Transfer of bacterial blight and blast resistance from the tetraploid wild rice Oryza minuta to cultivated rice,Oryza sativa

Summary Oryza minuta J. S. Presl ex C. B. Presl is a tetraploid wild rice with resistance to several insects and diseases, including blast (caused by Pyricularia grisea) and bacterial blight (caused by Xanthomonas oryzae pv. oryzae). To transfer resistance from the wild species into the genome of cultivated rice (Oryza sativa L.), backcross progeny (BC₁, BC₂, and BC₃) were produced from interspecific hybrids of O. sativa cv IR31917-45-3-2 (2n=24, AA genome) and O. minuta Acc. 101141 (2n=48, BBCC genomes) by backcrossing to the O. sativa parent followed by embryo rescue. The chromosome numbers ranged from 44 to 47 in the BC₁ progeny and from 24 to 37 in the BC₂ progeny. All F₁ hybrids were resistant to both blast and bacterial blight. One BC₁ plant was moderately susceptible to blast while the rest were resistant. Thirteen of the 16 BC₂ progeny tested were resistant to blast; 1 blast-resistant BC₂, plant 75-1, had 24 chromosomes. A 3 resistant: 1 susceptible segregation ratio, consistent with the action of a major, dominant gene, was observed in the BC₂F₂ and BC₂F₃ generations. Five of the BC₁ plants tested were resistant to bacterial blight. Ten of the 21 BC₂ progeny tested were resistant to Philippine races 2, 3, and 6 of the bacterial blight pathogen. One resistant BC₂, plant 78-1, had 24 chromosomes. The segregation of reactions of the BC₂F₂, BC₂F₃, and BC₂F₄ progenies of plant 78-1 suggested that the same or closely linked gene(s) conferred resistance to races 2, 3, 5, and 6 of the bacterial blight pathogen from the Philippines.     

Inheritance of dicrotophos resistance in greenhouse whitefly

We studied inheritance of resistance to dicrotophos in greenhouse whitefly,Trialeurodes vaporariorum Westwood (Homoptera, Aleyrodidae). Compared with females from a field-collected susceptible strains (S), females from a resistant strain (R) ofT. vaporariorum derived from heavily treated cotton fields had a 28-fold greater LC₅₀ to dicrotophos in laboratory bioassays. Concentration-mortality lines obtained from female progeny of reciprocal F₁ crosses (R_♀ XS_♂ and S_♀ XR_♂) were similar, suggesting that inheritance of dicrotophos resistance was autosomal and not influenced by maternal effects. Responses of F₁ female progeny were similar to those of the parental S strain, indicating that the resistance was partially recessive (degree of dominance, D, was −0.61). Mortality observed in female progeny obtained from a backcross (F_1♀ XR_♂) corresponded more closely to expectations derived from polygenic models than to expectations from a monogenic model. The estimated number of effective factors (sensu Lande, 1981) contributing equally to resistance was three.     

Combining transgenic and natural resistance to obtain broad resistance to tospovirus infection in tomato (Lycopersicon esculentum mill)

This study was undertaken to develop tomato plants with broad resistanceto tospoviruses which are a major limiting factor to tomato productionworldwide. A nontransgenic tomato line Stevens-Rodale (S-R), six transgenictomato lines expressing the nucleocapsid (N) protein gene of the lettuceisolate of tomato spotted wilt virus (TSWV-BL), and progeny of the crosses between S-Rand three of the transgenic lines homozygous for the N gene were evaluated fortheir resistance to tospovirus infection in greenhouse inoculation tests. S-Rhas the Sw-5 gene that confers resistance to several TSWVisolates. The six transgenic lines showed high levels of resistance wheninoculated with either TSWV-BL or a tomato isolate from Hawaii (TSWV-H).However, these same plants were highly susceptible to the Brazilian isolate ofgroundnut ringspot virus (GRSV-BR). Plants with the Sw-5gene were resistant to TSWV-BL and GRSV-BR, but were susceptible to TSWV-H.When inoculated with any of the three viruses, the F₁ progeny of thecrosses exhibited a susceptible, tolerant, or resistant phenotype with a higherproportion of the plants being either tolerant or resistant. When F₂progeny from F₁ resistant plants of each cross were inoculated withany of the three viruses, a higher proportion of tolerant and resistant plantswas observed compared to the F₁ progeny. Our results show thepotential to obtain broad resistance to tospoviruses by combining transgenicand natural resistance in a single plant.     

Inheritance and mechanism of resistance to herbicides inhibiting acetolactate synthase in Sonchus oleraceus L.

A biotype of Sonchus oleraceus L. (Compositae) has developed resistance to herbicides inhibiting acetolactate synthase (ALS) following field selection with chlorsulfuron for 8 consecutive years. The aim of this study was to determine the inheritance and mechanism of resistance in this biotype. Determination of ALS activity and inhibition kinetics revealed that K_m and V_max did not vary greatly between the resistant and susceptible biotypes. ALS extracted from the resistant biotype was resistant to five ALS-inhibiting herbicides in an in vitro assay. ALS activity from the resistant biotype was 14 19, 2, 3 and 3 times more resistant to inhibition by chlorsulfuron, sulfometuron, imazethapyr, imazapyr and flumetsulam, respectively, than the susceptible biotype. Hybrids between the resistant and a susceptible biotype were produced, and inheritance was followed through the F₁, F₂ and F₃ generations. F₁ hybrids displayed a uniform intermediate level of resistance between resistant and susceptible parents. Three distinct phenotypes, resistant, intermediate and susceptible, were identified in the F₂ generation following chlorsulfuron application. A segregation ratio of 121 was observed, indicative of the action of a single, nuclear, incompletely dominant gene. F₃ families, derived from intermediate F₂ individuals, segregated in a similar manner. Resistance to herbicides inhibiting ALS in this biotype of S. oleraceus is due to the effect of a single gene coding for a resistant form of the target enzyme, ALS.     

Decrease in tolerance to fenvalerate, in resistantHelicoverpa armigera after pupal diapause

Pyrethroid resistant and susceptible adults ofHelicoverpa armigera (Lepidoptera: Noctuidae) were screened for tolerance to pyrethroids after 6 wk or 12 wk pupal diapuse. Resistant larvae and F₂ larvae from a cross between resistant and susceptible parents (two replicates), were reared under conditions to induce pupal diapause. After eclosion, adults were tested in glass vials coated with the pyrethroid fenvalerate, at a dose (DD) that is known to discriminate between susceptible and heterozygous resistant individuals. In all diapause experiments, the frequency of resistance was lower in the test groups that had experienced diapause compared with the non-diapausing control group. The underlying cause of the decline is not certain but selective mortality of resistant versus susceptible individuals could not account for all the difference in two of the three experiments — tolerance to the pyrethroid, fenvalerate, is most likely to have declined either as a consequence of diapause or from the extended time of development associated with diapause. These results indicate that monitoring programs could underestimate pyrethroid resistance frequencies when usingH. armigera adults emerging from diapause.     

Mode of Inheritance of Amitraz Resistance in a Brazilian Strain of the Southern Cattle Tick, Boophilus microplus (Acari: Ixodidae)*

The southern cattle tick, Boophilus  microplus (Canestrini), has developed resistance to amitraz in several countries in recent years. A study was conducted at the USDA Cattle Fever Tick Research Laboratory in Texas to investigate the mode of inheritance of amitraz resistance with cross-mating experiments. The Muñoz strain, a laboratory reared acaricide-susceptible reference strain, was used as the susceptible parent and the Santa Luiza strain, originating in Brazil, was used as the resistant parent. A modified Food and Agriculture Organization Larval Packet Test was used to measure the levels of susceptibility of larvae of the parental strains, F₁, backcross, F₂, and F₃ generations. Results of reciprocal crossing experiments suggested that amitraz resistance was inherited as an incomplete recessive trait. There was a strong maternal effect on larval progeny’s susceptibility to amitraz in both the F₁ and the subsequent generations. The values of the degree of dominance were estimated at ?0.156 and ?0.500 for the F₁ larvae with resistant and susceptible female parents, respectively. Results of bioassays on larval progeny of the F₁ backcrossed with the resistant parent strain and that of the F₂ generations suggested that more than one gene was responsible for amitraz resistance in the Santa Luiza strain. Comparisons of biological parameters (engorged female weight, egg mass weight, and female-to-egg weight conversion efficiency index) indicated significant differences between different genotypes. The differences appeared to be heritable, but not related to amitraz resistance. Results from this study may have significant implications for the management of amitraz resistance.     

Inheritance of resistance to yellow mosaic virus in blackgram (Vigna mungo (L.) Hepper)

Summary The inheritance of resistance to mungbean yellow mosaic virus (MYMV) was studied in blackgram (Vigna mungo (L.) Hepper). The highly resistant donors Pant U-84 and UPU-2 and a highly susceptible line, UL-2, their F₁'s, F₂'s and backcrosses were grown with spreader located every 5 to 6 rows. The resistance was found to be digenic and recessive in all the crosses and free from cytoplasmic effect.     

Transgenic broccoli expressing aBacillus thuringiensis insecticidal crystal protein: Implications for pest resistance management strategies

We usedAgrobacterium tumefaciens to transform flowering stalk explants of five genotypes of broccoli with a construct containing the neomycin phosphotransferase gene and aBacillus thuringiensis (Bt) gene [CryIA(c) type] optimized for plant expression. Overall transformation efficiency was 6.4%; 181 kanamycin-resistant plants were recovered. Of the 162 kanamycin-resistant plants tested, 112 (69%) caused 100% morality of 1st-instar larvae of aBt-susceptible diamondback moth strain. Southern blots of some resistant transformants confirmed presence of theBt gene. Selected plants that gave 100% mortality of susceptible larvae allowed survival of a strain of diamondback moth that had evolved resistance toBt in the field. F₁ hybrids between resistant and susceptible insects did not survive. Analysis of progeny from 26 resistant transgenic lines showed 16 that gave segregation ratios consistent with a single T-DNA integration. Southern analysis was used to verify those plants possessing a single T-DNA integration. Because these transgenic plants kill susceptible larvae and F₁ larvae, but serve as a suitable host for resistant ones, they provide an excellent model for tests ofBt resistance management strategies.     

New RAPD markers of tomato spotted wilt virus (TSWV) resistance in Lycopersicon esculentum Mill

The selection of TSWV resistant individuals can be facilitated by molecular markers. RAPD analysis was carried out on three forms (Stevens × Rodade — resistant; Rey de los Tempranos — moderately tolerant; Potentat — susceptible) with the use of 271 primers. Out of 271 primers 28 generated stable polymorphism and so they were tested for linkage to resistance gene. Bulk segregant analysis (BSA) was applied to F₂ segregating progeny developed from resistant × susceptible parents. As a result, 5 primers enabling us to distinguish between resistant and susceptible forms were detected. Only one of them had previously been reported by Chague et al. (1996). The analysis should be repeated on a larger population to confirm the results obtained.     

Inheritance of Cry1Ac resistance and associated biological traits in the cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae)

The analysis of reciprocal genetic crosses between resistant Helicoverpa armigera strain (BH-R) (227.9-fold) with susceptible Vadodara (VA-S) strain showed dominance (h) of 0.65-0.89 and degree of dominance (D) of 0.299-0.782 suggesting Cry1Ac resistance as a semi-dominant trait. The D and h values of F₁ hybrids of female resistant parent were higher than female susceptible parent, showing maternally enhanced dominance of Cry1Ac resistance. The progeny of F₂ crosses, backcrosses of F₁ hybrid with resistant BH-R parent did not differ significantly in respect of mortality response with resistant parent except for backcross with female BH-R and male of F₁ (BH-R × VA-S) cross, suggesting dominant inheritance of Cry1Ac resistance. Evaluation of some biological attributes showed that larval and pupal periods of progenies of reciprocal F₁ crosses, backcrosses and F₂ crosses were either at par with resistant parent or lower than susceptible parent on treated diet (0.01 μg/g). The susceptible strain performed better in terms of pupation and adult formation than the resistant strain on untreated diet. In many backcrosses and F₂ crosses, Cry1Ac resistance favored emergence of more females than males on untreated diet. The normal larval period and the body weight (normal larval growth) were the dominant traits associated with susceptible strain as contrast to longer larval period and the lower body weight (slow growth) associated with resistance trait. Further, inheritance of larval period in F₂ and backcross progeny suggested existence of a major resistant gene or a set of tightly linked loci associated with Cry1Ac sensitivity.     

Inheritance of resistance to the two-spotted spider mite from L. pimpinellifolium (Jusl.) Mill. accession ‘TO-937’

The two-spotted spider mite (Tetranychus urticae Koch) is an important pest of tomato (Lycopersicon esculentum Mill.) crops in temperate regions as this spider mite has a very large capacity for population increase and causes severe tomato yield losses. There is no described tomato cultivar fully resistant to this pest, although resistant accessions have been reported within the green-fruited tomato wild species L. pennellii (Corr.) D’Arcy and L. hirsutum Humb. & Bonpl. We observed a L. pimpinellifolium (Jusl.) Mill. accession, ‘TO-937’, which seemed to be completely resistant to mite attacks and we crossed it with the susceptible L. esculentum cultivar. ‘Moneymaker’ to obtain a family of generations consisting of the two parents, the F₁, the F₂, the BC₁ to L. esculentum, and the BC₁ to L. pimpinellifolium. This family was evaluated for mite resistance in a polyethylene greenhouse using an experimental design in 60 small complete blocks distributed along 12 double rows. Each block consisted of five F₂ plants in one row and one plant of each of the two parents, the F₁, the BC₁ to L. esculentum, and the BC₁ to L. pimpinellifolium in the adjacent row. Plants at the 10–15 leaf stage were artificially infested by putting on them two pieces of French bean leaf heavily infested with T. urticae. After two months, evaluations of infestation were made by visual observation of mite nets and leaf damage. Plants that were free of signs of mite reproduction on the top half were considered as resistant, plants with silky nets only on their basal leaves, intermediate, and plants with mite reproduction on both basal and top canopies were scored as susceptible. Dominance for resistance appeared because all the ‘To-937’, BC₁ to L. pimpinellifolium, and F₁ plants were resistant. Not all ‘Moneymaker’ plants behaved as susceptible because 35% of plants were intermediate. In the BC₁ to L. pimpinellifolium and the F₂, most plants were scored as resistant, only 7 % BC₁ and 3 % F₂ plants were intermediate, and a single F₂ plant (0.3 %) was susceptible. With these figures, resistance seemed to be controlled by either four or two genes according to whether segregation in the BC₁ or in the F₂, respectively, were considered. These results could in part be explained because of appearance of negative interplot interference due to the high frequency of resistant genotypes within most of the generations. Therefore, the family was evaluated again but using a different experimental design. In the new experiment, 16 ‘TO-937’, 17 ‘Moneymaker’, 17 F₁, 37 BC₁ to L. pimpinellifolium, 38 BC₁ to L. esculentum, and 125 F₂ plants were included. Each of these test plants was grown besides a susceptible ‘Moneymaker’ auxilliary plant that served to keep mite population high and homogeneous in the greenhouse. Negative interplot interference was avoided with this design and all the ‘TO-937’, F₁, and BC₁ to L. pimpinellifolium plants were resistant, all ‘Moneymaker’ test plants were susceptible, and 52 % BC₁ to L. esculentum and 25 % F₂ plants were susceptible, which fitted very well with the expected for resistance governed by a single dominant gene. The simple inheritance mode found will favour sucessful introgression of mite resistance into commercial tomatoes from the very close relative L. pimpinellifolium.     

Genetic analysis of resistance to azinphosmethyl in the pear psylla Cacopsylla pyri

Laboratory selection with azinphosmethyl had little effect on the chemical resistance in Cacopsylla pyri strains, in comparison to the original wild populations. The resistance ratio relative to a susceptible strain varied from 10- to 40-fold depending on the generation studied. Crosses between two resistant strains and the susceptible strain show resistance to be autosomally inherited and semi-dominant in expression. Backcrosses between F1 and the susceptible strain were unable to distinguish unabigously between monogenic and polygenic inheritance. In the majority of experiments, however, the overall dose-response relationship for backcross progeny was consistent with a single gene hypothesis. Additional bioassays showed azinphosmethyl-resistant strain to cross-resistant monocrotophos and phosmet, but not carbamates, pyrethroids, amitraz or the organophosphates chlorpyrifos and mevinphos.     

Inheritance of resistance in maize silks to the corn earworm

The corn earworm,Helicoverpa zea (Boddie), is a perennial economic pest of field crops in the United States. Maize,Zea mays L., is the major host crop promoting the build-up of devastating corn earworm populations that limit full production of cotton, soybean, peanut, and grain sorghum. Resistance to the corn earworm in maize and in particular sweet maize, would provide an environmentally safe, economical method of control for this pest insect. Antibiotic effects of corn silks on this insect are: small larvae, extended developmental period, and reduced fecundity. Silks from individual maize plants of resistant and susceptible lines and progeny in six generations consisting of parents (P₁, P₂), F₁, F₂, and backcrosses BC_1.1 (F₁ × P₁) and BC_1.2 (F₁ × P₂) from each of four crosses were used to determine the genetic basis of the antibiotic resistance of silks to the corn earworm. In the cross of Zapalote Chico × PI340856, genes controlling resistance in the silks to the corn earworm larvae are dominant in PI340856 to those in Zapalote Chico. The cross of Zapalote Chico × GT114 involves parents differing in degree of resistance, and possibly differing for the genetic mechanism by which the resistance is inherited. The inheritance of resistance may involve non-additive (dominance and epistasis) genetic variance. A digenic 6-parameter model indicated (1) the resistance in this cross is controlled by more than one pair of genes and (2) some or all of the genes interact to cause non-allelic interaction. Thus, the resistance in this cross may be controlled by both dominant and recessive genes. The resistance of Zapalote Chico × CI64, an intermediate inbred, is influenced by additive gene effects. The digenic model adequately predicts all generation means of the cross of GT3 × PI340856 except for the F1. Thus, it appears that the additive-dominance model is not satisfactory for this cross involving susceptible and resistant parents. Generation mean analysis indicates that resistance to silk-feeding by corn earworm larvae is under genetic control, but gene action differs from one type of cross to another.     

Inheritance of powdery mildew (Erysiphe polygoni DC) resistance in mungbean (Vigna radiata L. Wilczek)

Detached mungbean (Vigna radiata L.Wilczek) leaves were inoculated with a conidial suspension of a local isolate (TI-1) of the powdery mildew pathogen (Erysiphe polygoni DC) under controlled environment conditions. Based on the latent period and severity of the infection, a rating scale of 0–5 was used to classify the host pathogen interactions. Reactions 0, 1 and 2 were considered resistant and referred to as R0, R1 and R2 while 3, 4 and 5 were classified as susceptible (S). RUM lines (resistant to powdery mildew) and their derivatives are crossed with several susceptible (reaction types 3–5) genotypes and the inheritance of the resistance was studied in the F₁, F₂ and F₃ generations. The results showed that powdery mildew resistance in mungbean is governed by two dominant genes designated as Pm-1 and Pm-2. When both Pm-1 and Pm-2 were present, an R0 reaction was observed after inoculation with TI-1. The resistant reaction was R1 when only Pm-1 was present and R2 in the presence of Pm-2. In the absence of both Pm-1 and Pm-2, susceptible reactions 3, 4 and 5 were observed.     

Biochemical genetics of powdery mildew resistance in pea

Summary A biochemical study on phenolic (total phenols and orthodihydroxy phenols) content and on the activities of phenol oxidizing enzymes (peroxidase and polyphenol oxidase) in pea cultivars resistant and susceptible to powdery mildew infection revealed that the resistant cultivars contained higher levels of phenolics and phenol-oxidizing enzymes than the susceptible ones. A further study of their F₁s, F₂s and backcross progenies suggested a high heritability for all biochemical traits. The correlation coefficients between the biochemical parameters and the disease index were also high. Both additive (d) and dominant () components were found to contribute to the inheritance of these constituents.Associate Professor (Genetics), Department of Basic Sciences     

Dominant pleiotropy controls enzymes co-segregating with paraquat resistance in Conyza bonariensis

Summary The genetics of paraquat-resistance in Conyza bonariensis was studied. Reciprocal crosses were prepared between resistant and sensitive individuals. The enzymes of the pathway that detoxifies superoxide to innocuous oxygen species involved in resistance were evaluated in the F₁ and F₂ generations. All F₁ plants were as resistant as the resistant parent, irrespective of parental sex, demonstrating dominance and excluding maternal inheritance. The activities of superoxide-dismutase, ascorbate-peroxidase and glutathione-reductase in the F₁ were constitutively as high as in the resistant parent. Resistance in the F₂ generation was distributed in a 31 ratio (resistant to sensitive). Leaves from F₂ plants were removed for a resistance assay and enzyme immuno-assays of single plants were performed. The high levels of superoxide-dismutase and glutathione-reductase, the two enzymes for which antibodies were available, were similar in resistant individuals to the levels in the resistant parent; the levels were low in the susceptible individuals. These results indicate either a very tight linkage, or more probably, that one dominant nuclear gene controls resistance by pleiotropically controlling the levels of enzymes of the activeoxygen detoxification pathway.     

Inheritance of heat-stable resistance to Meloidogyne incognita in Lycopersicon peruvianum and its relationship to the Mi gene

Summary The inheritance of heat-stable resistance to the root-knot nematode, Meloidogyne incognita (Kofoid and White) Chitwood, was studied in crosses between different accessions and clones of Lycopersicon peruvianum L. F₁, F₂ and BC₁ generations were evaluated for their index of resistance based on numbers of eggs and infective second-stage juveniles (J₂) per gram of root, and the segregation ratios were determined in experiments carried out at constant soil temperatures of 25 °C and 30 °C. L. peruvianum P.I. 270435 clones 3 MH and 2R2 and P.I. 126443 clone 1 MH, all heatstable resistant, were crossed with L. peruvianum P.I. 126440 clone 9 MH, which is susceptible at both 25 °C and 30 °C. All F₁ progeny were resistant at 25 °C and 30 °C; F₂ and BC₁ generations at 25 °C gave resistant: susceptible (RS) ratios of 151 and 31, respectively, which suggests that resistance is conditioned by two independently assorting genes. However, at 30 °C, RS ratios of 31 and 11 were observed for the F₂ and BC₁ generations, respectively. These results indicate that heat-stable resistance is conferred by a single dominant gene expressed at 30 °C, while the second resistance gene is heat unstable and not expressed at 30 °C. P.I. 270435 clones 2R2 and 3 MH and P.I. 126443 clone 1 MH were crossed with P.I. 128657 clone 3 R4 (source of gene Mi), which is resistant at 25 °C but susceptible at 30 °C. All of the F₁ progeny were resistant at 25 °C and 30 °C.TC₁ progeny of 270435-2 R2 x 128657-3 R4, 270435-3 MH x 128657-3 R4 and 126443-1 MH x 128657-3 R4 crossed with susceptible 126440-9 MH were all resistant at 25 °C and segregated in a 11 ratio at 30 °C. These results also suggest that the heat-stable resistance is monogenic and that it is non-allelic to gene Mi. The non-segregation of TC₁ progenies at 25 °C, suggests that the heat-unstable resistance factor in L. peruvianum P.I. 270435 clones 2 R2 and 3 MH and in P.I. 126443 clone 1 MH is allelic to or the same as gene Mi. We propose the symbol Mi-2 for the gene in P.I. 270435 that confers heat-stable resistance to M. incognita.     

Paternal plastid DNA can be inherited in lentil

Restriction fragment analysis was used to study the inheritance of chloroplast DNA (cpDNA) in F₁ progeny from crosses between Lens culinaris ssp. orientalis and L. culinaris ssp. culinaris. Twenty-five combinations of 11 restriction enzymes and three heterologous probes from Petunia hybrida cpDNA were used to screen six accessions of L.c. culinaris and one accession of L. c. orientalis for restriction fragment length polymorphisms (RFLPs). No variation in cpDNA was observed within the subspecies L. c. culinaris, but the L. c. orientalis accession was unambiguously distinguished from all six L. c. culinaris accessions by two RFLPs. Of ten F₁ progeny from L. c. orientalis x L. c. culinaris crosses, nine had only maternal cpDNA restriction fragments but one F₁ plant inherited cpDNA fragments from both parents. Nuclear DNA inheritance was biparental in all ten F₁ progeny.     

Resistance to powdery mildew (Oidium lycopersicum) in Lycopersicon hirsutum is controlled by an incompletely-dominant gene Ol-1 on chromosome 6

The inheritance of resistance to powdery mildew (Oidium lycopersicum) in Lycopersicon hirsutum was investigated by disease tests in segregating populations obtained by hybridising tomato (L. esculentum) cv Moneymaker with the wild relative L. hirsutum G1.1560. One incompletely dominant gene Ol-1 was found to largely control resistance to the disease. To map Ol-1, DNA pools from seven resistant and ten susceptible F₂ plants were analyzed for random amplified polymorphic DNA (RAPD). With 32 primers tested, one RAPD, primed with the sequence 5-GACGTGGTGA-3, was observed between the susceptible and the resistant bulks, which cosegregated with resistance in the F₂ population of L. esculentum × L. hirsutum G1.1560. This RAPD was mapped on chromosome 6 by using an F₂ (L. esculentum × L. pennellii) already mapped for 49 RFLPs. RFLP analysis of the F₂ from L. esculentum cv Moneymaker × L. hirsutum G1.1560 demonstrated that Ol-1 maps near the Aps-1 region on chromosome 6, in the vicinity of the resistance genes to Meloidogyne spp. (Mi) and to Cladosporium fulvum (Cf-2/Cf-5).     

Association of AFLP markers with downy mildew resistance in autotetraploid alfalfa

The amplified fragment length polymorphism (AFLP) assay is an efficient method for the identification of molecular markers useful in the improvement of numerous crop species. The identification of AFLP markers linked to disease resistance genes has been shown in segregating populations from crosses of inbred lines. The development of inbred lines in alfalfa is not possible, but existing breeding programs have produced populations selected for resistance to a single pest. Two such populations, UC-123 and UC-143, differing only in selection for resistance to downy mildew (Peronospora trifoliorum de Bary) isolate I-8, were used in this study. Thirty-six resistant plants from UC-143, and 36 susceptible plants from UC-123 were screened for DNA polymorphisms using fourteen AFLP primer combinations. Four AFLP fragment markers, ACACTC₂₀₈, ACACTC₁₅₀, ACACAT₂₁₆ and ACACTC₄₈₆, were found to be significantly associated with disease susceptibility or resistance. Resistant and susceptible plants were crossed in a diallel scheme and the progeny were screened for resistance to P. trifoliorum isolate I8. Two of the AFLP markers, ACACTC₂₀₈ and ACACTC₄₈₆ were significantly associated with resistance in the F₁ and S₁ progeny. The utilization of two populations, comprised of 36 resistant and 36 susceptible plants, for the identification of DNA fragments associated with disease resistance proved successful. Seventy-two plants is a very manageable number and provides a starting point for further refinement of marker-trait associations.