Molecular mapping of genic male-sterile genes ms15, ms5 and ms6 in tetraploid cotton

Two genic male sterile (GMS) lines, Lang-A conditioned by ms ₁₅ and Zhongkang-A conditioned by ms ₅ ms ₆ duplicate recessive genes in Gossypium hirsutum L., were chosen to map GMS genes. These two lines were crossed with Gossypium barbadense cv. 'Hai7124' to produce segregating populations. The ms ₁₅ gene was mapped on chromosome 12, and was flanked by two simple sequence repeat (SSR) markers, NAU2176 and NAU1278, with a genetic distance of 0.8 and 1.9 cM respectively. The ms ₅ and ms ₆ genes were mapped to one pair of homoeologous chromosomes, ms ₅ on chromosome 12 flanked by three SSR markers, NAU3561, NAU2176 and NAU2096, with genetic distances of 1.4, 1.8 and 1.8 cM, respectively, and ms ₆ on chromosome 26 flanked by two SSR markers, BNL1227 and NAU460, with a genetic distance of 1.4 and 1.7 cM respectively. These tightly linked markers with the ms ₁₅ , ms ₅ and ms ₆ genes can be used in the marker-assisted selection among segregating populations in a breeding programme, and provide the foundation for gene isolation by map-based cloning for these three genes.     

A CAPS marker TAO1902 diagnostic for the I-2 gene conferring resistance to Fusarium oxysporum f. sp. lycopersici race 2 in tomato

Fusarium oxysporum f. sp. lycopersici inhabits most tomato-growing regions worldwide, causing tomato production yield losses. A molecular marker linked to resistance would be useful for tomato improvement programmes. Thus, a cleaved amplified polymorphic sequence (CAPS) marker TAO1₉₀₂ was developed to identify tomato genotypes possessing the I-2 gene, which confers resistance to F. o. lycopersici race 2. The Rsa I or Fok I restriction fragments corresponded to the presence or absence of the I-2 allele in a segregating 100 F₂ progeny, tomato cultivars, 16 resistant and 20 susceptible to Fusarium wilt, respectively, lines and F₁ hybrids, representing various tomato gene pools. TAO1₉₀₂ may be helpful for selection of F. o. lycopersici -resistant tomato germplasm.     

Relationship of Root and Shoot Characters in Parent, F1 and F2 Populations of Rice

The direct and indirect contributions of root characters — root length, roots/plant, fresh and dry root weight on grain yield/plant were worked out from a 7 × 7 diallel set of rice hybrids. The materials were grown in pots with four replications. Path analysis was done at genotypic level of correlation.
The grain yield/plant showed positive correlation with all the root characters in parent, F₁ and F₂ except with roots/plant in F₁ population. Fresh root weight demonstrated positive direct effect on grain yield/plant in all the three generations. Roots/plant had highly positive direct effect in F₂. Direct effects were negative in respect of root length and dry root weight in F₂ generation.     

A codominant SCAR marker linked to the genic male sterility gene (ms1) in chili pepper (Capsicum annuum)

As one of the genic male sterility (GMS) materials in chili pepper ( Capsicum annuum L.), GMS1 has been used for commercial F₁ hybrid seed production. The male sterility of GMS1 is controlled by a recessive nuclear gene, named ms ₁ . In this study, we developed DNA markers linked to the ms ₁ locus using a combination of bulked segregant analysis and amplified fragment length polymorphism (AFLP) in a segregating sibling population. From the screening of 1024 AFLP primer combinations, the AFLP marker E-AGC/M-GTG (514 bp) was identified as being linked to the ms ₁ locus at a distance of about 3 cM. Based on internal sequencing analysis of the E-AGC/M-GTG marker between male fertile and sterile plants, we identified three small deletions with a size of altogether 42 bp in the male-fertile plant and developed a codominant sequence characterized amplified region (SCAR) marker. This SCAR marker may be valuable for marker-assisted breeding in the hybrid seed production system of chili pepper using the GMS1 line.     

Cytogenetic analysis of F1, F2 and BC1 plants from intergeneric sexual hybridization between Sinapis alba and Brassica oleracea by genomic in situ hybridization

By intergeneric sexual hybridization between Sinapis alba and Brassica oleracea , F₁, F₂ and BC₁ progeny plants were produced. S. alba plants (genome SS, 2n = 24) were pollinated with B. oleracea (genome CC, 2n = 18), and the fertile F₁ plants were pollinated with B. oleracea to obtain BC₁ plants. GISH analysis showed that 10 out of 12 F₁ plants had 12 S. alba chromosomes (one full S chromosome set) and nine B. oleracea chromosomes (one C chromosome sets), representing the expected hybrids. However, two F₁ plants had 12 S chromosomes and 18 C chromosomes (two C chromosome sets), indicating unexpected hybrids. A maximum of three trivalents between C and S chromosomes were identified at metaphase I of semi-fertile F₁ pollen mother cells (PMCs), which indicates homology and chromosome pairing between these two genomes. The C genome had obviously been doubled in two F₂ plants from selfed semi-fertile F₁ plants. BC₁ plants consisted of 18 C chromosomes and different numbers of one, five and six additional S chromosomes, respectively. Monosomic alien addition lines developed in the present study can be used for B. oleracea breeding and Sinapis alba gene mapping.     

Relationships between seed oil content and fatty acid composition in high stearic acid sunflower

The high stearic acid sunflower mutant CAS-3 is characterized by a low seed oil content, which might represent a constraint for the commercial production of high stearic acid sunflower oil. The objective of the present research was to investigate the relationships between fatty acid profile and seed oil content in CAS-3. Plants of CAS-3 were reciprocally crossed with plants of breeding line ADV-37, with high oil content and standard fatty acid profile. Oil content and fatty acid composition were measured in individual F₂ seeds and F₂ plants (F₃ seeds averaged). Both F₂ seeds and F₂ plants from the cross ADV-37 × CAS-3 had a significantly higher oil content than those from the reciprocal cross, which indicated the existence of cytoplasmic effects in the genetic control of the trait. A consistent negative correlation between oil content and palmitic acid and a positive correlation between oil content and oleic acid were detected both in F₂ seeds and F₂ plants. Conversely, no consistent correlation between oil content and stearic acid was observed, which suggested the feasibility of simultaneous selection for both traits.     

Identification of AFLP and RAPD markers linked to anthracnose resistance in grapes and their conversion to SCAR markers

Resistance to anthracnose or black spot ( Elsinoe ampelina ), a serious fungal pathogen in viticulture and table grape production, was investigated on 25 grape cultivars. Bioassays performed with culture filtrates produced by the pathogen revealed 14 resistant genotypes. In most plants resistance originated from Vitis labrucsa but also genotypes with V. rupestris and V. riparia  ×  V. rupestris background showed resistance. Genetic analysis was conducted in F₁, S₁ and BC₁ plants developed from various cultivars. In total, 326 F₁ plants were evaluated, 172 genotypes proofed to be resistant, whereas 154 were susceptible to anthracnose. A Mendelian segregation ratio of 1 : 1 (χ² = 0.30–0.65) indicating that anthracnose resistance is controlled by a single dominant gene. To facilitate the use of marker-assisted selection in grape-breeding PCR-based markers were developed by random amplified polymorphic DNA and amplified fragment length polymorphism in bulk segregant analysis. Finally, OPB 15₁₂₄₇ was developed as a sequence characterized amplified region marker being diagnostic for the locus of resistance to anthracnose in all resistant genotypes tested. Within the 25 grape cultivars OPB 15₁₂₄₇ is diagnostic in the genetic background of both V. labrucsa and V. rupestris and V. riparia  ×  V. rupestris .     

Production of intergeneric hybrids between Raphanm and Moricandia

Intergeneric F₁ hybrids between Raphanus sativus (2n = 18, RR) and Moricandia arvensis (2n = 28, MaMa) have been produced through ovary culture followed by embryo culture, when M. arvensis was used as a pistillate parent. Six BC₁ plants were also obtained through ovary culture followed by embryo culture in the backcross of an amphidiploid F₁, hybrid with R. sativus cv. 'Pink ball'. Two BC₁ plants were ses-quidiploids (2n = 32, MaRR), and the other BC₁, plants were hyperploid with 2n = 55, having MaMaRRR genomes. BC₂, seeds were obtained by conventional pollination in the successive backcross of two sesquidiploid BC₁, plants with R. sativus cv. 'Pink ball'. Their seed set percentages were 12.7% and 17.0%, respectively. These novel hybrid plants and derived progenies may be valuable materials for the genetic investigation and breeding of Brassiceae , including R. sativus.     

Inheritance of a weeping character and the low frequency of rooting from cuttings of the mulberry variety 'Shidareguwa'

A mulberry variety, Morus alba 'Shidareguwa', has a weeping habit and is used as an ornamental and landscape plant. This variety is known for being difficult to propagate by hardwood cuttings. To clarify the mode of inheritance of the weeping character and its relationship to the difficulty in cutting propagation, we crossed 'Shidareguwa' with a non-weeping variety 'Noi' that has a very high rooting ability. The phenotypic segregations exhibited by the F₁ and F₂ plants suggested that the weeping habit is controlled by a single recessive gene. In tests of the F₂ plants for cutting propagation, no obvious correlation was recognized between the weeping trait and the rooting ability of cuttings. As a result, we were able to develop new weeping mulberry strains with improved high rooting ability.     

A high-resolution map of the rice blast resistance gene Pi15 constructed by sequence-ready markers

The gene Pi15 for resistance of rice to Magnaporthe grisea was previously mapped to a ≈0.7-cM region on chromosome 9. To further define the chromosomal region of the Pi15 locus, a contig spanning the locus was constructed, in silico , through bioinformatics analysis using a reference sequence of the cultivar 'Nipponbare'. One simple sequence repeat marker adopted from the International Rice Microsatellite Initiative and six candidate resistance gene (CRG) markers, developed from gene annotation of the reference sequence of the contig, were used for linkage analysis in a mapping population consisting of 504 extremely susceptible F₂ plants. The Pi15 locus was delimited to a ≈0.5-cM region flanked by the markers CRG5 and CRG2 and co-segregated with the markers BAPi15₇₈₂, CRG3 and CRG4, which was physically converted to a 44-kb interval.     

Evaluation of the Heterosis on Leaf Photosynthesis of Remote-Cross F1 Rice (Oryza saliva L.)

The heterosis of leaf photosynthesis was studied on the main characters included in the range from CO₂exchange rate (CER) to enzymatic activity using a remote cross F, rice. The CER was significantly higher than those of the parental strains, showing a 111 % heterosis effect on average; at the same time strong heterosis was observed for the leaf area production and growth. Also stomatal and mesophyll conductances increased in the F₁ rice, which may contribute to the increase in CER. Chlorophyll content (Ch1), soluble protein content (SPC) and ribulose 1,5-bisphosphate (RuBP) carboxylase activity (RCA) were measured as the internal factors related to photosynthesis, and compared between the F₁, rice and the parents. For all these factors, the F₁ rice showed low values compared to the parents. Positive heterosis was not expressed here. On the other hand, the specific activity of RCA (RCA/SPC) increased in the F₁, rice, showing a 120% heterosis effect. This may be regarded as one of the main causes for the increase in CER of the F₁, rice. High CER expressed as heterosis concurrently with large leaf area production is one of the important findings in our study, and this may suggest a high possibility of further improvement in biomass production or yield of rice by gathering the advantageous elements into a hybrid plant.     

Identification and inheritance of a new gene for powdery mildew resistance in mungbean (Vigna radiata L. Wilczek)

Powdery mildew (PM) is one of the important foliar diseases of mungbean. Resistance sources have been identified in India and the inheritance studies showed that complete resistance (RO) was controlled by two dominant genes, Pm1 , Pm2 . The breakdown of complete resistance (RO) into moderate resistance (R2) by race-2 (Akola) has been reported. It is assumed that the change in resistance reaction is due to a mutation in the pathogen. The present investigation was carried out with a view to screen germplasm, cultivars and mutants for identification of complete resistance (RO) sources against race-2 and to study their inheritance. 'Mulmarada', a local mungbean cultivar from Maharashtra state of India was identified as a complete resistance (RO) source for race-2. The inheritance of Mulmarada's resistance (RO) was studied. The F₁ and the segregation in F₂ and F₃ showed that the complete resistance (RO) in 'Mulmarada' is controlled by a single dominant gene, which is different from the earlier identified Pm1 and Pm2 resistance genes. Mulmarada's resistance gene is designated as Pm3 for PM resistance.     

Inheritance of time of flowering in upland cotton under natural conditions

Time to flowering is an essential component of the adaptation and productivity of cotton ( Gossipium hirsutum ) in various agro-ecological zones. This article presents a study of the genetic control of this trait in two crosses obtained from different early-maturity parental lines. In each cross, multiple generations including P₁, F₁, P₂, B₁, B₂ and F₂ were evaluated under two natural field conditions in 2004 and 2005. The data on time to flowering in the F₂ populations had a continuous distribution but deviated from normality. A joint segregation analysis (JSA) revealed that time of flowering in upland cotton was controlled by a mixture of an additive major gene and additive-dominant polygenes. The first- and second-order genetic parameters were all calculated based on the mixture of major gene and polygene inheritance models using JSA. These results suggested that there was considerable genetic diversity and complexity in days to anthesis in upland cotton. This variation can be used to formulate the most efficient breeding strategy and to design cotton for a particular environment.     

The origin of the self-compatible almond 'Supernova'

The almond cultivar 'Supernova' is reported in the literature as a late-flowering self-compatible mutant, obtained by the irradiation of the early-flowering self-incompatible cultivar 'Fascionello'. Our work to investigate the molecular basis of this form of self-compatibility has called into question the origin of 'Supernova'. Test selfing was performed both on 'Supernova' and on the accession of 'Fascionello' from which it was derived –'Fascionello-Rome'. Both proved self-compatible. Amplification of S - RNase alleles, using consensus primers from the signal peptide region to the second conserved region of the S - RNase gene and primers specific for allele S _f , confirmed 'Supernova' and 'Fascionello-Rome' indeed have the same (in)compatibility genotype S ₁ S _f as the self-compatible cultivar 'Tuono'. Nine microsatellite markers were used to fingerprint the relevant accessions; 'Supernova' and 'Fascionello-Rome' were undistinguishable from 'Tuono'. Two Sicilian accessions, 'Fascionello' and 'Falso Fascionello', lacked S _f and differed at the SSR level, indicating no close relationship with 'Supernova'. Therefore we concluded that 'Fascionello-Rome' is actually the same as 'Tuono' and that 'Supernova' originates from 'Tuono' from which it derives its self-compatibility.     

Development and agronomic performance of common bean lines simultaneously resistant to anthracnose, angular leaf spot and rust

The common bean is affected by several pathogens that can cause severe yield losses. Here we report the introgression of resistance genes to anthracnose, angular leaf spot and rust in the 'carioca-type' bean cultivar 'Rudá'. Initially, four backcross (BC) lines were obtained using 'TO', 'AB 136', 'Ouro Negro' and 'AND 277' as donor parents. Molecular fingerprinting was used to select the lines genetically closer to the recurrent parent. The relative genetic distances between 'Rudá' and the BC lines varied between 0.0% and 1.99%. The BC lines were intercrossed and molecular markers linked to the resistance genes were used to identify the plants containing the genes of interest. These plants were selfed to obtain the F₂, F₃ and F₄ plants which were selected based on the presence of the molecular markers mentioned and resistance was confirmed in the F₄ generation by inoculation. Four F_4:7 pyramid lines with all the resistance genes showed resistance spectra equivalent to those of their respective donor parents. Yield tests showed that these lines are as productive as the best 'carioca-type' cultivars.     

Hybridization of Thinopyrum distichum and Secale cereale

An attempt was made to determine whether the salt tolerance genes of Thinopyrum distichum would be expressed in the presence of the rye genomes and if it would be possible to improve and utilize its hybrid with rye as a salt-tolerant fodder crop. Th. distichum florets were pollinated with diploid and tetraploid rye but hybrids were obtained only with tetraploid rye. The F₁ had 28 chromosomes and expressed high levels of salt tolerance. Fertility could not be induced through colchicines treatment but the plants produced some apomictic seeds during the summer months. The F₂ had 20-22 chromosomes and retained two complete or near-complete rye genomes. The F₂ plants responded better to colchicine treatment, however, the doubled hybrids remained male sterile. In summer, one F2 plant produced many apomictic seeds with chromosome numbers ranging from 14 to 16 (primarily rye chromosomes). The mechanism that caused the preferential loss of Thinopyrum chromosomes in each apomictic cycle remains unknown. The F₃ phenotypically resembled rye, had primarily reductional male meioses and produced seemingly functional pollen, but were self-sterile.     

Development of co-dominant amplified polymorphic sequence markers for resistance of sunflower to downy mildew race 730

The inheritance of the reaction of sunflower to downy mildew was investigated using resistant and susceptible near isogenic lines (NILs) and their F₃ families. Resistance to race 730 was evaluated using the whole seedling inoculation technique. Seventy-three F₃ families were inoculated, among which 54 families were resistant and 19 susceptible, fitting a 3 : 1 segregation ratio. F₃ families were also studied using several PCR markers. Ten markers at the Pl6 locus, specific for the resistant line, also segregated in F₃ families with a 3 : 1 ratio. The same segregation ratio occurred for microsatellite haplotypes that resembled the resistant parent, and were amplified with ORS 166 and ORS 1043. The only common fragment that was observed between resistant and susceptible parental lines was one of the TIR-NBS-LRR resistance gene analogue markers, having a restriction site. Two co-dominant cleaved amplified polymorphic sequence (CAPS) markers were obtained. The mapping data indicate that several dominant markers and two CAPS markers, developed here, completely co-segregate with the Pl6 gene conferring resistance to race 730. CAPS markers will facilitate efficient marker-assisted selection for sunflower resistance to downy mildew race 730.     

Identification and molecular tagging of a stripe rust resistance gene in wheat line P81

Stripe rust, caused by Puccinia striiformis f. sp. tritici (PST), is one of the most devastating diseases in common wheat ( Triticum aestivum L.). With the objective of identifying and tagging a new gene for resistance to stripe rust in wheat line P81, F₁, F₂ and F_2:3 populations from the cross 'Chuanmai 28'/P81 were inoculated with Chinese PST race CYR32 in greenhouse and field trials. P81 carried a single dominant gene for resistance (designated YrP81 ) to CYR32. Tests of allelism showed that YrP81 was different from Yr5 , Yr10 , Yr15 and Yr26 . Simple sequence repeat (SSR) and resistance gene-analogue polymorphism (RGAP) between the parents were used for genotyping the F₂ populations. YrP81 was closely linked to four SSR loci on chromosome 2BS with genetic distances of 18.3 cM ( Xwmc25 ), 1.8 cM ( Xgwm429 ), 4.1 cM ( Xwmc770 ) and 5.3 cM ( Xgwm148 ). Two RGAP markers RGA1 (NLRR/XLRR) and RGA2 (Pto kin4/NLRR-INV2) were also closely linked to YrP81 with genetic distances of 4.7 and 6.3 cM, respectively. The linkage map of YrP81 and molecular markers was established in the order Xwmc25 - RGA2 - RGA1 - Xgwm429 - YrP81 - Xwmc770 - Xgwm148 . Pedigree analysis, response patterns with Chinese PST races and associations with markers suggested that YrP81 is a novel stripe rust resistance gene. The PCR-based microsatellite and RGAP markers identified here could be applied in selection of YrP81 in wheat breeding.     

Correlation Analysis of Triticale Morphology, Chlorophyll Content and Productivity

In some lines and F₁ hybrids of winter hexaploid triticale ( xTriticosecale Witt.), significant positive associations were found amongst leaf anatomy, plant morphology, chlorophyll content, photochemical activity of chloroplasts, and photosynthetic or grain productivity. The data may be useful in practical triticale breeding.     

Phenotypic Yield Stability Depending on Plant Density and on Mean Yield per Plant of Winter Rapeseed Varieties and of Their F1 and F2-Generations

Among the common agricultural crops winter-rapeseed ( Brassica napus L.) is considered to be especially susceptible to varying environmental conditions. The commonly used stability statistics, which measure these fluctuations, are based on the yields of genotypes in different environments. An improvement of stability analysis may be obtained by an additional estimation of stability using yield components rather than absolute yield values. Relationships between stability of seed yield (SY) and stabilities of plant densities (PD) and of single plant yield (SPY) were examined for four winter-rapeseed varieties, their six F₁'s and their six F₂'s. Environmental variance was applied to estimate phenotypic stabilities. Furthermore, generations were compared for their stability in SY, PD and SPY. Regarding all genotypes and generations the correlation coefficients did not show any relation between stability of SY and stabilities of PD and of SPY (r = -0.04 and r = -0.17). Comparing generations the F₂ showed a higher stability of SY than varieties and F₁, But no consistent results were obtained regarding PD and SPY. The pattern of the heterotic effects of stability of yield components PD and SPY were different from the respective pattern of SY. Eg although possessing desired heterotic effects in stability of PD and SPY some crosses showed undesired heterotic effects in stability of SY. Therefore, a theory of multiplicative heterosis of stability should be developed in analogy to the corresponding theory for yield.