不结球白菜游离小孢子培养成胚影响因素的研究

以17个不同基因型的不结球白菜为供试材料,对影响小孢子胚胎发生频率的若干因素进行了研究。结果表明:在秋冬季日光温室内,供试植株主花序第一朵花开后第7~14天取样,即初花期是最佳取样时期;不同基因型间小孢子胚胎发生能力差异很大,在接种的17个基因型中,有12个诱导出胚,诱导成功率71%;其中102号产胚量最高,达到32.78个/蕾;在已产胚材料中,产胚量最高的基因型是产胚量最低的基因型的546倍;较易抽薹型材料的产胚能力大于易抽薹型和不易抽薹型。以B5有机成分代替NLN有机成分,能够提高产胚量,尤其是对难成胚基因型的小孢子胚胎发生有较好的促进作用,甘露醇对小孢子分裂无促进作用。     

利用小穗培养克服小麦与玉米杂种的幼胚败育

本研究的结果表明,小穗培养是克服小麦与玉米杂种幼胚败育的一个有效手段。让小麦与玉米的杂交穗在植株上生长14～16天时,获得杂种幼胚的频率仅为0.2％,而在授粉后2天采用小穗培养,则使获得杂种幼胚的频率提高到4.2％,获得杂种幼胚的效率提高21倍。     

四种鸢尾属植物种子休眠和萌发研究

燕子花(Iris laeviga ta)、玉蝉花(Iris ensa ta)、溪荪(Iris sanguinea)、黄菖蒲(Iris pseudacorus)是鸢尾科4种喜水湿植物。经萌发实验证明,除溪荪种子没有休眠外,其它3种种子都有休眠。经去除种皮、低温沙藏、赤霉素处理和胚培养4种处理,证明燕子花、玉蝉花、黄菖蒲中抑制种子萌发的主要因素是胚乳。不同种打破休眠的有效方法不同,其中50 mg/L GA3处理可提高黄菖蒲种子的发芽率,达到84.4%,而低温沙藏30 d可使玉蝉花种子的发芽率达到94.4%,低温沙藏90 d则可提高燕子花的发芽率达到87.8%。     

烟草原生质体再生植株的影响因子

原生质体培养和植株再生技术是植物生物工程的基础，植物原生质体的来源、酶解的条件、培养基的组成、培养方法等对原生质体的培养有很大的影响。本文着重对影响烟草原生质体培养再生植株的相关因素进行了综合论述，并讨论了原生质体培养技术在植物育种上的应用潜力。     

Continuous Plant Regeneration from Established Embryogenic Cell Suspension Cultures of Italian Ryegrass and Tall Fescue

The suitability of different protocols was compared for entire plant regeneration by somatic embryogenesis, of the forage plants Lolium multiflorum Lam. (Italian ryegrass) and Festuca arundinacea Schreb. (tall fescue). In the first protocol, miniature embryos were used as starting material, while mature seeds were retained in the other two. Whichever the considered protocol, undifferentiated calli were produced on Murashige and Skoog MS medium supplemented with 2,4-D. The calli were subcultured in the dark on solid MS agar medium, containing 5 mg/1 2,4-D (protocol 2) or on solid MS medium followed by transfer to a rotated liquid MS medium with 2 mg/1 2,4-D (protocol 1). In these conditions, induction of somatic embryogenesis occurred, and whole plants were regenerated during a limited lapse of time, upon transfer in the light, to MS medium supplemented with BAP but devoid of 2,4-D. The simultaneous elimination of 2,4-D and transfer to light appeared essential for full regeneration of the plants. Using this characteristic, an additional step was added to a new protocol (protocol 3) in which microcalli, cultured on liquid MS medium containing 5 mg/1 2,4-D, were transferred to the same medium with 2 mg/1 2,4-D, in the dark. In these conditions, the suspensions kept their embryogenic potential for months. In all cases, plantlets were successfully transferred into the soil. An evaluation of the somaclonal variation potential of the plants issued from each protocol is now underway.     

Effect of genotype,environment and carbohydrate on anther culture response in head cabbage (Brassica oleracea L. convar. capitata (L.) Alef.)

Summary The effect of genotype, growing conditions for donor plants and type and concentration of carbohydrate in the culture medium was investigated for anther culture of head cabbage (white cabbage, savoy cabbage, pointed-headed cabbage). Strong genotypic effects on embryo formation from the cultured anthers were shown as well as superior embryo formation from anthers of field grown donor plants compared to plants grown in the greenhouse. When comparing 7, 10 and 13% sucrose in the medium, embryo response increased with increasing sucrose concentration. With maltose, which was generally inferior to sucrose as carbohydrate source for anther culture, the embryo response did not increase with maltose concentration above 10 per cent.     

Anther Culture Response in Perennial Ryegrass (Lolium perenne L.)

20 diploid clones from 7 varieties, and 10 tetraploid clones from 3 varieties of Lolium perenne were tested in replicated anther culture experiments. Embryos or calluses, were obtained from all clones, and plants were regenerated from all clones except one. The total yield of plants (albino and green plants) ranged from 0 to 61 plants per 100 cultured anthers among genotypes, and there was a general tendency for tetraploic genotypes to be more responsive. Viable green plants were obtained from 5 diploid and 7 tetraploid clones representing 2 and 3 varieties, respectively. Their origin from reduced pollen was confirmed by a haploid chromosome number in some regenerants and by homozygosity for isozyme loci in spontaneously chromosome doubled plants produced from heterozygous diploid donor plants. A large number of the plants were successfully established in the soil. For most donor genotypes, green plants were rare exceptions, but two diploic clones consistently produced 2.3 and 3.8 green plants per 100 cultured anthers, respectively. Estimates of variance components from replicates with greenhouse and field-grown donor plants showed that genotypes accounted for about 73 per cent of the total variation in yield of embryos/calluses, while only 14—15 per cent of the total variation was due to replicates. Hence at present, emphasis should be placed on die selection of high-response genotypes in material of high agronomic potential.     

Genetic Variation for Regenerative Response in Immature Leaflet Cultures of the Cultivated Peanut,Arachis hypogaea*

Forty-seven genotypes of the cultivated peanut, Arachis hypogaea L., were screened for regenerative responses of immature leaflet cultures on two media varying in the auxin: cytokinin ratio. Statistical analysis revealed significant media and genotype differences for rhizogenesis plus media differences for callus proliferation but no differences for shoot formation. When genotypes were categorized according to botanical types, differences between subspecies were observed for rhizogenesis. Cultivars of the Virginia botanical type (subspecies hypogaea) were significantly different from either the Valencia or Spanish botanical types (both subspecies fastigiata). Overall, rhizogenesis varied from 0 to 70% among all genotypes. Shoot production was low (30%) while callus proliferation was extremely high (70%).     

寒地早粳花培育种研究进展

在花药培养技术上,通过49个组合400个幼穗1200个颖花的研究,提出了依据植株和颖花形态判断花粉发育时期的具体指标;通过低温预处理和双态双层法提高花药力的方法。在花培育种遗传规律的研究上,对株高和叶瘟抗性的结果表明:F2的分布倾向于晚熟、高秆及抗病,即倾向于亲本的显性性状,而H2的分布则均匀对称,其中早熟、矮秆及感病植株的比例较F2高。在选择效率上,花培育成的审定品种的选择效率为0.93%,是同期常规育成审定品种选择效率0.28%的3.32倍。通过上述研究培育出了8个花培新品种,并且在品质、抗性、产量等方面表现突出,成为寒地稻区主栽品种,并讨论了水稻花培育种今后的发展方向。     

Segregation distortion in doubled haploid lines of barley (Hordeum vulgare L.) detected by simple sequence repeat (SSR) markers

A total of 147 simple sequence repeat (SSR) markers (including86 barley and 61 wheat microsatellite markers) were tested for their segregation in a doubled haploid (DH) and an F₂ population of barley. The DH population consisted of 71 doubled haploid lines, developed from F₁ plants of a cross between Tadmor and WI2291using isolated microspore culture technique. A genetic linkage map consisting of 43 microsatellite markers was constructed using the DH population. Particularly on chromosome 4H microsatellite markers showed distorted segregation ratios. Segregation of DH lines based on molecular markers were compared with segregation of 92 F₂ lines from the same cross. The proportion of loci deviating from the expected monogenic segregation ratios in the DH population was significantly higher (19/43loci, 44%) than in the F₂ population (7/43 loci, 16%). The deviation was biased towards the WI2291 parent alleles. In line with this observation, WI2291 was found to perform better than Tadmor in regenerating green plantlets with the isolated microspore-culture technique. This revised version was published online in July 2006 with corrections to the Cover Date.