五唇兰雌配子体发育和胚胎发生的研究

五唇兰的胚珠倒生型,具薄珠心,两层珠被。胚囊发育为双孢子葱型,成熟胚囊８核。从传粉到受精约５０ｄ,正常双受精。胚具５－６细胞的胚柄,种子成熟时胚柄及胚乳核消失,成熟种子只具单层细胞的种皮和一个未分化的珠珠形胚。     

大花君子兰大小孢子发生与雌雄配子体发育的研究

该研究运用常规石蜡切片技术,对大花君子兰(Clivia miniata Regel)大、小孢子发生及雌、雄配子体发育进程进行解剖学观察分析,以探讨君子兰生殖生物学解剖特征,为君子兰种子发育和育种提供理论依据。结果表明：(1)大花君子兰花药4室,具分泌型绒毡层。(2)小孢子母细胞减数分裂的胞质分裂为连续型,小孢子四分体为左右对称型,成熟花粉为二细胞型。(3)倒生胚珠,双珠被,厚珠心和雌配子体发育为蓼型。(4)记录了雌雄配子体发育的对应关系,发现雄配子体发育趋于同步,雌配子体发育不同步。(5)开花散粉时,雌配子体尚有处于四核、八核胚囊的时期;成熟胚囊阶段,中央细胞的2个极核位于反足细胞端,反足细胞呈退化状态。具承珠盘结构。     

ScRALF3, a secreted RALF‐like peptide involved in cell–cell communication between the sporophyte and the female gametophyte in a solanaceous species

Small peptides have been shown to regulate numerous aspects of plant development through cell–cell communication. These signaling events are particularly important during reproduction, regulating gamete development and embryogenesis. Rapid alkalinization factor (RALF)‐like genes, a large gene family that encodes secreted peptides, have specific or ubiquitous expression patterns. Previously, five RALF‐like genes with potential involvement during reproduction were isolated from Solanum chacoense. Here, we show that ScRALF3 is an important peptide regulator of female gametophyte development. Its expression, which is auxin‐inducible, is strictly regulated before and after fertilization. Down‐regulation of ScRALF3 expression by RNA interference leads to the production of smaller fruits that produce fewer seeds, due to improper development of the embryo sacs. Defects include loss of embryo sac nuclei polarization, as well as an increase in asynchronous division, accounting for cellular dysfunctions and premature embryo sac development arrest during megagametogenesis. ScRALF3 is expressed in the sporophytic tissue surrounding the embryo sac, the integument and the nucellus, as revealed by in situ hybridization and GUS staining. As expected for a secreted peptide, fluorescence from an ScRALF3–GFP fusion construct is detected throughout the secretory pathway. Therefore, the ScRALF3 secreted peptide may be directly involved in the regulation of multiple aspects of cell–cell communication between the female gametophyte and its surrounding sporophytic tissue during ovule development.     

马哈利樱桃大孢子发生和雌配子体的发育

采用石蜡切片法对马哈利樱桃大孢子发生和雌配子体发育过程进行观察研究。结果表明:(1)马哈利樱桃雌配子体发育早期,在单室子房内可以看到2个倒生胚珠,但在后期其中一个退化,另一个发育为种子;其胚珠具双珠被,为厚珠心。(2)大孢子母细胞减数分裂形成直线型四分体,功能大孢子位于合点端;胚囊发育为蓼型,成熟胚囊为七细胞八核。(3)根据不同时间花的外部形态特征与内部解剖学对比的观察结果,在陕西关中地区,三月下旬是马哈利樱桃雌性生殖细胞分化和发育的重要时期,果园在此期间应加强肥水管理。     

Plasticity in Cell Division Patterns and Auxin Transport Dependency during in Vitro Embryogenesis in Brassica napus

In Arabidopsis thaliana, zygotic embryo divisions are highly regular, but it is not clear how embryo patterning is established in species or culture systems with irregular cell divisions. We investigated this using the Brassica napus microspore embryogenesis system, where the male gametophyte is reprogrammed in vitro to form haploid embryos in the absence of exogenous growth regulators. Microspore embryos are formed via two pathways: a zygotic-like pathway, characterized by initial suspensor formation followed by embryo proper formation from the distal cell of the suspensor, and a pathway characterized by initially unorganized embryos lacking a suspensor. Using embryo fate and auxin markers, we show that the zygotic-like pathway requires polar auxin transport for embryo proper specification from the suspensor, while the suspensorless pathway is polar auxin transport independent and marked by an initial auxin maximum, suggesting early embryo proper establishment in the absence of a basal suspensor. Polarity establishment in this suspensorless pathway was triggered and guided by rupture of the pollen exine. Irregular division patterns did not affect cell fate establishment in either pathway. These results confirm the importance of the suspensor and suspensor-driven auxin transport in patterning, but also uncover a mechanism where cell patterning is less regular and independent of auxin transport.     

The Putative Factors Involved in the Induction of Embryogenesis in Angiosperms

We quantified various endogenous cytokinins during wheat (Triticum aestivumL.) and dandelion (Taraxacum officinaleWeb.) ovary development. Wheat ovaries were studied at the following developmental stages: the mature embryo sac with eight nuclei (stage 1), the interphasic zygote 12 h and 24 h after fertilization (stage 2), and the onset of zygote division (stage 3). The dandelion ovaries were studied at the stage of the mature embryo sac (stage 1), in the interphase of the parthenogeneticaly developing ovule (stage 2), and during its first division (stage 3). The material was analyzed by the method of competitive solid-phase immunoenzyme assay (ELISA) using peroxidase-labeled anti-rabbit antibodies. The onset of embryogenesis in wheat and dandelion ovules was accompanied by the substantial rearrangement of their hormonal complexes, which preceded the morphogenetic processes leading to seed formation. This implies that the hormonal system of the whole maternal plant is involved in the induction of embryogenesis. The final stages of embryogenesis depend on the hormonal systems in the flower, ovary, and ovule.     

毛竹实时荧光定量PCR内参基因的筛选及成花基因PheTFL1表达分析

通过qRT-PCR对毛竹相关成花基因PheTFL1的表达进行研究,为毛竹开花机理的研究提供理论依据.从毛竹UBC18、PP2A和EF1α等9个候选内参基因中筛选出在叶、幼嫩花序、花序轴、枝、竹青等11个组织器官中都稳定表达的PP2A用于毛竹PheTFL1基因qRT-PCR结果的校正.结果显示:PheTFL1基因在开花竹叶、枝和竹青中低丰度表达,与未开花竹差异不显著,但在花和花序轴中高丰度表达;在实生苗叶和根中高丰度表达,在实生苗茎中低丰度表达.PheTFL1基因在具有分生能力的幼嫩组织中高丰度表达,说明其不仅参与花发育的调控,还参与了分生组织生长的调控.     

无距虾脊兰胚珠发育及种子形成研究

采用石蜡切片、半薄切片、扫描电镜技术对无距虾脊兰不同时期的子房(蒴果)进行研究.结果表明:(1)无距虾脊兰授粉后19d,胎座上分化出上万个胚珠原基,这些胚珠原基由1列细胞外包1层表皮细胞构成,其中胚珠原基内部顶端的细胞分化为孢原细胞,授粉后45 d,孢原细胞发育分化为大孢子母细胞.(2)无距虾脊兰成熟胚珠为倒生胚珠,双珠被,薄珠心,胚囊发育为蓼型,且胚珠的发育即便在同一个果实内也是不同步的.(3)受精后合子经过一次不均衡横裂形成基细胞和顶细胞;基细胞不参与胚体构成,分化为单细胞的胚柄,最后退化消失;顶细胞经多次分裂形成原球胚,胚胎发育类型为石竹型.(4)成熟种子呈纺锤形,由球形胚和内外双层种皮构成,双层种皮分别由内外珠被发育而来.     

濒危植物马蹄参的大孢子发生和雌配子体发育

为探讨马蹄参(Diplopanax stachyanthus Hand.-Mazz.)濒危机制与雌性生殖发育的关系,采用石蜡切片法观察马蹄参大孢子形成和雌配子体的发育过程。结果表明,马蹄参雌蕊单心皮,子房下位,1室,1枚胚珠。胚珠横生于短片状胎座上,具单珠被,厚珠心。单孢原细胞自珠心1~2层表皮细胞处分化;大孢子四分体为直线形。成熟胚囊中,2个极核在受精前融合为次生核,3个反足细胞不发达,较早退化;二核胚囊时期出现二核分裂不均且较小核退化消失的异常发育现象。因此,马蹄参雌配子体发育过程中出现异常现象是造成其结实率低的主要原因。     

EMBRYO SAC DEVELOPMENT IN THE CV. KS MALE-STERILE,FEMALE-STERILE LINE OF SOYBEAN (GLYCINE MAX)

Light microscopic observations were made on 22 ovules from fertile plants and 108 ovules from sterile plants of the cv. KS synaptic mutant, a highly male-sterile, female-sterile line of soybean [Glycine max (L.) Merr.] (2n = 2x = 40). Ovules of fertile siblings contained normal embryo sacs and embryos. Ovules from sterile plants contained various irregularities. The most consistent abnormality was the failure of the embryo sac to attain normal size. Small megasporocytes of irregular shape were seen; only one megasporocyte of normal shape and size was noted. No linear tetrads were found. However, two ovules contained nonlinear triads. A range from zero to 28 cells and nuclei, of various sizes, were identifiable in small megagametophytes and embryo sacs. Degeneration of these nuclei and cells was noted as early as the four-nucleate gametophyte stage. Other ovules contained degenerated nucellar centers without embryo sacs. Two ovules appeared to be normal. Late postpollination stages were marked by shrunken nucellus and integuments. The presence of pollen tube traces, endosperm, and aborting embryos in ovules of hand-pollinated flowers from sterile plants suggested that no incompatibility was involved. Degeneration of the gametophyte and embryo sac contents at many developmental stages indicated a wide array of effects, possibly resulting from meiotic irregularities similar to those seen in microsporogenesis of this mutant.     

罗汉果大孢子发生及雌配子体发育与花部形态、胚珠变化的关系

为弄清罗汉果(Siraitia grosvenorii)大孢子发生、雌配子体发育过程与花部形态特征、胚珠的关系,运用石蜡切片法对罗汉果子房进行了显微观察。结果表明,罗汉果的胚珠倒生,双珠被,厚珠心,大孢子四分体呈线型排列,合点端一个大孢子分化为功能大孢子,成熟胚囊为蓼型。花蕾形态、胚珠变化与大孢子发生、雌配子体的发育时期具有一定相关性,当子房长度为7.0 mm≤L<9.0 mm,珠心呈椭圆形时,约有45.83%的大孢子母细胞处于减数分裂时期。因此,依据罗汉果花部形态可有效确定大孢子发生与雌配子体发育的时期。     

八角莲大孢子发生和雌配子体形成

首次报道了八角莲(Dysosma versipellis (Hance)M.cheng)大孢子发生和雌配子体形成的过程.结果:双珠被,多为厚珠心胚珠,少数为假厚珠心,胚珠多为横生,少数为弯生;边缘胎座,子房一室,多胚珠,珠孔由两层珠被共同形成,呈"之"字形;多为单孢原,位于珠心表皮下:偶见2～3个孢原细胞位于珠心表皮下;大孢子母细胞有两种发生方式;直线形大孢子四分体,合点端的大孢子发育为功能大孢子,蓼型胚囊;成熟胚囊中,二个极核在受精前合并为次生核;三个反足细胞不发达,较早退化;"品"字形卵器极性明显,其中卵细胞与助细胞极性相反;助细胞发达,其丝状器在不同发育时期形态及大小不同,且具吸器功能.     

依兰(番荔枝科)雌配子体发育研究

荔枝科是被子植物的基部类群之一,依兰属是番荔枝科较为原始的类群,其有性生殖过程,特别是胚胎发生与发育的研究结果,可以补充被子植物胚胎学原始特征的相关基础资料。该研究利用常规石蜡切片技术,对依兰胚珠、大孢子和雌配子体的发生发育过程进行了观察。结果表明:依兰的胚珠为倒生胚珠、厚珠心、三层珠被,第三层珠被(中间珠被)发生在大孢子母细胞时期,于外珠被与内珠被之间、胚珠合点端两侧发生并隆起;雌配子体为蓼型。此外,依兰的个别胚珠中存在双雌配子体现象,且两个雌配子体均由大孢子母细胞发育而来,大小、形状相近,呈线形排列。该研究结果对于揭示原始被子植物胚胎发育特征具有重要意义。     

小叶兜兰的果实生长和雌配子体发育

为了解濒危兰科植物小叶兜兰(Paphiopedilum barbigerum Tang et Wang)胚珠和雌配子体的发育过程,采用常规石蜡切片技术对其果实的生长动态进行了研究。结果表明,授粉后60~75 d的蒴果内种子数量迅速增加,到授粉后120 d时种子充满整个蒴果。授粉后40 d的胎座上分化形成多数由1层表皮细胞包被1列细胞的胚珠原基;授粉后60 d时位于胎座指状结构末端处紧靠表皮细胞下方的孢原细胞分化为大孢子母细胞。之后,大孢子母细胞经过减数分裂和有丝分裂最终形成成熟胚囊;授粉后135 d胚囊发育成熟,附着在胎座上的种子个体分化明显。小叶兜兰胚囊的发育类型为双孢子葱型,胚珠为倒生胚珠,薄珠心,单珠被,成熟胚囊为8核。这为小叶兜兰的生殖生物学及繁殖体系的建立提供理论依据。     

延龄草(Trillium tschonoskii Maxim.)的胚胎学研究初报Ⅰ.——大孢子的发生及雌配子体的形成

本文描述延龄草(Trillium tschonoskii Maxim.)的大孢子发生,雌配子体的形成和雄配子体的形态。胚珠为倒生型,双珠被,厚珠心型。胎座为侧膜胎座向中轴胎座的过渡类型,胶囊发育为葱型的变异型。孢原细胞直接发生于幼胚珠的珠心表皮细胞之下,孢原细胞平周分裂,形成初生周缘细胞及初生造孢细胞。初生周缘细胞分裂先于初生造孢细胞,分裂结果与珠心表皮细胞共同形成了珠心组织。初生造孢细胞进一步发育,形成大孢子母细胞。大孢子母细胞经减数第一次分裂后,即出现壁,形成二分体。一般是珠孔端二分体细胞小于合点端二分体细胞,但偶尔也见到前者大于后者的情况。在二分体形成后珠孔端二分体细胞立即退化、或经减数第二次分裂后再退化(该次分裂多为斜向的)。合点端二分体细胞发育,经二核胚囊,四核胚囊,六核胚囊阶段至成熟胚囊。一般在珠孔端的周围淀粉粒丰富,并先于合点端的核进行分裂。珠孔端由二个助细胞,一个卵细胞构成卵器,助细胞具钩突,并具丝状器,两个极核。合点端常见多核仁的大核,成熟胚囊未见八核。成熟花粉粒为二细胞的,花药壁具变形绒毡层,花粉中充满淀粉粒。沼生目型胚乳。     

Localization of arabinogalactan-proteins in different stages of embryos and their role in cotyledon formation of <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> L.

Arabinogalactan proteins (AGPs) have been implicated in plant development including sexual plant reproduction. In this paper, the expression of AGPs and the effects of β-glucosyl Yariv reagent (βGlcY, which binds arabinogalactan proteins) in embryo development and cotyledon formation were investigated. Immunofluorescence assay displayed that the expression of AGPs labeled with antibody JIM13 was developmentally regulated. In early stages, AGPs were evenly distributed in the whole embryo, except for a short polar expression in the basal suspensor cell. In the globular stage of embryo, AGPs were condensed in the embryo proper (EP), apex of the EP, and at the juncture of the EP and suspensor. In heart-shaped embryo, APGs were only present at the juncture of the EP and suspensor. Immunogold labeling assay showed that the strong expression of AGPs at the juncture of the EP and suspensor was localized in the cell wall. Provision of βGlcY to the in vitro ovule culture medium caused delayed growth of embryos, cotyledon defect and abnormal venation pattern. Consequently, βGlcY induced the death of defective seedlings with the characteristics of deformed or irregular single cotyledon. Our results suggested that AGPs play functional roles in embryo development, cotyledon formation and seedling morphology establishment in Nicotiana tabacum L.     

粉叶小檗大孢子发生和雌配子体形成

采用石蜡切片方法对粉叶小檗（Berberis pruinosa Franch.)的大孢子发生和雌配子体形成过程进行了研究。主要结果如下：雌蕊1枚,子房单心皮,边缘胎座,2枚胚珠倒生,具双珠被,厚珠心,珠孔由内外两层珠被共同形成,呈“Z”字形;单孢原,位于珠心表皮下;直线形大孢子四分体,合点端的1个大孢子发育为功能大孢子,胚囊发育类型为蓼型;成熟胚囊中,2个极核在受精前融合为次生核;3个反足细胞不发达,较早退化;"品"字形卵器,其中助细胞发达且具丝状器。