Effect of Antibiotics on the Symptoms of Stunting Disease of Magnolia liliiflora Plants

Treatment of diseased magnolia plants with Oxytetracycline, Baytril or Tylan did not reduce the number of symptomatic plants, but promoted shoot growth, development of symptomless leaves and flower buds. The most efficient were 500 ppm Baytril, 200 ppm Tylan and 500 or 1000 ppm Oxytetracycline. Lower concentrations of Baytril and Oxytetracycline were less effective and higher concentrations of Tylan decreased the growth of magnolia shoots. All the tested antibiotic treated and untreated magnolias were shown by polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) to contain the AY (16SrI) phytoplasma and two also to contain a phytoplasma related to apple proliferation phytoplasma group (16SrX). The results indicate that Magnolia is a natural host of phytoplasmas belonging to the aster yellows and apple proliferation phytoplasma groups, and support the suggestion that phytoplasmas are the cause of magnolia stunting disease.     

Determination of first cleavage plane: the relationships between the orientation of the mitotic apparatus for first cleavage and the position of meiotic division-related structures in starfish eggs

In order to understand when the orientation of the first cleavage plane is fixed along the animal-vegetal axis in starfish eggs, the behavior of the sperm aster was examined by indirect immunofluorescence staining. After duplication, the sperm aster organizes the mitotic apparatus for first cleavage perpendicular to the cleavage plane. The sperm aster located in the egg periphery just after fertilization and moved to the site close to the animal pole rather than the egg center by meiosis II. At early metaphase II, duplication of the sperm aster was detected but the axis of the resultant sperm diaster randomly pointed. Subsequently, its axis had already turned perpendicular to the animal-vegetal axis before pronucleus fusion. These results indicate that the orientation processes of the sperm diaster consist of positioning before its duplication and successive determining its azimuth. Furthermore, the azimuth and position of the mitotic apparatus for first cleavage did not change by shifting or eliminating the meiotic division-related structures such as the germinal vesicle, meiotic spindle, and female pronucleus by micromanipulation. These results show that none of them determines the first cleavage plane. Therefore, we discuss the pointing mechanism of the first cleavage plane without the influence of these meiotic division-related structures.     

Asters play only a dispensable role in the induction of the cleavage furrow in the blastomeres of early Xenopus embryos

Kuroda et al. (2001) of our laboratory have previously revealed that exposure of early Xenopus embryos to 150 mm urethane results in complete suppression of formation of the asters and the cleavage furrow, as well as significant reduction of the size of the spindle in the blastomeres, allowing only 1 or 2 cycles of mitosis but not cytokinesis. In the course of closer examination of the effect of urethane on the cleavage of blastomeres of early Xenopus embryos, we unexpectedly discovered that exposure of early Xenopus embryos to 75 mm urethane did not prevent cell division at all, though asters were not detected in the blastomeres. Instead, they contained a spindle that appeared rather normal. They also formed the diastema, a thin yolk-free structure, which is considered to play an essential role in the induction of the cleavage furrow. Essentially the same results were obtained in the exposure of embryos to vinblastine, a well-known microtubule inhibitor: exposure of embryos to 20 micro g/mL vinblastine resulted in complete suppression of cleavage of the blastomeres, where formation of both the spindle and asters were perfectly suppressed. By contrast, exposure of embryos to 5 microg/mL vinblastine did not prevent cleavage in the blastomeres though asters were not detected, whereas the rather normal spindle was formed. Thus, there was a close correlation between the formation of the normal spindle, not asters, and that of the cell division furrow and the diastema in the blastomeres of early Xenopus embryos. We suggest that while the spindle plays an essential role, asters are likely to play only a dispensable role in the induction of the cleavage furrow in even very large cells like the blastomeres of early Xenopus embryos.     

Mitochondrial inhibitor sodium azide inhibits the reorganization of mitochondria‐rich cytoplasm and the establishment of the anteroposterior axis in ascidian embryo

In ascidian eggs, cytoplasmic and cortical reorganization, previously called ooplasmic segregation, occurs in two phases during the first cell cycle. In the second phase of reorganization, the mitochondria‐rich cytoplasm (myoplasm) moves to the future posterior side, concurrent with sperm aster migration along the egg cortex. Although this reorganization is the critical step for establishing the anteroposterior axis, its molecular mechanism is not fully understood. In this study, we showed that low concentrations of the mitochondrial inhibitor sodium azide (NaN₃), which showed the low toxicity in sperm, inhibited the second phase of reorganization without the microtubule depolymerization. In the NaN₃‐treated embryo, the sperm aster was not attracted to the cortex and altered its migration pathway; therefore, the myoplasm remained at the vegetal pole. Consequently, the anteroposterior axis was not established. Another mitochondrial inhibitor, oligomycin, did not affect these processes. These results suggest that NaN₃ inhibits unknown molecules that are important for the second phase of reorganization. Identifying the target molecule of NaN₃ will lead to a molecular understanding of cytoplasmic and cortical reorganization.     

The impact of plant associations on Macrosteles quadrilineatus management in carrots

Habitat diversification can influence the interactions of insects with plants and this can be used in agroecosystems for the management of pest populations. Plant diversification can be achieved through planting crops, such as trap crops, or by adjusting weed management. Aster leafhopper, Macrosteles quadrilineatus Forbes (Hemiptera: Cicadellidae), is a polyphagous species that uses cereals, vegetables, and weeds as host plants. The influence of weeds on M. quadrilineatus abundance was investigated experimentally in carrot [Daucus carota L. cv. Canada (Apiaceae)] field plots by adjusting the level of management of two groups of weeds (broadleaf and grass) and by comparing it to weed‐free plots. The preference of M. quadrilineatus for different cereal and weed species relative to carrots was tested in choice test assays. Habitat context influenced the abundance of M. quadrilineatus in the field experiments. The presence of border crops such as oat, rye, barley, wheat, and triticale did not significantly attract or repel this insect to carrot plots compared to the no‐border treatment. However, spelt‐bordered plots had 42% fewer M. quadrilineatus than three treatments, triticale, wheat, and barley, that had the highest insect abundance. The type of weed management affected M. quadrilineatus abundance in carrot plots, but not the frequency of herbicide application. Plots that had carrot growing with broadleaf‐weeds had about 59% fewer M. quadrilineatus compared with those growing with crabgrass or carrot alone. In the greenhouse choice tests, grasses (e.g., cereals) attracted and broadleaf‐weeds repelled M. quadrilineatus relative to carrots. In summary, carrot growers may be able to manage this pest by reducing the interaction of cereal cover crops with carrots and eliminating grassy weeds in commercial production fields.     

INFERRING FITNESS LANDSCAPES

Since 1983, study of natural selection has relied heavily on multiple regression of fitness on the values for a set of traits via ordinary least squares (OLSs), as proposed by Lande and Arnold, to obtain an estimate of the quadratic relationship between fitness and the traits, the fitness surface. However, well‐known statistical problems with this approach can affect inferences about selection. One key concern is that measures of lifetime fitness do not conform to a normal or any other standard sampling distribution, as needed to justify the usual statistical tests. Another is that OLS may yield an estimate of the sign of the fitness function's curvature that is opposite to the truth. We here show that the recently developed aster modeling approach, which explicitly models the components of fitness as the basis for inferences about lifetime fitness, eliminates these problems. We illustrate selection analysis via aster using simulated datasets involving five fitness components expressed in each of four years. We demonstrate that aster analysis yields accurate estimates of the fitness function in cases in which OLS misleads, as well as accurate confidence regions for directional selection gradients. Further, to evaluate selection when many traits are under consideration, we recommend model selection by information criteria and frequentist model averaging.     

Detection and Molecular Characterization of ‘Candidatus Phytoplasma asteris’ in European Hazel (Corylus avellana) in Poland

Stunted European hazel (Corylus avellana L.) plants showing leaf yellowing were observed in south‐eastern Poland. Phytoplasma‐specific primers P1/P7 and R16F2n/R16R2, as well as primers specific for aster yellows (16SrI), X‐disease (16SrIII) and apple proliferation (16SrX) groups were singly used in nested polymerase chain reaction (PCR) to amplify the 16S rDNA from 22 symptomatic and asymptomatic hazel plants. Restriction fragment length polymorphism with MseI, HhaI, RsaI and BfaI enzymes of the 16S rRNA gene fragments amplified with the primers R16F2n/R16R2 from three symptomatic hazel plants of cvs Katalonski, Webba and Halle revealed patterns identical to those from the AY1 strain related to ‘Candidatus Phytoplasma asteris’. The nucleotide sequence analysis confirmed this result. This is the first report of the natural occurrence of ‘Ca. P. asteris’ in European hazel in Poland.     

Effects of ‘Candidatus Phytoplasma asteris’ on the Volatile Chemical Content and Composition of Grindelia robusta Nutt.

Grindelia robusta, a perennial herb, contains an essential oil that is used as an antitussive, sedative, and analgesic agent. During the spring of 2007, ‘Candidatus Phytoplasma asteris’‐related phytoplasmas were identified in plants showing virescence and phyllody symptoms. The qualitative and quantitative composition of the oil of healthy and infected plants was compared by gas chromatography/mass spectrometry. Samples from six symptomatic and five asymptomatic plants tested by nested PCR followed by RFLP analyses confirmed the presence of ‘Ca. P. asteris’ in all symptomatic samples. The oils from healthy and infected plants, obtained by steam distillation, contained 42 components; that of healthy plants contained a higher concentration of monoterpenes, especially limonene and bornyl acetate, which were nearly 50% higher.     

Ypt32 recruits the Sec4p guanine nucleotide exchange factor,Sec2p,to secretory vesicles; evidence for a Rab cascade in yeast

SEC2 is an essential gene required for polarized growth of the yeast Saccharomyces cerevisiae. It encodes a protein of 759 amino acids that functions as a guanine nucleotide exchange factor for the small GTPase Sec4p, a regulator of Golgi to plasma membrane transport. Activation of Sec4p by Sec2p is needed for polarized transport of vesicles to exocytic sites. Temperature-sensitive (ts) mutations in sec2 and sec4 result in a tight block in secretion and the accumulation of secretory vesicles randomly distributed in the cell. The proper localization of Sec2p to secretory vesicles is essential for its function and is largely independent of Sec4p. Although the ts mutation sec2-78 does not affect nucleotide exchange activity, the protein is mislocalized. Here we present evidence that Ypt31/32p, members of Rab family of GTPases, regulate Sec2p function. First, YPT31/YPT32 suppress the sec2-78 mutation. Second, overexpression of Ypt31/32p restores localization of Sec2-78p. Third, Ypt32p and Sec2p interact biochemically, but Sec2p has no exchange activity on Ypt32p. We propose that Ypt32p and Sec4p act as part of a signaling cascade in which Ypt32p recruits Sec2p to secretory vesicles; once on the vesicle, Sec2p activates Sec4p, enabling the polarized transport of vesicles to the plasma membrane.     

Photosynthetic limitations of a halophyte sea aster (<Emphasis Type="Italic">Aster tripolium</Emphasis> L) under water stress and NaCl stress

To understand the mechanisms of salt tolerance in a halophyte, sea aster (Aster tripolium L.), we studied the changes of water relation and the factors of photosynthetic limitation under water stress and 300 mM NaCl stress. The contents of Na⁺ and Cl^- were highest in NaCl-stressed leaves. Leaf osmotic potentials (Ψ _s) were decreased by both stress treatments, whereas leaf turgor pressure (Ψ _t) was maintained under NaCl stress. Decrease inΨ _s without any loss ofΨ _t accounted for osmotic adjustment using Na⁺ and Cl^- accumulated under NaCl stress. Stress treatments affected photosynthesis, and stomatal limitation was higher under water stress than under NaCl stress. Additionally, maximum CO₂ fixation rate and O₂ evolution rate decreased only under water stress, indicating irreversible damage to photosynthetic systems, mainly by dehydration. Water stress severely affected the water relation and photosynthetic capacity. On the other hand, turgid leaves under NaCl stress have dehydration tolerance due to maintenance of Ψ _t and photosynthetic activity. These results show that sea aster might not suffer from tissue dehydration in highly salinized environments. We conclude that the adaptation of sea aster to salinity may be accomplished by osmotic adjustment using accumulated Na⁺ and Cl^-, and that this plant has typical halophyte characteristics, but not drought tolerance. Electronic Publication