Molecular evidence of biased inheritance of trifluralin herbicide resistance in foxtail millet

Inheritance of trifluralin herbicide resistance was investigated for phenotype and genotype (molecular identification) in 12 segregating populations derived from crosses between resistant green foxtail (Setaria viridis) and foxtail millet cultivars (S. italica). Combining a herbicide bioassay and a bidirectional allele‐specific polymerase chain reaction (PCR) assay, the trifluralin resistance in foxtail millet was proved to be conferred by a recessive α2‐tubulin gene mutation at Ile‐239, the allele transferred by green foxtail. However, a distorted segregation ratio in F₂ populations was confirmed, with 16.9% resistant plants on average instead of the 25% expected for a single gene recessive mutation. This was not due to non‐germinating seed and the distortion was also observed in the next generations, indicating a heritable phenomenon. After further crosses and self‐fertilization, however, more advanced generations did not show the same pattern of skewed inheritance as in the early generations. These results suggest the possibility of linkage between the α2‐tubulin gene and a modifier gene to explain the distorted segregation, which might be broken by crossing over.     

An Inheritance Study of Domestication in Foxtail Millet Using an Interspecific Cross

In order better to understand domestication processes, the inheritance of eight morphological characters and isozymes was studied in the F₂ progeny of an interspecific cross between a wild and 2 cultivated foxtail millet (Setaria viridis and S. italica). Two linkage groups were determined, one involving seed shedding and the phenol colour reaction of the grain. These two characters, as well as the pericarp pigmentation of the seed, appear to he highly specific for the difference between collections of wild and cultivated plants and probably indicate a genotype particularly suitable for plant domestication.     

Inheritance of sethoxydim resistance in foxtail millet, Setaria italica (L.) Beauv.

Interspecific hybridization between foxtail millet cultivars (Setaria italica) and a green foxtail (S. viridis) resistant to the herbicide sethoxydim were undertaken to breed foxtail millet for improved herbicide resistance. Parents, reciprocal F₁ hybrids, F₂ selfed derived populations and BC₁ backcross progeny were produced and analysed for mortality and fresh weight over a range of dosages. All resistant progeny were 700 times more resistant than susceptible cultivars and was symptom free under current field dosages. Segregations of resistant and susceptible progeny (3:1 in F₂ and 1:1 in BC₁) were not dependent upon dosage. Heterozygous individuals displayed the same magnitude of resistance as homozygous plants at twelve times the recommended field dosage. Results suggested that sethoxydim resistance in foxtail millet was controlled by a single, completely dominant, nuclear gene. This revised version was published online in July 2006 with corrections to the Cover Date.     

Breeding strategy in foxtail millet,Setaria italica (L.P. Beauv.), following interspecific hybridization

Summary An interspecific cross between foxtail millet Setaria italica and its wild relative Setaria viridis was undertaken in order to introduce from the latter triazine resistance in the cultivated plant. Four backcross generations obtained with S. italica as recurrent parent were studied. Results show that only two backcross generations associated with selection are enough to eliminate weedy characters and to return to the cultivated type. S. viridis could thus be an interesting source for improvement of foxtail millet without a complex and time consuming breeding strategy.     

Inheritance of the Powdery Mildew Resistance Gene Pm9 in Relation to Pm1 and Pm2 of Wheat

The inheritance of the powdery mildew resistance gene Pm9 originating from the hexaploid spring wheat cultivar ‘Normandie’ was analyzed in relation to Pm1 and Pm2. Two leaf segments of individual P_1?, P_2?, F_1? and F₂-plants of the cross ‘Normandie’ (Pm1, 2, 9) בFederation’ (no known Pm gene) were inoculated separately with two powdery mildew isolates. Using powdery mildew isolate No. 6 virulent for Pm1 and Pm2 but avirulent for Pm9, a 1 resistant (r): 3 susceptible (s) F₂-segregation was found for the Pm9 gene. Using powdery mildew isolate No. 3 virulent for Pm1 and Pm9 but avirulent for Pm2, a 3 (r): 1 (s) F₂-segregation was found for the Pm2 gene. Combining the data of both experiments (leaf segments of identical plants had been used), a 9 (sr): 3 (ss): 3 (rr): 1 (rs) segregation resulted for the F₂ of this cross: therefore, independent inheritance of the genes Pm2 and Pm9 can be concluded. Similarly, the cross ‘Mephisto’ (Pm1, 2, 9) בAmor’ (no known Pm gene) was analyzed. The Pm9 gene again showed a monogenically recessive inheritance, whereas Pm1 showed a monogenically intermediate segregation upon inoculation with powdery mildew isolate No. 9a virulent for Pm2 and Pm9 but avirulent for Pm1. Combining the single gene segregations, linkage between both genes was found among the progenies. A distance of 8.5 cM was calculated. Analyzing a set of spring wheat cultivars with seven defined powdery mildew isolates, the presence of Pm1, Pm2 and Pm9 in these lines was verified; in most cases, Pm1 occurred together with Pm9.     

Development and utilization of novel SSRs in foxtail millet [Setaria italica (L.) P. Beauv.]

Although the foxtail millet [Setaria italica (L.) P. Beauv.] is recently regarded as a model crop for studying functional genomics of biofuel grasses, its genetic improvement to some extent was limited due to the non‐availability of molecular markers, particularly the microsatellite markers and the saturated genetic linkage map. Considering this, we attempted to generate a significant number of microsatellite markers in cultivar ‘Prasad’. Two hundred and fifty‐six clones were sequenced to generate 41.82‐kb high‐quality sequences retrieved from genomic library enriched with dinucleotide repeat motifs. Microsatellites were identified in 194 (76%) of the 256 positive clones, and 64 primer pairs (pp) were successfully designed from 95 (49%) unique SSR‐containing clones. The 67.4% primer designing ability, 100% PCR amplification efficiency and 45.3% polymorphic potential in the parents of F₂ mapping population established the efficacy of genomic microsatellites. All the 64 microsatellite markers displayed high level of cross‐species amplification (~67%) in 10 millets and non‐millets species. These experimental findings suggest the utility and efficacy of SSRs in diverse genotyping applications, resolving QTLs, phylogenetic relationships and transferability in several important grass species.     

Inheritance of internal pigmentation in green cabbage (Brassica oleracea var. Capitata L.)

Summary Internal anthocyanin pigmentation (IP) in otherwise normally green cabbage occurs in a number of Oregon State University breeding lines. Extracted pigment, tested for spectral absorption and for color reactions with lead acetate and aluminium chloride, was similar but not necessarily identical to pigment extracted from red cabbage cultivar Redman. When IP line R52 was crossed with normal green line C70, the F₁, F₂ and backcross progenies indicated that IP at the intensity found in R52 was determined by a single factor in homozygous condition, with intermediate levels of IP expressed by the heterozygous genotypes. Modifying factors also appear to influence the level of IP. In the cross R52 (IP)×R51 (normal green), expression of IP in the F₁ was much reduced. The F₂ failed to fit the expected 3 IP: 1 green ratio due to an excess of green plants, but instead, closely fit a 9:7 ratio. This may have resulted from incomplete expression of IP because of modifiers, rather than from the effects of a second major gene. An allele at the A (anthocyanin) locus of B. oleracea is tentatively proposed and designated A ^IP or a ^IP pending further identification.Oregon Agricultural Experiment Station Technical Paper No. 4690.     

Inheritance of resistance to anthracnose caused by Colletotrichum capsici in Capsicum

Inheritance of resistance to anthracnose caused by Colletotrichum capsici (Syd.) Butler & Bisby was studied in interspecific Capsicum populations derived from a cross between a Thai elite cultivar Capsicum annuum L.‘Bangchang’ and a resistant line C. chinense Jacq.‘PBC932′. The resistance was assessed by measuring lesion area per fruit area (LFA) on detached chili fruits, using a laboratory‐based injection inoculation. Nil symptoms resembling the resistant parent ‘PBC932’ were also identified in the progeny F₂ and BC1 populations. Segregation of resistance (nil LFA) and susceptibility in the F₂ fitted a 1: 3 Mendelian ratio, indicating that resistance was responsible by a single recessive gene. The segregation of the trait in the testcrosses in both BC1s also confirmed the 1: 3 gene segregating model as found in the F₂.     

Inheritance of resistance to anthracnose (Colletotrichum capsici) at seedling and fruiting stages in chili pepper (Capsicum spp.)

Inheritance of resistance to anthracnose at fruiting and seedling stages was studied in two F₂ populations from a cross between Capsicum annuum cv. ‘Bangchang’ and Capsicum chinense‘PBC932’. The first F₂ was used to study anthracnose resistance at fruiting stage on mature green and ripe red fruit, and the second F₂ was used to study the resistance at both seedling and fruiting stages. Fruit inoculation was performed on detached fruit using a microinjector. Disease severity was assessed on a 0–9 scale at 7 days after inoculation. Seedling inoculation was performed using a drop method on detached leaves of 4‐week‐old seedlings. Disease severity was assessed on a 1–9 scale at 3 days after inoculation. The distribution of the disease scores of green and red fruit, and seedlings in both F₂ populations suggested a single gene model for each trait. Three different recessive genes were responsible for the three resistances from this cross. Linkage analysis suggested that the resistances at green and red fruit were linked (recombination frequency 0.25), and that the seedling resistance was not linked to the fruit resistances.     

Inheritance of black testa colour in lentil (Lens culinaris Medik.)

The inheritance of testa (seed coat) colour and interaction of cotyledon and testa colours were studied in seven crosses of lentil (Lens culinaris Medik.) involving parents with black, brown, tan or green testa and with orange, yellow or dark green cotyledons. Analysis of F₂ and F₃ seed harvested from F₁ and F₂ plants, respectively, revealed that although black testa is dominant over nonblack testa, its penetrance is not complete since both F₁ plants and heterozygous F₂ plants produced varying proportions of seeds with either black or nonblack testa. The F₂ populations of the crosses between parents with brown and tan, as well as brown and green, testa segregated in the ratio of 3 brown : 1 tan and 3 brown : 1 green, respectively, indicating monogenic dominance of brown testa colour over tan or green. The expression of testa colour was influenced by cotyledon colour when parents with brown or green testa are crossed with those having orange or green cotyledons. Thus F₂ seeds from these crosses with a green testa always had green cotyledons and never orange cotyledons. F₂ seeds from these crosses with a brown testa always had orange cotyledons and never green cotyledons. These results suggest diffusion of a soluble pigment from the cotyledons to the testa. This revised version was published online in July 2006 with corrections to the Cover Date.     

Inheritance of black leaf mold resistance in tomato

Summary Inheritance of black leaf mold (BLM) (caused by Pseudocercospora fuligena) resistance was studied in four crosses involving two resistant Lycopersicon accessions (PI134417, L. hirsutum and PI254655, L. esculentum) and four susceptible Asian Vegetable Research and Development Center tomato lines (CLN657BC₁F₂-267-0-3-12-7, CL143-0-10-3-0-1-10, CLN698BC₁F₂-358-4-13 and CL5915-93D₄-1-0-3). For each cross, six generations, i.e. P₁, P₂, F₁, F₂, BC₁F₁ and BC₁F₂ were evaluated following inoculations with isolate Pf-2 of P. fuligena. Chi-square analyses of the data based on the ratio of resistant to susceptible plants in the F₂ in three of four crosses gave a good fit to a segregation ratio of 1 R : 15 S, and BC₁F₂ data in three of four crosses gave an acceptable fit to the segregation ratio of 1 R : 63 S. The results indicate that resistance to BLM may be conditioned by two recessive genes acting epistatically in both PI134417 and PI254655.     

Crossability,fertility and cytogenetic studies in Solanum acaule × Solanum bulbocastanum

From 538 cross combinations made between 20 accessions of S. acaule and 28 clones of S. bulbocastanum 150 different F₁'s were obtained. Average berry set was 11%, average number of seeds per berry 0.7. The accessions of both parent species could be divided into distinct groups on the basis of the crossability. Of the 72 F₁-clones examined 59 were triploid and 13 were tetraploid. Evidence is presented that the 4x-F₁'s originated from unreduced gametes produced by two bulbocastanum accessions. Pollen stainability of the 6x-, 8x-, 4x- and 3x-F₁'s was 85, 29, 12 and <5% respectively. The genomic constitutions of the F₁'s adequately accounted for these fertility relations. Selfing of F₁'s was successful with 6x-F₁'s only. Crosses between the F₁'s and S. tuberosum, tuberosum-haploids and S. demissum did not succeed in spite of the large number of pollinations made. A breeding programme is recommended to be carried out within the hexaploid hybrid populations which aims at late blight resistance and crossability with S. tuberosum. The quadruple cross (acl × blb) × (acl × tbr) and the direct cross between S. tuberosum and S. bulbocastanum are discussed.     

Allelic relationships of genes controlling number of flowers per axis in chickpea

Genetic variation for number of flowers per axis in chickpea (Cicer arietinum L.) includes single-flower, double-flower, triple-flower and multi-flower traits. A double-flowered (DF) line ICC 4929, a triple-flowered (TF) line IPC 99-18 and a multi-flowered (MF) line JGM 7 were intercrossed in all possible combinations and flowering behavior of parents, F₁s and F₂s was studied to establish allelic relationships, penetrance and expressivity of genes controlling number of flowers per axis in chickpea. The F₁ from ICC 4929 (DF) × IPC 99-18 (TF) cross were double-flowered, whereas F₁ from ICC 4929 (DF) × JGM 7 (MF) and IPC 99-18 (TF) × JGM 7 (MF) crosses were single-flowered. The F₂ from ICC 4929 (DF) × IPC 99-18 (TF) cross gave a good fit to a 3:1 ratio for double-flowered and triple-flowered plants. The F₂ from ICC 4929 (DF) × JGM 7 (MF) cross segregated in a ratio of 9:3:3:1 for single-flowered, double-flowered, multi-flowered and double-multi-flowered plants. The F₂ from IPC 99-18 (TF) × JGM 7 (MF) cross segregated in a ratio of 9:3:4 for single-flowered, triple-flowered and multi-flowered plants. The results clearly established that two loci control number of flowers per axis in chickpea. The double-flower and triple-flower traits are controlled by a single-locus (Sfl) and the allele for double-flowered trait (sfl ^d ) is dominant over the allele for triple-flower trait (sfl ^t ). The three alleles at the Sfl locus has the dominance relationship Sfl > sfl ^d > sfl ^t . The multi-flower trait is controlled by a different gene (cym). Single-flowered plants have dominant alleles at both the loci (Sfl_ Cym_). The double-flower, the triple-flower and the multi-flower traits showed complete penetrance, but variable expressivity. The expressivity was 96.3% for double-flower and 76.4% for double-pod in ICC 4929, 81.2% for triple-flower and 0.0% for triple-pod in IPC 99-18, and 51.3% for multi-flower and 24.7% for multi-pod in JGM 7. Average number of flowers per axis and average number of pods per axis were higher in JGM 7 than double-flowered line ICC 4929 and triple-flowered line IPC 99-18. The results of this study will help in development of breeding strategies for exploitation of these flowering and podding traits in chickpea improvement.     

Results in the Development of Alloplasmic Carrots (Daucus carota sativus Hoffm.)

Alloplasmic forms of the cultivated carrot, Daucus carota sativus Hoffm., were created by substitution backcrosses. The wild Daucus forms D. muricatus L., D. gingidium L., D. c. gummifer Hook, and D. c. libanotifolia Wiinst. were used as cytoplasm donors. In the first two backcross generations disturbances in fertility and plant physiology did not occur. The plants of the generations BC₂F₂ and BC₂F₃ look like cultivated carrots. The alloplasmic forms were compared with cultivars for economic traits such as yield and yield components. The high content of total sugar in the alloplasmic forms was considerable. A new type of male sterility was found and described in a BC₃F₂ generation of the cross D. c. gummifer× D. c. sativus. Results of RFLP-analyses of mt-DNA and cp-DNA were discussed.     

Fusarium vascular wilt in lentil: Inheritance and identification of DNA markers for resistance

The inheritance of resistance to lentil (Lens culinaris Medik.) vascular wilt caused by Fusarium oxysporum f.sp. lentis was investigated in a cross between resistant (ILL5588) and susceptible (L692–16-l(s)) lines. F_2:4 progenies and F_6:8, F_6:9 recombinant inbred line (RIL) populations were assessed for their wilt reaction for three seasons in a well-established wilt-sick plot. Resistance to wilt was conditioned by a single dominant gene in the populations studied. The map location of the Fw locus was identified for the first time through linkage to a random amplified polymorphic DNA (RAPD) marker (OPK-15₉₀₀) at 10.8 cM. Two other RAPD markers (OP-BH₈₀₀ and OP-DI5₅₀₀) identified by bulked segregant analysis were associated in the coupling phase with the resistance trait, and another marker (OP-C04₆₅₀) was associated with repulsion. The DNA markers reported here will provide a starting point in marker-assisted selection for vascular wilt resistance in lentil.     

Linkage relationships of some genes for disease and insect resistance and semidwarf stature in rice

Summary Two-way classification of 400 F₃ families from the rice cross IR2153-159-1 x Babawee for plant stature and for resistance to brown planthopper, green leafhopper, and bacterial blight indicated that Glh 3 (dominant gene for resistance to green leafhopper) and bph 4 (recessive gene for resistance to brown planthopper) are linked with a map distance of 34 units. The bph 4 gene also appears to be linked with sd ₁ (recessive gene for semidwarf stature) although the linkage is less strong. However, bph 4 and Xa 4 (dominant gene for bacterial blight resistance) are inherited independently of each other. No segregation for susceptibility was observed among F₃ families of crosses between varieties having Bph 3 and bph 4 genes for resistance to brown planthopper. Apparently, Bph 3 and bph 4 are either allelic or closely linked.     

The inheritance of days to flowering in broccoli (Brassica oleracea var. italica)

Summary Crosses between an early flowering inbred broccoli (Brassica oleracea L. var. italica) HS140 and four later maturing inbred lines, S301, S310, s318, and S258, were studied to determine the inheritance of earliness as expressed by days to first open flower. Mean days to first flower for F₁ and F₂ were almost identical, and were close to the mean of the two parents in three crosses, indicating additive inheritance. In the cross HS140×S258, for which no F₁ plants were available, the F₂ mean was closer to the mean of the early parent, but this was likely due to a distortion of the data caused by the very late maturation of S258, the latest maturing parent in the study. Frequency distribution for parents and progenies supported the conclusions from parent and progeny mean values and indicated that days to maturity is a quantitative character, with mostly additive inheritance.Oregon Agricultural Experiment Station Technical Paper No. 7578.     

Characterization of segregation distortion on chromosome 3 induced in wide hybridization between indica and japonica type rice varieties

We previously surveyed chromosomal regions showing segregation distortion of RFLP markers in the F₂ population from the cross between a japonica type variety ‘Nipponbare’ and an indica type variety ‘Milyang23’, and showed that the most skewed segregation appeared on the short arm of chromosome 3. By comparison with the marker loci where distortion factors were previously identified, this region was assumed to be a gametophytic selection-2 (ga2) gene region. To evaluate this region, two near isogenic lines (NILs) were developed. One NIL had the ‘Nipponbare’ segment of this region on the genetic background of ‘Milyang23’ (NIL9-23), and the other NIL had the ‘Milyang23’ segment on the genetic background of ‘Nipponbare’ (NIL33-18). NIL9-23 and ‘Milyang23’, NIL33-18 and ‘Nipponbare’, and ‘Nipponbare’ and ‘Milyang23’ were respectively crossed to produce F₁ and F₂ populations. The F₁ plants of NIL9-23 × ‘Milyang23’ and NIL33-18 × ‘Nipponbare’ showed high seed fertility and the same pollen fertility as their parental cultivars, indicating that ga2 does not reduce seed and pollen fertility. Segregation ratio of a molecular marker on the ga2 region in the three F₂ populations was investigated to clarify whether segregation distortion occurred on the different genetic backgrounds. Segregation distortion of the ga2 region appeared in the both F₂ populations from the NIL9-23 and ‘Milyang23’ cross (background was ‘Milyang23’ homozygote) and the ‘Nipponbare’ and ‘Milyang23’ cross (background was heterozygote), but did notin the F₂ population from the NIL33-18 and ‘Nipponbare’ cross (background was ‘Nipponbare’ homozygote). This result indicates that ga2 interacts with a ‘Milyang23’ allele(s) on the different chromosomal region(s) to cause skewed segregation of the ga2 region. In addition, segregation ratio was the same between the F₂ populations from NIL9-23 × ‘Milyang23’ and ‘Nipponbare’ × ‘Milyang23’ crosses, suggesting that the both genotypes, ‘Milyang23’ homozygote and heterozygote, of gene(s) located on the different chromosomal region(s) have the same effect on the segregation distortion. This revised version was published online in July 2006 with corrections to the Cover Date.     

Apium graveolens PI 181714 is a source of resistance to Fusarium oxysporum f. sp. apii race 4 in celery (A. graveolens var. dulce)

Celery has little genetic diversity and is highly susceptible to the new fungal pathogen Fusarium oxysporum f. sp. apii (Foa) race 4. After screening an Apium graveolens germplasm collection for resistance to Foa race 4, we crossed celery cv. 'Challenger', which is Foa race 2-resistant but Foa race 4-susceptible and A. graveolens PI 181714, which is Foa races 2- and 4-resistant but non-celery type. After selfing F₁s, we screened the F₁S₁ for race 4-resistance and celery-type and then selfed selected F₁S₁. Greenhouse and field trials indicate that three selected F₁S₂ families (76–8-4, 76–8-27 and 76–8-36) are suitable as germplasm for celery breeders for resistance to Foa race 4. A F1S3 76–8–36-124 is either fixed or nearly so for resistance to Foa races 4 and 2. Furthermore, quantitative PCR indicates that PI 181714 is resistant, rather than tolerant, to Foa races 4 and 2, and that this resistance has been introgressed into F1S3 76–8–36-124.     

Inheritance of resistance to broomrape (Orobanche cumana Wallr.) race F in a sunflower line derived from wild sunflower species

Genetic resistance to broomrape (Orobanche cumana Wallr.) race F in sunflower line J1, derived from the wild perennial species Helianthusgrosseserratus Martens and Helianthus divaricatus L., has been reported to be controlled by dominant alleles at a single locus, Or6. However, deviations from this monogenic inheritance have been observed. The objective of the present study was to gain insight into the inheritance of resistance to broomrape race F in the sunflower line J1. F₁, F₂, F₃ and BC generations from crosses between J1 and three susceptible lines, P21, NR5 and HA821 were evaluated. F₁ hybrids showed both resistant (R) and moderately resistant (MR) plants, the latter having a maximum of five broomrape stalks per plant compared with >10 in the susceptible parents. This indicated incomplete dominance of the Or6 alleles. F₂ plants were classified as R, MR or susceptible (more than five broomrape stalks per plant). Three different segregation ratios were observed: 3 : 1, 13 : 3 and 15 : 1 (R + MR : S), suggesting the presence of a second gene, Or7, whose expression was influenced by the environment. A digenic model was confirmed, based on the evaluation of F_2:3 families.