Rooting of bean leaves and use in germplasm evaluation for common bacterial blight resistance

Summary A simple protocol for leaf rooting in beans (Phaseolus vulgaris L.) was developed and used to investigate the reaction of Xanthomonas campestris pv. phaseoli (Xcp), causal pathogen of common bacterial blight disease, in detached versus attached bean leaves. Trifoliate leaves of different sizes (one-third, two-thirds, and fully expanded), either with or without the pulvinus attached to the petioles, were excised from 20 day-old plants of six bean cultivars/lines. Leaf cuttings were cultured in potting medium and then incubated for 5 to 10 days under transparent polyethylene plastic cover in the greenhouse. Roots were readily initiated along the petioles of the leaf cuttings, whether the pulvinus was present or absent. All leaves which were two-thirds expanded and fully expanded developed roots 5 to 7 days after culture. Eighty to 90 percent of the leaves which were one-third expanded formed roots 8 to 10 days after incubation. Laminae of the rooted leaf cuttings were viable and green during the 2 to 3 months period in culture after removing the plastic cover. The common bacterial blight reactions were similar for inoculated attached leaves, detached rooted leaves (inoculated either after or prior to rooting), and moistened detached leaves incubated without rooting. The latter were only usable for evaluation of the Xcp reaction in growth chamber experiments but not under greenhouse conditions. The rooted leaves would be useful for screening bean lines for multiple disease resistance, especially if the pathogens require different environments for disease expression.Abbreviations CBB Common Bacterial Blight - Xcp Xanthomonas campestris pv. phaseoli (Smith) Dye     

Resistance to common bacterial blight in Phaseolus vulgaris L. recombinant inbred lines under natural infection of Xanthomonas axonopodis pv. phaseoli

Among the main causes of poor yield in common beans are fungal, viral and bacterial diseases. Common bacterial blight, caused by Xanthomonas axonopodis pv. phaseoli (Xap), is one of the major bacterial diseases leading to significant losses in Brazil. Chemical control is ineffective, therefore, the use of resistant varieties becomes an interesting alternative. The objective of the present work was to evaluate disease resistance under natural infection of the pathogen in 109 recombinant inbred lines (F₇) of P. vulgaris originated from the cross HAB-52 (susceptible — snapbean) × BAC-6 (resistant — common bean) in two different environments, as well as to calculate genetic parameters to assist in the selection of promising materials to be used in the CBB resistance breeding program. The data of the genetic parameters were compared to those calculated for the F₃ generation originated from the same cross. The heritability results for DI (disease index) and VI (variation index) in F₃ were 26.85% and 0.26, respectively, whereas in F₇ they were 91.77% and 1.36, respectively. These results demonstrate a potential to be explored for this advanced population, that in the future, along with other pathogen variability studies and tests in other environments, may provide more information regarding a more precise evaluation of promising genotypes to be used in common bean breeding programs aiming to obtain CBB resistant varieties. This revised version was published online in July 2006 with corrections to the Cover Date.     

Backcross breeding for improved resistance to common bacterial blight in pinto bean (Phaseolus vulgaris L.)

Common bacterial blight (CBB) caused by Xanthomonas campestris pv. phaseoli reduces common bean (Phaseolus vulgaris L.) yield and quality worldwide. Genetic resistance provides effective disease control; however. a high level of resistance is difficult to attain and does not exist in pinto bean, the most important dry bean market class in North America. Our objective was to determine if a backcross breeding approach with the aid of molecular markers linked to quantitative trait loci (QTL) for resistance to CBB in a donor parent could be used to attain higher levels of resistance to CBB in pinto bean. QTL conditioning CBB resistance from the donor parent XAN 159 were introgressed into the recurrent parent‘Chase’using classical backcross breeding and intermittent marker‐assisted selection.‘Chase’pinto bean is moderately resistant and the breeding line XAN 159 is highly resistant to Xanthomonas campestris. Marker assays confirmed the presence of independent QTL from GN no. 1 Sel 27 and XAN 159 in advanced backcross‐derived pinto bean lines with improved CBB resistance. Agronomic characteristics of‘Chase’were fully recovered in the backcross‐derived lines. An important QTL for CBB resistance from XAN 159 on linkage group B6 was not introgressed because tight linkage between this QTL and the dominant V allele that causes an unacceptable black‐mottled seed coat colour pattern in pinto bean could not be broken.     

Stability of the association of molecular markers with common bacterial blight resistance in common bean (Phascolus vulgaris L.)

Common bacterial blight (CBB) caused hy Xanthomonas campestrts pv. phaseoli is an important disease of common bean (Phaseolus vulgaris L.) throughout the world. Two random amplified polymorphic DNA (RAPD) markers (R7313 and R4865) linked to genes for CBB resistance, that were transferred to P- vulgaris by an interspecific cross with Phaseohus acutifoluis. Were identified in a previous study. The current study was conducted to examine the use of these markers for selecting CBB resistant material from 85 F_5,6, lines derived from crosses between two of the resistant lines used previously in the linkage study and susceptible breeding lines. The results showed that these two markers were located on the same linkage group and explained 22% (P = 0.0002) of the variation in response to CBB in the current population. Seventy per cent of the lines that had both markers were classified as resistant in a disease test of the F_5,6, lines, whereas 73% of the lines that had neither of the RAPD markers were susceptible. The results indicated that the marker-disease resistance associations remained stable in a plant breeding programme and that they can be used lor marker-assisted selection of CBB-resistant beans.     

An SSR marker in the nitrate reductase gene of common bean is tightly linked to a major gene conferring resistance to common bacterial blight

A simple sequence repeat (SSR) marker composed of a tetra nucleotide repeat is tightly linked to a major gene of common bean (Phaseolus vulgaris L.) conferring resistance to common bacterial blight (CBB) incited by Xanthomonas axonopodis pv. phasoli (Xap). This SSR is located in the third intron region of the common bean nitrate reductase (NR) gene, which is mapped to linkage group (LG) H7, corresponding to LG B7 of the bean Core map. Co-segregation analysis between the SSR marker and CBB resistance in a recombinant inbred line (RIL) population demonstrated a tight linkage between the NR gene-specific marker and the major gene for CBB resistance. In total, the marker explained approximately 70% of the phenotypic variation in the population. Because it is co-dominant, this SSR marker should be more efficient for marker-assisted selection (MAS) than dominant/recessive random amplified polymorphic DNA (RAPD) or sequence characterized amplified region (SCAR) markers that have been developed, especially for early generation selection. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effect of photoperiod and temperature on common blight disease of common beans (Phaseolus vulgaris L.)

Summary Although common blight disease is serious in many dry bean production areas, there is only limited information on the influence of photoperiod on the disease. Experiments were conducted in growth chambers and in the field (Nebraska, Dominican Republic, Puerto Rico) to investigate the influence of photoperiod × temperature and photoperiod, respectively, on the reaction of cultivars/lines to the causal bacterium Xanthomonas campestris pv. phaseoli. A split-plot design was used in all experiments except in the DR experiment where cultivars/lines were replicated within each photoperiod treatment. The disease reactions were more severe on cultivars/lines under short photoperiod and under higher temperature than under longer photoperiod and lower temperature in the growth chamber. Disease reactions were also more severe under the short photoperiod in the field experiments. No interactions were detected among these factors. These results have important implications for plant breeders in the evaluation of common blight disease reactions in different latitudes.     

Introgressing resistance to bacterial and viral diseases from the middle American to Andean common bean

The genetic base of cultivars within market classes of common bean (Phaseolus vulgaris L.) is narrow. Moreover, small- and medium-seeded Middle American cultivars often possess higher yield and resistance to abiotic and biotic stresses than their large-seeded Andean counterparts. Thus, for broadening the genetic base and breeding for higher yielding multiple stress resistant Andean cultivars use of inter-gene pool populations is essential. Our objective was to determine the feasibility of introgressing resistance to Been common mosaic virus (BCMV, a potyvirus), and the common [caused by Xanthomonas campestris pv. phaseoli (Xcp) and X. campestris pv. phaseoli var. fuscans (Xcpf)] and halo [caused by Pseudomonas syringae pv. phaseolicola (Psp)] bacterial blights from the Middle American to Andean bean, using gamete selection. Also, we investigated the relative importance of the use of a landrace cultivar versus elite breeding line as the last parent making maximum genetic contribution in multiple-parent inter-gene pool crosses for breeding for resistance to diseases. Two multiple-parent crosses, namely ZARA I = Wilkinson 2 /// ‘ICA Tundama’ / ‘Edmund’ // VAX 3 / PVA 773 and ZARA II = ‘Moradillo’ /// ICA Tundama / Edmund // VAX 3 / PVA 773 were made. From the F₁ to F₅ single plant selection was practiced for resistance to the common and halo bacterial blights in both populations at Valladolid, Spain. The parents and F₅-derived F₆ breeding lines were evaluated separately for BCMV, and common and halo bacterial blights in the greenhouse at Filer and Kimberly, Idaho in 2001. They were also evaluated for the two bacterial blights, growth habit, seed color and 100-seed weight at Valladolid in 2002. All 20 F₁ plants of ZARA I were resistant or intermediate to common and halo bacterial blights in the greenhouse, but their F₂ and subsequent families segregated for both bacterial blights. Segregation for resistant, intermediate, and susceptible plants for common bacterial blight occurred in the F₁ of ZARA II. Simple correlation coefficient for common bacterial blight between the F₁ and F₁-derived F₂ families was positive (r = 0.54 P < 0.05) for ZARA II. From the F₂ to F₅ the number of families resistant to both bacterial blights decreased in both populations. Only four of 20 F₁ plants in ZARA I resulted in seven F₆ breeding lines, and only one of 32 F₁ plants in ZARA II resulted in one F₆ breeding line resistant to the three diseases. None of the selected breeding lines had seed size as large as the largest Andean parent. The use of elite breeding line or cultivar as the last parent making maximum genetic contribution to the multiple-parent inter-gene pool crosses, relatively large population size in the F₁, and simultaneous selection for plant type, seed traits as well as resistance to diseases would be crucial for introgression and pyramiding of favorable alleles and quantitative trait loci (QTL) of interest between the Andean and Middle American beans.     

Interaction of common bacterial blight quantitative trait loci in a resistant inter‐cross population of common bean

Common bacterial blight (CBB), caused by Xanthomonas axonopodis pv. phaseoli, is an important disease of common bean (Phaseolus vulgaris L.). Genetic resistance is the most economically efficient, environmentally friendly and socially acceptable approach to control plant diseases including CBB. To examine the main and interaction effects of the previously identified CBB resistance quantitative trait loci (QTL) associated with markers BC420 (B) on Pv06, SAP6 (S) on Pv10 and PVctt001 (P) on Pv04, in presence of the major QTL associated with the marker SU91 (S_u) on Pv08, a resistant F_4:5 recombinant inbred line population of the reciprocal crosses between OAC Rex (bb/ss/PP/S_uS_u) and HR45 (BB/SS/pp/S_uS_u) was evaluated under artificial field inoculation in disease nurseries in 2009 and 2010. While, in presence of the CBB QTL on Pv08, the QTL on Pv06 of HR45 accounted for 37–46% of phenotypic variation in the field, the effect of QTL on Pv04 and Pv10 were not significant under field conditions, even in the absence of the Pv06 QTL. Broad‐sense heritability estimates of CBB resistance and the QTL associated with BC420 were high for CBB severity and the area under the disease progress curve, promoting the continued efforts in pyramiding the QTL on Pv06 and Pv08 in common background, which provide high levels of resistance.     

Qualitative and quantitative evaluation of cassava bacterial blight resistance in F1 progeny of a cross between elite cassava clones

Deployment of resistant varieties is one major approach to controlling cassava bacterial blight (CBB), caused by Xanthomonas axonopodis pv. manihotis (Xam). To understand the genetic determinism of resistance to CBB, the use of reliable parameters measuring resistance is necessary. In order to test a relevant method for evaluation of quantitative resistance for mapping QTL (quantitative trait loci), the response of 150 F₁ individuals, inoculated with four different Xam strains (CIO-84, CIO-1, CIO-136 and CIO-295), was assessed under controlled conditions. We used two types of evaluations at different intervals after inoculation, one based on a scale of 0 to 5 and the second based on the determination of the bacterial population in the vascular system. Both evaluation types revealed interaction between strains and F₁ genotypes. Population values at 3 and 6 cm from the point of inoculation showed a high level of correlation. By performing an association analysis, at 7 and 15 days after inoculation, a significant positive correlation between both evaluation types was obtained. However, the disease rating at 30 days did not correlate with bacterial populations at either 7 or 15 days after inoculation, except for one strain, CIO-84. Evaluation of the bacterial population in stem tissues is time and labour consuming, consequently, for a rapid and reliable assessment of CBB resistance for QTL analysis, we strongly recommend evaluation based on the use of a symptom scale.     

A major QTL for common bacterial blight resistance derives from the common bean great northern landrace cultivar Montana No.5

Knowledge of the evolutionary origin and sources of pest resistance genes will facilitate gene deployment and development of crop cultivars with durable resistance. Our objective was to determine the source of common bacterial blight (CBB) resistance in the common bean Great Northern Nebraska #1 (GN#1) and GN#1 Selection 27 (GN#1 Sel 27). Several great northern cultivars including GN#1, GN#1 Sel 27, and Montana No.5 (the female parent of the common x tepary bean interspecific population from which GN #1 and GN # 1 Sel 27 were derived) and known susceptible checks were evaluated for CBB reaction in field and greenhouse environments. These genotypes and CBB resistant and susceptible tepary bean including Tepary #4, the male parent and presumed contributor of CBB resistance toGN#1 and GN#1 Sel 27, were assayed for presence or absence of three SCAR markers tightly linked with independent QTLs conditioning CBB resistance. The parents and F₂ of Montana No. 5/GN #1 Sel 27 and Montana No.5/Othello(CBB susceptible) were screened for CBB reaction and SCAR markers. CBB resistance in Montana No.5 was comparable to that of GN#1 and GN#1 Sel27. The SAP6 SCAR marker present in GN#1 and GN#1 Sel 27 was also present in Montana No.5, and it co-segregated (R ² =35%) with the CBB resistance in the Montana No.5/Othello F₂ population. Although a few CBB resistant and susceptible transgressive segregants were found in the F₂ of MontanaNo.5/GN #1 Sel 27 and later confirmed by F₃ progeny tests, SAP6 SCAR marker was present in all progenies. None of the tepary bean specific CBB resistance-linked SCAR markers were present in GN#1, GN#1 Sel 27, or Montana No.5. A cluster analysis of 169 polymorphic PCR-based markers across three common bean and Tepary #4 indicated that GN#1, GN#1 Sel 27, and Montana No.5 were closely related, and not related at all with Tepary #4.Thus, these results clearly indicate Montana No.5, not Tepary #4, as the source of CBB resistance in GN#1 and GN#1 Sel 27. This revised version was published online in August 2006 with corrections to the Cover Date.     

Inheritance of resistance to Xanthomonas campestris pv phaseoli in tetary bean (Phaseolus acutifolius)

Summary The F1, F2 and F3 from two crosses within Phaseolus acutifolius were exposed to Xanthomonas campestris pv phaseoli to analyse the inheritance of resistance. The resistant parent, PI 319.443, gave a hypersensitive reaction in leaves and pods with small necrotic lesions. Based on the resistance of F1, the segregation in F2 and the reaction of F3 plants and lines, it is concluded that resistance in leaves and pods is governed by one dominant gene. Comparisons are made with the resistance to X. campetris in P. vulgaris.     

Marker-assisted selection of common beans for resistance to common bacterial blight: efficacy and economics

The possibility of using random amplified polymorphic DNA (RAPD) markers previously mapped in the common bean PC50/XANI59 population to select for resistance to common bacterial blight (CBB) in different populations was examined. Two out of 02 selected RAPD markers were polymorphic in HR56 and W0633d, the parental lines used in this experiment. Cosegregation analysis of the two polymorphic markers and disease reaction in a recombinant inbred (RI) population derived from HR67/W1744d confirmed that one of the two RAPD markers, BC420₉₀₀, was significantly associated with a major quantitative trait locus‐conditioning resistance to CBB in HR67. This locus accounted for approximately 51) of the phenotypic variation. The RAPD marker was transformed into a sequence characterized amplified region (SCAR) marker and used for selection in a different population derived from ‘Envoy’/HR67. Prediction for resistance to CBB with the BC420_.990 SCAR marker was 94.2% accurate in this population. A comparison between marker‐assisted selection (MAS) and conventional greenhouse screening showed that the cost of MAS is about one‐third less than that of the greenhouse test.     

Inheritance of resistance to bacterial blight disease of rice

Summary Inheritance of resistance to the Punjab isolate of Xanthomonas campestris pv. oryzae of bacterial blight disease of rice was studied in seven breeding lines resistant to the disease. The results revealed that resistance in breeding lines PAU 122-73-1-4-1, PAU 164-102-1-2-1-1-1, KJT 24, IR 5657-33-2-1-2 and IR 22082-41-2-2 was controlled by single dominant genes allelic to the dominant gene which confers resistance to the Punjab isolate in Patong 32. Resistance to the Punjab isolate in breeding lines IET 7172 and RP 2151-40-1 was found to be controlled by single recessive resistance genes allelic to one of the recessive resistance genes present in BJ 1. The two genes are independently inherited and are being used to develop bacterial blight resistant varieties.     

Comparison of leaf and pod disease reactions of beans (Phaseolus vulgaris L.) inoculated by different methods with strains of Xanthomonas campestris pv. Phaseoli (Smith) dye

Summary Common blight disease in beans (Phaseolus vulgaris), caused by the bacterium Xanthomonas campestris pv. phaseoli, reduces crop yield and seed quality. Information is needed on the variation of leaves and pods disease reaction to strains of the bacterium after different inoculation methods. Phaseolus vulgaris cultivars Red Kidney Charlevoix, GN Harris, GN 1140, and GN Emerson were inoculated with three different strains of Xanthomonas campestris pv. phaseoli at two inoculum concentrations (10⁸ and 10⁶ bacterial cells/ml) using water soaking, multiple needle, and razor blade inoculation on leaves, and razor blade scratch, dissecting needle, and razor blade cut inoculation on pods. Differential cultivar disease reactions of leaves, pods, or both to the bacterial strains were observed in some cases. Significant interactions among cultivars, inoculation methods, strains, and inoculum concentrations (leaves) were found. A rapid leaf chlorosis developed 6 to 7 days after inoculation. Strains of bacteria did not show specificity in inducing this reaction, but rapid leaf chlorosis was associated with high inoculum concentration and with the water soaking and multiple needle methods. Another experiment was conducted to count the number of living bacterial cells deposited in the leaf tissue after inoculation by different methods. The number of bacteria deposited by water soaking or multiple needle was higher than that deposited by razor blade.Published as Paper No. 8584, Journal Series, Nebraska Agricultural Experiment Station. Research was conducted under Project No. 20–36.     

Inheritance of resistance to Xanthomonas campestris pv. translucens in triticale

Summary Three triticale lines, Siskiyou, M2A-Beagle, and OK 77842 have been reported to possess resistance to bacterial leaf streak caused by Xanthomonas campestris, pv. translucens (Xct.). The three resistant lines were crossed to susceptible lines and crossed with each other. F₂, BC₁-F₁, BC₂-F₁ plants were inoculated with a mixture of two Xct strains. The segregation data indicate the presence of a single dominant gene in each of the three resistant lines to bacterial leaf streak. These three genes are either the same or closely linked herein designated as Xct₁.     

Interactions between QTL SAP6 and SU91 on resistance to common bacterial blight in red kidney bean and pinto bean populations

Breeding efforts to improve resistance in dry bean to common bacterial blight (CBB) have focused on applying marker assisted selection strategies. We examined the interaction between two independent QTL (quantitative trait loci), SAP6 and SU91, on the expression of resistance to CBB in a pinto bean F₂ population and dark red kidney bean F₂ population. The disease reaction for each F₂ population was scored at several time points after inoculation. The dominant SCAR markers SAP6 and SU91 were simultaneously genotyped as codominant markers using a multiplex real-time PCR assay. For both populations, at each time point plants having genotypes of (su91 su91//SAP6 SAP6); (su91 su91//SAP6 sap6), and (su91 su91//sap6 sap6) were significantly more susceptible to CBB than plants with any of the six other possible genotypes (SU91 _//_ _). Only two examples were observed across both populations and all time points in which mean disease reactions were significantly different between any of the six (SU91 _//_ _) genotypes. No significant differences were observed between SU91 SU91 and SU91 su91 genotypes at any time point for either F₂ population. Similarly, no significant differences in CBB reaction were observed among the three SAP6 genotypes for plants that had the su 91/su91 genotype. The results indicate CBB resistance in these two populations is primarily conditioned by the presence of at least a single copy of the SU91 QTL. The effects of pathogen isolate and population on discerning the influence of QTL on CBB resistance are discussed.     

Inheritance of resistance to black rot (Xanthomonas campestris pv. Campestris) in cauliflower (Brassica oleracea var. Botrytis)

Summary Black rot disease caused by Xanthomonas campestris pv. campestris is a limiting factor in the commercial production of the cauliflower crop. Crosses were attempted between SN 445, a mid season cultivar resistant to black rot and two highly susceptible commercial cultivars (Pusa Snowball-1 and K-1). Studies of the F₁'s, F₂'s and back crosses indicated that SN 445, carries a dominant gene imparting resistance to black rot.     

Segregation distortion of Brassica carinata derived black rot resistance in Brassica oleracea

Three segregating F₂ populations were developed by self-pollinating 3 black rot resistant F₁ plants, derived from across between black rot resistant parent line 11B-1-12 and the susceptible cauliflower cultivar ‘Snow Ball’. Plants were wound inoculated using 4 isolates ofXanthomonas campestris pv. campestris (Xcc) race 4, and disease severity ratings of F₂ plants from the three populations were scored. A total of 860 arbitrary oligonucleotide primers were used to amplify DNA from black rot resistant and susceptible F₂ plants and bulks. Eight RAPD markers amplified fragments associated with completely disease free plants following black rot inoculation,which segregated in frequencies far lower than expected. Segregation of markers with black rot resistance indicates that a single, dominant major gene controls black rot resistance in these plants. Stability of this black rot resistance gene in populations derived from 11B-1-12 may complicate introgression into B. oleracea genotypes for hybrid production. This revised version was published online in July 2006 with corrections to the Cover Date.     

A rapid screening technique to combine resistance to halo blight and bean common mosaic virus in Phaseolus vulgaris L.

Summary Seven bean lines (Phaseolus vulgaris L.) with differential resistance or susceptibility to race 2 of halo blight (Pseudomonas syringae pv. phaseolicola) and a necrosis-inducing isolate of bean common mosaic virus were inoculated with one or both pathogens in combination, to determine the feasibility of dual screening to identify resistance to both pathogens simultaneously. Dual screening yielded the same results as separate screenings. Neither pathogen affected the disease expression of the other. Simultaneously screening for resistance to both pathogens will shorten the recurrent screening-selection cycle of hybridization programs, and accelerate development of resistant cultivars.Abbreviations BCMV Bean Common Mosaic Virus - cvs Cultivars - HB Halo blight - Inoc. Pt. Inoculation point - NLL Necrotic local lesion - React Reaction - SVN Spreading veinal necrosis, System chloro-Systemic chlorosis - VN Vascular necrosis     

Genetic analysis of resistance to bacterial leaf streak caused by Xanthomonas campestris pv. undulosa in bread wheat

Summary The inheritance of resistance to bacterial leaf streak or black chaff of wheat was studied under field conditions, with an artificial epidemic of Xanthomonas campestris pv. undulosa. A complete series of crosses between five parents, differing in reaction to X. c. pv. undulosa, was generated. Disease was recorded at two different stages of growth. No evidence of cytoplasmic effect was found from the comparison between reciprocal F₁ crosses. The study of the F₃ generations attested that five genes were involved in resistance to bacterial leaf streak. Separate analyses carried out for the two scoring dates were mutually consistent: genotypes, number of genes, and their action and relative importance were verified. The genes differed in strength of expression of resistance. One of the two strongest genes, Bls1, is present in all three superior parents, Pavon 76, Mochis T88 and Angostura F88. Resistance was not complete, and proved to be stable over the season.