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.     

Somatic embryogenesis in pea (Pisum sativum L. and Pisum arvense L.): Diallel analysis and genetic control

Summary Six lines of Pisum were tested in vitro for their ability to produce somatic embryos from apices. Significant quantitative variation was observed. Inheritance of the ability to form somatic embryos was studied using a diallel cross among six different lines. About 80% of the observed genotypic variation was due to additive effects. There is a tendency for the favourable genes to be recessive. It appears that there are two genetic systems involved. Analysis of the distribution of F₃ families means from a cross among two extreme lines seems to indicate the presence of a few major genes in the control of somatic embryogenesis of pea.     

Wheat embryogenesis and haploid production in wheat × maize hybrids

Summary Embryogenesis was analyzed in wheat × maize hybrids using paraffin sectioning. Embryogenesis in wheat × maize hybrids is different from that in self-pollinated wheat plants. Development of the embryo is not accompanied by the formation of an endosperm. The endosperm nuclei remain free in the cytoplasm, fail to advance into the cellular stage, and degenerate at a later time. The antipodal cells quickly degenerate in the fertilized ovaries of wheat × maize hybrids similar to self-pollinated ovaries. The antipodal cells remain normal in unpollinated ovaries. The pre-embryo will abort if it is allowed to develop on the plant, because of a nutritional shortage in the absence of an endosperm. Therefore, embryo rescue is necessary for haploid production from a wheat × maize hybrids. Haploid polyembryos were obtained from spikelet culture of wheat × maize hybrids. The formation of polyembryos is due to the cleavage of the pre-embryo and the effect of 2,4-D. The frequency of haploid embryo production and plant regeneration is affected significantly by maize genotypes, but not by wheat genotypes. The concentration of 2,4-D affects only the size of the embryo.     

Intra-varietal variation for in vitro plant regeneration in the genus Trifolium

Summary Seedlings of Trifolium repens showed considerable variation with regard to the morphology and growth of their calli, and their ability for in vitro differentiation of shoots. One of the lines selected for regeneration in primary callus cultures also showed shoot formation from protoplasts. Somatic embryogenesis in callus cultures of T. pratense and T. arvense occurred only in selected seedling lines. This paper highlights the importance of screening a large number of plants within a cultivar of outbreeding species to achieve reproducible plant regeneration from tissue culture.     

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.     

Functional genomics of microspore embryogenesis

Isolated plant microspores, when stressed and cultured in vitro, can be diverted from their normal gametophytic pathway towards sporophytic development, with the formation of haploid embryos and ultimately doubled-haploid plants. This process is called androgenesis or microspore embryogenesis, and is widely used in plant breeding programmes to generate homozygous lines for breeding purposes. Protocols for the induction of microspore embryogenesis and the subsequent regeneration of doubled haploid (DH) plants have been successfully developed for more than 200 species. These practical advances stand in stark contrast to our knowledge of the underlying molecular genetic mechanism controlling this process. The majority of information regarding the genetic and molecular control of the developmental switch from gametophytic to sporophytic development has been garnered from four intensely studied (crop) plants comprising two dicotyledonous species, rapeseed (Brassica napus) and tobacco (Nicotiana tabacum), and two monocotyledonous species, wheat (Triticum aestivum) and barley (Hordeum vulgare). In these species the efficiency of microspore embryogenesis is very high and reproducible, making them suitable models for molecular studies. In the past, molecular studies on microspore embryogenesis have focussed mainly on the identification of genes that are differentially expressed during this developmental transition and/or early in embryo development, and have identified a number of genes whose expression marks or predicts the developmental fate of stressed microspores. More recently, functional genomics approaches have been used to obtain a broad overview of the molecular processes that take place during the establishment of microspore embryogenesis. In this review we summarise accumulated molecular data obtained in rapeseed, tobacco, wheat and barley on embryogenic induction of microspores and define common aspects involved in the androgenic switch.     

小孢子培养获得松花型花椰菜DH再生植株

对5个松花型花椰菜(Brassica oleraceavar.botrytisL.)杂种一代的小孢子培养研究表明,小孢子胎胚发生主要依赖于基因型,庆农65天的每花蕾胚状体产量最高,平均达15.5个。松花菜的胚状体萌发率一般在30%左右,并有效地获得了大量的DH再生植株。冷激预处理能显著地影响花椰菜小孢子的胚胎发生,但供体材料间存在不同的结果。结球期和开花结角期再生植株的生育期与育性出现较大分离,可育且能正常结角的比例约占全部小孢子再生植株的50%以上,因而不再需要加倍处理。     

植物生长调节剂在植物体细胞胚发生中的应用

合理应用植物生长调节剂是成功诱导体细胞胚的决定因素,也是影响体细胞胚高频率发生的主要因素。本文从生长素类、细胞分裂素类、赤霉素、脱落酸以及各类植物生长调节剂间组合使用等几个方面综述了外源植物生长调节剂在体细胞胚发生中的应用进展,为植物体细胞胚的诱导提供了一定的参考价值。     

INDUCTION OF SOMATIC EMBRYOS AND REGENERATION OF PLANTLETS FROM RICE ROOTS

<正> The number of somatic embryos and regenerated plantlets were directly induced from irra-diated rice roots of the variety Taipei 309(Oryza sativa L.).For the induction of somatic em-bryos,the suitable age of roots was no more than 7 days after seed culture on the medium and thesections near tip were more vigorous than those near base of a root;the number of somatic em-bryos induced by the media with 1.11mg/L2,4-D plus 0.47mg/L NAA and 2.21mg/L 2,4-Dplus 1.86mg/L NAA was much higher than that induced by 1.11mg/L 2,4-D alone;10～20 Gyand 40 Gy of γ irradiation were the most favourable for increasing somatic embryo number onthe media of 2.21mg/L 2,4-D plus 1.86mg/L NAA and 1.11mg/L 2,4-D plus 0.47mg/L NAAor 1.11mg/L 2,4-D alone respectively.The highest number of regenerated plantlets was fromthat cultured on the medium of 1.11mg/L 2,4-D plus 0.47mg/L NAA and irradiated with 30～40 Gy of γ rays.     

通过花药漂浮培养提高花椰菜小孢子胚胎发生率

通过花药漂浮培养,成功地诱导两个秋花椰菜基因型的小孢子胚胎发生.在液体培养基与固、液体双层培养基上,小孢子胚胎产量相仿,但后者褐色胚产生较多.在液体培养基中加入62.5mg/L的硝酸银对诱导小孢子胚胎发生效果较好.试验结果表明,采用花药漂浮培养方法可以明显提高花椰菜小孢子胚产量.