Studies on the Development of Gametophytes in Cornus officinalis (Cornaceae)

The floral bud of Cornus officinalis Sieb. et Zucc. began to differentiate at the end of April. In the beginning of November, female and male gametophytes reached their maturation. The flowers fell off in the following March. The wall of the microsporangium comprised epidermis, endothecium, two or three middle layers and a single layer of amoeboid tapetum with two nuclei. The extra-tapetal membrane was formed during the later stage of the development of anther. Meiosis of microspore mother-cell was normal and cytokinesis was of the simultaneous type. The tetrad was tetrahedral in shape. The mature pollen grains were 2-celled and 3-colporate. The ovule was unitegminous and tenuinucellate. During the development of the ovule, some special structures were formed, e. g. hypostase and obturator which originated from the integument. A single archesporium differentiated immediately below the nucellar epidermis. It functioned directly as the megaspore mother-celL This cell under went meiosis to form a linear tetrad. The chalazal megaspore was functional. The development of the embryo sac was conformed as the polygonum type. Two polar nuclei fused into the secondary nucleus and 'three antipodal cells degenerated soon after the embryo sac reached its maturation, at that time the female gametophyte had become an embryo sac which consisted of only four cells each with a nucleus just before two months of blooming. The nuclei of some synergids located in the chalazal part of the cells. Contrarily, the micropylar past of the synergids were occupied by a large vacuole. The secondary nucleus was usually located in the chalazal part of the embryo sac.     

Change of Superoxide Dismutase Activities During Somatic Embryogenesis in Asparagus officinalis L.

The superoxide dismutase activities in callus and somatic embryoids at different developmental stages of Asparagus officinalis L. were determined by means light of induced oxidation-reduction of nitro blue tetrazolium. It was shown that the superoxide dismutase activity was higher in callus than in somatic embryoids at different developmental stages. The activity increased with growth, differentiation and maturation of somatic embryoids during somatic embryogenesis. The relations between superoxide dismutade activity and somatic embryogenesis were discussed.     

HPLC法测定山茱萸注射液中马钱素和莫诺甙的含量

山茱萸注射液中有多种有效成分,采用高效液相色谱法对山茱萸注射液中的马钱素(Loganin)和莫诺甙(Morroniside)进行分离定量.色谱柱:Kromasil-100℃_(18)柱.流动相:0.05mol/L磷酸二氢钠缓冲液-乙腈(6.4:1).马钱素和莫诺甙的平均回收率和相对标准偏差分别为99.79%、1.51(n=6)和99.73%、2.75%(n=6).结果满意,可作为山茱萸主射液质量控制的标准方法.     

The effect of triacontanol on micropropagation of balm, Melissa officinalis L.

 Triacontanol, a long-chain primary alcohol was found to be an effective growth regulator in the micropropagation of balm, Melissa officinalis. In both the multiplication and the rooting phase, concentrations of 2, 5, 10 and 20 μg triacontanol per liter were applied. After 4 weeks of culture, the fresh weight of shoots was measured in the multiplication phase and root formation, photosynthetic activity, chlorophyll content and the fresh and dry weights of shoots were analyzed in the root induction phase. In the multiplication phase, 5 μg/l triacontanol was found to be the optimal concentration, while in the rooting phase 2 μg/l was the most effective. Triacontanol increased the number and length of roots, and it enhanced shoot growth, fresh weight, and the chlorophyll content, but it had no effect on the dry weight and the photosynthetic activity of the plants. Results of our work demonstrate that triacontanol can be applied as an effective growth regulator in the tissue culture of balm. Received: 3 December 1997 / Revised: 24 February 1998 / Accepted: 26 February 1999     

Somatic embryogenesis and in vitro rosmarinic acid accumulation in Salvia officinalis and S. fruticosa leaf callus cultures

The effect of explant age, plant growth regulators and culture conditions on somatic embryogenesis and rosmarinic acid production from leaf explants of Salvia officinalis and S. fruticosa plants collected in Greece was investigated. Embryogenic callus with numerous spherical somatic embryos could be induced on explants derived from both species and cultured for 3 weeks on a Murashige and Skoog (MS) medium supplemented with 1.8–18 μm 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (Kin) or 10.5–21 μm 1-naphthalenacetic acid and 6-benzyladenine. Only explants from young plants (with six to eight leaves) responded to the culture treatments and, in general, low light intensities (50 μmol m^–2 s^–1) favoured callus formation and induction of somatic embryos. Somatic embryos were further developed on the same medium. Heart- and torpedo-shaped embryos (1–2 mm long) were subcultured on a growth-regulator-free MS medium for maturation. Maximum rosmarinic acid accumulation in S. officinalis and S. fruticosa callus cultured on 4.5 μm 2,4-D and 4.5 μm Kin was 25.9 and 29.0 g/l, respectively. Received: 17 January 1997 / Revision received: 26 May 1997 / Accepted: 30 June 1997     

A Calcium/Calmodulin-Dependent Nitric Oxide Synthase, NMDAR2/3 Receptor Subunits, and Glutamate in the CNS of the Cuttlefish Sepia officinalis

Chemical, biochemical, and immunohistochemical evidence is reported demonstrating the presence in the brain of the cuttlefish Sepia officinalis of a Ca2+-dependent nitric oxide synthase, NMDAR2/3 receptor subunits, and glutamate, occurring in neurons and fibers functionally related to the inking system. Nitric oxide synthase activity was concentrated for the most part in the cytosolic fraction and was masked by other citrulline-forming enzyme(s). The labile nitric oxide synthase could be partially purified by ammonium sulfate precipitation of tissue extracts, followed by affinity chromatography on 2',5'-ADP-agarose and calmodulin-agarose. The resulting activity, immunolabeled at 150 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis by antibodies to rat neuronal nitric oxide synthase, depended on NADPH and tetrahydro-L-biopterin, and was inhibited by N(G)-nitro-L-arginine. NMDAR2/3 subunit-immunoreactive proteins migrating at 170 kDa could also be detected in brain extracts, along with glutamate (whole brain: 0.32 +/- 0.03 micromol of glutamate/mg of protein; optic lobes: 0.22 +/- 0.04; vertical complex: 0.65 +/- 0.06; basal lobes: 0.58 +/- 0.04; brachial lobe: 0.77 +/- 0.06; pedal lobe: 1.04 +/- 0.08; palliovisceral lobe: 0.86 +/- 0.05). Incubation of intact brains with 1.5 mM glutamate or NMDA or the nitric oxide donor 2-(N,N-diethylamino)diazenolate-2-oxide caused a fivefold rise in the levels of cyclic GMP, indicating operation of the glutamate-nitric oxide-cyclic GMP signaling pathway. Immunohistochemical mapping of Sepia CNS showed specific localization of nitric oxide synthase-like and NMDAR2/3-like immunoreactivities in the lateroventral palliovisceral lobe, the visceral lobe, and the pallial and visceral nerves, as well as in the sphincters and wall of the ink sac.     

Modeling habitat distribution of Cornus officinalis with Maxent modeling and fuzzy logics in China

Aims Predicting suitable habitat distribution is an effective way to protect rare or endangered medicinal plants. Cornus officinalis is a perennial tree growing in forest edge and its air-dried pericarp is one of the traditional Chinese medicines (TCM) with significant medicinal values. In recent years, C. officinalis has undergone severe degeneration of its natural habitat owing to growing market demands and unprecedented damage to the forests. Moreover, the degeneration of suitable habitat has threatened the supply of medicinal materials, and even led to the extinction of some engendered medicinal plant species. In this case, there is a great risk to introduce and cultivate medicinal plants if planners determine the suitable cultivation regions based on personal subjective experience alone. Therefore, predicting suitable potential habitat distribution of medicinal plants (e.g. C. officinalis) and revealing the environmental factors determining such distribution patterns are important to habitat conservation and environmental restoration.Methods In this article, we report the results of a study on the habitat distribution of C. officinalis using maximum entropy (Maxent) modeling and fuzzy logics together with loganin content and environmental variables. The localities of 106 C. officinalis in China were collected by our group and other researchers and used as occurrence data. The loganin content of 234 C. officinalis germplasm resources were tested by high-performance liquid chromatography (HPLC) and used as content data. 79 environmental variables were selected and processed with multicollinearity test by using Pearson Correlation Coefficient (r) to determine a set of independent variables. The chosen variables were then processed in the fuzzy linear model according to the cell values (maximum, minimum) of localities with estimated loganin content. The SDMtoolbox was used to spatially rarefy occurrence data and prepare bias files. Furthermore, combined Maxent modeling and fuzzy logics were used to predict the suitable habitat of C. officinalis. The modeling result was validated using null-model method.Important findings As a result, six environmental factors including tmin3, prec3, bio4, alt, bio12 and bio3 were determined as key influential factors that mostly affected both the habitat suitability and active ingredient of C. officinalis. The highly suitable regions of C. officinalis mainly distribute in a 'core distribution zone' of the east-central China. The statistically significant AUC value indicated that combined Maxent modeling and fuzzy logics could be used to predict the suitable habitat distribution of medicinal plants. Furthermore, our results confirm that ecological factors played critical roles in assessing suitable geographical regions as well as active ingredient of plants, highlighting the need for effective habitat rehabilitation and resource conservation.     

阿里红黄酮对衰老模型小鼠的抗衰老作用

目的：研究阿里红黄酮对衰老模型小鼠的抗衰老作用。方法：将小鼠随机分为6组（n=12）：正常对照组,衰老模型组,阳性对照组,低、中、高剂量阿里红黄酮组。除正常对照组外,其余各组均采用D-半乳糖颈背部皮下注射建立亚急性衰老小鼠模型,用不同剂量的阿里红黄酮（100、200和400 mg/（kg·d））灌服小鼠6周后,计算衰老模型小鼠脑指数及脾脏指数、胸腺指数,测定其脑组织丙二醛（MDA）含量及谷胱甘肽过氧化物酶（GSH-Px）活性、肝组织过氧化氢酶（CAT）及超氧化物歧化酶（SOD）活性。结果：阿里红黄酮3个剂量组可不同程度的升高衰老模型小鼠的脑指数、脾脏指数、胸腺指数、脑组织中的GSH-Px活性及肝组织中的CAT、SOD活性,降低其脑组织中的MDA含量。结论：阿里红黄酮可能通过提高机体的抗氧化能力而达到抗衰老作用。     

高山植物黄管秦艽cDNA 文库构建与表达序列标签(EST) 分析

黄管秦艽( Gentiana officinalis) 是一种重要的藏药高山植物, 本研究构建了该物种开花期的cDNA 文库。经检测达到中等cDNA 文库水平, 文库滴度为1 . 2×107 pfu􊄯ml , 重组率95.9% , 插入片段平均长度大于500 bp。对343 个随机挑选的重组克隆进行部分测序, 获得的ESTs 经编辑后共有181 条有效序列。经生物信息学方法分析181 条表达序列标签(EST) 代表144 个单克隆序列, 其中55 个与已鉴定的基因同源, 35 个序列与未鉴定的EST 匹配, 54 个未找到同源序列; 后两者共有89 个EST 序列未发现功能相似的蛋白。对已鉴定的EST进行功能分析发现, 相关基因主要编码以下蛋白: 与蛋白表达相关的占35%; 光合作用相关的占22%; 新陈代谢相关的占18%; 抗性相关的占11%; 质膜运输和细胞分裂相关的分别占5% ; 染色体变化和细胞信号转导的分别占2%。根据有效EST 序列设计引物, 通过RT-PCR 进一验证了所得EST 的准确性。这些研究结果为将来研究黄管秦艽的功能基因以及该物种与相关物种的群体遗传学、进化生物学等方面提供了基础。