. The specific RR metabolites identified in each DH line, the possible mechanisms of resistance in each and their use as potential biomarkers are discussed.     

A Quantitative Method to Screen Common Bean Plants for Resistance to Bean common mosaic necrosis virus

ABSTRACT A quantitative method to screen common bean (Phaseolus vulgaris) plants for resistance to Bean common mosaic necrosis virus (BCMNV) is described. Four parameters were assessed in developing the quantitative method: symptoms associated with systemic virus movement, plant vigor, virus titer, and plant dry weight. Based on these parameters, two rating systems (V and VV rating) were established. Plants from 21 recombinant inbred lines (RILs) from a Sierra (susceptible) x Olathe (partially resistant) cross inoculated with the BCMNV-NL-3 K strain were used to evaluate this quantitative approach. In all, 11 RILs exhibited very susceptible reactions and 10 RILs expressed partially resistant reactions, thus fitting a 1:1 susceptible/partially resistant ratio (chi(2) = 0.048, P = 0.827) and suggesting that the response is mediated by a single gene. Using the classical qualitative approach based only on symptom expression, the RILs were difficult to separate into phenotypic groups because of a continuum of responses. By plotting mean percent reduction in either V (based on visual symptoms) or VV (based on visual symptoms and vigor) rating versus enzyme-linked immunosorbent assay (ELISA) absorbance values, RILs could be separated clearly into different phenotypic groups. The utility of this quantitative approach also was evaluated on plants from 12 cultivars or pure lines inoculated with one of three strains of BCMNV. Using the mean VV rating and ELISA absorbance values, significant differences were established not only in cultivar and pure line comparisons but also in virus strain comparisons. This quantitative system should be particularly useful for the evaluation of the independent action of bc genes, the discovery of new genes associated with partial resistance, and assessing virulence of virus strains.     

Insights on gene expression response of a characterized resistant genotype of Solanum commersonii Dun. against Ralstonia solanacearum

Solanum commersonii is a wild species related to the cultivated potato. Some S. commersonii genotypes have been proven to be resistant to the pathogenic bacteria Ralstonia solanacearum, which causes damage in potato and other economically important crops. Here an expression analysis of the response of a resistant S. commersonii genotype against R. solanacearum was performed using microarrays. The aims of this work were to elucidate the molecular processes involved in the interaction, establish the timing of the response, and contribute to identify genes related to the resistance. The response to the treatment was already initiated at 6 h post-inoculation (hpi) and was established at 24 hpi; during this period, a high number of genes was differentially expressed and several candidate genes for the resistance of S. commersonii to R. solanacearum were identified. At an early stage, the photosynthetic process was highly repressed and several genes encoding proteins related to reactive oxygen species (ROS) production were differentially expressed. The induction of ERF and ACC-oxidase genes related to the ethylene pathway and PR1 related to the salicylic acid pathway suggested the induction of both pathways, and back up the previously reported hemibiotrophic nature of the pathogen. Five genes related to plant defence and observed to be differentially expressed at the first two time points were validated by real time PCR. This work gives a glimpse to the molecular processes involved in S. commersonii resistance and identifies the species as a valuable genetic source for potato breeding against bacterial wilt.     

Molecular Mapping of the rps1.a Recessive Gene for Resistance to Stripe Rust in BBA 2890 Barley

ABSTRACT Stripe rust, caused by Puccinia striiformis f. sp. hordei, is one of the most important diseases of barley in the south-central and western United States. Growing resistant cultivars is the best approach for controlling the disease. The barley genotype BBA 2890 has all-stage resistance against all races of P. striiformis f. sp. hordei (PSH) identified thus far in the United States. The resistance in BBA 2890 is controlled by a single recessive gene, rps1.a. The objectives of this study were to identify resistance gene analog polymorphism (RGAP) markers for the all-stage resistance gene rps1.a, to map the gene on a barley chromosome using chromosome-specific simple sequence repeat (SSR) markers, and to determine the presence or absence of the flanking RGAP markers for the gene in 24 barley genotypes. Seedlings of the parents and 200 F(8) recombinant inbred lines (RILs) were tested for resistance to pathogen races PSH-14, PSH-48, and PSH-54 in the greenhouse in 2005. Genomic DNA was extracted from the parents and 150 F(8) RILs. The RGAP technique was used to identify molecular markers for the rps1.a gene. Twelve primer pairs generating repeatable polymorphic bands were selected for genotyping the 150 F(8) RILs. A genetic linkage group was constructed for the resistance gene with 13 RGAP markers and four chromosome-specific SSR markers. The four SSR markers mapped the gene on the long arm of barley chromosome 3H. The closest RGAP marker for the resistant allele was within a genetic distance of 2.1 centimorgans (cM). The closest marker for the susceptible allele was 6.8 cM away from the locus. The two closest RGAP markers for the resistant allele detected polymorphisms in 67 and 71% of the 24 barley genotypes when used individually, and detected polymorphism in 88% of the genotypes when used in combination. This information should be useful in incorporating the resistance gene into barley cultivars and in pyramiding the gene with other resistance genes for superior stripe rust resistance.     

Alteration of gene expression profile in the roots of wild diploid Arachis duranensis inoculated with Ralstonia solanacearum

Bacterial wilt caused by Ralstonia solanacearum is a serious disease of peanut (Arachis hypogaea) in China. However, the molecular basis of peanut resistance to R. solanacearum is poorly understood. Arachis duranensis, a wild diploid species of the genus Arachis, has been proven to be resistant to bacterial wilt, and thus holds valuable potential for understanding the mechanism of resistance to bacterial wilt and genetic improvement of peanut disease resistance. Here, suppression subtractive hybridization (SSH) and macroarray hybridization were employed to detect differentially expressed genes (DEGs) in the roots of A. duranensis after R. solanacearum inoculation. A total of 317 unique genes were obtained, 265 of which had homologues and functional annotations. KEGG analysis revealed that a large proportion of these unigenes are mainly involved in the biosynthesis of phytoalexins, particularly in the biosynthetic pathways of terpenoids and flavonoids. Subsequent real‐time polymerase chain reaction (PCR) analysis showed that the terpenoid and flavonoid synthesis‐related genes showed higher expression levels in a resistant genotype of A. duranensis than in a susceptible genotype, indicating that the terpenoids and flavonoids probably played a fundamental role in the resistance of A. duranensis to R. solanacearum. This study provides an overview of the gene expression profile in the roots of wild Arachis species in response to R. solanacearum infection. Moreover, the related candidate genes are also valuable for the further study of the molecular mechanisms of resistance to R. solanacearum.     

Natural variation of Medicago truncatula resistance to Aphanomyces euteiches

We analysed the resistance variation in 14 natural populations of Medicago truncatula from Tunisia to Aphanomyces euteiches infection. The reaction of M. truncatula lines to A. euteiches infection varied from susceptibility to full resistance. Of the overall level of phenotypic variation, 65.4 % was found to occur within populations. Principal component analysis showed a high spread of lines belonging to the same population, indicating no clear structure in the Tunisian M. truncatula populations and supporting the hypothesis of gene flow among populations. Likewise, there was no association between local resistance composition and the geographical distances between populations, ruling out isolation by distance as an explanation. Furthermore, significant correlations were observed between quantitative traits and ecological factors consistent with the local adaptation hypothesis. A cluster analysis of the populations showed the presence of three groups. The first group comprised the populations originating from the centre of the country, containing the main resistant lines. The second group included the populations collected in the south and the mountain region of Thala and contained the main partially resistant lines. The third group comprised the populations sampled from the north regions and saline soils and included the main susceptible lines. Overall, we found that the natural M. truncatula lines were more likely to be susceptible (71.3 %) than resistant (28.7 %) to A. euteiches attack. Nevertheless, many resistant lines exhibiting new reaction patterns to A. euteiches attack were identified in the natural populations and these can be used for the identification of potentially new resistance genes.     

Narrow sense heritability estimates of bacterial leaf spot resistance in pseudo F2 (F1) population of mulberry (Morus spp.)

Bacterial leaf spot incited by Xanthomonas campestris pv. mori is a devastating foliar disease of mulberry reported globally. Host plant resistance is the most sustainable and economic control measure but so far unexplored. Highly heterozygous plant behaviour and scant genetic information of bacterial leaf spot resistance limits a targetted breeding approach in mulberry. In the present research eight pseudo-F₂(F₁)full-sib progenies derived from selected resistant and susceptible sources were evaluated symptomatically for bacterial leaf spot resistance under natural disease occurrence in 2008 and 2009. Significant variation for bacterial leaf spot resistance was observed in the parents and progenies. Broad sense heritability estimate (0.9) indicates that selection of resistant genotypes can be useful for exploitation in future advanced breeding programs for mulberry. High narrow sense heritability estimates (0.76)[2008] and (0.79)[2009] suggest additive gene effects for the disease resistant trait. The continuous frequency distribution of diseases severity across the progenies indicates that bacterial leaf spot resistance in mulberry may be inherited quantitatively.     

Identification of allele specific AFLP markers linked with bacterial wilt [Ralstonia solanacearum (Smith) Yabuuchi et al.] resistance in hot peppers (Capsicum annuum L.)

Even though the bacterial wilt is identified as the most destructive disease in hot peppers world-wide, robust molecular markers that facilitate marker assisted selection are absent till date. Kerala Agricultural University (India) has released two hot peppers named Ujwala and Anugraha which show high level field resistance to this pathogen. The variety Anugraha was developed through backcross breeding between a high yielding but highly susceptible variety Pusa Jwala with the highly resistant Ujwala, using Pusa Jwala as a recurrent parent. Thus, Pusa Jwala and Anugraha are near isogenic lines (NILs) differing for the resistance to bacterial wilt only and the resistance is governed by a homozygous recessive (rr) gene action. The F₁s of Anugraha × Pusa Jwala were selfed to generate the segregating F₂ population. The F₂ population has been field screened, 10 highly susceptible and 10 most resistant plants were identified and DNA from these plants were bulked separately. Bulked segregant analysis with AFLP primer combination EcoACT + MseCAC was done using the DNA from donor parent Ujwala, susceptible parent Pusa Jwala, resistant parent Anugraha, bulked susceptible F₂ and bulked resistant F₂ plants. On resolution using capillary electrophoresis system in genetic analyzer, the AFLP products have yielded three polymorphic bands (103, 117, and 161 bp) which were linked with the resistant recessive allele and three polymorphic bands (183, 296, 319 bp) linked with the dominant susceptible allele of the bacterial wilt resistance gene. The results were confirmed through co-segregation analysis in most resistant and susceptible plants of F₂ segregating population.     

Genetic analysis of partial resistance to bacterial leaf streak (Xanthomonas campestris pv. cerealis) in wheat

Genetic variability of partial resistance to bacterial leaf streak was investigated in hexaploid winter wheat ( Triticum aestivum. ), using 16 parental genotypes and 48 pure lines (F10) derived from a composite cross programme. Two experiments were undertaken in a controlled growth chamber. Seeds of all genotypes were grown under controlled conditions using a randomized block design with three replications. Each replication consisted of a row of 20 seedlings of each parent and pure line. An Iranian strain of bacterial leaf streak was used for the inoculation of 12-day-old seedlings. In a third experiment, eight genotypes from parents and F10 pure lines representing a large variability for partial resistance were inoculated with four other Iranian strains of bacterial leaf streak. A large genetic variability was observed amongst the 64 genotypes for partial resistance to the disease. Partial resistance heritability estimates were rather high (70%), indicating that the resistance factors may be transmitted by crossing. Amongst all genotypes investigated, 'DC²-30-N2' and 'IBPT-66' displayed the highest partial resistance to the disease. Significant correlations between strains in the third experiment show that a genotype resistant or susceptible to one strain will have similar reactions with other strains. No significant genetic gain was observed for partial resistance in the best pure line of the 48 lines studied, when compared with the best parental line. Increasing the number of pure lines is likely to result in the identification of genotypes that might prove to be more resistant.