Genetic Variability and Interrelationship Among Opium and its Alkaloids in Opium Poppy (Papaver Somniferum L.)

Summary The plant species Papaver somniferum L. (opium poppy) is grown commercially in several countries under secure area to produce opium and its major alkaloids i.e. morphine, codeine, thebaine, narcotine and papaverine for pharmaceutical industries. In recent years the global demand for these alkaloids and its derivatives are growing. Seeing the global demand there is urgent need to develop high opium yielding varieties integrated with high specific alkaloids. However, a breeding programme focused on developing improved varieties requires knowledge about the genetic variability for the concern traits. Therefore the present investigation was undertaken to examine phenotypic and genotypic variability, broad sense heritability, genetic advance under selection and interrelationship of traits. A total of 122 accessions were found to be highly variables for all the traits studied. Broad sense heritability ranged from 74.18% (for opium yield/plant) to 99.00% (for papaverine content). Expected genetic advance was varied between 33.91% (for morphine content) and 257.46% (for papaverine content). Genetic correlation analysis revealed that opium yield is negatively correlated with morphine and papaverine content. Among alkaloids codeine, thebaine, narcotine and papaverine showed positive correlation among themselves. The correlations among alklaloids were justified based on the biosynthetic pathway of opium alkaloids.     

Genetic divergence in parental genotypes and its relation with heterosis, F1 performance and general combining ability (GCA) in opium poppy (Papaver somniferum L.)

Summary Genetic divergence in 110 population (20 parents and 90 F₁ hybrids) of opium poppy was studied by multivariate analysis. All the entries were grouped in 14 clusters. Out of 20 parents, ten were accumulated in one cluster and rest 10 distributed over seven clusters. Estimates of cluster distance showed that maximum intra-cluster distance was in -cluster IX (25.62) followed by cluster I (22.41), V (22.23) and IV (21.06). The inter cluster distances varied from 16.62 (between cluster XII and XIV) to 195.10 (between cluster IV and IX). The cluster analysis indicated substantial diversity among the parental genotypes, which had the potential to release considerable variation in their crosses. The first four principal components (PCs) with eigen values >1 accounted for 68% of variation among the population. The first and second components could account for 29 and 20% of the total morphological and alkaloidal variations, respectively. The PC1 was closely related to yield and yield related traits. The characters contributing greatest positive weight on PC1 were opium yield, plant height, stem diameter, capsule weight/plant and seed yield/plant. The possibilities of relationship between genetic divergence, F₁ performance, heterosis and GCA of parents have been explored.     

Traits related to drought resistance in common bean

Common bean (Phaseolus vulgaris L.) is grown in regions where water deficits during reproductive development significantly reduce yield. The purpose of this study was to evaluate the association of specific phenological and physiological traits with drought resistance in common bean. Five genotypes were grown under and near a rain shelter in 1988, and an additional 16 progeny lines were included in 1990. Drought stress determined by the drought intensity index was severe (0.78) in 1988 and more moderate (0.63) in 1990. Water stress reduced the expression of most traits with the exception of days to flower and leaf moisture retention capacity. Seed yield among genotypes was reduced from 22 to 71% due to drought. Yield under stress was correlated with yield under nonstress in 1990 and negatively correlated with the drought susceptibility index in 1988. Yield components which exhibited the largest differential genotypic responses to stress were pod and seed number, whereas seed size was more stable. Genotypic variation was detected in all the partitioning indexes, chiefly harvest index and relative sink strength, and the heritability estimates for these traits were high. The limited genetic variability observed among water relations traits and their role in water conservation would restrict their potential use in the selection for drought resistance in common bean. The differential correlations between phenological, biomass and partitioning traits and the indexes for yield and drought susceptibility would suggest that the most effective approach in breeding for drought resistance in common bean would be based first on selection for high geometric yield followed by selection among the high-yielding individuals for low to moderate levels of the drought susceptibility index. This revised version was published online in July 2006 with corrections to the Cover Date.     

Types and components of resistance to Fusarium head blight of wheat

Resistance of wheat to Fusarium head blight caused by Fusarium graminearum and F. culmorum was identified in natural epidemics in 1985 and 1987 as well after artificial inoculations (1983–1988 and 1984–1987). Out of 25 genotypes tested, five were identified with no significant difference in head blight scores, but differing significantly in yield after artificial inoculation, i.e. tolerance differences were detected at different resistance levels. Some genotypes that were similar in yield or head blight scores differed in seed infection severity. Genotypes with awns were more susceptible to head blight when tested under natural epidemic condition in the field; but this trait did not influence head blight severity in artificial inoculations. Dwarf genotypes were more severely infected by head blight than tall genotypes under natural conditions, but genotypes of different plant height classes were similarly susceptible after artificial inoculations. In the early generations of a breeding programme resistance measured by visual evaluation of artificial inoculation is the most important way to screen. If selection of dwarf and awned genotypes cannot be avoided, the higher susceptibility caused by awns and dwarfness under natural epidemic conditions can be decreased by a higher level of physiological resistance, as variability in physiological resistance is available. In later generations, traits like percentage of seed infection or tolerance can be identified by additionally measuring yield reduction. Stability of disease reaction appears to be connected with resistance level, the most resistant genotypes are the most stable, and the most susceptible ones tend to have more unstable reactions in different epidemic conditions.     

Contrastant banana accessions for resistance to the burrowing nematode,based on molecular markers RAPD

This work aimed to proceed molecular characterization of seven banana accessions (Borneo, Grand Naine, 1304-06, 4249-05, 0337-02, 0323-03 and 4279-06) resistance to the nematode Radopholus similis. These accessions were selected taking in account the reproduction factor (RF) among 26 banana genotypes from a working collection belonging to Embrapa Mandioca e Fruticultura Tropical. The genomic DNA of the seven accessions was extracted, and 36 decamere primers had been used to obtain RAPD markers. The resulting markers were converted into a matrix of binary data. From that matrix the genetic distance between the accessions were estimated, for further clustering and graphic dispersion analyses. From a total of 521 RAPD markers generated, 420 (81%) were polymorphic, including 140 (27%) potentially promising for application on works related to genetic mapping of the resistance to R. similis. OPE-15, OPH-17, and OPG-09 were the primers that contributed to the highest number of bands promising for genetic mapping of resistance (12, 8, and 8, respectively). The genetic distances between accessions ranged from 0.106 to 0.455, with the longest one observed between cv. Borneo and the genotype 4279-06, considered as highly susceptible and resistant, respectively, to the nematode according to the RF. The graphic dispersion distinguished three groups of accessions, and most of resistant genotypes clustered together in the same group. The most contrastant genotypes for resistance (Borneo and 4249-05) were separated by a genetic distance of 0.374, and possessed a total of 114 polymorphic bands promising for genetic mapping of resistance. In addition, the results of pathogenicity tests were congruent with those obtained by RAPD analyses.     

Heritability of, and genetic correlations among, forage and seed yield traits in Ladino white clover

Sixteen white clover genotypes and their half-sib progenies were grown in pure stand. Each clover progeny was also grown in a mixed stand that also included three grass varieties of different species. Dry matter (DM) yield was measured over 2 years and seed yield at the second year. Competitive ability of clover families was defined as the ratio between mixed stand and pure stand for clover DM yield. Narrow-sense heritability from parent-offspring regression of pure stand data was high to moderate (h²0.50) for DM yield, seed yield and most of their components. Persistence as predicted by stolon density showed negative genetic correlations with seed yield (r_g= -0.70) and DM yield (r₂= -0.60), whereas seed yield and DM yield tended towards a negative correlation (r₂=0.45). Sizes of different vegetative or reproductive organs were generally correlated positively. Evidence was provided that the set of parent genotypes represents well the genetic variation available within the Ladino gene pool. The results highlighted the difficulty of combining relatively short-term DM yield, persistence and seed yield into a unique plant type. Differences in competitive ability emerged only in the second year, better ability being related mainly to petiole length among the traits assessed in pure stand.     

Seed quality attributes of food-grade soybeans from the U.S. and Asia

Diversity of food-grade soybeans is critical for utilization of genetic resources in cultivar development, germplasm enhancement, and end-product commercialization. The objective of this study was to assess seed quality attributes and phenotypic variability among 54 U.S. and 51 Asian food-grade cultivars and breeding lines. The results showed greater genetic diversity of protein content, calcium content, seed hardness, and seed size uniformity than other quality traits in both small- and large-seeded genotypes evaluated in this study. Among the small-seeded soybeans, the U.S. genotypes were more diverse and exhibited higher swell ratio and oil content but lower stone seed ratio and protein content than Asian accessions. Among the large-seeded accessions, U.S. genotypes had higher stone seed ratio and oil content but lower swell ratio and protein content, and were less diverse than Asian genotypes. The characterization of diverse food-grade soybeans will facilitate parent selection in specialty soybean breeding.     

Inheritance pattern of mungbean yellow mosaic disease resistance and gene action for different traits in mungbean (Vigna radiata (L.) Wilczek) under protected and unprotected field conditions

Understanding of mungbean yellow mosaic disease resistance is crucial to develop resistant varieties to combat worldwide threat of the disease epidemics. This study was aimed to determine nature of disease resistance in terms of number of genes governing it and genetics of related traits. Experimental site was located on 31.43°N and 73.06°E with an elevation of 186 m, and evaluation trials were conducted during spring season due to high disease epidemics in this season. Two contrasting genotypes, that is, NM 6‐68‐2 (resistant) and NM 1‐32‐1 (highly susceptible), were crossed to raise six populations, that is, P₁, P₂, F₁, F₂, BC₁ and BC₂ for evaluation under protected and unprotected field conditions using chi‐square test and generation mean and variance analysis. It was discovered that disease resistance was governed by two major genes with additive effects. Disease resistance can easily be incorporated through backcrossing and direct selection following hybridization. Direct selection should be practised at earlier generation for plant height, chlorophyll contents and number of seeds per pod due to preponderance of additive effects whereas at later generations for seed yield per plant, 100 seed weight, harvest index, number of pods per plant and pod length due to involvement of duplicate epistasis.     

Development of non-narcotic (opiumless and alkaloid-free) opium poppy, Papaver somniferum

A mutation breeding programme using gamma rays and ethyl methane sulphonate (EMS) was carried out for genetic conversion of narcotic ‘opium poppy’ into non-narcotic ‘seed poppy’. Two families contained 31 latexless (opiumless) and 23 partial latex-bearing plants which were identified in the M² and confirmed in the M³ generations by the ‘ray-pluck’ method. Thin-layer chromatography (TLC)-densitometry of powdered capsule hulls (straw) from 22 representative plants also revealed that none of the five opium alkaloids (morphine, codeine, thebaine, papaverine and narcotine) was present in the straw. Six plants also possessed high seed yield (4.0–5.66 g/capsule vs. 3.39 g for control) and seven had a high seed oil content (50.7-53.5%). The best mutant genotype, LL-34 of the family C¹-Comb-113-2, with 5.66 g/capsule seeds containing 52.6% oil was designated as cv. ‘Sujata’. This, perhaps the world's first opiumless and alkaloid-free seed poppy cultivar, offers a cheap and permanent (fundamental) solution to the global problem of opium-linked social abuse. Simultaneously, it serves as a safe and potential food crop with protein-rich seeds and healthy unsaturated seed oil.     

Genetic variability and interrelationship of traits in the industrial oil crop Vernonia galamensis

Vernonia galamensis is a wild plant from the family Asteraceae which is endemic to East Africa and has the potential to become a new oil crop for industrial uses. Its seed oil is rich in vernolic acid, a fatty acid of high interest for oleochemical applications. However, a breeding program for Vernonia galamensis cultivars with high seed and oil yields requires knowledge about the genetic variability of traits that influence seed and oil production. This study was undertaken to examine phenotypic and genotypic variability, broad-sense heritability, genetic advance under selection and interrelationships of agronomic and seed quality traits. A total of 122 Vernonia accessions, 115 collected from different regions of Ethiopia and seven introduced, were grown at two locations in Ethiopia (Alemaya and Babile), in 2001/2002 and were analyzed for 20 traits including phenology, yield, yield components, and seed quality with special emphasis on fatty acid composition. The collections exhibited significant variation for all traits except for days to emergence. Genotypes and locations interacted significantly (P 0.01) for all traits. Broad-sense heritability estimates ranged from 11% (for days to emergence) up to 79% (for days to maturity). Expected genetic advance was between 1.3% (for days to emergence) and 44.8% (for seed oil yield). Genetic correlation analysis revealed that seed yield per plant is highly and positively correlated with seed weight and head number; highly significant and negative correlations (r = –0.59, –0.82, –0.85, and –0.89) were found between vernolic acid and palmitic, stearic, oleic, and linoleic acid, respectively. Highly significant positive correlations (r = 0.55, 0.44, and 0.36) were observed between vernolic acid and oil content, meal protein content and seed oil yield, respectively. Path-coefficient analysis indicated seed weight and secondary head number to be the most important components of seed yield per plant. Vernolic acid, oleic acid and linoleic acid had positive direct effects and stearic acid had a negative direct effect on oil content. The direct positive effect of oleic acid on oil content was, however, compensated by the negative indirect effects of stearic and vernolic acid resulting in a negative correlation (r = –0.60) between oleic acid and oil content. These observations will support the selection of accessions with high seed and oil yield, high meal protein contents, and high vernolic acid content.     

Inheritance of resistance to Acanthoscelides obtectus in a wild common bean accession crossed to commercial bean cultivars

Summary The bean weevil Acanthoscelides obtectus, is an important storage pest of common beans in Latin America and Africa. A few wild bean accessions from Mexico have been identified as highly resistant to the weevil. One accession, G 12952, was crossed to two susceptible bean cultivars differing in seed size. Reciprocal F1 and F2 individual seed were evaluated for days to adult emergence (DAE) and emerged adult weight. Maternally inherited seed size affected resistance measurements only in the F1 reciprocal crosses, however, the overall resistance level of the F1 was more similar to that of the susceptible cultivars. The F2 showed a continuous, but skewed distribution from low to high DAE. Very few F2 individuals had the resistance level of G 12952. When the frequency distributions were divided into discrete categories based on parental response, resistance was found to be inherited as two recessive complementary genes. The F3 generation showed an overall lowering of resistance levels compared to their original F2 evaluations. However, none of the lines classified as resistant (50 DAE) in the F2, fell into the susceptible category in the F3, indicating that the resistant genotypes were relatively stable as expected with recessively inherited traits. Modifying genes from the commercial parents may be responsible for general lowering of resistance. Seed size was negatively correlated with adult weight but not with DAE. The unique resistance of the wild bean accessions is discussed in relation to its inheritance. The results and obstacles encountered in the A. obtectus breeding program at CIAT are described.     

Comparative yield component analysis in <Emphasis Type="Italic">Gossypium hirsutum</Emphasis> parents using fibre quality grouping

The usual practice of estimating association and direct and indirect effects among various traits in Gossypium hirsutum is with ungrouped genetic material based on staple length and strength. During 2003–2004 and 2004–2005 genotypic and phenotypic association among ten characters were estimated in G. hirsutum genotypes: (i) set-1: 10 genotypes of low fibre strength (≤196.2 mN/tex) and medium staple length (≤25.0 mm), (ii) set-2: 10 genotypes of high fibre strength (≥235.44 mN/tex) and long staple length (≥28 mm) and (iii) set-3: 20 genotypes of set-1 and 2. The differences in the estimates of associations, direct and indirect effects for different characters in fibre quality groups were examined. The complete analysis of variance for the characters under study viz; days to 50% flowering, plant height (cm), number of monopodial and sympodial branches per plant, seed cotton yield per plant (g), boll weight (g), total number of bolls per plant, lint %, 2.5% span length (mm) and fibre strength (mN/tex) indicated highly significant genotypic differences for genotypes, years and genotype × years interaction. Variation within medium staple length and low strength (set-1), high staple length and high strength genotypes (set-2) and their interaction with year were also significant. Medium staple length and low strength (set-1) versus high staple length and high strength component (set-2) was significant for all the traits indicating significant variability between the sets. A true relationship and direct selection were observed for days to 50% flowering, number of monopodial and sympodial branches per plant with seed cotton yield in the three sets suggesting that separation of genetic material based on staple length and fibre strength for improvement of seed cotton yield through these traits is not required. Magnitude and direction of direct effect and association with yield of the other traits; plant height, number of total bolls per plant, boll weight, lint %, staple length and fibre strength differed between the sets. This study concluded that more successful planning of a breeding programme can be made through path coefficient analysis if the genetic material is grouped based on staple length and fibre strength. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Response of white clover genotypes to evaluation environments of dense and spaced planting, and implications for selection

Twelve genotypes were grown for two years asmonocultures in the evaluation environments (EE) ofspaced planting (SP) and dense planting (DP). Drymatter (DM) yield, seed yield and their contributingtraits were recorded. The environments were comparedfor: i) genotype performance, and selection efficiencyfor the target environment of DP, and ii) estimates ofgenetic correlations between traits. DP was obtainedby close transplanting of clonal cuttings. EE × Ginteraction occurred for DM yield over harvests, andstolon density, petiole length, inflorescenceproduction and number of florets per inflorescence averagedover repeated measurements. Interaction effectsleading to relatively higher DM yield in SP tended tobe associated positively with those for the othertraits and featured the genotypes with greater abilityto colonize bare soil by stolon production. Theinteraction effects decreased with time and generallydisappeared (p < 0.05) by the end of the experiment,when the environments reached a similar DM yield andstolon density. EE × G interaction also occurred forseed yield (p < 0.08), recorded at the second year.Indirect selection in SP for performance in DP waspredicted to be less efficient for DM yield (–40%),seed yield (–23%) and the other traits subject to EE ×G interaction, compared with direct selection in DP.Evaluation under SP led to identification of severalgenetic correlations that were not confirmed under DP.DP compared with the traditional SP evaluationtechnique is more labour-demanding but allows for i)increasing the selection efficiency and/or decreasingthe time needed for a reliable evaluation, and ii)obtaining a more reliable estimation of geneticcorrelations for ordinary agricultural environments.     

Genetic and histological characterization of downy mildew resistance at the cotyledon stage in <Emphasis Type="Italic">Raphanus sativus</Emphasis> L.

Downy mildew (DM; Hyaloperonospora brassicae) is one of the most serious diseases in radish and responsible for high production losses. The objectives of this study were (1) to identify new sources of DM resistance at the cotyledon stage; (2) to determine the inheritance of the resistance; and (3) to clarify the histological and cellular response involved in DM infection. We screened 200 radish accessions for DM resistance at the cotyledon stage. Radish accessions Rd001, Rd004, Rd048 and Rd150 showed a complete resistance response (hypersensitive reaction), and accession Rd193 exhibited a partial resistance with confined sporulation. The highly susceptible accession Rd197 was then crossed with all resistant genotypes for a genetic analysis. F₂ segregation analysis suggested that cotyledon resistance in Rd001, Rd004, Rd048 and Rd150 is conferred by a single dominant gene while resistance in Rd193 is polygenic. Genetic data suggests Rd193 resistance might be conferred by two dominant genes with complementary action, althought further experiments are needed to confirm this hypothesis. To conduct a histological study using light and fluorescence microscopy, we collected cotyledons from three genotypes with either differing levels of resistance (Rd004 and Rd193) and high susceptibility (Rd197) at ten different time points, from 3 h to 7 days post inoculation. Tissue necrosis was responsible for the inhibition and reduction of H. brassicae growth in Rd004 and Rd193 within 24 h of inoculation. For susceptible accession Rd197, the first evidence of necrotic cells appeared 48 h post inoculation. Callose deposition associated with intercellular H. brassicae growth occurred 3 h post inoculation for all genotypes and was the lowest in resistant accession Rd004. Cotyledon resistance to DM identified in radish is of high commercial interest and has the potential to be used in breeding programs.     

Morpho-physiological parameters associated with iron deficiency chlorosis resistance and their effect on yield and its related traits in groundnut

Iron deficiency chlorosis (IDC) causes a significant reduction in yield of groundnut grown in calcareous and alkaline soils in India. The main aim of the study was to assess genotypic differences for morpho-physiological parameters associated with IDC resistance across different stages and their effect on yield and its related traits. The factorial pot experiment was comprised of two major factors, i) soil-Fe status [normal-Fe, deficit-Fe], and ii) genotypes [five] with differential IDC response, constituting 10 treatments. They were assessed for five morpho-physiological parameters associated with IDC resistance across five crop growth stages and also yield and its related traits. Associations between these traits were also estimated. Under deficit-Fe conditions, IDC resistant genotypes recorded significantly lower visual chlorosis rating (VCR), higher SPAD values, active Fe, chlorophyll content, peroxidase activity, and high yield compared to susceptible ones. Between normal- to deficit-Fe soils, resistant compared to susceptible genotypes showed no change in VCR scores; a lower reduction in SPAD, chlorophyll, active Fe, peroxidase activity, and pod yield. Under deficit-Fe conditions, high yield among resistant genotypes could be attributed to higher seed weight, number of pods and haulm yield, while contrasting reduction in main stem height and number of primaries. The results indicate that for initial large-scale screening of groundnut genotypes for IDC resistance, SPAD values are most ideal while active Fe could be utilized for confirmation of identified lines.     

Cercosporiosis resistance in coffee germplasm collection

Cercosporiosis, or brown eye spot, is currently one of the main diseases of the coffee tree. It is caused by Cercospora coffeicola Berk. & Cooke. Nevertheless, genetic resistance to this disease has not yet been explored in any depth. Our objectives (a) were evaluate the response of 124 accessions from the germplasm collection of the Minas Gerais State (GC), Brazil, and eight commercial cultivars of C. arabica to cercosporiosis and (b) determine the best way to perform early progenies selections via controlled greenhouse experiments. Three controlled greenhouse experiments (1–3) were run in different seasons to determine the best way to proceed the selection. The seedlings were inoculated with a four isolates mixture obtained from different regions. The experimental data were analyzed individually (1–3), in a joint analysis, and as repetitions of a randomized complete block design. In each analysis we estimated genetic parameters and E-BLUP (empirical best linear unbiased predictor) genotypic values of the access. There was genetic variability to C. coffeicola resistance among the coffee tree accessions germplasm collection. Therefore, genetic improvements could be obtained by selection. Experimental repetitions in different seasons increase the selection efficiency and reliability of resistant genotypes with low cercosporiosis severity. The genotype Sarchimor MG 8840 showed the highest resistance level followed by Guatenano and the Timor Hybrid UFV 377-34, Timor Hybrid UFV 376-14 BE 5, and Wush–Wush × Timor Hybrid UFV 366-08.     

Genetic progress of upland rice (Oryza sativa L.) lines for disease resistance

The rice crop is affected by diseases throughout its cycle, impacting negatively on grain yield and quality. The control of the disease impact can be accomplished via crop breeding, using highly multiple resistant genotypes. This study aimed to evaluate the efficiency of multiple-character and specific selection of multiple resistance to major culture-associated diseases (neck blast, leaf scald and grain discoloration) in rice lines of the Upland Rice Genetic Breeding Program. The experiments were conducted in 35 sites during 12 agricultural years, where 124 lines were evaluated for the severity of fungal diseases, under natural field conditions. Multiple parameters were calculated based on the diseases´ scores: genetic, phenotypic and environmental variances, heritability, selection gain, renewal rate, and genetic and renewal progress. Genetic variance for the disease resistance was identified in the population, and the selection gain for multiple-character selection was of 3.16 year⁻¹ throughout the breeding process with a renewal rate of over 35%. The programme has showed efficiency in selecting multiple resistant genotypes to the mentioned diseases, highlighting genotypes with high potential for market release.     

Development of EST‐SSR markers for diversity and breeding studies in opium poppy

All publicly available opium poppy expressed sequence tag (EST) sequences, totalling 20 885, were assembled into unigenes and examined for simple sequence repeats (SSRs). Nearly 19% of the 14 957 unigenes contained SSRs with 4% harbouring more than one SSR. Average density of the SSRs was 1 SSR per 3.6 kb of non‐redundant EST sequence. Trinucleotide SSRs were most frequently identified (39%), and many of the most prevalent motifs were AT‐rich. Flanking primers were designed for 86% of the SSRs and 67 primer pairs were tested on 37 opium poppy accessions and seven related species. All markers were transferable to the related species. Polymorphism information content (PIC) values for the markers were intermediate for comparisons within opium poppy (average of 0.27) and slightly higher for comparisons across species (average of 0.29). The markers were found to be useful for diversity analysis as they successfully distinguished among Turkish opium poppy accessions and land races.     

Bru1 gene and potential alternative sources of resistance to sugarcane brown rust disease

Brown rust, caused by the fungus Puccinia melanocephala, is responsible for important yield losses in sugarcane production globally and it is therefore an important objective to introduce resistance to this disease in breeding programs. A major gene, Bru1, has been shown to confer resistance to P. melanocephala strains from different parts of the world and two molecular markers, R12H16 and 9O20-F4, closely associated to this gene have been previously reported. The usefulness of these molecular diagnostic markers in order to predict a rust resistant phenotype under natural high pressure inoculums conditions was analyzed. A total of 129 sugarcane accessions were evaluated under field infection for resistance or susceptibility to brown rust and subsequently screened for presence or absence of the two Bru1 diagnostic markers. A total of 49 genotypes (38 %) were phenotyped as resistant to brown rust but only eight (16.3 %) of them were harboring the Bru1 gene. To determine overall frequency of the Bru1 in the local sugarcane germplasm collection, 190 additional genotypes were examined. Presence of Bru1, as determined by the diagnostic markers, was detected in only 7 % of the genotypes evaluated. In conclusion, Bru1 diagnostic markers enable positive selection for brown rust resistance in sugarcane and moreover allowed detecting at least one additional source(s) of resistance. Interestingly, whilst only little genetic variability of rust resistance independent of Bru1 has been reported previously, this alternative genetic resource(s) found in our local germplasm constitutes the predominant one and should be helpful in order to amplify the narrow genetic basis for brown rust resistance in sugarcane.