Clonal segmentation – The development of physiological independence within stolons of Glechoma hederacea L. (Lamiaceae)

The youngest parts of clonal plants benefit from substantial physiological support from older parts, but the extent to which this physiological dependence persists through time is poorly understood. The development of autonomy among connected subunits was therefore analysed in the clonal species Glechoma hederacea. The stolons of a series of clonal fragments with differing numbers of primary ramets were severed at a fixed point relative to the four oldest primary ramets. The subsequent growth of both parts of the severed fragments was compared with that of a series of intact fragments.The growth of apical stolon portions that included five or more rooted primary ramets at the time of severing was unaffected by severing. Apical portions with three or fewer rooted ramets at the time of severing produced fewer new primary ramets than equivalent parts of intact fragments, while apical portions with four or fewer rooted ramets produced less above-ground mass than equivalent apical portions of intact clonal fragments. Basal portions of clonal fragments severed when there were one or two rooted ramets in the apical portion produced more secondary ramet mass than equivalent parts of intact fragments. The gain in mass of secondary ramets in the basal portions of severed fragments matched the reduction in mass of secondary ramets in the apical portions. However, severing caused an overall loss of mass when apical portions had three or fewer rooted ramets at the time of severing, because the mass of primary ramets in basal portions did not increase following severing. Severing had little impact on the allometry of the apical portions. The relationship between mass in secondary ramets and mass in primary ramets was similar in the apical portions of severed and intact clonal fragments. None of the severing treatments increased the total mass of secondary ramets, suggesting that apical dominance in this species only affects branches very close to the apex.These observations, combined with existing knowledge of vascular architecture in G. hederacea, demonstrate that, whether or not physical connections persist between ramets, growing stolons rapidly develop into physiologically autonomous segments. This may be a characteristic of species that exploit disturbed, spatially heterogeneous habitats through rapid multiplication of ramets connected by long, aerial runners or stolons.     

Further studies on branch buds in mosses; “Pseudoparaphyllia” and “scaly leaves”

The development of so-called foliose pseudoparaphyllia in the species of theClimacium-type branch development was studied with a paraffin sectioning method and SEM. Scaly leaves (or scale-like leaves) and “foliose” pseudoparaphyllia proved to originate as leaves by segmentation of an apical cell of a branch initial in the very first stage of development into a branch bud. Branch buds of two species among the six species examined develop linear-lanceolate appendages which serve to protect the buds as well as scaly leaves. These appendages originate in the peripheral part of the epidermal layer of buds, and therefore they can be homologous to trichomes previously reported for species with branch primordia. Emendation of two terms are proposed; every organ originating from buds as leaves that are effective for bud protection should be called scaly leaves, while those which homologous to trichomes should be called pseudoparaphyllia. Both terms are used here in a narrower sense than before: There might befilamentous orfoliose scaly leaves, andfilamentous orfoliose (linear-to broad-lanceolate) pseudoparaphyllia. A scheme is given to show the pattern of branch development and the manner in which branch buds (or primordia) are protected.     

中国安徽省祁门县大鲵属一新种（两栖纲：隐鳃鲵科）

中国大鲵(Andrias davidianus)是世界上最大的两栖动物,曾在中国广泛分布。前期的3项分子遗传学研究表明：黄山的中国大鲵种群存在独特的遗传分化,是一个未被描述的物种。这3项研究中的分子数据均基于本文通讯作者1995年之前采自中国安徽省黄山市祁门县的原生大鲵标本。本文基于形态学比较,将1995年以前本文通讯作者在祁门县采集的11个大鲵标本描述为大鲵属一新种：祁门大鲵(Andrias cheni sp. nov.)。我们测量了11个标本的25个形态特征,并与大鲵属的其他物种进行比较。祁门大鲵与大鲵属其他物种可通过以下特征进行区分：(1)头躯扁平,头长略大于头宽,头宽/头长(HW/HL)比为0.80 ~ 0.98;(2)头部和下颌相对光滑,有不规则排列的小结节;(3)颈部侧褶与体褶不连续;(4)第三指比第一指长,指式Ⅰ< Ⅳ < Ⅲ < Ⅱ;(5)四肢粗而短,后肢比前肢长,前肢长/后肢长(FLL/PLL)比为0.78 ~ 0.96;(6)尾部肌肉发达,基部粗大,后部和上部逐渐变平,尾长约为总长度的1/3。经形态比较,本研究将祁门大鲵描述为新种,大鲵属物种增加到5个。本文提供了相对完备的形态数据,可为后续大鲵野生谱系的资源保护、人工种群遗传管理和增殖放流等提供参考。     

Lack of B-cell participation in acute lymphocyte choriomeningitis disease of the central nervous system.

The pattern of acute lymphocytic choriomeningitis (LCM) virus-induced central nervous system (CNS) disease was studied in BALB/c mice rendered deficient in B cells and incapable of producing a humoral immune response. The CNS disease in mice treated from birth with goat anti-mouse μ-chain globulin (μ-suppressed mice) was clinically and histopathologically indistinguishable from that of controls. This result demonstrates that neither B cells nor antibody participate in the pathogenesis of acute LCM of the CNS. These observations support the view that the host cell-mediated response to viral antigens is solely responsible for acute CNS disease induced by the intracerebral inoculation of LCM virus.     

Interpretation of light scattering associated with prolonged neural activity and temperature changes

Changes in light scattering from lobster giant axon which accompany the action potential were observed during periods of prolonged stimulation and as a function of temperature. At an initial temperature of 10°C most (more than 90%) axons produced positive light scattering signals which increased in amplitude when the temperature was lowered. At 2 and 5°C approximately half of the axons produced positive scattering signals. The remaining half produced negative scattering signals which became positive when the temperature was raised to 10°C. The amplitude of the negative signals followed sigmoid transition to positive values as a function of time. The time and temperature dependence of the signal are interpreted in terms of differential changes between the indices of refraction of the membrane matrix and the open or closed early activation channel.     

Factors influencing changes in trait correlations across species after using phylogenetic independent contrasts

Comparative interspecific data sets have been analyzed routinely by phylogenetic methods, generally using Felsenstein’s phylogenetic independent contrasts (PIC) method. However, some authors have suggested that it may not be always necessary to incorporate phylogenetic information into statistical analyses of comparative data due to the low influence of shared history on the distribution of␣character states. The main goal of this paper was to undertake a comparison of results from non-phylogenetic Pearson correlation of tip values (TIPs) and phylogenetic independent contrasts analyses (PICs), using 566 correlation coefficients derived from 65 published papers. From each study we collected the following data: taxonomic group, number of species, type of phylogeny, number of polytomies in the phylogenetic tree, if branch length was transformed or not, trait types, the original correlation coefficient between the traits (TIPs) and the correlation coefficient between the traits using the independent contrasts method (PICs). The slope estimated from a regression of PIC correlations on TIP correlations was lower than one, and a paired t-test showed that correlations from PIC are significantly smaller than those obtained by TIP. Thus, PIC analyses tend to decrease the correlation between traits and usually increases the P-value and, thus, favoring the acceptance of the null hypothesis. Multiple factors, including taxonomic group, trait type and use of branch length transformations affected the change in decision regarding the acceptance of the null hypotheses and differences between PIC and TIP results. Due to the variety of factors affecting the differences between results provided by these methods, we suggest that comparative methods should be applied as a conservative approach to cross-species studies. Despite difficulties in quantifying precisely why these factors affect the differences between PIC and TIP, we also suggest that a better evaluation of evolutionary models underlying trait evolution is still necessary in this context and might explain some of the observed patterns.     

Light controllable siRNAs regulate gene suppression and phenotypes in cells

Small interfering RNA (siRNA) is widely recognized as a powerful tool for targeted gene silencing. However, siRNA gene silencing occurs during transfection, limiting its use is in kinetic studies, deciphering toxic and off-target effects and phenotypic assays requiring temporal, and/or spatial regulation. We developed a novel controllable siRNA (csiRNA) that is activated by light. A single photo removable group is coupled during oligonucleotide synthesis to the 5′ end of the antisense strand of the siRNA, which blocks the siRNA's activity. A low dose of light activates the siRNA, independent of transfection resulting in knock down of specific target mRNAs and proteins (GAPDH, p53, survivin, hNuf2) without stimulating non-specific effects such as regulated protein kinase PKR and induction of the interferon response. We demonstrate survivin and hNuf2 csiRNAs temporally knockdown their mRNAs causing multinucleation and cell death by mitotic arrest, respectively. Furthermore, we demonstrate a dose-dependent light regulation of hNuf2 csiRNA activity and resulting phenotype. The light controllable siRNAs are introduced into cells using commercially available reagents including the MPG peptide based delivery system. The csiRNAs are comparable to standard siRNAs in their transfection efficiency and potency of gene silencing. This technology should be of interest for phenotypic assays such as cell survival, cell cycle regulation, and cell development.     

Characterization of an alkylresorcinol synthase that forms phenolics accumulating in the cuticular wax on various organs of rye (Secale cereale)

Alkylresorcinols are bioactive compounds produced in diverse plant species, with chemical structures combining an aliphatic hydrocarbon chain and an aromatic ring with characteristic hydroxyl substituents. Here, we aimed to isolate and characterize the enzyme that forms the alkylresorcinols accumulating in the cuticular wax on the surface of all above‐ground organs of rye. Based on sequence homology with other type‐III polyketide synthases, a candidate alkylresorcinol synthase was cloned. Yeast heterologous expression showed that the enzyme, ScARS, is highly specific for the formation of the aromatic resorcinol ring structure, through aldol condensation analogous to stilbene synthases. The enzyme accepts long‐chain and very‐long‐chain acyl‐CoA starter substrates, preferring saturated over unsaturated chains. It typically carries out three rounds of condensation with malonyl‐CoA prior to cyclization, with only very minor activity for a fourth round of malonyl‐CoA condensation and cyclization to 5‐(2′‐oxo)‐alkylresorcinols or 5‐(2′‐hydroxy)‐alkylresorcinols. Like other enzymes involved in cuticle formation, ScARS is localized to the endoplasmic reticulum. ScARS expression patterns were found correlated with alkylresorcinol accumulation during leaf development and across different rye organs. Overall, our results thus suggest that ScARS synthesizes the cuticular alkylresorcinols found on diverse rye organ surfaces.     

Relationship between induction of aryl hydrocarbon hydroxylase and de novo synthesis of cytochrome P-448 (P1-450) in mice

Phenobarbital, 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT), benzpyrene, 3-methylcholanthrene (3-MC) or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) were administered i.p. for 1 or 3 days to genetically “responsive” (C57BL/6J) and genetically “non-responsive” (DBA/2J) mice. 3-MC or benzpyrene stimulated aryl hydrocarbon hydroxylase (AHH) activity in C57BL/6J (B6) mice but not in DBA/2J (D2) mice. TCDD induced AHH activity in both B6 and D2 mice. Time-course studies showed that in the first 12 h after a single injection of 3-MC to B6 mice there was no shift in the reduced cytochrome P-450-CO complex absorption spectra from 450 to 448 nm, although AHH activity increased 4–5 times over (above) that of the control group. The relationship between induction of AHH activity by polycyclic hydrocarbons in B6 mice and the concomitant synthesis of cytochrome P-448 is discussed.