水曲柳体细胞胚与合子胚发生的细胞学研究

水曲柳(Fraxinus mandshurica)属木犀科(Oleaceae)白蜡树属,是我国东北重要珍贵硬阔树种之一,主要分布于小兴安岭、长白山、辽宁东部山地等地区, 以材质优良而著称.由于长期不合理的采伐利用,目前可利用的资源急剧减少,已被列为国家三级保护植物(傅立国,1992).进行水曲柳体细胞胚胎发生的研究,在资源保护、树种快繁和基因工程育种上有其重要的现实意义.     

草坪草体胚发生及应用于现代育种

综述了草坪草体细胞胚诱导对高频组培再生体系和高效遗传转化体系建立的重要性,及影响体胚发生的因素,体胚的发生在遗传育种中的应用。     

Monitoring genetic stability inQuercus serrata Thunb. somatic embryogenesis using RAPD markers

Genetic stability of propagules regeneratedvia somatic embryogenesis is of paramount importance for its application to clonal forestry. Random amplified polymorphic DNA (RAPD) markers were used to determine the genetic stability in somatic embryogenesis ofQuercus serrata Thunb. (Japanese white oak). Forty samples from an embryogenic line, consisting of regenerated plantlets, somatic embryos, and embryogenic calli, were examined using 54 decanucleotide primers. A total of 6520 clear reproducible bands obtained from these studies exhibited no aberration in RAPD banding pattern among the tested samples. Our results show that somaclonal variation is absent in our plant propagation system. The genetic stability is discussed in terms of the origin of somatic embryos.     

Morphology of somatic embryogenesis and plantlet formation in tissue cultures of lantern tree (Koelreuteria bipinnata var. integrifoliola)

Somatic embryogenesis of Koelreuteria bipinnata var.integrifoliola was observed,plantlet formation in different types of somatic embryos was studied and the effect of abnormal embryos on plantlet formation was identified.Results show that somatic embryos of K.bipinnata var.integrifoliola include normal embryos,embryos with abnormal cotyledons,vitrified embryos,albino embryos,secondary embryos,linked embryos,embryos with abnormal growing points and embryos with expanding hypocotyl.After 40 d of callus cultur...     

Somatic embryogenesis from cell suspension cultures of aspen clone

Suspension cultures initiated from callus derived from petiole explants of aspen hybrid (Populus tremuloides × P. tremula) produced somatic embryos. Callus was induced on a MS medium supplemented with 5 mg·L–1 2,4-D and 0.05 mg·L–1 zeatin under light conditions. Embryogenic calli were obtained when a subsequent subculture of calli was suspended in the same basal me-dium with 10 mg·L–1 2,4-D. The highest number of globular embryos were induced from embryogenic calli by cell suspension cul-ture in a MS liquid medium supplemented with 10 mg·L–1 2,4-D. Genotype and 2,4-D concentration were vital to the induction of embryogenic calli producing competent cells. Embryogenic calli for each genotype were heterogeneous. Green calli with gel-like consistency could yield more competent cells than light yellow embryogenic calli. However, some globular embryos broke into slices and some developed abnormally after one month of culture under the same or other hormonal conditions.     

Somatic embryogenesis of Pinus sylvestris

Embryogenic cultures (EC) of Scots pine (Pinus sylvestris L.) were initiated from immature embryos. Whole ovules were used as expiants. The responsive period for initiation began just after fertilization and remained throughout the development of first stages of early embryos. The main part of the embryogenic cultures were initiated by the time of cleavage polyembryony. Strong correlation was obtained between degree days and the responsive period. During subsequent years the experiments were repeated in Finland and Sweden. In all cases the responsive period for initiating embryogenic cultures was the same, about two weeks after fertilization.

In 1991–1993, a total of 138 clones of elite pine trees were tested for their ability to initiate embryogenic cultures. Of these, 33% were responsive under our experimental conditions. Based on about 300 ovules per clone the number of embryogenic lines induced by the responding clones varied from 0.2% to 9.0% of the expiants.

Several nutrient media were found to be suitable for initiation and proliferation of ECs. About half of the cell lines responded to abscisic acid by producing maturing embryos. The embryos reached full maturity in cultures of only a few cell lines. Some of the embryos that produced roots were planted in soil and transferred to a greenhouse.     

Somatic Embryogenesis and Plantlet Regeneration in Callus Cultures Derived from Mature Zygotic Embryos of Slash Pine

1TheworkwassupportedbyChineseNationalHighTechnologyDevelopmentProgram"863project"IntroductionSomaticembryogenesishasagreatpotentiaIforrapidProPaationofsuPCriorconifcrspccics.ThefirstreportonsomaticembryogenesisandplantlctrcgencrationinconifersPeCieswasach…     

杉木成熟合子胚器官发生和体胚发生

以杉木成熟合子胚为起始外植体,研究基本培养基、不同种类的细胞分裂素及生长素对杉木器官发生和体胚发生及不定芽生根的影响,并用石蜡切片法观察不定芽和体胚的发生和发育过程.结果表明:DCR 6-BA 1.0mg·L-1 TDZ 0.002 mg·L-1 NAA 0.1 mg·L-1对成熟合子胚诱导不定芽最有效;DCR 6-BA 1.0 mg·L-1 TDZ 0.003mg·L-1 NAA 0.1 mg·L-1是成熟合子胚诱导体胚的最佳培养基;不定芽在DCR基本培养基中可有效伸长;1/4MS IBA 0.2 mg·L-1 NAA 0.1 mg·L-1对不定芽生根最有效.石蜡切片观察表明:不定芽有2种发生方式,一种是表皮及表皮下的3～4层细胞分生组织化发育而成,另一种是外植体内部的小维管束脱分化发育而成.2种发生方式均为直接器官发生途径.     

刺五加体细胞胚状体发生及成苗的研究

以刺五加试管苗叶片为试材,将叶片接种于含有不同种类和浓度激素的培养基上进行培养,研究不同激素对刺五加胚性愈伤组织诱导、胚状体发生及生根的影响。结果表明：诱导叶片胚性愈伤组织发生的最适培养基为MS＋6-BA 1.5 mg.L-1＋2,4-D 1.0 mg.L-1,胚状体生长发育的最适培养基为MS＋6-BA1.0 mg.L-1＋IBA0.5 mg.L-1,生根的最适培养基为1/2MS＋NAA0.4 mg.L-1。     

落叶松体细胞胚胎发生研究进展

综述了欧洲落叶松、日本落叶松、欧日和日欧杂种落叶松、西部落叶松和华北落叶松体细胞胚胎发生的研究进展，对胚性培养物的超低温保存、原生质体培养和基因转化等技术进行了介绍，并对落叶松体细胞胚胎发生的研究趋势作了展望。     

Zygotic and somatic embryo morphogenesis in Pinus pinaster: comparative histological and histochemical study

We compared morphogenesis and accumulation of storage proteins and starch in Pinus pinaster Ait. zygotic embryos with those in somatic embryos grown with different carbohydrate sources. The maturation medium for somatic embryos included 80 microM abscisic acid (ABA), 9 g l(-1) gellam gum and either glucose, sucrose or maltose at 44, 88, 175 or 263 mM in the presence or absence of 6% (w/v) polyethylene glycol (PEG) 4000 MW. Maturation medium containing 44 or 88 mM of a carbohydrate source produced only one or no cotyledonary somatic embryos per 0.6 g fresh mass of culture. The addition of PEG to the basal maturation medium resulted in a low yield of cotyledonary somatic embryos that generally showed incomplete development and anatomical abnormalities such as large intercellular spaces and large vacuoles. High concentrations of maltose also induced large intercellular spaces in the somatic embryonic cells, and 263 mM sucrose produced fewer and less developed cotyledonary somatic embryos compared with 175 mM sucrose, indicating that the effect of carbohydrate source is partially osmotic. Zygotic embryos had a lower dry mass than somatic embryos at the same stage of development. Starch granules followed a similar accumulation pattern in zygotic and somatic embryos. A low starch content was found in cotyledonary zygotic embryos and in somatic embryos developed in the presence of 175 mM maltose or 263 mM glucose. In zygotic embryos and in PEG-treated somatic embryos, protein bodies appeared later and were smaller and fewer than in well-developed somatic embryos grown without PEG. We propose that storage protein concentration might be a marker of embryo quality.     

Somatic embryogenesis inCephalotaxus harringtonia embryo-megagametophyte co-culture

Somatic embryogenesis was initiated fromCephalotaxus harringtonia (Forbes) K. Koch embryo culture. Explants consisted of embryo and megagametophyte halves both cut longitudinally. They were removed aseptically from mature seeds and grown together on a solid Murashige and Skoog modified medium supplemented with 5 mg·l ⁻¹ 2,4-dichlorophenoxyacetic acid. Embryogenic cultures started from callus after three or more months on the primary medium. The embryogenic callus originated from the suspensor region of the embryo. All chromosome counts made in the cells of the embryonic structures demonstrated a diploid stage, which suggest that they originated from zygotic embryo tissue. The early stages of somatic embryogenic development were achieved,i.e., formation of small clusters consisting of an embryonal region made up of isodiametric meristematic cells. A more advanced stage was reached in some cultures in which the distal embryonal end of the embryo appeared smooth and opaque. The ultrastructural characteristics of the embryos, the two types of embryo cells, embryonal and suspensor cells, as well as their contents were similar to those already reported in the case of somatic embryogenesis of other conifers.     

Plant regeneration by somatic embryogenesis in Eucalyptus spp.: current status and future perspectives

Forest tree improvement programs benefit from the emergence of new biotechnological strategies that complement plant developmental biology and discovery of genes associated with complex multigenic traits. Recently, significant progress has been made in the area of plant regeneration via somatic embryogenesis (SE) for economically important tree species (e.g. pines). These advances have opened up new scenarios for deployment of new high-performance clonally replicated planting stock to forest plantations and may also be a valuable tool for the development of efficient gene transfer techniques. Although high rates of plant propagation from axillary shoot proliferation can be achieved easily in many Eucalyptus species, even higher multiplication rates through SE have been recorded in other tree species. If the clonal propagation of Eucalyptus through SE proves to be an effective propagation method, it has the potential to meet the increasing industrial demands for high-quality uniform materials and to rapidly capture the benefits of breeding programs. Since 2002 a reproducible protocol for SE induction from mature zygotic embryos of E. globulus has been available. However, for SE to be useful in E. globulus improvement programs, the frequency of SE initiation, maturation, germination and acclimatisation needs to be improved and controlled. If this technology could be extended to elite germplasm, it would become an economically feasible tool for large-scale production and delivery of improved planting stock. This is one of the greatest current challenges in Eucalyptus tissue culture. In this review we update the most important aspects of SE in Eucalyptus with particular emphasis on E. globulus. We highlight both genetic control and the influence of different environmental factors in the SE process (e.g. medium composition, antioxidants, light and plant growth regulators), from induction to plant acclimatisation in both primary and secondary SE.     

火炬松体细胞胚胎发生和干化体细胞的氧化物酶活性(英文)

培养于附加2,4-D、BA和KT的愈伤组织诱导培养基上的火炬松成熟合子胚在培养3-9周后形成白色、半透明、有光泽的粘性愈伤组织。这类愈伤组织形成于成熟合子胚的子叶,但当用NAA或者IBA代替愈伤组织诱导培养基中的2,4-D时,它的诱导频率明显降低。这种粘性愈伤组织在分化培养基上形成体细胞胚。体细胞胚经过去50μm ABA和8.5%PEG600处理后成为耐干化胚。扫描电镜观察表明,萌发处理36小时后,耐干化胚恢复到干化处理之前的状态且大小和形态正常,而不耐干化胚不能恢复到干化处理之前的状态且表面撕裂。过氧化物酶活性的分析结果表明,耐干化胚有更高的过氧化物酶活性。耐干化胚的高过氧化物酶活性可能与催化H2O2的分解和保护体细胞胚免受氧化的伤害有关。     

Rooting of Pinus radiata somatic embryos: factors involved in the success of the process

In vitro conditions of the culture media, plant growth regulators and culture containers may cause anatomical and physiological changes that have negative effects on rooting and ex vitro acclimatization of somatic plantlets. The control of these factors could contribute to the improvement of somatic embryogenesis systems in conifers, especially in pines. The influence of macronutrient concentrations, explant type and culture containers in Pinus radiata D. Don in vitro somatic embryo rooting were analyzed. The highest rooting percentage was observed using half-strength macronutrient concentrations, complete micronutrients and vitamins of Quoirin and Lepoivre medium. Although the use of glass culture vessels was the best to increase the efficiency of the somatic embryogenesis process in terms of rooting, the use of ventilated containers resulted in a significant increase in the percentage of plants able to be planted in the field.     

Influence of osmotic pressure on somatic embryo maturation in Pinus densiflora

The effect of an osmoticum, polyethylene glycol (PEG), on somatic embryo production was examined using embryogenic cells of Pinus densiflora. In the basal medium containing 30 μM abscisic acid and 6% maltose, the quality of the embryos formed was poor even though somatic embryos were produced. The addition of PEG with molecular weight of 4000 or 8000 significantly enhanced the development of both the quality and quantity of somatic embryos. Furthermore, higher levels of a constant osmotic pressure with PEG 8000 in a range from about 300 to 450 mmol/kg could remarkably enhance the morphogenesis of somatic embryos and their number of embryos produced. A higher stable osmotic pressure with an appropriate molecular weight of PEG is a key factor for the production of good quality somatic embryos in P. densiflora.     

黑松合子胚和体细胞胚发育阶段的形态特征比较

为掌握黑松合子胚和体细胞胚发育进程,以未成熟合子胚为外植体,建立了黑松体细胞胚胎发生体系。对合子胚和体细胞胚发育阶段的形态特征进行观察和比较,结果表明,合子胚和体细胞胚的发育进程均可划分为8个阶段,两者同一阶段的形态特征大致相同,只有第2阶段和第4阶段有细微差别。     

Effect of parent genotype on somatic embryogenesis in Scots pine (Pinus sylvestris)

Controlled crosses of seven Scots pine (Pinus sylvestris L.) trees produced 49 families that included both reciprocals and selfings. Embryogenic cultures were initiated from immature megagametophytes and after 6 months in maintenance culture, mature somatic embryos were produced from the surviving 166 lines. The effect of parent genotypes on the cultures was evaluated at initiation of the tissue culture period, after 6 months in maintenance culture and at embryo maturation. The effect of the maternal parent was most pronounced at culture initiation. After 6 months in tissue culture, the maternal effect had decreased and the effects of both parents were significant. By the somatic embryo maturation stage, the maternal effect was still considerable but the paternal effect was no longer detectable. There was little correlation between the ranking of mothers and fathers, indicating that the maternal effect was caused by factors other than the paternal effect. No mother x father interaction was found, indicating that mothers successful at initiation and after 6 months in tissue culture, pollinated by any of the successful fathers, produced somatic lines and mature somatic embryos.