Abnormalities of female gametophyte development in apomictic bluegrass forms

The structure of female gametophytes was studied at different developmental stages in apomictic bluegrass forms (Poa badensis Haenke, P. chaixii Vill., and P. pratensis L.). The range of structural abnormalities of undifferentiated and mature embryo sacs has been described. Possible mechanisms underlying their appearance are discussed and schematic linear arrangement of differentiation zones in the embryo sac are considered.     

The MIDASIN and NOTCHLESS genes are essential for female gametophyte development in Arabidopsis thaliana

Female gametophyte development in Arabidopsis thaliana follows a well-defined program that involves many fundamental cellular processes. In this study, we report the involvement of the Arabidopsis thaliana MIDASIN1 (AtMDN1) gene during female gametogenesis through the phenotypic characterization of plants heterozygous for an insertional mdn1 mutant allele. The MDN1 yeast ortholog has previously been shown to encode a non-ribosomal protein involved in the maturation and assembly of the 60S ribosomal subunit. Heterozygous MDN1/mdn1 plants were semisterile and mdn1 allele transmission through the female gametophyte was severely affected. Development of mdn1 female gametophyte was considerably delayed compared to their wild-type siblings. However, delayed mdn1 female gametophytes were able to reach maturity and a delayed pollination experiment showed that a small proportion of the female gametophytes were functional. We also report that the Arabidopsis NOTCHLESS (AtNLE) gene is also required for female gametogenesis. The NLE protein has been previously shown to interact with MDN1 and to be also involved in 60S subunit biogenesis. The introduction of an AtNLE-RNA interference construct in Arabidopsis led to semisterility defects. Defective female gametophytes were mostly arrested at the one-nucleate (FG1) developmental stage. These data suggest that the activity of both AtMDN1 and AtNLE is essential for female gametogenesis progression.     

Identification of genes expressed in the angiosperm female gametophyte

Until recently, identification of gene regulatory networks controlling the development of the angiosperm female gametophyte has presented a significant challenge to the plant biology community. The angiosperm female gametophyte is fairly inaccessible because it is a highly reduced structure relative to the sporophyte and is embedded within multiple layers of the sporophytic tissue of the ovule. Moreover, although mutations affecting the female gametophyte can be readily isolated, their analysis can be difficult because most affect genes involved in basic cellular processes that are also required in the diploid sporophyte. In recent years, expression-based approaches in multiple species have begun to uncover gene sets expressed in specific female gametophyte cells as a means of identifying regulatory networks controlling cell differentiation in the female gametophyte. Here, recent efforts to identify and analyse gene expression programmes in the Arabidopsis female gametophyte are reviewed.     

Identification of genes expressed in the Arabidopsis female gametophyte

The angiosperm female gametophyte typically consists of one egg cell, two synergid cells, one central cell, and three antipodal cells. Each of these four cell types has unique structural features and performs unique functions that are essential for the reproductive process. The gene regulatory networks conferring these four phenotypic states are largely uncharacterized. As a first step towards dissecting the gene regulatory networks of the female gametophyte, we have identified a large collection of genes expressed in specific cells of the Arabidopsis thaliana female gametophyte. We identified these genes using a differential expression screen based on reduced expression in determinant infertile1 (dif1) ovules, which lack female gametophytes. We hybridized ovule RNA probes with Affymetrix ATH1 genome arrays and validated the identified genes using real-time RT-PCR. These assays identified 71 genes exhibiting reduced expression in dif1 ovules. We further validated 45 of these genes using promoter::GFP fusions and 43 were expressed in the female gametophyte. In the context of the ovule, 11 genes were expressed exclusively in the antipodal cells, 11 genes were expressed exclusively or predominantly in the central cell, 17 genes were expressed exclusively or predominantly in the synergid cells, one gene was expressed exclusively in the egg cell, and three genes were expressed strongly in multiple cells of the female gametophyte. These genes provide insights into the molecular processes functioning in the female gametophyte and can be used as starting points to dissect the gene regulatory networks functioning during differentiation of the four female gametophyte cell types.     

Genetic analysis of female gametophyte development and function.

The female gametophyte is an absolutely essential structure for angiosperm reproduction. It produces the egg cell and central cell (which give rise to the embryo and endosperm, respectively) and mediates several reproductive processes including pollen tube guidance, fertilization, the induction of seed development, and perhaps also maternal control of embryo development. Although much has been learned about these processes at the cytological level, specific molecules mediating and controlling megagametogenesis and female gametophyte function have not been identified. A genetic approach to the identification of such molecules has been initiated in Arabidopsis and maize. Although genetic analyses are still in their infancy, mutations affecting female gametophyte function and specific steps of megagametogenesis have already been identified. Large-scale genetic screens aimed at identifying mutants affecting every step of megagametogenesis and female gametophyte function are in progress; the characterization of genes identified in these screens should go a long way toward defining the molecules that are required for female gametophyte development and function.     

矮沙冬青雌配子体及胚胎发育研究

矮沙冬青子房单心皮1室,边缘胎座,弯生胚珠,胚珠具双珠被、厚珠心。大孢子孢原细胞发生于珠心表皮下,大孢子母细胞减数分裂形成直线排列的四分体,合点端大孢子具功能,并按蓼型胚囊发育,雌配子体成熟于4月中旬。双受精后,胚乳发育为核型。在矮沙冬青大孢子发生、雌配子体和胚胎发育过程中未发现异常现象,因此认为矮沙冬青濒危不存在雌性生殖结构与发育过程异常的内在因素。     

Overseeding endophytic perennial ryegrass into stands of Kentucky bluegrass to manage bluegrass billbug (Coleoptera: Curculionidae)

Studies were conducted to determine the utility of overseeding endophyte enhanced varieties of perennial ryegrass, Lolium perenne L., into existing stands of Kentucky bluegrass, Poa pratensis L., to manage bluegrass billbug, Sphenophorus parvulus Gyllenhal, larval populations and their damage. Two commercial varieties of endophyte enhanced perennial ryegrass (Repell II and Triple Play) were overseeded at two different rates in the field. S. parvulus larval populations, their damage, and turfgrass composition were monitored over the course of 3 yr. Significantly fewer larvae were found in plots overseeded with Repell II, whereas Triple Play had no significant effect on larval population density. Endophyte infection was confirmed in approximately 58% of all Repell II shoots, whereas only approximately 31% of Triple Play shoots were infected. Regression analysis, using a segmented model, demonstrated that larval populations declined as the proportion of endophyte-infected perennial ryegrass increased. However, no additional reductions in larval population density were achieved in stands containing >40% endophyte-infected perennial ryegrass. Visual damage varied with cultivar, seeding rate, and year, but generally decreased as the proportion of endophyte-infected perennial ryegrass increased. Although seeding rate had an initial effect on turfgrass composition, these differences were not detected after the first year of the study. Results indicate overseeding existing stands of Kentucky bluegrass with endophytic varieties of perennial ryegrass can reduce S. parvulus larval populations and their damage. However, plant genetic characteristics (i.e., different cultivars) may be overriding determinants of many aspects of plant performance including establishment, tolerance to insect feeding, and expression of endophyte-mediated resistance.     

草地早熟禾草坪土壤水分动态与根系生长分布

对草地早熟禾草坪土壤水分动态和根系生长发育状况进行研究,结果发现不同土壤层次水分变化有所不同,0～15cm变化最大,15～30cm次之,30cm以下土层水分变化不大;草地早熟禾的根系生长呈现双峰曲线模式,5月中旬和8月中下旬总根量处于峰值;其主体根系主要分布在0～30cm土层内,占总根量的85%以上;根重密度随土层深度呈指数衰减关系,0～30cm土层下降幅度较大,30cm以下土层根重密度相差不大;在0～30cm土层内不同层次根量占总根量的比例在不同时期亦有差异,春秋季节10～20cm和20～30cm土层内根量比例较大,说明此时期主体根系分布在较深的土层;综合分析认为草地早熟禾草坪主要利用土壤浅层水分,在降雨较少的春秋季节,根系较深,适宜深层灌溉,在降雨频繁的夏季,根系较浅,适宜浅层灌溉。     

Megasporogenesis, female gametophyte development and embryonic development of Ambrosia L. in China

The megasporogenesis, female gametophyte development and embryonic development of Ambrosia artemisiifolia L. and Ambrosia trifida L. of genus Ambrosia L. in China were studied using conventional paraffin section technology and optical microscopy. The results show that both A. artemisiifolia L. and A. trifida L. have a bilobed pistil stigma, two carpels, one chamber, basal placenta, unitegmic, tenuinucellate, anatropous ovule, and well-developed integumentary tapetum. Megaspore mother cells are directly developed from archesporial cells originated from the nucellar cells under the nucellar epidermis and further undergo meiosis to form linear tetrads. The megaspore at the chalazal end develops into a functional megaspore and the other three megaspores are degraded. The development of embryo sac is monosporic type. After three consecutive mitosis, mononucleate embryo sac becomes a mature embryo sac with two synergids and one egg cell at the micropylar end, a central cell at the center and three antipodal cells at the chalazal end. Most antipodal cells are mononucleate or binucleate, only few are trinucleate. The embryonic development process contains four stages: globular embryo, heart-stage embryo, torpedo-stage embryo and mature embryo. The development of endosperm is cellular type.     

南方山荷叶大、小孢子发生与雌、雄配子体发育

首次报道了南方山荷叶大、小孢子发生和雌、雄配子体形成。主要结果如下:花药4室,花药壁发育为基本型,由7层细胞组成,由外到内依次为表皮、药室内壁、3层中层和2层绒毡层;药室内壁有带状加厚现象,绒毡层细胞多具双核,为腺质型绒毡层;多孢原,发生于表皮下;小孢子母细胞减数分裂为同时型,小孢子四分体多为四面体型,少数为十字交叉形、直线形、左右对称形或T形;成熟花粉粒为2-细胞型,偶见3-细胞型;成熟花粉粒呈圆形,具单萌发孔。雌蕊1枚,子房单心皮,纵切面呈瓶状;1室,边缘胎座,具4～5枚发育不同步的横生胚珠。胚珠具双珠被,厚珠心;珠孔由两层珠被共同形成,在一直线上或呈之字形。多孢原,位于珠心表皮下。直线形四分体,合点端的1个大孢子发育为功能大孢子,胚囊发育为蓼型。成熟胚囊中,3个反足细胞大而明显,但宿存时间短,较早退化。     

The development of female gametophyte and antipodal embryo formation in Sedum fabaria

In Sedum fabaria, the ovule is anantropus, bitegmic and crassinucellate. The development of the nucellus conforms to the Sedum type. The development of the embryo sac is of the Allium type. The antipodal cells in unfertilized embryo sac occasionally divide and one of them forms four-celled structures resembling embryos and remaining once elongate in the form of haustoria. The entry of the pollen tube is porogamous. After division the primary endosperm nucleus forms two cells: the apical one develops into cellular endosperm according to the Acre type and the basal one acts as the endosperm haustorium of the Sempervivum type. The embryogeny corresponds to the Caryophyllad type. This revised version was published online in July 2006 with corrections to the Cover Date.     

AGAMOUS‐LIKE13, a putative ancestor for the E functional genes,specifies male and female gametophyte morphogenesis

Arabidopsis AGL13 is a member of the AGL6 clade of the MADS box gene family. GUS activity was specifically detected from the initiation to maturation of both pollen and ovules in AGL13:GUS Arabidopsis. The sterility of the flower with defective pollen and ovules was found in AGL13 RNAi knockdown and AGL13 + SRDX dominant‐negative mutants. These results indicate that AGL13 acts as an activator in regulation of early initiation and further development of pollen and ovules. The production of similar floral organ defects in the severe AGL13 + SRDX and SEP2 + SRDX plants and the similar enhancement of AG nuclear localization efficiency by AGL13 and SEP3 proteins suggest a similar function for AGL13 and E functional SEP proteins. Additional fluorescence resonance energy transfer (FRET) analysis indicated that, similar to SEP proteins, AGL13 is able to interact with AG to form quartet‐like complexes (AGL13–AG)₂ and interact with AG–AP3–PI to form a higher‐order heterotetrameric complex (AGL13–AG–AP3–PI). Through these complexes, AGL13 and AG could regulate the expression of similar downstream genes involved in pollen morphogenesis, anther cell layer formation and the ovule development. AGL13 also regulates AG/AP3/PI expression by positive regulatory feedback loops and suppresses its own expression through negative regulatory feedback loops by activating AGL6, which acts as a repressor of AGL13. Our data suggest that AGL13 is likely a putative ancestor for the E functional genes which specifies male and female gametophyte morphogenesis in plants during evolution.     

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.     

功能雄性不育茄子差异基因及代谢途径分析

以茄子功能雄性不育系(S13)和可育系(F142)为材料,对花蕾不同发育时期的细胞学特征进行观察,发现S13花药中环形细胞簇的解体时期比F142延迟2d,开花当天F142的裂口组织及内绒毡层的细胞解体,而S13中无此现象发生.转录组测序分析表明,F142和S13开花前8天、5天和开花当天的花药共有1 436个差异表达基...