Production of 2n pollen of Asiatic hybrid lilies by nitrous oxide treatment

Tetraploid varieties of lilies have superior agronomic traits such as large flowers and resistance to physiological disorders. In the present study, we attempted to induce 2n pollen of Asiatic hybrid lilies by arresting the meiotic process with nitrous oxide (N₂O) gas. To determine which meiotic stage is optimal for induction of 2n pollen, plants with attached buds at different meiotic stages were treated with N₂O for 24 h in a pressure-tolerant cylinder. A few 2n pollen grains were induced using plants with anthers in prophase I, whereas mixed pollen grains of differing size were produced using plants undergoing meiotic metaphase predominantly in anthers. Although normal lily pollen grains are elliptical, nitrous oxide exposure induced giant pollen grains that appeared spherical. Flow cytometry analysis showed that the giant pollen grains were diploid. When mixed pollen that included normal and giant pollen was crossed to tetraploid cultivars, the resulting seedlings were tetraploid and aneuploid, indicating that the giant pollen grains were diploids that could generate tetraploid seedlings through fusion to diploid eggs supplied from a tetraploid female parent. Thus, treatment with N₂O is useful for the production of 2n lily pollen and may provide a new approach for tetraploid lily breeding.     

Screening for Fusarium resistance in seedling populations of Asiatic hybrid lily

A test to select Fusarium resistant seedlings of the Asiatic hybrid lily is described. Young seedlings of 28 populations, obtained from an incomplete diallel between eight parents with different levels of Fusarium resistance, were tested for resistance. Significant differences in Fusarium resistance between and within populations were detected. The average percentage of selected seedlings ranged from 34% in resistant × resistant crosses to 2% in susceptible × susceptible crosses. Although resistant descendants were obtained in susceptible × susceptible crosses, using at least one resistant parent produced higher percentages of resistant seedlings. The resistance level of the parents correlated highly with the general combining ability for Fusarium resistance based on the seedling test. For eight populations, seedlings selected for Fusarium resistance and non-tested (control) seedlings of the same cross were compared, after propagation, in a clonal test. Variation between and within populations, found at seedling level, was confirmed at clonal level. A positive selection response was found for all eight populations. In the seedling test, approximately 18% of the seedlings were selected as resistant of which 15% (2.7% of seedlings tested) appeared to be susceptible escapes. Comparison between selection at seedling level and at clonal level indicated that approximately 25% of the seedlings tested were missed (rejected resistant plants) in the seedling test. The practical use of a seedling test for Fusarium resistance in lily breeding programs is discussed.     

Low light intensity and flower bud abortion in Asiatic hybrid lilies. I. Genetic variation among cultivars and progenies of a diallel cross

Summary When Asiatic lilies are forced during the winter period, additional light is needed to prevent flower bud abscission and flower bud blasting. Growth room experiments showed considerable differences among eight lily cultivars in their response to low light. Among these cultivars, Connecticut King and Enchantment appeared to be the most sensitive to low light conditions while Uncle Sam and Scout were the least sensitive.Analysis of an incomplete diallel cross between nine cultivars showed significant differences in general combining ability (GCA) for flower bud abortion, number of buds, forcing time, plant height and leaf scorch under winter greenhouse conditions. Enchantment had the highest positive GCA for bud abortion, indicating a high sensitivity to low light conditions. Uncle Sam had the lowest GCA. Pirate and Scout appeared to have a high GCA for sensitivity to leaf scorch. Connecticut King was found to be a probable source of cytoplasmic male sterility. The data suggest that male sterility is associated with better response to low light conditions.     

N2O induces mitotic polyploidization in anther somatic cells and restores fertility in sterile interspecific hybrid lilies

Fertile plants undergoing male gametogenesis can be treated with nitrous oxide (N₂O) gas to obtain 2n male gametes. N₂O treatment is also expected to restore the fertility of interspecific hybrids through meiotic restitution or mitotic amphidiploidization. However, this technique has few applications to date, and it is un-known how N₂O treatment restores fertility in sterile hybrids. To establish optimal N₂O treatment conditions and determine its cytological mechanism of action, we treated various sized floral buds with N₂O gas at different anther developmental stages from fertile and sterile hybrid lilies. N₂O treatment using the optimal 1–4 mm floral buds induced mitotic polyploidization of male archesporial cells to produce 2n pollen in fertile hybrid lilies. In sterile hybrid lilies, N₂O treatment doubled the chromosome number in male archesporial cells followed by homologous chromosome pairing and normal meiosis in pollen mother cells (PMC), resulting in restoration of pollen fertility. Backcrossing the resultant fertile pollen to Lilium × formolongi produced many triploid BC₁ plants. Thus N₂O treatment at the archesporial cell proliferating stage effectively overcame pollen sterility in hybrid lilies, resulting in fertile, 2n pollen grains that could produce progeny. The procedure presented here will promote interspecific or interploidy hybridization of lilies.     

Occurrence of 2n gametes in the F1 hybrids of Oriental × Asiatic lilies (Lilium): Relevance to intergenomic recombination and backcrossing

Cytological modes of the origin of 2n gametes were investigated in six different genotypes of F₁ hybrids between Oriental and Asiatic (OA) lilies (Lilium, 2n = 2x = 24). Chromosome pairing between the parental genomes was very low, the average frequency range from 0.3 to 1.2 bivalents per cell among the genotypes. Within a genotype the frequency of bivalents varied from 0 to 6 in some cases. The normally occurring haploid pollen grains were totally sterile. In contrast, in different genotypes, variable percentages of 2n pollen were found and shown to be fertile as estimated from pollen germination. A cytological analysis of Metaphase I and subsequent stages of meiosis using genomic in situ hybridization (GISH) revealed that there was intergenomic recombination between the alien genomes. Following Metaphase I stage, three different types of abnormal cytological events led to the formation of 2n pollen: (i) Post-Metaphase I division (PMI), (ii) Post-Metaphase II division (PMII) and (iii) Asymmetric Cytokinesis of the pollen mother cell followed by chromosome division. All three cytological events led to first division restitution (FDR) gametes. Based on in vitro pollen germination it was proved for two genotypes that 2n pollen was viable only during the first day of anthesis. It was possible to use 2n pollen successfully for backcrossing. Implications of 2n pollen for intergenomic recombination in BC₁ progenies are discussed.     

A treatment to improve the vase life of cut tulips: Effects on tepal senescence, tepal abscission, leaf yellowing and stem elongation

The vase life of cut tulips (Tulipa spp.) is limited by a combination of leaf yellowing, tepal senescence, and tepal abscission. In many cultivars, moreover, high rates of stem elongation result in stem bending during vase life. In tests with the cvs. Apeldoorn and Frappant, stem bending could be prevented by treatment with ethylene or ethephon. However, these treatments resulted in poor flower opening. The ethephon treatment also resulted in precocious tepal abscission. The negative effect of ethephon on flower opening was overcome by a treatment with gibberellic acid (GA₃). GA₃ also somewhat delayed early leaf yellowing, but did not prevent the early tepal abscission induced by ethephon. The latter problem was overcome by a treatment with benzyladenine (BA). In addition, BA effectively delayed leaf yellowing and also delayed tepal senescence. However, BA produced browning of the lower stem end. This was prevented by the inclusion of calcium ions in the solution. The combination of chemicals (ethephon, GA₃, BA, and calcium ions) was similarly successful in a large number of other tulip cultivars tested. After adaptation of the concentrations the four chemicals were also effective if given as a pulse treatment shortly after harvest.     

Ethanol vapor and saprophytic yeast treatments reduce decay and maintain quality of intact and fresh-cut sweet cherries

The objective of this study was to evaluate the use of an ethanol vapor release pad and a saprophytic yeast Cryptococcus infirmo-miniatum (CIM) to reduce decay and maintain postharvest quality of intact or fresh-cut sweet cherries (Prunus avium) cv. Lapins and Bing. Intact or fresh-cut fruit were packed in perforated clamshells (capacity 454 g) and stored at 1, 10 or 20 °C for up to 21, 14 and 8 d, respectively. For ethanol treatment, a pad made with silica gel powder containing 10 g ethanol and covered with perforated film, which allows ethanol vapor to diffuse gradually, was attached to the upper lid of the clamshells. Ethanol treatment caused accumulation of ethanol in the packaging headspace, about 10 μL L⁻¹ with little change within 14 d at 1 °C, 23 μL L⁻¹ at d 1 and decreased to 15 μL L⁻¹ at d 10 at 10 °C, and 26 μL L⁻¹ at d 1 and decreased to 13 μL L⁻¹ at d 3 at 20 °C. Ethanol content in fruit was less than 9 mg kg⁻¹ in all the control fruit, and increased to 16, 34 and 43 mg kg⁻¹ in ethanol-treated fruit at 1, 10 and 20 °C, respectively. Nonetheless, a sensory taste panel did not perceive any flavor difference from the ethanol treatment. The ethanol treatment retarded softening, darkening, and acid decrease in fruit as well as discoloration of the stems, and extended shelf-life of intact cherries. Ethanol reduced brown rot (Monilinia fructicola) in fresh-cut cherries stored at 20 °C, but not at 1 and 10 °C. A pre-packaging dip in CIM completely controlled brown rot in inoculated fresh-cut cherries stored at 1 °C, and in naturally infected cherries at 20 °C.     

Effect of meta-topolin on leaf senescence and rooting in Pelargonium × hortorum cuttings

Pelargonium (Pelargonium x hortorum) is grown as potted or bedding plants for their colourful, showy flowers and scented foliage. Absence of senescence symptoms in the leaves of Pelargonium cuttings, their capacity to initiate roots and continued growth of initiated roots is an important quality attribute. The effects of postharvest treatments with meta-topolin (mT) and thidiazuron (TDZ) to ‘Katinka’ Pelargonium cuttings were investigated. Leaves treated with mT or TDZ for 5 d had higher leaf chlorophyll contents than untreated controls. Exposing cuttings to mT had no effect on the rooting proportion (%) and average root diameter. Similarly, 0.05 mM mT had no effect on number of roots per cutting. However, mT slightly reduced root length, root surface area and total volume of the roots. TDZ severely inhibited adventitious root formation, thus it reduced all the root parameters investigated. In conclusion, mT is very active in retarding leaf senescence, and combined with the observed ease of rooting of cuttings after mT treatment, this treatment is a suitable alternative to TDZ in delaying the onset of leaf yellowing in ornamental crops.     

The combination of curing with either chitosan or Bacillus subtilis CPA-8 to control brown rot infections caused by Monilinia fructicola

Recently, it has been reported that brown rot in peaches and nectarines can be effectively controlled by exposing fruit to 50 °C for 2 h and 95-99% relative humidity (RH). This treatment was effective at reducing infections that had become established in the field. However, it did not provide protection for further Monilinia fructicola infections, indicating that fruit was susceptible to subsequent infections after the treatment process and before cool storage. Chitosan and Bacillus subtilis (strain CPA-8) were evaluated for their ability to prevent M. fructicola infections and for their ability to complement the heat treatment. Two chitosan concentrations (0.5% or 1%) were applied at three temperatures (20, 40 or 50 °C) for 1 min to wounded and unwounded fruit that were artificially inoculated with M. fructicola. One percent chitosan applied at 20 °C had a preventive effect against further M. fructicola infections on heat-treated fruit that had been previously inoculated: brown rot incidence was reduced to 10%, in comparison with the control (73%). However, chitosan applied to wounded fruit had a poor preventive effect. The antagonist, B. subtilis CPA-8, had a preventive effect in controlling M. fructicola infections: the incidence of brown rot was reduced to less than 15% for both varieties evaluated (‘Baby Gold 9’ and ‘Andros’ peaches), in comparison with the control fruit (higher than 98%). In contrast, when fruit were stored at 0 °C, this preventive effect was not detected. These findings indicate that heat-treated fruit can be protected from subsequent fruit infection after heat treatment by use of chitosan or B. subtillis CPA-8, thereby providing packinghouses with an effective biologically based, combined approach to the management of postharvest brown rot.     

Characterization and genetic mapping of eceriferum-ym (cer-ym), a cutin deficient barley mutant with impaired leaf water retention capacity

The cuticle covers the aerial parts of land plants, where it serves many important functions, including water retention. Here, a recessive cuticle mutant, eceriferum-ym (cer-ym), of Hordeum vulgare L. (barley) showed abnormally glossy spikes, sheaths, and leaves. The cer-ym mutant plant detached from its root system was hypersensitive to desiccation treatment compared with wild type plants, and detached leaves of mutant lost 41.8% of their initial weight after 1 h of dehydration under laboratory conditions, while that of the wild type plants lost only 7.1%. Stomata function was not affected by the mutation, but the mutant leaves showed increased cuticular permeability to water, suggesting a defective leaf cuticle, which was confirmed by toluidine blue staining. The mutant leaves showed a substantial reduction in the amounts of the major cutin monomers and a slight increase in the main wax component, suggesting that the enhanced cuticle permeability was a consequence of cutin deficiency. cer-ym was mapped within a 0.8 cM interval between EST marker {"type":"entrez-nucleotide","attrs":{"text":"AK370363","term_id":"326489160","term_text":"AK370363"}}AK370363 and {"type":"entrez-nucleotide","attrs":{"text":"AK251484","term_id":"151420132","term_text":"AK251484"}}AK251484, a pericentromeric region on chromosome 4H. The results indicate that the desiccation sensitivity of cer-ym is caused by a defect in leaf cutin, and that cer-ym is located in a chromosome 4H pericentromeric region.     

Seventh supplementary list of wheat varieties classified according to their genotype for hybrid necrosis and geographical distribution of Ne-genes

Summary The seventh supplement of the genotype hybrid necrosis in Triticum is presented. This list contains 414 Ne ₁-, 179 Ne ₂- and 662 non-carriers. The total number of varieties tested is 4629 viz. 1298 (28.0%) Ne-, 1031 (22.3%) Ne ₂- and 2300 (49.7%) non-carriers.The various aspects reported in literature and those studied by me are discussed.The geographical distribution of Ne-genes and their alleles indicates two Ne ₁ ^w -areas, one Ne ₁ ^m -area, probably two Ne ₂ ^ms.s -areas and four non-carrier-areas.     

甘蓝型油菜黄化突变体的光合特性及叶绿素荧光参数分析

调查油菜自发黄化突变体(NY)、野生型(NG)及其正反交后代材料(F₁和rF₁)的光合色素含量、光合特性、叶绿素荧光参数及农艺性状,分析五叶期各参数的变化规律。表明,突变体叶绿素a、叶绿素b、类胡萝卜素和总叶绿素均大幅减少,其中叶绿素b减幅最大;净光合速率显著降低,胞间CO₂浓度升高,但气孔导度与野生型等相当,表明光合速率不受气孔限制;光补偿点和光饱和点升高,暗呼吸速率与野生型等相当,表观量子效率和光补偿点处量子效率显著降低;CO₂补偿点、光呼吸速率和羧化效率均显著降低,CO₂饱和点则显著升高;突变体的荧光参数,包括F_o、F_m、F_v/F_m、F_v'/F_m'、Φ_PSII、q_p、NPQ和ETR均显著降低,说明光合色素含量降低导致PSII反应中心捕光能力弱和光化学转化效率低,使叶片光合速率降低。突变体的黄化持续时间较长,对生长发育产生影响较大,单株籽粒产量只有野生型的57.09%,但与正常材料组配F₁的光合特性和农艺性状均能恢复到正常水平。     

Evaluation of wild Arachis species for abiotic stress tolerance: I. Thermal stress and leaf water relations

The Indian groundnut cultivars have a narrow genetic base. Hence, it was of interest to investigate the genetic variability among wild Arachis species and their accessions for tolerance to thermal stress. A wide variation was observed in leaf morphological characters such as colour, shape, hairiness, length and width and thickness (SLA). The temperature and time required for 50% leaf injury was worked out with limited number of genotypes and was found to be 54°C for 50 min. Among 36 genotypes (having SLA in the range of 66 and 161 cm² g⁻¹) screened, the inherent potential for cold as well as heat tolerance in terms of relative leaf injury (RI) was observed. Thus, based on RI-values, A. glabrata 11824 and A. paraguariensis 12042 were identified as heat-tolerant and cold-tolerant genotypes, respectively while A. appresipila 11786 was found to be susceptible to both heat and cold. Correlation between SLA and RI values for heat (r = 0.38, P < 0.05) and cold (r = 0.52, P < 0.05) tolerance was positive, indicating that thicker the leaf the lower the injury or higher the tolerance. Among six species and 13 accessions, comprising both heat-tolerant and heat- susceptible genotypes, the concentrations of various leaf chemical constituents such as total protein, phenols, sugars, reducing sugar, amino acids, proline, epicuticular wax load and chlorophyll varied significantly. The epicuticular wax load ranged between 1.1 and 2.5 mg dm² among 13 A.glabrata accessions. These accessions were categorized into two groups, i.e. high-wax (range: 2.0–2.5 mg dm²) and low-wax types (range: 1.1–1.6 mg dm²). The high-wax type showed a higher diffusion resistance (dr) as compared to low-wax type; though the transpiration rate (tr) in high-wax type was moderate (between 9.5 and 11.6 μg cm⁻² s⁻¹). Genetic variability in parameters such as canopy temperature, dr and tr was also distinct. The fully turgid leaves with relative water content ≥91%, showed leaf water potential (ψ_leaf) between −0.7 and −1.2 MPa. Results indicated that the plants with thicker leaves are better protected from heat injuries. Further, epicuticular wax load seems to help in maintaining stomatal regulation and leaf water relations, thus affording adaptation to wild Arachis species to thrive under water-limited environments. The sources of tolerance, as identified in this study, could be utilized to improve thermal tolerance of the groundnut cultivars by intra-specific hybridization, following either conventional breeding using embryo rescue techniques, if required or utilizing biotechnological tools.     

Gametocidal effects and resistance to wheat leaf rust and stem rust in derivatives of a Triticum turgidum ssp. durum/Aegilops speltoides hybrid

Summary Genes for leaf rust and stem rust resistance and segregation distortion (Gc), that seemed to derive from an Aegilops spetroides ssp. ligustica accession, were transferred to common wheat. While the advanced backcrosses had normal meioses and 42 chromosomes, high levels of male and female sterility, abnormal endosperm development and chromosome aberrations were evident. These effects were more pronounced in Gc-heterozygotes than in homozygotes. Gametes without Gc genes did not survive, and the Gc-associated defects were always inherited with the resistance. Since the resistance genes were effective against local pathotypes of the leaf rust and stem rust pathogens, an attempt was made to disrupt the Gc-system through irradiation, treatment with the mutagen N-nitroso-N-methyl-urea or growing the material at elevated temperatures. A very low frequency of the treated material showed slightly better fertility and seed development. However, these effects did not persist in subsequent generations and were apparently not strong enough to allow the recovery of segregates which had lost the Gc gene(s).