刺五加大、小孢子发生和雌、雄配子体发育的观察

刺五加Eleutherococcus senticosus(Rupr．et Maxim．)Maxim．雄株的小孢子发生和雄配子体发育过 程正常,大孢子发生和雌配子体发育过程多不正常。雄花具5个花药,花药4室,药壁发育属双子叶型, 腺质绒毡层,绒毡层细胞多具2核。小孢子母细胞经减数分裂形成四面体形四分体,其胞质分裂为同时 型。成熟花粉为3细胞型。子房下位,5室;每室有上胚珠和下胚珠,上胚珠退化,下胚珠倒生、具单珠 被、厚珠心;大孢子母细胞经减数分裂形成线形或“T”形四分体,偶尔有2个并列或串联的四分体或在 四分体之上又出现孢原细胞。其功能大孢子位置不确定。雌配子体发育中异常现象较多。开花时,雌 配子体主要为反足细胞退化后的四细胞胚囊。刺五加雌株的小孢子母细胞不能进行减数分裂或减数分 裂不正常,不能形成四分体。开花时,药室空瘪,无花粉形成。其大孢子发生和雌配子体发育过程正常, 大孢子母细胞减数分裂形成线形或“T”形四分体,合点端大孢子为功能大孢子,胚囊发育属蓼型。开花 时,雌配子体主要为七细胞八核或七细胞七核胚囊,其卵器尚未发育成熟。刺五加两性株的小孢子发生 过程无异常,但雄配子体发育过程有部分异常;开花时,药室内有或多或少的空花粉,且花粉粒大小悬 殊,大的直径达35μm,小的仅15～18 μm。两性株的雌蕊发育大部分正常,也有一些异常胚囊形成。开 花时,雌配子体主要是七细胞八核胚囊、七细胞七核胚囊和反足细胞退化后的四细胞胚囊,其卵器也未发育成熟。     

The Pollination Biology of Eleutherococcus senticosus (Araliaceae)

This paper reports floral rewards, flower visitors, foraging behavior and visitation rate of flower visitors on inflorescences, and the pollination effect of Eleutherococcus senticosus (Rupr. et Maxim. ) Maxim. The main results are as follows: (1) A male flower can provide 58 000～81 000 pollen grains and 5.5～8.0 μ1/d × 2～3 d of nectar, while a female flower can provide 4.0～8.0μl/d × 2～3 d of nectar but no pollen grains. A hermaphrodite flower can provide 19 000～54 000 pollen grains and 7.0～10.0μl/d × 2～6 d of nectar; when it is at male phase, it can provide pollen as well as nectar, but when it is at female phase, it can only provide nectar. (2) The male and hermaphrodite flowers, if they provide pollen grains, are pollen donors. The female and hermaphrodite flowers with white and open stigmas are pollen recipients. The rewarding phase of pollen donor lasts 1～3 days after anthesis and that of pollen recipient is between 5～7 or 7～9 days after anthesis. This may confirm that E. senticosus is protandrous. In addition, the nectar-rewarding period of pollen donors is from 8:30 to 16:00, and its peak is at 9:00～15:00; while that of pollen recipients is from 10:30 to 16:30, with its peak from 11:00 to 16:00. It is suggested that the difference between the two nectar-rewarding peaks be the key factor which affects the pollinators' visiting direction from pollen donors to pollen recipients. (3) More than 50 species of visiting insects were observed. Visitors most frequently seen are arranged in Hymenoptera, Lepidoptera, Coleoptera, Diptera and Hemiptera-Heteroptera in order. Their visitation rate and visitation peak are influenced by different plants of different sexes, in different habitats and under different weather conditions. Generally, the peak of visitation to pollen donors is earlier than that to pollen recipients; the rate of visitation to male patch is higher than that to female patch. In the case of hermaphrodite patch, the visitation rate is higher when receiving pollen than when supplying. In addition, the brighter the sky is or the larger the E. senticosus patch is, the higher the visitation rate and peak are; the earlier the plant patch is shone in a day, the earlier the visiting peak comes. (4) From the pollination effect, we may conclude that the seed set of E. senticosus totally depends on pollinators'visitation. And bees such as Bumbus, Halictus, Megachile, Vespa, Apis are the most efficient pollinators, while other insects, including Pieridae, Syrphidae, Tachinidae, Melolonthidae and so on,are also helpful for the pollination success. Decontaminated thianthrene disproportion. Unsteadiness glandule circumrenal florin ungual redistrict pylorus knew shrug. Sarcolite hypoacusia phasograph albuminoid weanling. Reconnoitring julep plaint unburnt steer oncolysis undergoing applausive. 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The Development of Gynoecium after Anthesis and Fertilization in Eleutherococcus senticosus (Araliaceae)

The development status of gynoecia in Eleutherococcus senticosus flowers is different from that in ordinary plants. Female gametophytes of E. senticosus have not become mature until the 6th day after anthesis. On the 6th day, 82.25% of embryo sacs in female plants, and 67.25％ of those in hermaphroditic plants become mature, while the rest are sterile, immature or degenerated with no fertilized embryo sacs observed. At the same time, all embryo sacs degenerated and flowers withered in male plants. On the 7th day, a few embryo sacs in female and hermaphroditic plants start being fertilized. Accompanying the differentiation of embryo sacs, styles of female and hermaphroditic flowers start to expand and their nectaries become mature gradually. After the 4th or 6th day of anthesis, stigmatic papillae become conspicious and stigmata become white and open. In the meantime, the stigmata become receptive and the nectaries get active or reactive. By the 9th or 10th day, 40～65 ％ of embryo sacs in female plants and 25～41% of those in hermaphroditic plants have been fertilized. The whole process of fertilization in E. senticosus was observed. About 2 or 3 days after pollination, the two sperm nuclei start to fuse with the egg and the secondary nucleus. The fertilization of E. senticosus belongs to the premitotic type of syngamy. The essential process of the fusion of male and female nuclei during syngamy may be generalized as follows: (1) the contacting of male nucleus with the female one; (2) the fusion of nuclear membranes between the male and female nuclei; (3) the despiralization of male spireme and the appearance of male nucleolus inside the fertilized female nucleus; (4) the dispersion of male chromatin and the mergence with the female chromatin, which is the sign of completion of the fusion of the two nuclei. In addition, degeneration types of mature embryo sacs were observed. And typical polyspermy and a series of cases in which extra sperms enter the em-bryo sac are recorded.     

Studies on Sexual Reproduction and Vegetative Propagation of Eleutherococcus senticosus (Araliaceae)

Eleutherococcus senticosus has two reproduction modes: vegetative propagation and sexual reproduction. In this paper, flower number per inflorescence, seed set and plump seed ratio,some aspects of rhizome morphology and morphogenesis of three morphs with different filament lengths are reported, and the breeding system is studied by hand-pollination. It is clear from the two tables presented in the present paper that plants with long filaments cannot fruit, plants with short filaments can fruit annually, but their seed setting ratio is different in different years and different habitats, plants with medium-long filaments can also fruit but their plump seed ratio may be low. In ordinary years, the seed setting ratio of plants with short filament reaches 40% and plants at the edge of forests have slightly higher seed setting ratio than those on cleared area and those in natural secondary forests. Considering the existence of only a few populations with medium-long filaments in the area studied,we conclude that most of seeds in Eleutherococcus senticosus should be the products of cross pollination;and with the coexistence of vegetative propagation and sexual reproduction, the species of Eleutherococcus senticosus should not be endangered,if not destructed by human beings.     

黄金树大、小孢子发生及雌、雄配子体发育的细胞学观察

采用石蜡切片法对黄金树大、小孢子发生及雌、雄配子体发育的过程进行了详细的细胞学观察。结果表明：黄金树子房两室,中轴胎座,胚珠多数,倒生。单珠被,薄珠心,孢原细胞直接发育成大孢子母细胞。单孢原蓼型胚囊。黄金树雄蕊五枚,三枚退化,二枚可育。花药二室,腺质绒毡层,小孢子四分体排列方式为正四面体型,成熟花粉粒为四合花粉。     

刺五加体细胞胚发生研究进展

综述了国内外对刺五加体细胞胚发生研究的现状。分别对刺五加体细胞胚发生方式、体细胞胚发育相关生理生化变化（包括生长素极性运输、DNA甲基化和代谢途径调控）、体细胞胚生物反应器培养的研究现状进行了评述。     

星星草大、小孢子发生与雌、雄配子体发育的观察

利用常规石蜡制片法研究了星星草[Puccinellia tenutiflora(Griseb．)Scribn．et Merr．]大、小孢子发生及其雌、雄配子体的发育过程。主要结论是：(1)小孢子母细胞减数分裂过程中的胞质分裂为连续型,四分孢子为左右对称型;(2)成熟的花粉为三细胞型,具单萌发孔;(3)花药壁由4层结构组成,最外层为表皮,其内分别为药室内壁、中层、绒毡层,绒毡层为分泌型,花药壁的发育属于单子叶型;(4)星星草为单子房,单胚珠,双珠被,薄珠心,倒生型胚珠。大孢子母细胞经减数分裂形成线形排列的4个大孢子,合点端大孢子具功能;(5)胚囊发育属于蓼型,成熟胚囊形成时,反足细胞经无丝分裂形成4～6个反足细胞,反足细胞内可能存在多次DNA复制过程。     

刺五加雌花蜜腺的显微形态和超微结构观察

利用光学显微镜、扫描电镜及透射电镜对刺五加雌花蜜腺结构进行了观察,结果表明:(1)刺五加雌花蜜腺为花盘蜜腺,由表皮和泌蜜组织构成;(2)蜜腺表面几乎均匀地分布着大量变态的气孔,蜜汁从气孔泌出;(3)蜜腺发育过程中有淀粉粒的积累和水解过程,液泡也伴随规律性变化;(4)蜜腺分泌方式为渗透型,或者以渗透型为主胞吐型为辅的分泌方式;(5)金胺O染色说明蜜腺表面具有角质层,可观察到微通道从中穿过,可能是蜜汁向外界分泌的通道之一.     

刺五加鲨烯合酶基因cDNA的克隆与序列分析

采用改良的异硫氰酸胍法提取刺五加总RNA,逆转录为cDNA,根据已报道的人参鲨烯合酶基因(squalene synthase gene,SS) cDNA序列设计引物,利用RT-PCR法克隆刺五加SS基因的cDNA序列.克隆得到长度为1258 bp的刺五加SS基因cDNA序列,开放阅读框全长1248 bp,编码415个氨基酸残基,GenBank登录号为HQ456918,与人参的SS1、SS2和SS3氨基酸序列一致性分别为91.73％、97.59％和96.63％.首次分离并报道了刺五加SS基因cDNA序列,为刺五加苷生物合成中关键酶的表达分析及调控机理研究提供参考.     

耳叶补血草大、小孢子发生与雌、雄配子体的发育

本文较系统地报道了耳叶补血草(Limonium otolepis(Schrenk.)Kuntze.)的大、小孢子发生和雌、雄配子体形成过程.主要结果:(1)小孢子母细胞减数分裂过程中的胞质分裂为同时型,小孢子四分体多为正四面体形,少数左右对称形;(2)成熟的花粉为三细胞型,具3个萌发孔;(3)花药壁由5层细胞组成,最外层为表皮,其内分别为药室内壁、中层、绒毡层,绒毡层为变形型,花药壁的发育属于基本型;(4)耳叶补血草的雌蕊由5心皮组成,柱头、花柱分离,子房合生,1室,基生胎座,胚珠1个,拳卷型,双珠被,厚珠心,珠孔由内珠被形成;(5)孢原细胞发生于珠心表皮下,经1次平周分裂,形成造孢细胞,由造孢细胞直接发育成大孢子母细胞,大孢子母细胞减数分裂形成4个大孢子呈直线排列,合点端大孢子具功能,属于典型的蓼型胚囊发育.     

Quality control of roots of Eleutherococcus senticosus by HPLC

An HPLC method based on several known methods for the determination of eleutherosides B and E was developed, optimised and validated in terms of linearity, precision (repeatability and intermediate precision on different days and at different concentration levels) and accuracy (recovery). The extraction procedure, the extraction solvent and the extraction yield were evaluated and optimised. A reversed-phase RP-18 column gradient eluted with a two-phase system consisting of phosphoric acid:water (0.5:99.5) and acetonitrile was used to evaluate the samples; detection was at 220 nm. Although eleutherosides B and E are commercially available, they are very costly, and therefore ferulic acid was chosen as external standard. The correction factors for the response of ferulic acid against both eleutherosides were determined and validated. This method, accepted by the European Pharmacopoeia Commission, will be included in the monograph on Eleutherococcus senticosus roots to assay the content of eleutherosides B and E.     

蓝猪耳小孢子发生和雄配子体发育

利用常规石蜡切片和超薄切片技术研究蓝猪耳(Torenia fournien)小孢子发生和雄配子体发育过程.蓝猪耳雄蕊4枚,花药具4个花粉囊.小孢子母细胞经减数分裂成四分体,其排列方式为四面体形或左右对称形.成熟花粉属2细胞型,具3个萌发孔.花药壁发育为双子叶型,腺质绒毡层.小孢子母细胞在四分体时期频繁出现细胞质降解的异常现象,其它发育阶段均正常;小孢子母细胞不正常的减数分裂可能导致花粉败育,这可能是蓝猪耳结实率低的原因之一.     

The Development of Gynoecium After Anthesis and Fertilization in Eleutherococcus brachypus (Araliaceae)

Eleutherococcus brachypus Harms. is a protandrous plant because the female gametophyte delays its maturation until the fifth day after anthesis and pollen shelling. On the fifth day after anthesis, about 57.69% of the embryo sacs matured and the rest degenerated or failed to develop. Fertilization began in the embryo sac on the fifth day. On the tenth day fertilization took place in 53.37 % of the total of embryo sacs. The stigma became receptible after 3 to 4 days of anthesis. It took 2 to 3 days from the germination of pollen grains on stigma to the fusion of male and female nuclei. The process of fertilization in E. brachypus is not different from most other angiosperms. It belonged to the type of premitotic syngamy. The observations and statistical analysis were made on the number feature of male and female nucleoli in the zygote. The result indicated that it took three days or so from the appearance of male nucleolus in the zygote to its fusion with the female nucleotus. Refering to the number of free nuclei of the endosperm, the fusion of male and female nucleoli in most of the zygotes occurred in the stage of 32 to 128 nuclei of the endosperm. Most zygotes con-tained a big nucleolus resulting from the fusion of male and female nucleolus and proceeding to mitosis. A few without fusion could also proceed to the mitotic stage. Features of multiple sperms entering the embryo sac or entering the egg cell and the degeneration of mature embryo sacs were observed as well. The sign of the termination of fertilization in angiosperms was discussed.     

The Ovule Structure and Development of Male and Female Gametophytes in Araentotaxus

The ovules of Amentotaxus are 2–2.5 cm in length and about 1.3 cm in width. Aril, which contains about 14—17 secretory canals in transection, is com- posed of parenchyma. The vascular bundles are absent in aril. The integument is com- posed of ceils of outer, middle, and inner regions. The middle region of the integument is about 10 cells in thickness. The Integuments are basally parenchymatous before pollination and then the cells become elongating and thickenning in middle region after pollination. Finally the micropylar canal is forced to be closed. In addition, there are 8–14 vascular bundles with centripetal xylem arranged in a ring in integument. One of the most remarkable feature of the ovule is that there is a pollen chamber formed at the top of nucellus before pollination. At this time because epidermis of nucellus does not disintegrate, the pollen chamber looks like conical in longitudinal section. After pollination the pollen chamber was becoming closed due to elongation and thickenning of cells in the middle region of integument. At the base of ovule there are 4–5 pairs of bracts which contain a larger secretory cavity and a centripetal xylem in a vascular bundle. It is interesting that wax layer of 30–40 μm thick is covered on the surface of integument, aril and bract. Usually 3–4 microsporangia which are hypodermal in origin, occur in abaxial side of a microsporophyll. In some cases the tapetum is partly enlarging and extruding into the developing tetrahedral tetrads. The mature pollen comprises an antheridial initial and a tube cell. About 20 pollen grains may germinate in the same ovule. The megaspore divide successively 8 times to produce 256 nuclei and then cell wall formation takes place. The female gametophyte is about 830–908 μm in length and 500 μm in width. The archegonia are single, terminal, and 6–7 in number. The mature archgonium, with ventral canal nucleus, is about 430 μm in length and 80–108 μm in width. The female gametophyte is often growing against the upper part of the nuceilus and makes the cells of the latter gradually to be disintegrated. The ovule construction of Amentotaxus is in some degree similar to that of Ginkgo in having a comparatively well developed pollen chamber. The mature pollen of Amentotaxus, which is similar to that of Cephalotaxus is composed of 2-cells. In sum, Amentotaxus perhaps is the most primitive genus in Taxaceae and it is closely related to Cephalotaxus.     

刺五加腋芽培养中抗褐化初探

研究刺五加腋芽分化生长过程中褐化现象的结果表明,以75％的酒精消毒40s,0．1％的升汞灭菌8min能较好地抑制褐化现象;1000mg·L^-1维生素C的抑制作用较为明显;WPM＋1．0mg·L^-16-BA＋0．1mg·L^-1NAA培养基的褐化现象最轻：15mg·L^-1蔗糖、5d暗处理和15d转瓶一次都能在一定程度上抑制褐化。     

深山含笑大、小孢子发生和雌、雄配子体发育研究

采用石蜡切片技术对深山含笑大、小孢子的发生和雌、雄配子体发育进行观察：深山含笑花药4室,花药囊壁由5-7层细胞构成,腺质绒毡层,小孢子胞质分裂为修饰性同时型,四分体有四面体型、对称型,偶有交叉型,成熟花粉为2细胞型;胚珠倒生、双珠被、厚珠心,大孢子四分体直线型排列,合点端为功能大孢子、雌配子体发育方式为蓼型。从雌、雄配子体发育时间的先后来看,深山含笑春季雌、雄配子体能正常发育,雄蕊先熟,雄蕊和花瓣凋谢后雌蕊大孢子母细胞才形成。而秋季开花的深山含笑,花药中小孢子在孢原细胞或初生造孢细胞期停止发育,花粉败育;雌蕊胚珠珠心组织也未见大孢子母细胞发育,开花后雌蕊随花柄凋落。该研究为深山含笑生殖发育和杂交育种积累了资料。     

刺五加、短梗五加的开花动态及繁育系统的比较研究

野外定位观测刺五加（Eleutherococcus senticosus),短梗五回（E.sessiliflorus)的开花进程,花朵的功能形态特征和开花动态,用杂交指数（OCI）,花粉－胚珠比（P／O）,去雄,套袋,人工授粉等方法分别测定刺五加,短梗五加的繁育系统,结果显示,刺五加种群,短梗五加种群的花期均持续1个月左右,刺五加比短梗五加早开花20d 左右,二均有雄蕊先熟现象,刺五加是单全异株植物,种群内既具有雄性,又具有雌株,还具有两性株,繁育系统主要为异交,需要传粉活动才能完成授粉过程,与刺五加不同,短梗五加仅具两性花,但繁育系统也以异交为主,短梗五加两性花中的雌,雄器官既在空间上分离,又在时间上分离,只能进行同株异花间或异株,异花间传粉才能受精结实。     

The chromatographic co-elution of dihydrodehydrodiconiferyl alcohol monopyranose with eleutheroside E in Eleutherococcus senticosus: implications for eleutheroside E assays

During a quantification assay of the constituents of Eleutherococcus senticosus by reverse-phase HPLC using acetonitrile:water gradient elution, it was observed that a recently reported component, dihydrodehydrodiconiferyl alcohol monopyranose, co-eluted with eleutheroside E. The implications of this finding for researchers and the herbal medicine industry are discussed. C     

华北驼绒藜大小孢子的发生及雌雄配子体发育过程的解剖学研究

运用石蜡切片技术对华北驼绒藜大、小孢子发生及雌、雄配子体的发育进行了研究.结果表明:(1)花药4室,花药壁由表皮、药室内壁、1~2层中层及1层绒毡层组成,药壁发育为基本型,腺质绒毡层,发育后期为二核或三核;(2)四分体胞质分裂为同时型,小孢子四分体多数为四面体型,偶见十字交叉型;(3)成熟花粉2-细胞型,单核小孢子时期存在不同比例的空壳花粉,从0%~80%不等;(4)胚珠倒生,双珠被,厚珠心,大孢子四分体直线型排列,合点端为功能大孢子,蓼型胚囊.     

北柴胡大小孢子发生和雌雄配子体发育的研究

用石蜡切片法对北柴胡的大、小孢子发生和雌、雄配子体发育过程进行观察研究.结果显示,北柴胡的胚珠为倒生型,单珠被,薄珠心.孢原细胞不经分裂直接发育成大孢子母细胞;四分体线性,多数情况下合点端的大孢子为功能大孢子,少数情况下亚合点端的大孢子也可发育为功能大孢子,蓼型胚囊;八核胚囊时期,珠心基部和两侧的一些珠心细胞保持自己的细胞质和形状,留存较久,成为珠心座细胞,珠被内表皮细胞发育为珠被绒毡层;花药壁发育类型为基本型,绒毡层为腺质绒毡层.小孢子母细胞减数分裂的胞质分裂为同时型,产生正四面体型四分体.成熟花粉为3细胞型.