Somatic embryogenesis and histological analysis from zygotic embryos in Vitis vinifera L. ＇Moldova＇

We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape ＇Moldova＇ （Vitis vinifera U ＇Moldova＇）. Primary calli were initiated on Nitsch and Nitsch （NN） medium supplemented with 1.0 mg·L^-1 2,4-D and 0.5 mg·L^-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg·L^-1 6-BA and 2 mg·L^-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg·L^-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dynamic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic embryos, including typical epiderma, cotyledon primordium and vascular tissue.     

Changes in root growth and relationships between plant organs and root hydraulic traits in American elm （Ulmus americana L.） and red oak （Quercus rubra L.） seedlings due to elevated CO2 level

The relationships between plant organs and root hydrological traits are not well known and the question arises whether elevated CO2 changes these relationships. This study attempted to answer this question. A pseudo-replicated experiment was conducted with two times 24 American elm(Ulmus americana L.) and 23 and 24 red oak(Quercus rubra L.) seedlings growing in ambient CO2(around 360 μmol·L-1) and 540 ± 7.95 μmol·L-1 CO2 in a greenhouse. After 71 days of treatment for American elm and 77 days for red oak,14...     

Dynamic changes of telomeric restriction fragment (TRF) lengths in cells during the developmental process from embryos to seedlings and a comparison with the embryonal calli in Ginkgo biloba L.

Telomeres are the structures that locate at the terminals of linear eukaryotic chromosomes. They can play essential roles in many cellular processes. The terminal location of Arabidopsis-type TTTAGGG tandem repeats were thought to be highly conserved. The terminal location of Ginkgo biloba L. consisting of TTTAGGG tandem repeats, were confirmed by Bal31 exonuclease degradation and Southern blotting. By comparing telomeric restriction fragment (TRF) lengths at different developmental stages from embryos to seedlings, a fluctuant tendency towards variation was found in these samples. The TRF length of embryos was also compared with that of embryonal calli and an upward trend was discovered in callus culture. The results suggest that there should be a telomerase mechanism or/and ALT mechanism for the maintenance of telomere length.     

Changes induced by osmotic stress in the morphology,biochemistry, physiology,anatomy and stomatal parameters of almond species （Prunus L. spp.） grown in vitro

We investigated the influence of different levels of osmotic stress on growth and development in selected wild almond species(eight Prunus spp.) grown in vitro. The study, while endorsing the efficacy of in vitro screening of auxiliary buds of wild almond for osmotic stress tolerance, showed species variability in its response to osmotic stress. Osmotic stress reduced growth and development of all the species. However, the putative tolerant Prunus spp. showed better performance than the putative susceptible genotypes. On average there was an 80% decrease in shoot dry weight at-1.2 MPa. Reduction in shoot weight was more common in osmotic stress-susceptible species in the section labeled ‘Euamygdalus'. The tolerant Prunus species produced smaller changes in biochemical responses than the sensitive cultivars for malondialdehyde content, catalase activity, relative permeability of protoplast membranes, and net photosynthetic rate. The tolerant species maintained cell integrity better than drought sensitive species. Wild almond species in the section labeled ‘Spartioides'(Prunus arabica(Olivier) Neikle, Prunus glauca (Browicz) A.E. Murray, Prunus scoparia Spach) and ‘Lycioides'(Prunus lycioides Spach, Prunus reuteri Bossi. et Bushe) were best adapted to osmotic stress. Increase in chlorophyll concentration and leaf thickness under high osmotic stress can be considered as preliminary selection parameters for osmotic stress tolerance in Prunus spp. The study confirmed the efficacy of the in vitro method for screening of large number of genotypes for osmotic stress tolerance in wild almond species.     

An efficient method for clonal propagation and in vitro establishment of softwood shoots from epicormic buds of teak （Tectona grandis L.）

Softwood shoots were produced from 40 cm long stem segments placed horizontally in flat trays containing sterilized sand under natural light or shade conditions for subsequent rooting and micropropagation studies in teak(Tectona grandis L.) . Higher number of shoots(6.17) per log was produced under natural light as compared to shade conditions. Forcing was also better in natural light as compared to shade in terms of shoot length,number of nodes or leaves. For rooting,2-4 cm long softwood shoots were excise...     

Somatic embryogenesis and histological analysis from zygotic embryos in Vitis vinifera L. ''Moldova''

We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape 'Moldova' (Vitis vinifera L. 'Moldova'). Primary calli were initiated on Nitsch and Nitsch (NN) medium supplemented with 1.0mg'L-1 2,4-D and 0.5 mg'L-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg-L-1 6-BA and 2 mg.L-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg.L-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dy-namic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic em-bryos, including typical epiderma, cotyledon primordium and vascular tissue.     

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.     

火炬松体细胞胚胎发生体系的优化

目的 建立和优化火炬松体胚发生技术体系,为火炬松优良基因型体胚繁育及遗传转化体系建立提供技术支撑。 方法 以火炬松优良基因型的未成熟合子胚为材料,从基因型、球果采集期、基本培养基、植物外源激素组合及浓度等方面对胚性愈伤组织诱导条件进行优化;选取增殖效果好、具有胚性胚柄细胞团（ESM）的胚性愈伤组织开展体胚成熟的正交试验,选择发育正常的子叶胚进行萌发,8周后移栽。 结果 基因型、球果采集期、基本培养基和外源激素(PGR)组合及浓度等因子均对胚性愈伤组织诱导有不同程度的影响。火炬松6种基因型中荣山52胚性愈伤组织诱导率最高,达63.33%;处于裂生多胚期的合子胚为外植体的最佳采样期（7月20日左右）;6种基本培养基中,在DCR基本培养基上胚性愈伤组织诱导率最高,达40.00%;不同PGR组合及浓度中,以0.5 mg·L−1 6-BA+1.0 mg·L−1 2,4-D + 2.0 mg·L−1 NAA + 5.0 mg·L−1 ABA的胚性愈伤组织诱导率最高,达50.00%。胚性愈伤组织在基本培养基MLP上培养14 d后,转入含有ABA 8.0 mg·L−1、PEG 8000 140 g·L−1以及麦芽糖40.0 g·L−1的体胚成熟分化培养基上,体胚发生数平均为411个·g−1,体胚萌发率可达93.33%;体胚苗移栽成活率可达86.30%。 结论 本研究优化了火炬松胚性愈伤组织诱导与体胚发生技术体系,体胚诱导率显著提升,为火炬松良种规模化繁育及遗传转化提供技术支撑。     

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.     

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。     

葡萄叶片SPAD值与测定位置及光合色素含量的关系

为了实时、准确、无损地检测葡萄叶片中叶绿素的含量水平,以评价葡萄生长发育与营养状况,确保栽培的科学和施肥的合理,以‘夏黑’‘巨玫瑰’‘户太8号’‘阳光玫瑰’‘金手指’‘摩尔多瓦’这6个品种葡萄叶片为试验材料,采用叶绿素测定仪测定了6个葡萄品种叶片不同位置的SPAD值,使用分光光度计测定了葡萄叶片光合色素含量,探寻了葡萄叶片SPAD值的最适测量位点,分析了SPAD值与光合色素含量的关系,并建立了SPAD值与光合色素含量的线性函数拟合方程。结果表明:6个品种葡萄叶片的SPAD值,叶片顶部的均最高,中部的均次之,基部的均最小;其中,葡萄叶片中部的SPAD值最具有代表性。6个品种葡萄叶片的SPAD值与叶绿素a、叶绿素b、总叶绿素和类胡萝卜素含量均显著相关,其与叶绿素a含量的相关性最高;在不同品种葡萄叶片的SPAD值与叶绿素a、叶绿素总含量、类胡萝卜素含量的线性函数拟合方程中,‘户太8号’的拟合方程决定系数值最高,其叶绿素a、叶绿素总含量、类胡萝卜素含量的决定系数值分别为0.903 5、0.867 2、0.749 9;在叶绿素b与葡萄叶片SPAD值的拟合方程中,‘金手指’的拟合方程为y=0.034 7x-0.084 0,其决定系数R2值(0.773 4)最高。单一品种葡萄叶片的SPAD值与光合色素含量的回归模型的拟合准确率比6个葡萄品种汇总拟合回归模型的要高。     

Somatic embryogenesis from immature and mature zygotic embryos ofCryptomeria japonica I: Embryogenic cell induction and its morphological characteristics

Embryogenic cells (ECs) of sugi (Cryptomeria japonica D. Don) were induced from immature and mature zygotic embryos cultured on different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D)-containing modified Campbell and Durzan medium. The rate of induction of ECs varied depending on the stage of embryos collected. The highest percentage of induction (35%) was obtained with immature zygotic embryos collected July 18 and July 30, 1997, when 1 M of 2,4-D was added to the induction medium. The ECs easily proliferated when subcultured in a medium of the same composition as the induction medium within 3 weeks. Morphological characteristics of nonembryogenic cells and embryogenic cells of different developmental stages were studied under an inverted fluorescence microscope.Part of this work was presented at the 109th annual Japanese Forestry Society meeting at Tochigi, April 1998     

刺槐未成熟合子胚的体细胞胚胎发生和植株再生

以刺槐不同胚龄的未成熟合子胚为外植体,采用混合正交试验设计,研究幼胚胚龄和不同外源激素种类及质量浓度对刺槐胚性愈伤组织的诱导和体细胞胚分化、萌发的影响.结果表明:开花后8周(55天左右)是胚性愈伤组织和体胚诱导的最佳外植体取材时期;MS+2,4-D 5.0 mg·L-1 +BA0.5 mg·L-1是诱导胚性愈伤组织的最佳培养基,出愈率最高为95.42％±0.02％;MS +NAA0.5 mg·L-1 +BA0.5 mg·L-1+谷氨酰胺250 mg·L-1+水解酪蛋白500 mg·L-1是体细胞胚诱导和分化的最佳培养基,直接体细胞胚发生率最高为92.40％±0.12％,通过愈伤组织诱导体细胞胚发生的频率最高为90.73％ ±0.49％.一旦形成球形胚,将培养物转接到不含任何生长调节剂的MS培养基上,体细胞胚经成熟萌发可进一步形成完整小植株.