Overcoming the self-incompatibility of Lilium longiflorum Thunb. by application of flower-organ extract or temperature treatment of pollen

Summary The application of flower organ extracts to stigmas and the temperature treatment of pollen were tried to overcome self-incompatibility of Lilium longiflorum cv. Georgia.Substances in stigma, style, ovary and anther were extracted with ethanol and fractionated with ethylacetate into the acidic, basic and aqueous fractions. The extracs melted in a small volume of distilled water were applied to stigmas prior to self-pollination. Hinomoto stigma extract, self-pollinated and cross-pollinated Georgia stigma extracts of high concentrations and Georgia anther extract of high concentration were effective in overcoming the self-incompatibility and resulted in a high percentage of fruit set and many normal seeds. Extracts from Hinomoto ovary, style and anther were ineffective, except a basic fraction, which was very slightly effective.Pollen was treated with 40°C for 60 or 90 minutes and 50°C for 30 or 60 minutes, and a half of each lot was followed by –20°C for 24 h, prior to self-pollination. All treatments were effective, especially at 40°C for 60 minutes or 50°C for 30 minutes, and 40°C for 90 minutes or 50°C for 60 minutes followed by –20°C for 24 h.     

麝香百合的抗热生理指标初探

以田间抗热表现不同的麝香百合品种White Forest和新铁炮百合基因型O2-28、O1-13作为试验材料进行38℃高温处理，测定百合叶片的可溶性蛋白含量、过氧化物酶活性、过氧化氢酶活性、蒸腾强度4个生理指标的变化，结果显示百合的可溶性蛋白含量均下降,抗热性越强的基因型下降幅度越缓慢。酶活性与抗热性正相关，抗热基因型的两种酶活性提高的幅度大大高于热敏感基因型。蒸腾强度的变化没有明显规律，与百合抗热无相关性。     

A reproductive calendar of Lilium longiflorum Thunb. cv. Gelria

Summary The development and timing of different processes in the floral biology and sexual reproduction of Lilium longiflorum were studied and brought together into a chronological reproductive calendar. Care must be taken when emasculating flowers later than 4 days before anthesis (DBA), because the stomium of the anther was open from 8 DBA and the pollen grains appear to be mature from 3–2 DBA. The receptivity period of the pistil lasts for a rather long period, but before wilting of an unpollinated flower as determined by a change of the colour of the perianth from white into brown, the flower had already lost its receptivity. Because L. longiflorum pollen grains can easily be stored, the pistil receptivity should thus determine the pollination schedule. The relationships between pollen tubes and pistil are discussed with respect to plant breeding.Abbreviations DAA Days After Anthesis - DAP Days After Pollination - DBA Days Before Anthesis     

商陆抗病毒蛋白基因在麝香百合中的转化和表达

以麝香百合叶片愈伤组织为受体,利用根瘤农杆菌介导法将美洲商陆蛋白（PAP）基因和抗卡那霉素筛选基因以共转化的方式转入百合叶片愈伤组织中,然后在含有MS培养基中筛选愈伤组织并得到再生植株,在建立的农杆菌转化百合的遗传转化体系中,获得49株再生植株,经PCR检测表明PAP基因已经转移到有抗性愈伤组织再生出百合植株中。     

麝香百合花蕾组织原位杂交体系的建立

本研究以麝香百合未开放的花蕾为实验材料,以百合花发育相关的B功能基因LLGLO1的特异片段为探针,研究了花蕾切离时间对RNA结构完整性的影响,探讨了蛋白酶K消化浓度及时间对材料内蛋白质消化及材料结构完整性的影响.研究结果表明:花蕾离体1.5 h内RNA仍然保存完整,花蕾离体2hRNA开始降解,因此,花蕾取材后应在1.5...     

Incompatible pollen tube growth and protein composition in styles of Japanese pear following high temperature treatments

Summary Immersing the excised-prepollinated styles of self-incompatible Japanese pear, Pyrus serotina Rehd., into distilled water at 45°C for 1.5–2.5 min promoted the growth of self-pollen tubes; both the number of styles penetrating the pollen tubes through styles to cut ends and of pollen tubes protruding from cut ends were increased. The subsequent growth of the protruded pollen tubes on agar media was also enhanced by the treatments. Treatments at 40°C did not promote the growth of self-pollen tubes and those at 50°C led the styles to necrosis. In the stylar proteins analyzed by isoelectricfocusing polyacrylamide gel electrophoresis, several changes in banding pattern were found following the heat treatment; one of the bands was a glycoprotein that increases in quantity in developing styles as self-incompatibility of the style becomes stronger (Hiratsuka et al., 1986). This protein band was stained more weakly by Coomassie Brilliant Blue-G250 dye. Antigenic substances in the styles examined by immunodiffusion seemed not to be denatured by high temperature treatments.Abbreviations IEF-PAGE isoelectricfocusing polyacrylamide gel electrophoresis - CBB Coomassie Brilliant Blue     

Development of diploid and triploid interspecific hybrids between Lilium longiflorum and L. concolor by ovary slice culture

Ovary slice culture, after cut-style pollination, was used to develop interspecific hybrids between Lilium longiflorum and L. concolor. Reciprocal crosses between diploid cultivars (2n = 2x = 24) were carried out. On the days 30, 35, 40 and 45th after pollination (DAP), ovaries were sliced and cultured on a modified hormone-free Murashige-Skoog (M–S) medium without NH4NO3, supplemented with 6% sucrose, 50 mg/1 yeast extract and 0.25% gelrite at pH 6.3. For the L. longiflorum × L. concolor cross, ovule germination was found to be best at 30 DAP. After transfer to a M–S (half-strength) medium supplemented with 1.5% sucrose and 0.25% gelrite at pH 5.8, diploid and triploid hybrid plants were established. In contrast, ovules from the L. concolor × L. longiflorum cross did not germinate. The hybridity of the plantlets obtained was verified by karyotype and isozyme analysis. The importance of the ovary slice culture technique as a tool to develop new hybrids between incompatible lilly plants is discussed.     

Production of diploid and triploid interspecific hybrids between Lilium concolor and L. longiflorum by in vitro ovary slice culture

Successful hybridization between Lilium concolor and Lilium longiflorum has not been reported but ovary slice culture technique, after cut-style pollination has now been used to produce diploid and triploid interspecific hybrids between these species. Reciprocal crosses between diploid cultivars (2n= 2x= 24) were conducted. 5, 10, 15, 20, 25, 30, 35, 40, and 45 days after pollination (DAP), ovaries were sliced and cultured on a modified Murashige and Skoog (MS) medium without growth regulators and NH₄NO₃, supplemented with 6% sucrose, 50 mg/1 yeast extract and 0.25% gelrite at pH 6.3. For the L. concolor × L. longiflorum cross, embryo germination was found to be best at 20 DAP, while for the L. longiflorum × L. concolor at 25 DAP. After transfer to a MS (half-strength) medium supplemented with 1.5% sucrose, 0.25% gelrite and 0.2% active charcoal at pH 5.8, diploid and triploid hybrid plants were developed. All regenerated plants were identified as hybrids on the basis of karyotype and isozyme analyses. Ovary slice culture technique as a method of producing polyploids is discussed.     

Breakdown of self-incompatibility in perennial ryegrass at high temperature and its uses in breeding

Summary The maximum temperature (34°C) at which perennial ryegrass plants from one breeding population successfully shed pollen was determined and the seed set of ten clones taken from the same population and self-pollinated both in an unheated glasshouse and at a constant 34°C during anthesis was compared. Seed set by selfing of all ten clones was markedly increased by heat treatment, from a mean of 2.3% to 30.7%. Heat treatment of anthers, but not stigmas, greatly enhanced pollen tube growth on stigmas from the same genotype. Germination of progeny was not affected by the heat treatment. Heat treatment of ten clones from a breeding population with contrasting morphology and growth rate was not so successful. Only one clone shed pollen freely at 34°C and showed a marked improvement in subsequent seed set. The advantages to the breeder of routinely producing full-sib families of elite perennial ryegrass parents by selfing are outlined.     

Effects of low temperature storage and sucrose pulsing on the vase life of Lilium cv. Brindisi inflorescences

Lilium cv. Brindisi inflorescences were stored at 2.5 °C for 5, 10, 15 or 20 d, comparing dry storage with storage of the stem ends in water. Prior to storage, inflorescences were treated with 20 or 100 g L⁻¹ sucrose in water, for 20 h at 20 °C. After storage the inflorescences were individually placed in water at 20 °C. The floral buds were still closed at the end of cold storage. In experiments carried out in summer, the time to bud opening was hastened by storage at 2.5 °C in water, more so after a longer period of cold storage. The time to tepal senescence after cold storage in water decreased with the time of storage. The time to tepal abscission was about 1 day longer than the time to tepal senescence. Repeat experiments in late fall and winter additionally showed early leaf yellowing after cold storage. Compared to the experiments in summer, more desiccated floral buds were found in the fall. Pulse treatment with 100 g L⁻¹ sucrose prior to cold storage reduced the number of desiccated buds. However, leaf yellowing was aggravated by the 100 g L⁻¹ sucrose pulse treatment. Compared to cold storage in water, dry storage at 2.5 °C further hastened the time to bud opening and also further hastened tepal senescence and abscission. Dry storage also produced more buds that desiccated or opened poorly. Sucrose treatment (100 g L⁻¹) alleviated the effects of dry storage on tepal senescence and bud desiccation. The data showed that lily cv. Brindisi inflorescences are prone to chilling injury, but can be stored, depending on the treatment, for 5–10 d, during most of the year.     

Levels of self-incompatibility in wallflower,Cheiranthus cheiri L.

Summary Approximately 1000 plants from 30 wallflower cultivars were selfpollinated in the open flower and bud stage. Only a very small number of plants failed to set seed and the averages ranged from 3.3 to 11.0 seeds per siliqua. Cultivars with mixed colours appeared to show most self-incompatibility and it is suggested that they form un-evolved populations in terms of the incompatibility system by comparison with their related single-colour cultivars.S1 progenies tended to remain at the same self-incompatibility level as their parents although some segregation occurred with approximately the same number of plants from low to high self-incompatibility and vice-versa.S1 progenies showed no apparent inbreeding depression, as is the case also for their descendants, but there was a tendency for progeny heterogeneity to be associated with high parental self-incompatibility while homogeneous inbreds were most often derived from more self-compatible parents.     

甘蓝SI相关基因BoCDPK14的克隆与分析

甘蓝自交不亲和性(self-incompatibility,SI)是柱头对相同单倍型的花粉产生的排斥或抑制反应。钙依赖蛋白激酶(calcium-dependent protein kinase, CDPK)是植物面对逆境信号时参与抗逆反应的重要元件。本文通过甘蓝自花授粉0～60min的柱头转录组数据分析,成功地筛选到一个受自花授粉诱导上调表达的基因BoCDPK14,该基因与拟南芥中参与植物逆境信号传导的钙依赖蛋白激酶基因高度同源。BoCDPK14基因开放阅读框1599bp,编码一种具有533个氨基酸残基的亲水性蛋白,可在大肠杆菌胞质中被诱导表达,其相对分子质量为60.4kD,表明BoCDPK14为活性胞质蛋白。该基因起始密码子上游2000bp的核苷酸序列中含有胁迫反应、激素反应、代谢调节等应答元件。BoCDPK14在甘蓝柱头、花粉、花蕾、花瓣和叶片中表达,且柱头中的表达量低于花粉。荧光定量PCR结果证实,BoCDPK14在0～60min的表达变化趋势与转录组分析结果一致。通过酵母双杂交发现,BoCDPK14蛋白激酶结构域与谷氨酸受体通道蛋白BoGLR2.8d存在相互作用,表明BoCDPK14可能是参与SI反应过程的新蛋白。本研究结果表明BoCDPK14可能作为Ca~(2+)信号元件参与甘蓝柱头响应花粉刺激的分子过程,这为甘蓝自交不亲和的进一步研究和利用提供了新内容。     

Selection for high self-incompatibility in inbred lines of brussels sprouts

Summary An attempt to improve the self-incompatibility of an advanced inbred line of Brussels sprouts by selection has been unsuccessful. The apparent plant-to-plant variation in self-incompatibility has little or no genetic basis. It results from environmental effects and from intrinsic physiological differences between flowers on a single plant. A temperature rise from 17°C to 26°C markedly increases the self-compatibility of the lines tested, but no evidence was found for end-of-season compatibility. The best methods for producing inbreds with high and consistent self-incompatibility are discussed.     

生长素参与淡黄花百合珠芽发育调节

百合是一类重要的经济/观赏植物,包含百合属的多个物种或杂交品种。珠芽繁殖是多种百合的重要营养繁殖途径,但其发育的分子机制少有研究。本研究利用转录组测序技术对来自淡黄花百合(Lilium sulphureum) 5个部位的混合样本进行测序获得6.64 Gb数据,组装得到51 450个Unigene,32 784个Unigene在NR等功能数据库中得到注释(其中487个与生长素代谢及信号转导相关基因同源)。利用表达谱测序技术分别对5个组织样品进行测序共得到46 897个Unigene,其中337个与生长素代谢及信号转导相关基因同源的Unigene在珠芽源器官、发育早期或后期珠芽之间差异表达。结果表明淡黄花百合珠芽发育早期依赖于活跃运输、低失活及逐渐增强的自主合成提高内部生长素含量,后期伴随珠芽内生长点分化形成其自主合成和失活代谢变得更加活跃;生长素浓度的变化引起大量信号转导相关基因表达量发生变化从而调节珠芽的发育。本研究结果为进一步探究生长素在珠芽发育过程中的调节作用提供了依据。     

不同百合品种间杂交亲和性研究

以国外引进的性状优良的栽培品种7个作为亲本进行杂交,摸索不同品种间交配亲和性规律,提高育种效率。试验结果表明：4种杂种系不同品种百合系内杂交,后代平均有胚率最高的为OO杂种系（43.63%）,其次为AA杂种系（25.35%）。以OO杂种系的‘多顿’、‘蒙特祖玛’为母本,与其他杂种系杂交,后代平均有胚率高与其他组合,亲和性好。‘多顿’ב穿梭’这个组合的结实率为50%,有胚率为50.8%,杂交亲和性最好。     

百合染色体细胞学研究进展

百合属绝大多数种是2n=24的二倍体,其核型具有稳定性,一般为3B型,存在少数多倍化现象,但广泛存在B染色体,百合染色体核型的差异正是环境因素和结构变异共同作用的结果。通过百合属C-带带型中单套染色体条带数及特征染色体可以清晰地区分形态学相似的百合属植物,但采用尿素法进行G带带纹的鉴定有更高的分辨率。原位杂交技术已运用于百合属植物的区分和杂种后代的鉴定,而GISH较FISH更适用于杂种百合的鉴定。     

Overcoming the self-incompatibility of Raphanus sativus by application of plant hormones,amino acids and vitamines,and by temperature treatment of pollen

Summary Overcoming self-incompatibility by application of three kinds of plant hormones, sucrose, 3 kinds of amino acids and 2 kinds of vitamines was tested in cvs. Honbashi-taibyo Minowase (H-Mino) and Minowase (Mino) of Raphanus sativus. Effects differed between the cultivars. In H-Mino, BA (100 mg/l) and glutamic acid, folic acid and nicotinic acid (500 mg/l) resulted in higher fruit set and higher number of seeds per pollinated flower. In Mino, BA and NAA (100 mg/l) and glutamic acid and glycine (500 mg/l) induced a high number of seeds per pollinated flower. These chemicals, however, induced parthenocarpic fruit set, especially GA3. From the observation of pollen on stigmas washed with glutamic acid, it appeared that the pollen-tube penetrated into a papilla cell after 1 hour and openings of papillae and detached pollen grains and tubes were found after 2 hours as the result of successful pollentube penetration of papillae. Pollen was heated at 50°C for 30, 45 or 60 minutes, at 60°C for 15, 30 or 45 minutes and at 70°C for 10, 20 or 30 minutes prior to self-pollination. In H-Mino, 60 and 70°C were effective, and expecially 60°C for 15 or 30 minutes resulted a higher percentage fruit set and more seeds per fruit. In Mino, although 50–70°C were effective, the mean number of seeds per pollinated flower was lower than in H-Mino.     

Influence of timing of cross-versus self-pollination on self-incompatibility in Brussels sprout inbreds

Summary The impact of cross pollination 30 h, 4 h, and immediately prior to self-pollination, and self-pollination 2, 4, 8, 12, 24 and 30 h prior to cross-pollination was assessed for pollen tube number per style, seed number per siliqua, and proportion of self-seed (sibs) per siliqua, in two inbred lines of brussels sprout. Pollination procedure had a marked effect on the amount of sib-and hybrid seed produced. Cross pollination 30 h prior to self-pollination produced significantly greater numbers of sib progeny, as did self-pollination 8–12 h before cross pollination. Prior application of self pollen reduces the number of pollen tubes in the style. The results of this experiment suggest that self-incompatibility in brussels sprout could better be assessed by self-pollination with a subsequent cross pollination 8 h later, and the determination of amounts of sib and F₁ hybrid seed per siliqua.     

Molecular mechanism of the S-RNase-based gametophytic self-incompatibility in fruit trees of Rosaceae

Self-incompatibility (SI) is a major obstacle for stable fruit production in fruit trees of Rosaceae. SI of Rosaceae is controlled by the S locus on which at least two genes, pistil S and pollen S, are located. The product of the pistil S gene is a polymorphic and extracellular ribonuclease, called S-RNase, while that of the pollen S gene is a protein containing the F-box motif, SFB (S haplotype-specific F-box protein)/SFBB (S locus F-box brothers). Recent studies suggested that SI of Rosaceae includes two different systems, i.e., Prunus of tribe Amygdaleae exhibits a self-recognition system in which its SFB recognizes self-S-RNase, while tribe Pyreae (Pyrus and Malus) shows a non-self-recognition system in which many SFBB proteins are involved in SI, each recognizing subset of non-self-S-RNases. Further biochemical and biological characterization of the S locus genes, as well as other genes required for SI not located at the S locus, will help our understanding of the molecular mechanisms, origin, and evolution of SI of Rosaceae, and may provide the basis for breeding of self-compatible fruit tree cultivars.