Differences in growth rates among cohorts of Encrasicholina punctifer and Engraulis japonicus larvae in the coastal waters off Tanshui River Estuary, Taiwan, as indicated by otolith microstructure analysis

The hatching dates of Encrasicholina punctifer and Engraulis japonicus larvae collected in the coastal waters off Tanshui River Estuary during the fishing seasons of 1992 and 1993 indicated that these two anchovies had protracted spawning seasons, which resulted in multiple recruitment cohorts. Encrasicholina punctifer larvae recruited to the estuary from October to March, while the majority of E. japonicus larvae came in March-May and to a lesser extent in October and November. The E. punctifer larvae on arrival to the estuary were 17·4–35·6 mm in length, 167ndash;89 days old and had growth rates of 0·4–1·0 mm day⁻¹, E. japonicus larvae were 12·1–32·7 mm in length, 19–62 days old and had growth rates of 0·7–0·9 mm day⁻¹. Growth rates were significantly different among cohorts and positively correlated to water temperature.     

秦岭产珠子参叶的达玛烷型皂甙研究(1)

从陕西省秦岭产珠子参(Panax japonicus C.A.Meyer var.major(Burk.)Wu etFeng)的叶中分离到十个新的达玛烷型四环三萜皂甙,经光谱测定和化学降解,其中四个的化学结构分别为珠子参甙(majoroside)F_1(1)、F_2(2)、F_3(3)和F_4(4)。同时,还分离到已知的人参甙(ginsenoside)Rd(5)、Re(6)、Rg_1(7)、Rg_2(8)和F_2(9)。     

Life history,distribution and molecular phylogenetics of the Upward-Mouth Spikefish Atrophacanthus japonicus (Teleostei: Tetraodontiformes: Triacanthodidae)

Ninety-six juvenile specimens (37–54 mm standard length; L_S) of the rarely collected Upward-Mouth Spikefish Atrophacanthus japonicus (Triacanthodidae) were obtained from the stomachs of three Yellowfin Tuna Thunnus albacares collected off Guam in the Mariana Islands in the central Pacific Ocean. These specimens extend the range of A. japonicus eastward into Oceania. We review the systematic characters of the monotypic genus Atrophacanthus and present colour photographs of freshly collected specimens. The diet of the juvenile specimens of A. japonicus consisted of thecosome pteropods and foraminiferans. We present a range map of A. japonicus based on all known specimens and show that specimen size is related to whether specimens were collected in the pelagic zone or on the bottom. Our results support that, compared to all other Triacanthodidae, A. japonicus has an unusually extended pelagic larval and juvenile period, up to 54 mm L_S, before settling to the bottom as adults. Lastly, we provide a multilocus phylogeny addressing the phylogenetic placement of Atrophacanthus based on eight of 11 triacanthodid genera and six genetic markers. Our results reveal that Atrophacanthus is the sister group of Macrorhamphosodes and they provide new insights about the evolutionary history of the family.     

Loop‐mediated isothermal amplification (LAMP) for the identification of invasive Aedes mosquito species

Invasive Aedes mosquito species (Diptera: Culicidae) are of public health concern in Europe because they are either recognized or potential vectors of pathogens. Loop‐mediated isothermal amplification (LAMP) is a rapid and simple method for amplifying DNA with high specificity and efficiency, with the technique having potential for application in the field, including in high‐throughput format. Specific LAMP assays based on rDNA internal transcribed spacers 1 or 2 sequences, considering intraspecies variability at these loci, were developed for Aedes aegypti, Aedes albopictus, Aedes japonicus, Aedes koreicus and the indigenous Aedes geniculatus. No such assays could be developed for Aedes atropalpus and Aedes triseriatus because both loci were too short to serve as target. The assays rely on the clearly visible colour change from violet to sky blue after successful amplification. Sensitivity of egg detection was confirmed with ratios of up to one mosquito egg in 99 other eggs. Simple sample preparation of adults or eggs by mechanical homogenization in water required an additional heat treatment or centrifugation step to avoid non‐specific colour changes. Thus, further technical improvements are needed to render these assays truly field‐applicable, which would greatly facilitate surveillance of these invasive mosquito species and allow for prompt implementation of control measures.     

Targeted discovery and identification of novel nucleoside biomarkers in Apostichopus japonicus viscera using metabonomics

In this study, we investigated the metabonomic profiles of Apostichopus japonicus using an LC-MS-based method in conjunction with multivariate data analysis. Based on the PLS-DA model, 85 differential metabolites (VIP value >1.0) were obtained from viscera and body wall samples. The MS/MS and NMR experiments were used for the qualitative identification of the characteristic peaks. Sphingoid-based nucleoside analogues were the main components in Chinese A. japonicus viscera. Our findings demonstrate that A. japonicus viscera contain a large number of compounds that may have applications as nutraceuticals or pharmaceuticals.     

日本囊对虾4个地理群体线粒体16S rRNA序列及遗传结构分析

对我国东南沿海日本囊对虾(Marsupenaeus japonicus)的4个地理群体广东群体(GD)、台湾群体(TW)、福建群体(FJ)和浙江群体(ZJ)的线粒体16S rRNA基因片段进行PCR扩增,对产物进行测序后分析。经比对获得470bp的核苷酸分析序列,发现了16个变异位点,得到了10种单倍型。广东、台湾、福建和浙江群体的核苷酸多样性依次分别为0.0008、0.0010、0.0051、0.0015,各群体均存在独有的单倍型和共有单倍型。群体遗传距离分析表明各群体间保持着一定的遗传差异,其中福建群体与其他群体之间存在着较远的遗传距离并保持了较高的遗传多样性。另外,利用其423bp的16S rRNA同源序列探讨了对虾科6个属共12种对虾的系统进化关系,囊对虾属与沟对虾属亲缘关系较近聚为一支,其他4个属的10种对虾聚为一支。     

日本囊对虾Kunitz型蛋白酶抑制剂在毕赤酵母中的表达纯化及活性分析

SKPI(shrimp Kunitz-type protease inhibitor)是日本囊对虾(Marsupenaeus japonicus)体内的一个小分子多肽, 含有一个Kunitz型结构域, 属于丝氨酸蛋白酶抑制剂。目前已知丝氨酸蛋白酶抑制剂在节肢动物免疫系统中起着非常重要的作用, 为了了解SKPI在对虾天然免疫系统中的作用, 首先对其进行了重组表达。从日本囊对虾肝胰腺中扩增skpi的cDNA片段, 插入改造后的pPIC9K酵母表达载体, 获得的重组质粒转化至毕赤酵母GS115进行表达。由于改造的pPIC9K载体加入了6-His标签, 因此利用Ni?Sepharose?High?Performance对SKPI进行了高效纯化。初步的活性研究表明, 重组表达的SKPI能特异性地抑制胰蛋白酶的水解活性。     

Development and characterization of simple sequence repeat (SSR) markers in the water lotus (Nelumbo nucifera)

Simple sequence repeat (SSR) markers were developed in the water lotus (Nelumbo nucifera Gaertn.) from an SSR-enriched genomic library. Of the SSR markers tested, 11 primer pairs produced clearly distinguishable DNA banding patterns. Forty-three alleles were detected with the 11 markers. The allele number per locus ranged from 2 to 5 with an average of 3.9. Polymorphism values ranged from 0.11 to 0.66 with an average of 0.51. These primers were also applicable to another Nelumbo species, Nelumbo lutea (Willd.) Pers. (American lotus) and hybrids between N. nucifera and N. lutea. These results indicate that the SSR markers developed in this study are informative and will be useful for genetic analysis in Nelumbo species.     

Characterization of expressed sequence tag‐derived microsatellites from the kuruma prawn,Marsupenaeus japonicus

The kuruma prawn, Marsupenaeus japonicus, is a commercially important benthic marine crustacean in East Asia. Understanding the species’ population structures will be very important for its proper stock assessment and management strategy. Herein, 13 polymorphic microsatellite loci originating from expressed sequence tag libraries of M. japonicus were isolated and characterized. Number of alleles per locus ranged from two to 15, observed and expected heterozygosities ranged from 0.250 to 0.906 and from 0.310 to 0.932, respectively. The adequate level of variability within the population renders these microsatellite loci ideal markers for population genetic analysis of M. japonicus.     

Isolation,culture and regeneration of protoplasts from birdsfoot trefoil (Lotus corniculatus)

A method was developed for rapid plant regeneration from protoplasts of birdsfoot trefoil (Lotus corniculatus L. cv. Leo). Green cotyledons from in vitro grown seedlings were preplasmolyzed in CPW salts containing 13% mannitol (CPW 13 M) for 1 h prior to the enzyme treatment. The enzyme formula consisted of 2% (w/v) Onozuka Cellulase R-10, 1% (w/v) Macerase and 0.1% (w/v) Pectolyase Y-23 in CPW 13 M. This method produced high yields of viable protoplasts after purification. The procedure is reproducible and takes approximately 2.5 months from protoplast isolation to plantlet establishment in a greenhouse. More than 100 plantlets were grown in soil. Two somaclonal variants, a chimeric plant for chlorophyll production and an albino cell line, have been obtained by this procedure.