异质性水分环境中克隆整合对活血丹生物量分配及叶片结构特征的影响

喀斯特石漠化环境有着高度的生境异质性,异质性生境中土被不连续,土壤瘠薄,岩溶漏斗上的土壤保水性差,严重制约着喀斯特植被的生长及分布。为探究克隆植物在喀斯特地区的适应策略,本研究以喀斯特黄色石灰土为基质,选用克隆植物活血丹（Glechoma longituba）,以一个节间连接的两个分株为材料,保持节间连接或切断,种植于相邻花盆中,并施以不同浇水量,以明确不同水分可用性水平下克隆整合对活血丹生物量积累、生物量分配、叶片气孔及叶片组织特征的影响。结果显示,克隆整合显著促进活血丹生物量的积累及对根、叶的生物量分配;增加了活血丹叶气孔导度,降低了气孔指数;叶海绵组织受克隆整合影响较小,但栅栏组织及栅海比（栅栏组织/海绵组织）表现为非整合分株高于整合分株。本研究表明,克隆整合可增加活血丹胁迫分株对根、叶的投资,并以更佳的叶气孔、组织适应策略提高其在喀斯特生境中的生存与适应。     

灰杨克隆繁殖、自身养分与土壤养分季节变化及其耦合关系的林缘——林内差异

阐明灰杨(Populus pruinosa Schrenk)克隆特征变化及受外界环境影响,是增强灰杨林生态服务功能的基础。本研究在中国新疆塔里木河上游灰杨河岸林,沿垂直河道方向在林缘(距河道200 m)、林内(距河道400 m)生境设置样方,于2014年4～10月,每隔20天调查两生境不定芽、未出土和出土克隆分株数量,测定横走侧根及土壤养分含量。结果表明:(1)横走侧根全氮、全磷、全钾、有机碳含量的变化趋势与不定芽和未出土分株数在4～6月逐渐增加、7～10月逐渐减少变化趋势基本一致,与土壤有机质、碱解氮、速效磷及速效钾含量在4～6月逐渐升高或保持最高水平,在7～10月逐渐降低的变化趋势同步;横走侧根碳氮比在4～6月初逐渐降低,随后呈现增大又减小的趋势;氮磷比在4～6月逐渐减小,7月显著减小后逐渐增大(林缘)或基本保持不变(林内)。(2)不论是林缘还是林内,不定芽数量受到横走侧根全氮和氮磷比的影响,未出土分株数受到横走侧根全氮、有机碳含量、碳氮比的影响;林缘不定芽数量与土壤有机质、碱解氮及速效钾含量相关紧密,未出土分株数与土壤碱解氮和速效磷显著相关,土壤养分也与横走侧根全氮、碳氮比、氮磷比紧密相关,而在林内受到土壤养分影响的主要是横走侧根全氮和氮磷比。(3)林内的不定芽、未出土及出土克隆分株数量显著低于林缘,土壤养分含量通过影响横走侧根养分含量而影响不定芽、未出土分株、出土分株数量。灰杨是杨属为数不多的国家保护植物,具有重要的生态服务功能,我们的结果对基于克隆繁殖的灰杨林保护策略提供基础数据支持。     

Molecular discrimination and phylogeographic patterns of clones of the parthenogenetic gecko Lepidodactylus lugubris in the Japanese Archipelago

Vertebrates usually reproduce sexually in which males and females contribute their offspring genome and produce genetically diverse offspring. However, some of them are asexual without genetic contribution from males. The nocturnal gecko, Lepidodactylus lugubris, is all females and reproduces parthenogenetically. This gecko is known to consist of diploid and triploid clones in the tropical and subtropical regions, which can be identified by their dorsal marking patterns, ploidy, and protein polymorphism. This gecko is also distributed in the southern parts of Japan, and several clones have been reported. In this study, we investigated the origins and genetic diversity of Japanese L. lugubris by clonal discrimination using microsatellite and mitochondrial DNA analyses. A total of 748 individuals were collected from 21 islands of five island groups (Ogasawara, Okinawa, Miyako, Yaeyama and Daito Islands) and 17 clones were distinguished genetically. Mitochondrial cyt b sequences of these clones suggested that they were all closely related and differentiated recently. Clonal diversity was much higher (14 clones) in the Daito Islands than in the other island groups in which only one or two clones coexisted. Judging from the dorsal marking patterns and ploidy known so far, six clones were cosmopolitan and may be colonized from the outside of Japan. However, other 11 clones were endemic to the Daito Islands and explained by possible hybridization between the one female diploid clone and one male diploid clone because other 9 clones were triploid and all had the combinations of polymorphic microsatellite alleles of these female and male diploid clones. Although the males have never been recorded in the Daito Islands, males might appear in the past. These findings contribute to understanding of clonal diversity and dynamics of asexually reproducing animals. If diploid parthenogenetic geckos can produce triploid clones by mating with the diploid males, clonal diversity would increase rapidly in a small island, and such newly produced triploid clones would expand widely.     

Nutrient addition does not increase the benefits of clonal integration in an invasive plant spreading from open patches into plant communities

• One benefit of clonal integration is that resource translocation between connected ramets enhances the growth of the ramets grown under stressful conditions, but whether such resource translocation reduces the performance of the ramets grown under favourable conditions has not produced consistent results. In this study, we tested the hypothesis that resource translocation to recipient ramets may reduce the performance of donor ramets when resources are limiting but not when resources are abundant.
• We grew Mikania micrantha stolon fragments (each consisting of two ramets, either connected or not connected) under spatially heterogeneous competition conditions such that the developmentally younger, distal ramets were grown in competition with a plant community and the developmentally older, proximal ramets were grown without competition. For half of the stolon fragments, slow‐release fertiliser pellets were applied to both the distal and proximal ramets.
• Under both the low and increased soil nutrient conditions, the biomass, leaf number and stolon length of the distal ramets were higher, and those of the proximal ramets were lower when the stolon internode was intact than when it was severed. For the whole clone, the biomass, leaf number and stolon length did not differ between the two connection treatments. Connection did not change the biomass of the plant communities competing with distal ramets of M. micrantha.
• Although clonal integration may promote the invasion of M. micrantha into plant communities, resource translocation to recipient ramets of M. micrantha will induce a cost to the donor ramets, even when resources are relatively abundant.

     

Effects of parental light environment on growth and morphological responses of clonal offspring

• Environments experienced by parent ramets of clonal plants can potentially influence fitness of clonal offspring ramets. Such clonal parental effects may result from heritable epigenetic changes, such as DNA methylation, which can be removed by application of DNA de‐methylation agents such as 5‐azacytidine.
• To test whether parental shading effects occur via clonal generation and whether DNA methylation plays a role in such effects, parent plants of the clonal herb Alternanthera philoxeroides were first subjected to two levels of light intensity (high versus low) crossed with two levels of DNA de‐methylation (no or with de‐methylation by application of 5‐azacytidine), and then clonal offspring taken from each of these four types of parent plant were subjected to the same two light levels.
• Parental shading effects transmitted via clonal generation decreased growth and modified morphology of clonal offspring. Offspring responses were also influenced by DNA methylation level of parent plants. For clonal offspring growing under low light, parental shading effects on growth and morphology were always negative, irrespective of the parental de‐methylation treatment. For clonal offspring growing under high light, parental shading effects on offspring growth and morphology were negative when the parents were not treated with 5‐azacytidine, but neutral when they were treated with 5‐azacytidine.
• Overall, parental shading effects on clonal offspring performance of A. philoxeroides were found, and DNA methylation is likely to be involved in such effects. However, parental shading effects contributed little to the tolerance of clonal offspring to shading.

     

Life‐history evolution under fluctuating density‐dependent selection and the adaptive alignment of pace‐of‐life syndromes

We present a novel perspective on life‐history evolution that combines recent theoretical advances in fluctuating density‐dependent selection with the notion of pace‐of‐life syndromes (POLSs) in behavioural ecology. These ideas posit phenotypic co‐variation in life‐history, physiological, morphological and behavioural traits as a continuum from the highly fecund, short‐lived, bold, aggressive and highly dispersive ‘fast’ types at one end of the POLS to the less fecund, long‐lived, cautious, shy, plastic and socially responsive ‘slow’ types at the other. We propose that such variation in life histories and the associated individual differences in behaviour can be explained through their eco‐evolutionary dynamics with population density – a single and ubiquitous selective factor that is present in all biological systems. Contrasting regimes of environmental stochasticity are expected to affect population density in time and space and create differing patterns of fluctuating density‐dependent selection, which generates variation in fast versus slow life histories within and among populations. We therefore predict that a major axis of phenotypic co‐variation in life‐history, physiological, morphological and behavioural traits (i.e. the POLS) should align with these stochastic fluctuations in the multivariate fitness landscape created by variation in density‐dependent selection. Phenotypic plasticity and/or genetic (co‐)variation oriented along this major POLS axis are thus expected to facilitate rapid and adaptively integrated changes in various aspects of life histories within and among populations and/or species. The fluctuating density‐dependent selection POLS framework presented here therefore provides a series of clear testable predictions, the investigation of which should further our fundamental understanding of life‐history evolution and thus our ability to predict natural population dynamics.     

High efficiency Agrobacterium‐mediated site‐specific gene integration in maize utilizing the FLP‐FRT recombination system

An efficient Agrobacterium‐mediated site‐specific integration (SSI) technology using the flipase/flipase recognition target (FLP/FRT) system in elite maize inbred lines is described. The system allows precise integration of a single copy of a donor DNA flanked by heterologous FRT sites into a predefined recombinant target line (RTL) containing the corresponding heterologous FRT sites. A promoter‐trap system consisting of a pre‐integrated promoter followed by an FRT site enables efficient selection of events. The efficiency of this system is dependent on several factors including Agrobacterium tumefaciens strain, expression of morphogenic genes Babyboom (Bbm) and Wuschel2 (Wus2) and choice of heterologous FRT pairs. Of the Agrobacterium strains tested, strain AGL1 resulted in higher transformation frequency than strain LBA4404 THY‐ (0.27% vs. 0.05%; per cent of infected embryos producing events). The addition of morphogenic genes increased transformation frequency (2.65% in AGL1; 0.65% in LBA4404 THY‐). Following further optimization, including the choice of FRT pairs, a method was developed that achieved 19%–22.5% transformation frequency. Importantly, >50% of T0 transformants contain the desired full‐length site‐specific insertion. The frequencies reported here establish a new benchmark for generating targeted quality events compatible with commercial product development.     

The Promises and the Challenges of Biotransformations in Microflow

The challenges of transition toward the postpetroleum world shed light on the biocatalysis as the most sustainable way for the valorization of biobased raw materials. However, its industrial exploitation strongly relies on integration with innovative technologies such as microscale processing. Microflow devices remarkably accelerate biocatalyst screening and engineering, as well as evaluation of process parameters, and intensify biocatalytic processes in multiphase systems. The inherent feature of microfluidic devices to operate in a continuous mode brings additional interest for their use in chemoenzymatic cascade systems and in connection with the downstream processing units. Further steps toward automation and analytics integration, as well as computer‐assisted process development, will significantly affect the industrial implementation of biocatalysis and fulfill the promises of the bioeconomy. This review provides an overview of recent examples on implementation of microfluidic devices into various stages of biocatalytic process development comprising ultrahigh‐throughput biocatalyst screening, highly efficient biocatalytic process design including specific immobilization techniques for long‐term biocatalyst use, integration with other (bio)chemical steps, and/or downstream processing.