Morphological and molecular investigation of polymorphism and cryptic species in tanaid crustaceans: implications for tanaid systematics and biodiversity estimates

A combination of traditional taxonomic procedures and molecular techniques has provided new insight into the problems of cryptic species and sexual and ontogenetic polymorphism in the Tanaidacea. Using polymerase chain reaction and DNA markers, three cryptic species of Paratanais were identified. PCR primers were used to amplify the divergent internal transcribed spacers (ITS) of these species. Restriction digestion of the amplified rDNA generated species specific DNA banding. Male and five female stages of Paratanais maleticus sp. nov. and two other new species, P. malign us and P. perturbatius , are described. Morphological variation, both sexual and ontogenetic, was found in several generic characters of Paratanais and required the diagnosis to be modified. The identification of three undescribed cryptic species from a single microhabitat in a well studied, shallow water and easily accessible locality, demonstrate that the biodiversity of tanaid crustacean is significantly underestimated.     

Ceramide Inhibits Cell Proliferation through Akt/PKB Inactivation and Decreases Melanin Synthesis in Mel‐Ab Cells

Ceramide is a bioactive sphingolipid that mediates a variety of cell functions. However, the effects of ceramide on cell growth and the melanogenesis of melanocytes are not known. In the present study, we investigated the actions of cell‐permeable ceramide and its possible role in the signaling pathway of a spontaneously immortalized mouse melanocyte cell line, Mel‐Ab. Our results show that C₂‐ceramide inhibits DNA synthesis in Mel‐Ab cells and G361 human melanoma cells in a dose‐dependent manner. Cell cycle analysis confirmed the inhibition of DNA synthesis by a reduction in the S phase. To investigate the ceramide signaling pathway, we studied whether C₂‐ceramide is able to influence extracellular signal‐regulated kinase (ERK) and/or Akt/protein kinase B (PKB) activation. We demonstrated that phosphorylated Akt/PKB is decreased by C₂‐ceramide, whereas phosphorylated ERK was only slightly affected. Therefore, the C₂‐ceramide‐induced inactivation of Akt/PKB may be closely related to the reduced cell proliferation of Mel‐Ab cells. Furthermore, we assessed the effects of C₂‐ceramide on the pigmentation of Mel‐Ab cells. The results obtained showed that the melanin content of cells was significantly reduced by C₂‐ceramide at concentrations in the range of 1–10 μM, and that the pigmentation‐inhibiting effect of C₂‐ceramide is much greater than that of kojic acid at 1–100 μM. In addition, we found that the activity of tyrosinase is reduced by C₂‐ceramide treatment. Our results demonstrate that C₂‐ceramide reduces the pigmentation of Mel‐Ab cells by inhibiting tyrosinase activity.     

日本松干蚧的重要天敌——隐斑瓢虫的初步研究

隐斑瓢虫是近年来在杭州地区发现的日本松干蚧的一种重要捕食性天敌。该虫在杭州一年发生四代,其年发生规律基本上和日本松干蚧的生活周期相吻合,而且能在松林中形成比较稳定的群落。根据测定,隐斑瓢虫对日本松干蚧各虫态的捕食能力较强,一头成虫的日平均捕食量为:显露若虫13.33头,雌成虫4.27头,卵囊3.67个(每个卵囊内平均有卵265.5粒),雄蛹28.27只;一头幼虫全期(或2—4龄和3—4龄)分别能捕食显露若虫39.20—105.56头,雌成虫40.80—47.79头,卵囊39.38—41.79个,雄蛹220.50只。它在引进辽宁省沈阳和旅大地区后,不仅仍能正常的捕食日本松干蚧(显露若虫),其成虫和幼虫喂以各种蚜虫均能正常生殖和发育,并且一年可以繁殖三代:冬季,只要在0℃以上的温度条件下,给予少量的糖水等作补充营养,亦能安全的越冬。据此,作者认为隐斑瓢虫在浙江杭州和辽宁,均具有作为控制日本松干蚧的发生来加以饲养繁殖与利用的价值。     

Evidence for non-random spatial positioning of migrating smolts (Salmonidae) in a small lowland stream

1. The ontogenetic development of anadromous salmonids includes downstream emigration of immature individuals from freshwater towards the marine environment. Although this migration of juvenile salmonids (smolts) may be associated with severe mortalities, only limited attention has been paid to the spatial positioning of smolts in small streams. 2. Using a novel approach, this study examined the vertical and horizontal positioning of brown trout and Atlantic salmon smolts while performing downstream migration in a small lowland stream. 3. Pre‐smolts of indigenous and hatchery‐reared (F1) brown trout (Salmo trutta), and two different populations of Atlantic salmon (S. salar), were tagged with passive integrated transponder (PIT) tags and subsequently released upstream of an antenna array consisting of five circular swim‐through PIT antennas. Antennas were positioned in order to determine whether the migrating smolts were bottom or surface oriented, and if they were oriented towards the mid‐channel or the stream bank. 4. During the smolt emigration period, data describing both the detection of the migrating fish and the amount of water passing through the antennas were collected. This was accomplished in order to determine if the fish were performing active positioning behaviour independently of the vertical and horizontal discharge distributions in the stream. 5. The results showed that the smolts migrated in a non‐random spatial pattern independently of the stream discharge distributions. Vertically, the indigenous brown trout and the Atlantic salmon demonstrated a preference for the bottom orientated positions. In contrast, the distribution of the F1 brown trout was not different from the discharge distribution. The latter observation suggests random vertical positioning, which may be indicative of inferior migratory performance. Horizontally, all tested smolt populations strongly preferred the mid‐channel positions. 6. The discharge‐corrected preferences for certain spatial positions suggest that smolt emigration is not entirely a matter of passive displacement in lowland streams. 7. Anthropogenically altered channels may inhibit or delay downstream emigration of smolts resulting in increased mortalities. Given that the smolts in this study actively selected spatial positions in the mid‐channel and near the bottom, it is suggested that deep, mid‐channel furrows may be used to help guide migrating smolts past adverse habitats in lowland streams.     

A NEW TECHNIQUE FOR ASSESSING THE CONTACT TOXICITY OF MOLLUSCICIDES TO SLUGS

A technique is described to accurately assess the dermal contactdose-mortality response of molluscicides. The technique involvescarbon dioxide anaesthesia of laboratory reared Deroceras reticulatum(Müller) followed by application of test substances insolution to the dorsal surface. The LD₅₀ of the substances testedwere as follows: Sodium pentachlorophenate, 55.9 ng.mg^–1,Copper sulphate, 124.1 ng.mg^–1, Methiocarb, 314.6 ng.mg^–1and Potassium permanganate, 418.9 ng.mg^–1 body weight. (Received 18 August 1988; accepted 17 December 1988)     

Miscanthus×giganteus plant regeneration: effect of callus types,ages and culture methods on regeneration competence

The perennial rhizomatous grass, Miscanthus×giganteus is an ideal biomass crop due to its rapid vegetative growth and high biomass yield potential. As a naturally occurring sterile hybrid, M. ×giganteus must be propagated vegetatively by mechanically divided rhizomes or from micropropagated plantlets. Plant regeneration through somatic embryogenesis is a viable approach to achieve large‐scale production of plantlets in tissue culture. Effect of the callus types, ages and culture methods on the regeneration competence was studied to improve regeneration efficiency and shorten the period of tissue culture in M. ×giganteus. Shoot‐forming calli having a yellow or white compact callus with light‐green shoot‐like structures showed the highest regeneration frequency. Percentage of shoot‐forming callus induction from immature inflorescence explants was 41% on callus induction medium containing 13.6 μM 2,4‐d and 0.44 μM benzyladenine (BA). The use of a regeneration medium containing 1.3 μM NAA and 22 μM BA was effective at shortening the incubation period required for plantlet regeneration, with 69% of total regenerated plantlets obtained within 1 month of incubation on regeneration medium. Embryogenic‐like callus morphotype could maintain regeneration competency for up to 1 year as suspension cultures. Field grown regenerated plants showed normal phenotypic development with DNA content and plant heights comparable to rhizome propagated plants. Winter survival rates of the regenerated plants planted in 2006 and 2007 at the University of Illinois South Farm, Urbana‐Champaign, Illinois, were 78% and 56%, respectively.     

Large seasonal changes in Q10 of soil respiration in a beech forest

We analyzed one year of continuous soil respiration measurements to assess variations in the temperature sensitivity of soil respiration at a Danish beech forest. A single temperature function derived from all measurements across the year (Q₁₀ = 4.2) was adequate for estimating the total annual soil respiration and its seasonal evolution. However, Q₁₀'s derived from weekly datasets ranged between three in summer (at a mean soil temperature of 14 °C) and 23 in winter (at 2 °C), indicating that the annual temperature function underestimated the synoptic variations in soil respiration during winter. These results highlight that empirical models should be parameterized at a time resolution similar to that required by the output of the model. If the objective of the model is to simulate the total annual soil respiration rate, annual parameterization suffices. If however, soil respiration needs to be simulated over time periods from days to weeks, as is the case when soil respiration is compared to total ecosystem respiration during synoptic weather patterns, more short‐term parameterization is required. Despite the higher wintertime Q₁₀'s, the absolute response of soil respiration to temperature was smaller in winter than in summer. This is mainly because in absolute numbers, the temperature sensitivity of soil respiration depends not only on Q₁₀, but also on the rate of soil respiration, which is highly reduced in winter. Nonetheless, the Q₁₀ of soil respiration in winter was larger than can be explained by the decreasing respiration rate only. Because the seasonal changes in Q₁₀ were negatively correlated with temperature and positively correlated with soil moisture, they could also be related to changing temperature and/or soil moisture conditions.