Sexual dimorphism in white campion: deletion on the Y chromosome results in a floral asexual phenotype

White campion is a dioecious plant with heteromorphic X and Y sex chromosomes. In male plants, a filamentous structure replaces the pistil, while in female plants the stamens degenerate early in flower development. Asexual (asx) mutants, cumulating the two developmental defects that characterize the sexual dimorphism in this species, were produced by gamma ray irradiation of pollen and screening in the M1 generation. The mutants harbor a novel type of mutation affecting an early function in sporogenous/parietal cell differentiation within the anther. The function is called stamen-promoting function (SPF). The mutants are shown to result from interstitial deletions on the Y chromosome. We present evidence that such deletions tentatively cover the central domain on the (p)-arm of the Y chromosome (Y2 region). By comparing stamen development in wild-type female and asx mutant flowers we show that they share the same block in anther development, which results in the production of vestigial anthers. The data suggest that the SPF, a key function(s) controlling the sporogenous/parietal specialization in premeiotic anthers, is genuinely missing in females (XX constitution). We argue that this is the earliest function in the male program that is Y-linked and is likely responsible for "male dimorphism" (sexual dimorphism in the third floral whorl) in white campion. More generally, the reported results improve our knowledge of the structural and functional organization of the Y chromosome and favor the view that sex determination in this species results primarily from a trigger signal on the Y chromosome (Y1 region) that suppresses female development. The default state is therefore the ancestral hermaphroditic state.     

A floral third whorl-specific marker gene in the dioecious species white campion is differentially expressed in mutants defective in stamen development

The CCLS4 gene of white campion is specifically expressed in anther epidermis and endothecium from pre- to post-meiotic stages. We report on a detailed in situ analysis of the gene's expression and show that it is a marker of the floral third whorl. The gene is expressed (1) in the anther (epidermis, parietal cells and the derived endothecium) in normally developing stamens and (2) in distinct sub-domains of third-whorl epidermis in mutants exhibiting aberrant states of parietal differentiation. Our results suggest that CCLS4 may fulfil different functions during pre- and post-meiotic anther development and reveal the complex role parietal cells may play during early stamen formation.     

Ethylene perception is involved in female cucumber flower development

It is well established that ethylene promotes female flower development in cucumber. However, little is known about how the gaseous hormone selectively affects female flowers, and what mechanism it uses. Previously, we found organ‐specific DNA damage in the primordial anther of female cucumber flowers. This finding led to a hypothesis that ethylene might promote female flower development via the organ‐specific induction of DNA damage in primordial anthers. In this study, we tested this hypothesis first by demonstrating ethylene induction of DNA damage via the ethylene signaling pathway using cucumber protoplasts. Then, using representative component genes of the ethylene signaling pathway as probes, we found that one of the ethylene receptors, CsETR1, was temporally and spatially downregulated in the stamens of stage‐6 female cucumber flowers, especially along with the increase of the nodes. Furthermore, by constructing transgenic Arabidopsis plants with organ‐specific expression of antisense CsETR1 under the control of an AP3 promoter to downregulate ETR1 expression in the stamens, we generated Arabidopsis ‘female flowers’, in which the abnormal stamens mimic those of female cucumber flowers. Our data suggest that ethylene perception is involved in the arrest of stamen development in female cucumber flowers through the induction of DNA damage. This opens up a novel perspective and approach to solve the half‐century‐long puzzle of how gaseous ethylene selectively promotes female flowers in the monoecious cucumber plant.     

Stamen abscission and water balance in Metrosideros flowers

Cymules (3-flowered units borne on single pedicels) were cut from inflorescences of Metrosideros collina J.R. & G. Forst. cv. Tahiti and used to test the effects of ethephon and ethylene on stamen abscission in the presence of silver thiosulphate (STS) and 1-methylcyclopropene (1-MCP), and to test the effects of holding solutions on cymule water balance and the progression of floral development. Flower bud and stamen abscission occurred in response to 0.5–5.0 and 0.1 μl l⁻¹ ethylene, respectively. Ethylene effects were partially negated by scrubbing exogenous ethylene, and more completely negated by STS (2.0 m M ). 1-MCP caused greater ethylene production and inhibited stamen abscission for only 1–2 days after treatment. Ethephon (10-10 000 mg l⁻¹) induced stamen wilting rather than abscission, an effect that was not negated by STS. Stamen wilting was negatively correlated with stamen relative water content, and the increase in stamen wilting was generally reduced by treatments that enhanced cymule mass. Stamen wilting was least using a 100 g l⁻¹ sucrose pulse or holding solutions containing 30–40 g l⁻¹ sucrose, with hydroxyquinoline citrate (200 mg l⁻¹) maintained at pH 5. Our results indicate that 1-MCP may be relatively ineffective in blocking the effects of ethylene on the abscission of organs, such as the stamens of M . collina , which are highly sensitive to this hormone.     

Stamen development in <Emphasis Type="Italic">Arabidopsis</Emphasis> is arrested by organ-specific overexpression of a cucumber ethylene synthesis gene <Emphasis Type="Italic">CsACO2</Emphasis>

Cucumber (Cucumis sativus L.) has served as a model to understand hormone regulation in unisexual flower development since the 1950s and the role of ethylene in promoting female flower development has been well documented. Recent studies cloned the F-locus in gynoecious lines as an additional copy of the ACC synthase (ACS) gene, which further confirmed the role of ethylene in the promotion of female cucumber flowers. However, no direct evidence was generated to demonstrate that increases in endogenous ethylene production could induce female flowers by arresting stamen development. To clarify the relationship between ethylene production and stamen development, we overexpressed the ethylene synthesis cucumber gene CsACO2 to generate transgenic Arabidopsis, driven by the organ-specific promoter P _AP3 . We found that organ-specific overexpression of CsACO2 significantly affected stamen but not carpel development, similar to that in the female flowers of cucumber. Our results suggested that increases in ethylene production directly disturb stamen development. Additionally, our study revealed that among all floral organs, stamens respond most sensitively to exogenous ethylene. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Characterization of an ethylene-inducible, calcium-dependent nuclease that is differentially expressed in cucumber flower development

? Production of unisexual flowers is an important mechanism that promotes cross-pollination in angiosperms. We previously identified primordial anther-specific DNA damage and organ-specific ethylene perception responsible for the arrest of stamen development in female flowers, but little is known about how the two processes are linked. ? To identify potential links between the two processes, we performed suppression subtractive hybridization (SSH) on cucumber (Cucumis sativus L.) stamens of male and female flowers at stage 6, with stamens at stage 5 of bisexual flowers as a control. ? Among the differentially expressed genes, we identified an expressed sequence tag (EST) encoding a cucumber homolog to an Arabidopsis calcium-dependent nuclease (CAN), designated CsCaN. Full-length CsCaN cDNA and the respective genomic DNA sequence were cloned and characterized. The CsCaN protein exhibited calcium-dependent nuclease activity. CsCaN showed ubiquitous expression; however, increased gene expression was detected in the stamens of stage 6 female flowers compared with male flowers. As expected, CsCaN expression was ethylene inducible. It was of great interest that CsCaN was post-translationally modified. ? This study demonstrated that CsCaN is a novel cucumber nuclease gene, whose DNase activity is regulated at multiple levels, and which could be involved in the primordial anther-specific DNA damage of developing female cucumber flowers.     

Expression of ASK1-like genes in arrested stamens of female Silene latifolia plants

Silene latifolia is a dioecious plant in which sex is determined by heteromorphic sex chromosomes. In female plants, stamen development is arrested before microspore mother cells are formed. In this study, we isolated four cDNAs (SlSKP1-1 to 4) encoding ASK1-like protein as expression markers to reveal when expression levels are reduced in arrested stamens of female flowers. Expression patterns of the SlSKP1 genes were analyzed by in-situ hybridization. We use the flower development classification of Grant et al. (in Plant J 6:471–480, 1994). SlSKP1 genes were highly expressed in primary parietal cells and primary sporogenous cells that develop into microspore mother cells in male flowers. Expression levels started to be reduced in the external stamens of the female flowers when stamen development was arrested at stage 7. Although microspore mother cells could not be developed in female flowers and SlSKP1 expression may be unnecessary in arrested stamens, SlSKP1 genes were still expressed in sporogenous cells of degenerated stamens at stage 8. Parietal cells stopped differentiating earlier than sporogenous cells in arrested stamens. These results suggest that not all types of cell are arrested simultaneously at a particular stage of stamen development during stamen suppression of S. latifolia.     

MORPHOGENESIS OF THE STAMENLESS-2 MUTANT IN TOMATO. I. COMPARATIVE DESCRIPTION OF THE FLOWERS AND ONTOGENY OF STAMENS IN THE NORMAL AND MUTANT PLANTS

The single gene recessive mutant stamenless-2 (sl₂/sl₂) differs phenotypically from the normal (+/+) only in the stamen structure. Stamens of the mutant plants were laterally free, twisted, shorter, paler in color, possessed abnormal pollen, and bore naked external ovules (E.O.) on the adaxial surface near the junction of anther and filament. Mutant plants grown in the field during summer produced flowers in which a number of carpel-like organs (‘carpelloid stamens‘) with few or no E.O. replaced the stamens. On the other hand, plants grown in the greenhouse during winter possessed flowers with greater number of yellow and pubescent stamens and many E.O. Study of stamen ontogeny revealed that at initiation (up to 100 μ in length) stamen primordia of normal and mutant plants resembled each other. Thereafter the development of stamens in the two genotypes could be distinguished.     

两性花的雄蕊运动:多样性和适应意义

雄蕊运动指雄蕊在自身能量支持下发生的主动运动,不包括雄蕊在访花者触碰下造成的被动位移。该文总结了雄蕊的应激运动、快速猛烈弹射、缓慢运动以及级联运动等4种主要类型,分析了这些运动类型的系统分布及繁殖适应意义等方面的研究进展。雄蕊的应激运动由访花者或其他外力诱发,可能起到促进散粉和实现繁殖保障的作用;雄蕊快速猛烈的弹射运动可将花粉猛然撒向空中或访花者身上,促进了花粉的风媒或虫媒散布;缓慢运动的雄蕊可能通过在不同花期改变雄蕊的空间位置和雌雄异位程度来调节繁殖策略,或主动将雄蕊花药移至特定部位(如柱头表面)实现自交;雄蕊逐一、依次发生的级联运动较为复杂,主要分布在刺莲花科、梅花草科、旱金莲科和芸香科中,目前还缺乏实验研究;但根据"花粉呈现理论"以及其他类型的雄蕊运动研究结果,雄蕊的级联运动可以将花粉分批呈现给不同的传粉者,通过不同传粉者的分别传粉来提高花粉的输出;而且可避免已散粉雄蕊对即将散粉雄蕊的干扰,同时可能也降低了雌雄功能干扰和(或)花内自交。在芸香(Ruta graveolens)中,级联运动之后的雄蕊还会在花末期再同时向花中央运动;这种多向、多次运动方式是目前发现的最复杂的雄蕊运动类型。雄蕊运动领域值得今后开展进一步实验研究的方向主要有:1)雄蕊运动尤其是级联运动对雌雄功能干扰(性别间干扰)、雄蕊与雄蕊的"性别内干扰"等植物繁殖格局的影响;2)雄蕊运动与雌雄异熟、雌雄异位等花部特征的相互作用;3)雄蕊运动复杂类型的生理与发育机制。     

Isolation of early genes expressed in reproductive organs of the dioecious white campion (Silene latifolia) by subtraction cloning using an asexual mutant

The dioecious white campion (Silene latifolia) has been chosen as a working model for sexual development. In this species, sexual dimorphism is achieved through two distinct developmental blocks: inhibition of carpel development in male flowers, and early arrest of anther differentiation in female flowers. The combined advantages of the dioecious system and the availability of a sexual mutant lacking both male and female reproductive organs have been exploited in a molecular subtraction approach using male and asexual flower buds. This resulted in the cloning of 22 cDNA clones expressed in stamens at distinct stages of development. Fourteen of these clones corresponded to genes whose expression was detected in pre-meiotic stamens, a stage of development for which very little information is presently available. Furthermore, the absence of similarities with database sequences for ten clones suggests that they represent novel genes. Functional analysis of each clone will enable their positioning within the reproductive organ developmental pathway(s). In parallel, these clones are being exploited as developmental markers of early differentiation within the flower.     

Silver Ions Increase Auxin Efflux Independently of Effects on Ethylene Response

Silver nitrate and aminoethoxyvinylglycine (AVG) are often used to inhibit perception and biosynthesis, respectively, of the phytohormone ethylene. In the course of exploring the genetic basis of the extensive interactions between ethylene and auxin, we compared the effects of silver nitrate (AgNO₃) and AVG on auxin responsiveness. We found that although AgNO₃ dramatically decreased root indole-3-acetic acid (IAA) responsiveness in inhibition of root elongation, promotion of DR5-β-glucuronidase activity, and reduction of Aux/IAA protein levels, AVG had more mild effects. Moreover, we found that that silver ions, but not AVG, enhanced IAA efflux similarly in root tips of both the wild type and mutants with blocked ethylene responses, indicating that this enhancement was independent of ethylene signaling. Our results suggest that the promotion of IAA efflux by silver ions is independent of the effects of silver ions on ethylene perception. Although the molecular details of this enhancement remain unknown, our finding that silver ions can promote IAA efflux in addition to blocking ethylene signaling suggest that caution is warranted in interpreting studies using AgNO₃ to block ethylene signaling in roots.     

大戟科麻疯树属三种植物花器官发生

利用扫描电子显微镜观察了大戟科Euphorbiaceae麻疯树属Jatropha麻疯树J. curcas L.、佛肚树J. podagrica Hook.和棉叶麻疯树J. gossypifolia L.花器官发生。结果表明: 麻疯树、佛肚树和棉叶麻疯树花萼原基均为2/5型螺旋发生。在同一个种不同的花蕾中, 花萼的发生有两种顺序: 逆时针方向和顺时针方向。远轴面非正中位的1枚先发生。5枚花瓣原基几乎同时发生。雄花中雄蕊两轮, 外轮对瓣, 内轮对萼。研究的3种麻疯树属植物雄蕊发生方式有两种类型: 麻疯树亚属麻疯树的5枚外轮雄蕊先同时发生, 5枚内轮雄蕊后同时发生, 佛肚树亚属佛肚树和棉叶麻疯树雄蕊8-9枚, 排成两轮, 内外轮雄蕊同时发生。雌花的3枚心皮原基为同时发生。麻疯树属单性花, 雌花的子房膨大而雄蕊退化, 雄花的雄蕊正常发育, 子房缺失。根据雄蕊发生方式, 支持将麻疯树属分为麻疯树亚属subgen. Jatropha和佛肚树亚属subgen. Curcas。     

Allocation to floral structures in Thalictrum pubescens (Ranunculaceae), a cryptically dioecious species

Females of Thalictrum pubescens produce stamens that contain sterile pollen, whereas males are both functionally and morphologically unisexual. This study examines the investment in stamen production by females of T. pubescens by comparing the female structures with those of their fully functional male counterparts. Stamens from females had the same biomass and contained the same amount of nitrogen and phosphorus as stamens from males. Anther size was the same in males and females, but filaments were longer in stamens from males. Females produced more pollen per anther than males, and pollen size was the same in both sexes. Within flowers, there was a positive correlation between the amount of pollen per anther and the length of anthers in males, but not in females. This would be expected if males growing in better environmental conditions or with greater vigour invested more resources in pollen production, thereby increasing fitness. Females, who receive no fitness benefits from increased pollen production, did not show this pattern. There was also evidence of a trade-off within female flowers between the number of stamens and the number of pistils. This trade-off was noted in conditions when variance among plants was reduced, namely in the field during a year when flower size was particularly small and in a previous glasshouse study. Therefore, it appears that when environmental variance is low, stamens are produced at the expense of producing more pistils, and hence seeds. In conclusion, stamen production does not appear to be inconsequential to females of Thalictrum pubescens.     

The mechanism of stamen movement in <Emphasis Type="Italic">Chimonanthus praecox</Emphasis> (Calycanthaceae): differential cell growth rates on the adaxial and abaxial surfaces of filaments after flower opening

In flowers of Chimonanthus (Calycanthaceae), stamen movements have been known for 150 years. Stamens are initially recurved when flowers open. Subsequently, within approximately 48 h, the stamens gradually turn upright and eventually enclose the gynoecium. However, the exact mechanism involved in this process is still unresolved. In this study, we found that in recurved stamens at the female stage the cell lengths in the epidermal and hypodermal tissue of the abaxial surfaces were significantly smaller than those of the adaxial surfaces. In erect stamens, however, no significant difference in cell length between the hypodermal tissues of both surfaces was found. Although the difference in cell length between the epidermal tissues for erect stamens was significant, it was smaller than that in recurved filaments. We found no variations in cell number between the epidermal tissues or between the hypodermal tissues among recurved and erect stamens. Thus, we conclude that in Chimonanthus the differential cell growth rates between the adaxial and abaxial surfaces of filaments could account for the gradual inward stamen movement following flower opening rather than cell division. Furthermore, application of indole-3-acetic acid (IAA) and gibberellic acid (GA₃) to intact flowers in vitro or the wounding of stigmas significantly promoted the stamen erection rate. Surprisingly, we did not observe any effect on this movement following hand-pollination. Different patterns of stamen movement in Calycanthaceae and their implications for reproductive biology are also discussed.     

The forkhead-associated domain 2 (FHA2) in Arabidopsis plays a role in plant fertility by regulating stamen development

The forkhead-associated (FHA) domain is involved in protein–protein interaction by recognizing a phosphothreonine epitope on target proteins. In this study, we investigated in planta functions of the Arabidopsis FHA domain 2. AtFHA2 was mainly localized in the nucleus. Arabidopsis fha2 null mutants grew normally during the vegetative stage, but had severely reduced fertility during reproductive stage. The reduced fertility was mainly caused by defective stamen filament elongation, while female flower parts of the mutants were fertile. Additionally, the mutants had fewer stamens than the wild type and the vegetative organs of the mutants, such as cotyledons and leaves, had increased ploidy. These results suggest that AtFHA2 may play a role in a signaling pathway for the control of plant organ development.     

Correlation between male and female reproduction in the subdioecious herb Astilbe biternata (Saxifragaceae)

Genotypic trade-offs between male and female reproduction are commonly assumed in theoretical studies of the evolution of gender specialization. Although these trade-offs are supported by higher seed production of females than hermaphrodites in natural populations of gynodioecious species, comparisons between male and female reproductive allocation among hermaphrodite individuals under controlled conditions are rare. We assessed phenotypic and genotypic correlations between stamen and fruit production in fruiting males of the near-dioecious herb Astilbe biternata. In the field, we found a significant negative phenotypic correlation between stamen production and fruit production within individuals that produced both stamens and fruit as well as higher fruit set in females than fruiting males. The negative correlation between fruit and stamen production that was observed in the field was also apparent across clonally propagated genotypes. These results suggest that negative genetic correlations between male and female reproduction may limit the independent evolution of fruit and stamen production in A. biternata.     

水蕹菜（Ipomoea aquatica）对含银废水的净化功能研究

 本文通过对水蕹菜净化含银废水的室内外静、动态试验研究得出：(1)水蕹菜对离子态银(Ag+)的去除速率高于对络合态银(Ag(S2O3)2)3-的去除速率即KAg+>K[Ag(S2O3)2]3-;(2)水蕹菜对银的净化率R与污水停留时间t呈指数函数关系(R=AeB/t,A、B为常数,B<0),其表面去除负荷Ps则与t呈幂函数关系(Ps=AtB B<0);(3)试验证明水蕹菜是生物净化含银废水的优良品种之一。     

Sex Determination by Sex Chromosomes in Dioecious Plants

Abstract: Sex chromosomes have been reported in several dioecious plants. The most general system of sex determination with sex chromosomes is the XY system, in which males are the heterogametic sex and females are homogametic. Genetic systems in sex determination are divided into two classes including an X chromosome counting system and an active Y chromosome system. Dioecious plants have unisexual flowers, which have stamens or pistils. The development of unisexual flowers is caused by the suppression of opposite sex primordia. The expression of floral organ identity genes is different between male and female flower primordia. However, these floral organ identity genes show no evidence of sex chromosome linkage. The Y chromosome of Rumex acetosa contains Y chromosome-specific repetitive sequences, whereas the Y chromosome of Silene latifolia has not accumulated chromosome-specific repetitive sequences. The different degree of Y chromosome degeneration may reflect on evolutionary time since the origination of dioecy. The Y chromosome of S. latifolia functions in suppression of female development and initiation and completion of anther development. Analyses of mutants suggested that female suppressor and stamen promoter genes are localized on the Y chromosome. Recently, some sex chromosome-linked genes were isolated from flower buds of S. latifolia.     

花内雄蕊分化及其适应意义

对花内雄蕊存在显著分化的现象进行了分析与归纳, 总结了花内雄蕊分化的各种主要形式及其繁殖适应意义。“花内雄蕊分化”是指花内雄蕊与雄蕊之间存在显著分化的现象, 这一概念可以把二强雄蕊、四强雄蕊和异型雄蕊等以往单独进行研究的相关性状结合起来, 并明确区分了几种新的花内雄蕊分化形式, 以期更准确全面地认识这些相关性状的适应意义与进化。该文将花内雄蕊分化区别为花丝的分化、花药的分化、雄蕊合生的分化、雄蕊运动的分化、退化雄蕊5大类。花丝的分化主要是花丝长度的分化, 如四强雄蕊、二强雄蕊和单强雄蕊; 花药分化主要指花药颜色、花药与花粉粒大小和花药开裂时间等的分化; 雄蕊合生的分化主要体现为花内部分雄蕊合生而部分雄蕊离生; 雄蕊运动的分化指的是花内雄蕊在运动时间或方式上存在差异, 造成雄蕊处于不同的成熟阶段和位于不同的空间位置; 退化雄蕊则是花内部分雄蕊失去了生产花粉的繁殖功能, 通常也发生了花药形态上的巨大改变。异型雄蕊不仅存在花丝和花药的形态分化, 还存在着明显的功能分化, 是分化程度很高的一类特殊的花内雄蕊分化形式。一些特殊的繁育系统, 如异长花柱和镜像花柱等在种内不同个体上存在着不同形式的花内雄蕊分化。花内雄蕊分化在花内造成了多个不同的花药位置, 在很大程度上影响了雌雄异位程度, 对植物自交与异交水平、花内雌雄功能干扰等有着潜在作用; 花内雄蕊分化形成的多个不同空间位置的雄蕊还增加了对多种访花者的吸引与适应潜力, 有可能影响到访花者的类型与访花行为, 得以适应多种传粉者。此外, 花内雄蕊分化可将花粉逐渐分批次分发给访花者, 提高花粉散布效率, 可看成是“花粉呈现理论”所指的花粉装配与分发机制之一。现有的实验研究发现, 花内雄蕊分化能够吸引传粉者、保护正常花药和花粉、促进花粉散发(降低花粉竞争)、实行延迟自交和降低花内雌雄功能干扰等。花内雄蕊分化还缺少系统研究, 有些雄蕊分化形式如单强雄蕊和雄蕊运动的分化还没有针对性的实验揭示其适应意义, 鸭跖草科和某些豆科植物的雄蕊三型分化等现象也缺少进化适应意义的研究。花内雄蕊分化对植物雌性和雄性适合度可能不同的影响、如何与访花者相互作用、如何与其它花部特征一起影响植物繁殖过程等, 可能是这一领域值得今后优先研究的课题。