Generation of tribenuron‐methyl herbicide‐resistant OsCYP81A6‐expressing rapeseed (Brassica napus L.) plants for hybrid seed production using chemical‐induced male sterility

Chemical‐induced male sterility (CIMS) is a method for hybrid rapeseed (Brassica napus L.) production. Some sulphonylurea herbicides such as tribenuron‐methyl (TBM) are used as chemical hybridization agents (CHAs) in CIMS systems. However, the male parents must be protected from herbicide injury with a shield during spraying of the female parents with CHAs to induce male sterility. Thus, using herbicide‐resistant rapeseed lines as the male parents can significantly simplify the seed production procedure and reduce the cost in hybrid seed production. A rice cytochrome P450 hydroxylase, OsCYP81A6, has been previously characterized to confer resistance to bentazon and sulphonylurea herbicides. We demonstrate here that the introduction of OsCYP81A6 renders rapeseed plants resistant to TBM. Compared with wild‐type plants, the transgenic plants displayed normal stamen development and male fertility when treated with 0.05 mg/l of TBM, the dose used for inducing male sterility in hybrid seed production. These results indicate that the OsCYP81A6‐expressing rapeseed plants can be used as the male parents for hybrid rapeseed production using CIMS.     

Inheritance and mapping of a restorer gene for the rapeseed cytoplasmic male sterile line 681A

A novel cytoplasmic male sterility‐fertility restoration system has been developed in rapeseed (Brassica napus). The cytoplasmic male sterile line 681A was derived from a spontaneous male sterile mutant in a newly released double‐low rapeseed cultivar ‘Xiangyou 13′. The restorer line 714R was identified in the interspecific progeny from a B. napus×B. juncea‐cross. Genetic analysis showed that fertility restoration for 681A cytoplasmic male sterility was controlled by a single dominant nuclear gene which might originate from B. juncea. The RAPD marker S1039_‐520 was found to be linked to the restorer gene in F₂ progeny of 681A × 714R with a recombination frequency of 5.45%.     

Bud pollination and hybrid seed production in detergent-induced male sterile plants of Brassica juncea

The efficacy of a synthetic detergent (Surf Excel) as a potential chemical hybridizing agent in Brassica juncea was studied. Foliar sprays with various concentrations of the detergent caused reductions in plant height, number of branches and leaves per plant, size of leaves, anther size, pollen per flower, ovules per flower, pollen fertility, fruits per plant, fruit size, seeds per fruit, total yield per plant and 100 seed weight as compared with those of untreated plants. The style in the floral buds of plants sprayed with different concentrations of Surf Excel elongated and so the receptive stigma protruded from the buds which facilitated cross‐pollination by honey bees. The plants sprayed once with 2% Surf Excel exhibited an elongated style with a raised receptive stigma and 100% pollen sterility without causing a significant reduction in total yield.     

Pleiotropic effects of the ea7 photoperiod response gene on the morphology and agronomic traits in barley

The photoperiod‐insensitive barley mutant ‘Atsel’, carrying the recessive gene ea₇, was studied together with the donor variety ‘Atlas’ (wild‐type, Ea₇) under different daylengths with the aim of analysing pleiotropic effects. Grown under long and short photoperiods ‘Atsel’ flowered about 10 days and 34 days, respectively, earlier than ‘Atlas’. The significantly shorter life‐cycle of the photoperiod‐insensitive mutant resulted in several changes of plant morphology. Tillering, plant height, number of leaves and number of internodes were reduced. A lower number of florets per main spike was observed for ‘Atsel’, but only in the long photoperiod experiment. Finally, photoperiod insensitivity combined with a lower grain yield per plant was most pronounced under long‐day treatment. The data are comparable with results obtained from single chromosome recombinant lines of wheat that have differences in their photoperiod response caused by the genes Ppd1 or Ppd2.     

Do floral resources influence pollination rates and subsequent fruit set in pear (Pyrus communis L.) and apple (Malus x domestica Borkh) cultivars?

Pear and apple are among the main fruit crops worldwide. These species can be planted in mixed orchards, and they both depend on insect pollination for fruit set. As pollinating insects are attracted by the floral resources, we investigated nectar and pollen production and chemical composition in four pear (‘Concorde’, ‘Conférence’, ‘Doyenné du Comice’, ‘Triomphe de Vienne’) and five apple (‘Braeburn’, ‘Golden Reinders’, ‘Jonagored’, ‘Pinova’, ‘Wellant’) cultivars commonly grown in Belgium. We also investigated whether insect flower visitation rate and pollination efficiency are linked to floral resource quantity and quality. The pear cultivars flowered one week before the apple cultivars in early spring, and their flowers were about six times less visited by insects. The visitors foraged more on the pollen of the pear trees and the nectar of the apple trees. Pear flowers produced higher volumes of nectar than apple flowers (1.3–3.2 μl vs. 0.4-0.6 μl), but with lower sugar concentration (9.6%-10.8% vs. 28.3%-36.4%). Pear flowers also produced fewer pollen grains per anther than apple flowers (2425–4937 vs. 3284–7919), but these had higher polypeptide (346–362 μg/mg vs. 216–303 μg/mg), amino-acid (40–77 μg/mg vs. 12–18 μg/mg) and phytosterol (21–47 μg/mg vs. 15–43 μg/mg) concentrations. The foraging behavior of the insects is thus better explained by nectar and pollen quality rather than quantity. Despite the differences in flower visitation rates, pollination of both species resulted in valuable fruit production.     

Effects of Temperature and Photoperiod on Days to Flowering, Yield and Yield Components of Lupinus albus (L.) under Field Conditions

A better understanding of the agronomic importance of planting date and the influence of cold temperatures and photoperiod during germination and plant growth may lead to better management strategies for cultivation of the sweet white lupin (Lupinus albus). The effects of planting date (temperature and photoperiod) were determined on the number of days to flowering, yield and yield components of four early to medium and one late sweet white lupin genotype in a field trial at Potchefstroom, South Africa, planted during February 1996 to January 1997. Moisture stress was avoided through regular irrigation. Duration of the developmental phases planting date to emergence, emergence to floral initiation, initiation to first flower, duration of flower and days to physiological and harvest maturity was related to field measurements of temperature and photoperiod. Differences in the main determinants of yield, i.e. seeds per pod, pods per plant, single seed mass (SSM), plant and pod height and yield, were measured. Results showed that both temperature and photoperiod influence the growth and development of the Lupinus albus genotypes ‘Esta’, ‘Hantie’, ‘Tifwhite’, ‘Kiev’ and ‘LAL 186’. Temperature influences include the effect of vernalization at seedling emergence. Minimum grass temperatures under 5 °C at emergence are effective for vernalization. However, after grass temperatures at emergence increased again from June to December, to gether with an increase in the photoperiod length, ‘Tifwhite’ as well as the other genotypes still flowered earlier, confirming that these cultivars are long‐day plants, which is in accordance with controlled‐environment data. Cool vernalizing temperatures thus not only influence obligate vernalization requiring genotypes such as ‘Tifwhite’, but also influence the non‐obligate genotypes studied. Plan‐ting date had a significant influence on pods per plant, single seed mass (SSM) and seed yield. In all trials laterplanting, from June to November, decreased SSM and seed yield. The highest seed yield of 1.5 t ha^?1 was obtained for the 10 June planting date and the lowest average yield of 0.450 t ha^?1 for the 5 November planting date.     

Creating favourable morphological and yield variations for rapeseed by interspecific crosses between Brassica rapa and Brassica oleracea

Brassica napus is a most important oilseed grown worldwide with a limited genetic background, due to the short history of speciation, domestication and cultivation. To create novel germplasm for rapeseed breeding, we made interspecific crosses followed with chromosome doubling between B. rapa and B. oleracea to generate novel B. napus with favourable agronomic traits. The resynthesized (S0) hybrids were confirmed by SSR and cytogenetic analysis, and the fertility was increased from 32.7% in S0 generation to ~97.31% in S1 generation. The plant shapes of the progeny were dramatically improved compared to the diploid parents and B. napus cv. ‘Yangyou 6’, especially for the branch initiation height, branch number and pod number. The single‐plant yield was significantly improved in S1 progeny for the variations in branching sites and number. Significant improvement in plant shape and yield was observed on S2 generation compared to the local elite commercial open‐pollinated cultivar, which would be further fixed by intensive selection and pyramiding breeding. Such variation is of great value for breeding rapeseed with improved plant architecture and harvest index.     

Improved application of tribenuron-methyl as a chemical hybridizing agent with forchlorfenuron for rapeseed hybrid breeding

Chemicals or agents that can induce plant male sterility are considered chemical hybridizing agents (CHAs) and are quite useful for plant hybrid breeding. Tribenuron-methyl (TBM) can effectively induce male sterility in rapeseed plants. However, applications of TBM in rapeseed breeding practices have not been reported. Here, treatment with the CHA referred to as CHA-TBM, which contained 0.1–0.9 mg/L TBM, induced male sterility in all 22 tested rapeseed inbred lines. However, the highest induced male sterile rates in the tested rapeseed lines were only approximately 54–57% together with poisoned rates of approximately 36–39%, which were resulted from two treatments of CHA-TBM containing 0.5 mg/L TBM at an interval of 9 days beginning at the floral bud stage (FBS) of 2.0–3.5 mm. Such male sterile rates and poisoned rates caused by CHA-TBM indicated that TBM is not an ideal CHA for rapeseed hybrid breeding. The results of many screening experiments showed that forchlorfenuron can alleviate the toxic effects of TBM on rapeseed plants. Two treatments with the CHA referred to as CHA-TBM-FCF, which contained 0.6 mg/L TBM and 2.1 mg/L forchlorfenuron, at an interval of 9 days beginning at the FBS of 2.0–3.5 mm induced male sterility in approximately 96–98% of plants of all the 22 tested rapeseed inbred lines and poisoned only approximately 2–4% of the tested plants. The CHA-TBM-FCF treatment did not affect pistil fertility of the rapeseed plants, and had no carry-over influence on stamen fertility of the rapeseed hybrids produced with this CHA. Some rapeseed hybrids were successfully produced using CHA-TBM-FCF (e.g., Qinrong 1, Qinrong 2, Qinyou 19, Qinyou 33, Qinzayou 4 and Qinzayou 5). The hybridity values of the F1 seeds of the rapeseed hybrids produced using CHA-TBM-FCF were determined to be approximately 93–98% with the simple sequence repeat (SSR) marker techniques. Together, the results indicate that CHA-TBM-FCF containing TBM and forchlorfenuron at suitable concentrations is an ideal CHA for rapeseed hybrid breeding.     

A major quantitative trait locus for resistance to Turnip Yellows Virus (TuYV, syn. beet western yellows virus, BWYV) in rapeseed

Turnip yellows virus (TuYV) is responsible for a recognizable loss of yield in European winter oilseed rape cultivation. To map genes involved in TuYV resistance, a double haploid population was established by crossing a resynthesized rapeseed line (R54) as donor for TuYV resistance with an elite rapeseed line (‘Express’). Resistance was determined with 10 plants per line by double antibody sandwich‐enzyme‐linked immunosorbent assay. After screening 17 primer combinations (Pstl/Msel and EcoRI/Msel), 143 amplified fragment length polymorphism markers were mapped to 20 linkage groups representing 15 chromosomes of the rapeseed genome. Quantitative trait loci (QTL) were mapped using the composite interval mapping approach. As a result, one major quantitative trait locus was found on linkage group MS17, explaining up to 50% of the phenotypic variation. Because no other factors displaying a significant effect on the expression of resistance could be identified, a simple mode of inheritance for TuYV resistance is suggested, thus enabling marker‐assisted selection during rapeseed breeding.     

Characterization of different cytoplasmic male sterility systems in Indian mustard (Brassica juncea L. Czern & Coss)

Six cytoplasmic male sterility (CMS) systems, viz. moricandia, ogura, oxyrrhina, siifolia, tournefortii and trachystoma of Indian mustard (Brassica juncea L.) were characterized for agronomic and floral characteristics. Introgression of alien cytoplasm caused alterations in different floral traits in ogura, siifolia, tournefortii and trachystoma CMS systems. Varied response to different genetic backgrounds of CMS lines indicated the presence of cytoplasmic–nuclear interaction in alteration of floral traits. On the basis of floral characteristics, CMS systems could be grouped into distinct classes. Siifolia, tournefortii and trachystoma CMS lines had narrow petals, while moricandia, ogura and oxyrrhina had wider petals, which were distinguishable on the basis of visual observations. The ratio between length and width of petals were >2.0 in wide petal group but <2.0 in narrow petal group. Further, the relative position of anther and stigma, which was estimated as the ratio between stamen and style length could differentiate the CMS systems. Stamens were longer than styles in oxyrrhina, equal in moricandia and shorter in ogura, siifolia, tournefortii and trachystoma male sterile lines. Non‐viable pollen grains were present in moricandia and oxyrrhina systems, but absent in other systems. In tournefortii and trachystoma, few flowers showed petaloid corolla. All male sterile lines, except trachystoma, which showed crooked siliqua formation were at par with their respective maintainers for flower initiation, plant height, primary branches, seeds per siliqua, seed yield, harvest index, oil and protein content. In general, flower senescence and maturity occurred earlier in male sterile lines than in their respective maintainer lines.     

Molecular mapping of a dominant genic male sterility gene Ms in rapeseed (Brassica napus)

Rs1046AB is a genic male sterile two‐type line in rapeseed that has great potential for hybrid seed production. The sterility of this line is conditioned by the interaction of two genes, i.e. the dominant genic male sterility gene (Ms) and the suppressor gene (Rf). The present study was undertaken to identify DNA markers for the Ms locus in a BC₁ population developed from a cross between a male‐sterile plant in Rs1046AB and the fertile canola‐type cultivar ‘Samourai’. Bulked segregant analysis was performed using the amplified fragment length polymorphism (AFLP) methodology. From the survey of 480 AFLP primer combinations, five AFLP markers (P10M13₃₅₀, P13M8₄₀₀, P6M6₄₁₀, E7M1₂₃₀ and E3M15₁₀₀) tightly linked to the target gene were identified. Two of them, E3M15₁₀₀ and P6M6₄₁₀, located the closest, at either side of Ms at a distance of 3.7 and 5.9 cM, respectively. The Ms locus was subsequently mapped on linkage group LG10 in the map developed in this laboratory, adding two additional markers weakly linked to it. This suite of markers will be valuable in designing a marker‐assisted genic male sterility three‐line breeding programme.     

Untersuchungen zum Einfluß der Bestandesdichte auf das Ertragspotential von Rapspflanzen (Brassica napus L. var. napus)*)

Investigations on the effect of number of plants/area on the yield potential of rapeseed plants (Brassica napus L. var. napus) Based on field experiments with winter rapeseed, the effect of number of plants per area on yield potential and components of yield were investigated. The dependence of yield from number of plants per area before and after winter was analyzed by manipulating the number of plants immediately after sowing and in spring before beginning of growth. The number of plants per area was varied between 10 plants/m² till 185 plants/m². The results show that the yield potential is partly dependend on the growing conditions before winter as influenced by the number of plants per area; the yield potential declines with increasing number of plants/area before winter; affected are the number of siliquae producing lateral shoots, the number of siliquae per shoot and seeds per siliqua. The growing conditions before winter, in this way control the capacity of the rapeseed plant to utilize environmental factors in the main growing period. 50–100 plants/m² before winter and 45–90 plants/m² after winter are within the compensation capacity of rapeseed plants.     

油菜新品种（组合）98P37-21稻田免耕适宜播种量和移栽密度研究初探

为了探索油菜新品种（组合）98P37-21高产、高效栽培技术,在稻板田中开展不同播种量和移栽密度试验,研究其对油菜生长发育、主要经济性状、产量及其构成因素的影响,并对直播和移栽油菜效益进行了比较。结果表明：直播油菜4.5~7.5 kg/hm²播种量范围内（密度27.3万株/hm²~38.7万株/hm²）,产量随着播种量的增加呈先增后减的趋势;移栽油菜9.0万株/hm²~15.0万株/hm²密度范围内,产量随着移栽密度的增加而增加,直播油菜以6.0 kg/hm²产量最高,移栽油菜以15.0万株/hm²产量最高,但高密度增产优势不明显,表明密度过大也不利于高产,甚至可能降低产量。随着播种量或移栽密度的增加,主茎总叶数、绿叶数、根颈粗、叶面积、一次有效分枝数、全株角果数、每角粒数、千粒重、单株地上部干重、单株产量呈现下降的趋势,而株高、分枝高度变高,叶面积指数变大。移栽油菜主要经济性状指标优于直播油菜,直播油菜产量和产值明显要低,但直播油菜节省劳力和减少投入,净收入却比移栽油菜多1735.7元/hm²,效益明显要高。     

Heritability and genetic correlation in daylily selection

Daylily (Hemerocallis × hybrida Hort.) is a popular ornamental plant and hybrid cultivars are frequently selected for medium size, large number of flowers per stem, and flower color diversity. However, daylily selection is limited due to the lack of information on genetic parameters and their correlation to phenotypic characteristics. This study aimed at estimating genetic parameters and genetic correlations among full sib families of 13 daylily cultivars using REML/BLUP (restricted maximum likelihood/best linear unbiased prediction) and to select families with high genotypic values for plant height and number of floral buds per stem. Crosses were performed among those cultivars and 24 families were obtained. In the third year, plant height, leaf length and width, stem number, number of floral buds per stem, and number of floral buds per plant were evaluated. The estimates of average heritability of families were superior to the estimates of restrict individual heritability for all characters evaluated. High heritability estimates were obtained for plant height, leaf length, leaf width, stem number, and number of floral buds per plant. Selected families showed genetic gain of 19.80% for plant height, 32.48% for stem number, and 38.64% for number of floral buds per plant.     

Identification of genetic variation in Brassica napus seeds for tocopherol content and composition using near‐infrared spectroscopy technique

Tocopherol is an essential fat‐soluble nutrient for humans. Increasing the tocopherol content in Brassica napus seeds can add value to rapeseed vegetable oil; this has become an important breeding target. However, there is no efficient and non‐destructive method for selecting rapeseed accessions with high tocopherol contents. Here, we report the first near‐infrared reflectance spectroscopy (NIRS)‐based technique for predicting rapeseed tocopherol content. Individual seed tocopherol compositions were estimated from 373 rapeseed genotypes of different origins. This method and chemical methods produced comparable predicted values of the tocopherol constituents in the seeds. Three equations were generated for the prediction of tocopherol content by using a modified partial least squares (MPLS) model. The total tocopherol content for the determination coefficient of cross‐validation (R²_cv) (0.74), determination coefficient (RSQ) (0.76) and one minus the ratio of unexplained variance to total variance (1‐VR) (0.65) values indicates a strong correlation between the calibration and validation sets. Overall, our model confirmed the NIRS method as feasible for predicting tocopherol content in rapeseed and as an efficient screening tool for future breeding programs.     

Heredity of phenylpropenes in sweet basil (Ocimum basilicum L.) chemotypes and their distribution within an F2 population

Phenylpropenes play an important role in plant defense against animals and microorganisms, and in attracting pollinators and insects. We report the genetic inheritance of methyl chavicol and eugenol following a cross between the sweet basil varieties ‘Perrie’ and ‘Cardinal,’ eugenol and methyl chavicol chemotypes, respectively. Methyl chavicol was detected only in ‘Cardinal,’ accounting for more than 95% of the total phenylpropenes. Eugenol was most abundant in ‘Perrie,’ accounting for more than 99% of the total phenylpropenes. Eugenol, chavicol and methyl chavicol were detected in F₁ hybrids at intermediate levels (10%–52%) without statistical differences (p > .05) for any compound among the F₁ progeny arising from the different crossed pairs. The F₂ progeny segregated into three groups, 23%–25% to a eugenol chemotype, 23%–25% to a methyl chavicol chemotype, and the remaining (~50%) into an intermediate mixture of the two compounds. This distribution fitted a segregation ratio of 1:2:1 (χ² = 1.71; p = .4249), suggesting that the phenylpropene phenotype is regulated by a single bi‐allelic gene with incomplete dominance. A putative association with biosynthesis enzymes is discussed.     

PCR primers for the estimation of contamination by seeds with normal cytoplasm in rapeseed lots bearing male-sterility-inducing Ogu-INRA cytoplasm

A polymerase chain reaction (PCR)-based test was developed to detect seeds bearing the ‘so-called’ normal rapeseed cytoplasm in seed lots with an OGU-INRA type cytoplasm. The test is based on the amplification of the orfB region of male fertile rapeseed mitochondrial DNA (mtDNA). The amplification reaction uses total nucleic acids of young seedlings, extracted in bulk. After the sequencing of the orfB gene region in the normal Brassica mtDNA, primers were designed for its amplification by PCR. Although the specificity of amplification for the male fertile (mf) rapeseed cytoplasm is partly impaired by the presence of tiny amounts of this fragment in the mtDNA of male sterile (ms) plants, this test proved to be applicable for the estimation of the level of contamination in seed lots in reconstituted mixes as well as in real lots.     

Pyramiding of insect- and disease-resistance genes into an elite indica, cytoplasm male sterile restorer line of rice,'Minghui 63'

The wild‐rice‐derived dominant gene Xa21 conferring multi‐race resistance to bacterial blight and a fused Bt gene cry1Ab/cry1Ac conferring resistance to lepidopteran insects were individually introduced into the same genetic background of an elite indica cytoplasm male sterile (CMS) restorer line ‘Minghui 63′. The line showed the desirable insect‐ and disease‐resistant phenotypes. To maximize the effect, the two genes were also pyramided into the same recipient plant of ‘Minghui 63’ by marker‐assisted selection. After being subjected to natural infestation of leaf‐folders and yellow stem borers and inoculation of Xoo strain mixtures, the pyramiding line and its derived hybrids showed high levels of resistance against both insect damage and disease. Furthermore, data from field trials demonstrated that the hybrids made by crossing this pyramiding line with the CMS lines ‘Zhenshan 97A’ and ‘Maxie A’ retained a similar level of yield under conditions without chemical spray, indicating that the pyramiding genes have a yield‐stabilizing effect on the recipient line and its hybrids.     

Influence of rare alleles of β‐carotene hydroxylase and lycopene epsilon cyclase genes on accumulation of provitamin A carotenoids in maize kernels

Vitamin A deficiency is one of the major health problems worldwide. Traditional yellow maize possesses very low provitamin A (proA) concentration in endosperm. The influence of rare alleles of β‐carotene hydroxylase (crtRB1) and lycopene epsilon cyclase (lcyE) genes capable of enhancing proA concentration was studied in four BC₂F₂ populations generated using subtropical inbreds and CIMMYT‐HarvestPlus lines. The occurrence of severe segregation distortion for crtRB1 gene was observed, while lcyE gene was segregated as per Mendelian ratio. Genotype with favourable allele of crtRB1 (CC) had a significant effect on β‐carotene (BC) (7.9‐fold), β‐cryptoxanthin (BCX) (twofold) and proA (5.5‐fold) accumulation, compared to unfavourable genotype (C⁺C⁺). Genotype with favourable allele of lcyE (LL) showed 2.1‐fold, 1.6‐fold and twofold significant enhancement in BC, BCX and proA, respectively, over unfavourable genotype (L⁺L⁺) in pooled analysis. Of the nine genotypes, double homozygote (CC/LL) had the highest mean BC (12.60 μg/g), BCX (4.44 μg/g) and proA (14.82 μg/g), and combined effect was significantly better than individual gene effects or any other combinations. The information generated here would be useful in designing strategy for proA enrichment in subtropical maize.     

Identification and Inheritance of Fertility Restorer Genes for 'tour' CMS in Rapeseed (Brassica napus L.)

Eighteen genotypes of Brassica napus were crossed to a cytoplasmic male sterile (CMS) line of B. napus BO 15 carrying B. tournefortii cytoplasm (‘tour’ cytoplasm). Fourteen genotypes were found to be stable maintainers of the ‘tour’ CMS. Of the remaining four genotypes, GSL-1 and ‘Asahi-natane’ were found to be heterozygous and ‘Mangun’ and ‘Yudal’ were homozygous for the restorer gene. Analysis of the F₁ and F₂ progenies of (CMS) BO 15 בMangun’ and (CMS) BO 15 בYudal’ showed that fertility restoration is controlled by a single dominant gene. The availability of a number of stable maintainer lines and the simple inheritance pattern of fertility restorer gene makes ‘tour’ CMS a useful system for hybrid seed production in rapeseed.