First Report of Pseudomonas lurida Causing Bacterial Leaf Spot on Miscanthus sinensis

Miscanthus spp. are large perennial wetland grasses that are receiving considerable attention as bioenergy crops. In late summer 2011, leaf spot symptoms were observed in a field of Miscanthus sinensis in Jeongseon, Gangwon province, Korea. Bacterial strains that belonged to the γ‐Proteobacteria genus Pseudomonas were isolated from the affected leaves. By phylogenetic analysis and phenotypic characterization, the representative strain MDM‐03 was identified as Pseudomonas lurida. Healthy M. sinensis leaves inoculated with MDM‐03 developed leaf spots similar to those observed in field. Bacteria re‐isolated from the leaf lesions were identical to the original strain MDM‐03 based on their cultural characteristics and 16S rDNA sequencing. This is the first report of bacterial leaf spot in Miscanthus sinensis.     

Resistance of Hibiscus esculentus L. and Vigna sinensis (L.) Endl. to Pseudocercospora and plant peroxidase activity in relation to infection

Five cultivars of Hibiscus esculentus L. and six cultivars of Vigna sinensis (L.) Endl. were tested for their relative resistance to Pseudocercospora spp. Differences in susceptibility to the leaf spot pathogen were observed in both host and non-host interactions. The H. esculentus cv. South Sea and V. sinensis cv. Purple Mart were most susceptible to Pseudocercospora abelmoschi and P. cruenta, respectively. The H. esculentus cv. Pure Luck was most tolerant to P. abelmoschi while the V. sinensis cv. KY Bush was highly resistant to P. cruenta. A distinct host-specific interaction was observed among the different species of Pseudocercospora. Cultivar specific interactions were most pronounced between V. sinensis and P. cruenta. A direct correlation was observed between the variation in peroxidase activity in the soluble fraction of inoculated leaves and resistance to infection in H. esculentus and V. sinensis cultivars. The soluble fraction of inoculated leaves had higher peroxidase activity than either mitochondrial or chloroplast extracts.     

Description of 21 microsatellites for the Chinese bitterling,Rhodeus sinensis Günther, 1868

Chinese bitterling (Rhodeus sinensis) is a freshwater fish endemic to China. During the past several decades, natural populations of this species have become endangered due to habitat fragmentation and water pollution. To help conserve and restore R. sinensis, Illumina Miseq sequencing was used to identify 21 polymorphic microsatellites. The number of alleles varied from 3 to 13, and the observed heterozygosity and expected heterozygosity ranged from 0.3067 to 0.9145 and 0.3845 to 0.9521, respectively. The polymorphism information content ranged from 0.3173 to 0.9105. These novel microsatellites will be useful in both conservation and management of R. sinensis.     

皂荚刺发育过程中形态与结构观察研究

以皂荚实生苗为研究材料,利用扫描电镜、体式显微镜和石蜡切片法观察记录皂荚刺的发育过程,以明确皂荚刺发育过程的形态与结构变化,为皂荚遗传改良、定向栽培以及开发利用提供理论依据和技术指导。结果表明：(1)皂荚刺为枝刺,起源于皂荚幼苗叶腋内分生组织,发育过程与枝条的发育相似。(2)皂荚刺结构呈椭圆形,其木质部具有环纹导管,且木质部生长速度大于韧皮部。(3)皂荚刺的发育时期分为8个时期,依次为无刺期(催芽处理后2 d)、刺原基期(3 d)、鳞叶基本完成期(7 d)、刺分化期(8 d)、刺基本结构形成期(14 d)、刺开始木质化期(30 d)、刺开始褐化期(75 d)和刺完全褐化期(165 d)。     

PHENOL OXIDASE ACTIVITY IN ANOPHELES SINENSIS AND AEDES ALBOPZCTUS EXPOSED TO MICROFILARIAE OF BRUGIA MALAYI

Abstract Phenol oxidase (PO activity was investigated in cell-free hemolymph collected from Anopheles sinensis and Aedes albopictlcs female adults intrathoracically inoculated with Brugia dayi microfilariae (mff), inoculated with saline alone, or from uninoculated mosquitoes. The activity of PO from uninoculated 1-day-old mosquitoes was significantly greater than those of mosquitoes on 3, 12 and 16 day. PO activity in mff-inoculated A. sinensis at 24 h postinoculation (PI) was 2–3 times higher as compared with uninoculated or saline-inoculated mosquitoes. Inoculation of B. malayi mff into A. albopictus elicited ?-fold increase in PO activity at 24 h PI as compared with uninoculated mosquitoes. Immune-activated levels of PO activity in A. alboplctus was significantly higher as compared with those seen in A. sinensis. The relationship of observed differences in PO activity to differences in immunological capability between A. sinensis and A. albopictus is discussed.     

Isolation and characterization of polymorphic microsatellite markers of Anopheles sinensis,a malaria vector mosquito in the East Asia region

Eleven polymorphic dinucleotide (GA and CA) microsatellites were isolated and characterized in the mosquito Anopheles sinensis; this species is distributed over the East Asia region and is a primary vector of malaria, particularly in Korea. The number of alleles per locus ranged from 4 to 13. The observed heterozygosity (H_O) and expected heterozygosity (H_E) varied from 0.30 to 0.89 and from 0.59 to 0.90, respectively. These microsatellites could be useful in studying the evolution of the widely distributed A. sinensis in diverse environments.     

广东省中山市五桂山土沉香空间遗传结构

为了解五桂山的土沉香(Aquilaria sinensis)种群空间遗传结构,利用18个微卫星位点对143株胸高直径大于5 cm的土沉香个体基因型进行了检测。结果表明,五桂山土沉香种群的观测杂合度和期望杂合度均为0.534;近交系数为0.000 1,说明五桂山土沉香遗传多样性并不低,且种群处于随机交配状况。空间遗传结构分析结果表明,五桂山土沉香缺乏空间遗传结构,不同遗传背景的个体相互混杂,导致相邻个体遗传差异较大。这可能是五桂山土沉香种群发育过程中自疏所致,也可能是这一地区土沉香为人工种植,而非自然发育形成而造成的。     

Molecular variation of Spiranthes sinensis (Orchidaceae) in Japan, with special reference to systematic treatment of seasonally differentiated groups and a dwarf form, f. gracilis, from Yakushima Island

Molecular variations of Spiranthes sinensis Ames var. australis (R.Br.) H. Hara et Kitam. ex Kitam. in Japan were examined to evaluate the validity of the seasonally differentiated groups and a dwarf form of the species, which is endemic to Yakushima Island, Japan. Sequence differences in the plastid trnL-F locus clearly distinguished Japanese S. sinensis var. australis from S. sinensis var. sinensis collected from Ryukyu. In contrast, the trnL-F sequence of S. sinensis var. australis from Sabah, Malaysia, clearly differed from that of Japanese S. sinensis var. australis, suggesting genetic heterogeneity of Spiranthes sinensis var. australis in Asia. Moreover, a molecular analysis based on the sequences of nuclear ITS1 regions indicated that there are two major groups of S. sinensis var. australis in Japan, with a geographic distribution boundary on Kyushu Island. However, the trnL-F and ITS1 sequences did not support the genetic differentiation of the seasonally differentiated groups or the dwarf form from the other Japanese individuals. Based on these molecular data, the systematic treatment of physiological and morphological variations in the Japanese population of S. sinensis. var. australis is discussed.     

The allopolyploid origin of Spiranthes hongkongensis (Orchidaceae)

Morphological and ecological similarities between Spiranthes hongkongensis Hu et Barretto and S. sinensis (Pers.) Ames suggest a close evolutionary relationship. The hypothesis that S. hongkongensis evolved through natural hybridization between S. sinensis and S. spiralis was tested using isozyme electrophoresis. A total of 28 isozyme loci were interpreted in the diploid species S. sinensis and S. spiralis, and “fixed heterozygosity” owing to gene duplication was observed at ten of these loci in S. hongkongensis. Isozyme phenotypes provided strong evidence for an allotetraploid origin of S. hongkongensis, with S. sinensis and most likely S. spiralis as its diploid progenitors. Chromosomal counts of S. hongkongensis gave a tetraploid number of ≈60. The allotetraploid species is self-fertile, and completely isolated by sterility barriers from S. sinensis. Premating isolation also exists between S. hongkongensis and S. sinensis due to earlier flowering and autogamy in S. hongkongensis, which may account for the rarity of natural hybrids in the sites of sympatry. The role of the autogamous breeding system in the establishment and spread of S. hongkongensis is discussed.     

Biomass yield in a genetically diverse Miscanthus sinensis germplasm panel evaluated at five locations revealed individuals with exceptional potential

To breed improved biomass cultivars of Miscanthus ×giganteus, it will be necessary to select the highest‐yielding and best‐adapted genotypes of its parental species, Miscanthus sinensis and Miscanthus sacchariflorus. We phenotyped a diverse clonally propagated panel of 569 M. sinensis and nine natural diploid M. ×giganteus at one subtropical (Zhuji, China) and five temperate locations (Sapporo, Japan; Leamington, Ontario, Canada; Fort Collins, CO; Urbana, IL; and Chuncheon, Korea) for dry biomass yield and 14 yield‐component traits, in trials grown for 3 years. Notably, dry biomass yield of four Miscanthus accessions exceeded 80 Mg/ha in Zhuji, China, approaching the highest observed for any land plant. Additionally, six M. sinensis in Sapporo, Japan and one in Leamington, Canada also yielded more than the triploid M. ×giganteus ‘1993‐1780’ control, with values exceeding 20 Mg/ha. Diploid M. ×giganteus was the best‐yielding group at the northern sites. Genotype‐by‐environment interactions were modest among the five northern trial sites but large between Zhuji, and the northern sites. M. sinensis accessions typically yielded best at trial sites with latitudes similar to collection sites, although broad adaptation was observed for accessions from southern Japan. Genotypic heritabilities for third year yields ranged from 0.71 to 0.88 within locations. Compressed circumference was the best predictor of yield. These results establish a baseline of data for initiating selection to improve biomass yield of M. sinensis and M. ×giganteus in a diverse set of relevant geographies.     

皂荚种质资源SCoT遗传多样性分析及指纹图谱的构建

采用SCoT标记分析了18个皂荚种质的遗传多样性,并采用UPGMA法对18个皂荚种质进行了聚类分析。在此基础上,通过筛选出的多态性条带构建了18个皂荚种质的SCoT指纹图谱。扩增结果表明:从51个SCoT引物中筛选了15个引物进行PCR扩增,共扩增出226条带,其中多态性条带216条,多态性比率为96.61%。各引物多态性信息含量(PIC)、观测等位基因数(Na)、有效等位基因数(Ne)、Nei’s基因多样性指数(H)和Shannon’s信息指数(I)的平均值分别为0.875 9、1.964 9、1.440 1、0.272 6、0.426 1。18个皂荚种质的遗传相似系数在0.491 4~0.938 1之间,表明供试材料之间具有较丰富的遗传多样性。聚类分析结果表明:在遗传相似系数为0.60处可将18个皂荚种质分为3组,其中野皂荚单独为一组,山皂荚和皂荚-T聚为一组,其它皂荚材料聚为一组。利用3个引物扩增的8个多态性位点构建了18份皂荚种质资源的DNA指纹图谱,可以将其区分并精准鉴定。该研究结果为皂荚种质的鉴定和新品种选育提供了一定的理论依据。     

孑遗蕨类中华水韭根系发育的研究

以发育良好的中华水韭植株为材料,用常规半薄切片法,详细观察了根的发育及其根系的特征,并讨论了水韭根的内外两种起源途径、不同粗细根的结构功能、分支及横隔的生物学意义,为探讨古老维管植物的系统演化及濒危原因提供形态学依据。结果显示:(1)中华水韭成熟植株没有主根,有近百条浅棕色管状根,既有内起源根,也有外起源根,且每条根都有多回二歧分支。(2)二歧分支的发生一般频率密集且为不均等分裂,根越细分支越早,而且分支频率越密集;二歧分支处都有由2~3层细胞组成的永久性横隔,即使在老根内这种横隔也始终存在。(3)子根尖的部分皮层细胞能够直接转化成表皮,子根尖粗细不等。研究认为,中华水韭有茎起源根和根托起源根,皮层细胞具有特殊的分化能力,横隔是古生态的高能环境所造成的适应性结构,其根系具有旺盛的再生能力及高度的复杂性。     

Success of classical biocontrol agent Torymus sinensis within its expanding range in Europe

Torymus sinensis Kamijo (Hymenoptera, Torymidae), a classical biocontrol agent of chestnut gall wasp Dryocosmus kuriphilus (Hymenoptera, Cynipidae), was released in Croatia, Slovenia and Hungary in 2015. Following the introduction, the research of parasitism rates and population genetic indices on 40 different sites was performed to monitor and evaluate the success of establishment. The observed parasitism rates were unexpectedly high and negatively correlated with the distance from westernmost locality, while population genetic indices showed that populations of T. sinensis did not suffer from bottleneck‐induced founder effect phenomenon. Moreover, lack of genetic differentiation demonstrates that all populations share similar genetic structure, which could be shaped only by high levels of gene flow. We conclude that T. sinensis established viable and genetically diverse populations and successfully spread naturally from Italy across Slovenia to Croatia and Hungary.     

中华水韭叶舌和缘膜的发生及其发育进程研究

采用石蜡切片技术,以人工培养的中华水韭幼苗的最初几枚叶至成熟植株的叶为实验材料,连续解剖观察其叶舌和缘膜的发生、发育进程,并分析其发育进程与孢子囊和叶片的关系.结果显示:(1)中华水韭叶舌与叶片在其个体发育早期来自于同一原基,但叶舌最初的发育速度快于叶片.(2)中华水韭的苗龄达到15枚叶时开始有孢子囊发生,此时的叶舌下方有明显的缘膜结构.(3)当中华水韭的孢子体达到30枚叶片以上时,早期产生于植株外围的孢子囊已经发育成熟,可以清楚地区分出大、小孢子囊,其中在已经成熟的大孢子叶上,叶舌相对于孢子囊的长度变短,下唇萎缩,缘膜消失;成熟小孢子叶的叶舌比大孢子叶的叶舌长,上翻程度大,下唇萎缩程度不如大孢子叶明显,缘膜也退化消失.研究认为,缘膜是水韭系统发育早期的普遍结构,而演化后期一些地区的缘膜则显著退化甚至消失;对于系统发育初期的中华水韭,其叶舌与叶片的差异并不像现代水韭那么明显.     

Genetic diversity of six populations of Atractomorpha sinensis and Atractomorpha peregrina in Shanxi Province, China

Ten enzymes (AAT, CK, G3PDH, HEX, IDH, LDH, MDH, ME, PGI, PGM) were examined using horizontal starch gel electrophoresis to estimate the levels of genetic variation within and among six natural populations of two grasshopper species Atractomorpha sinensis and A. peregrina from Shanxi, China. The collecting sites were geographically distant from each other from south to north: Quwo district, Linfen city; Xiangyuan county, Changzhi; Jinyuan district, Taiyuan city; Yuanping county, Xinzhou city and Fanshi county of Xinzhou.A. sinensis showed 43 alleles at 16 loci but A. peregrine showed 39 alleles at 15 loci (Idh-1 was deficient). The zymograms showed that some common alleles were shared at several loci in these two species (Aat-1-b, Aat-2-b, G3pdh-a, Ck-1-b and Ldh-b). However, Hex-1-a, Hex-2-a, Hex-3-a, Idh-2-b, Mdh-2-b, Mdh-1-f, Pgi-b, Pgm-b had common alles in A. sinensis and Hex-1-b, Hex-2-b, Hex-3-b, Idh-2-a, Mdh-2-a, Mdh-1-d, Pgi-a, Pgm-c were of high frequency in A. peregrine instead. Most of the observed genotype frequencies were found to significantly deviate from the Hardy-Weinberg expectations in both species. A tendency of clinal distribution of allele frequency was observed at three loci. The frequency of the moderately migrating allele Me-c (0.318–0.740) in A. peregrina, Hex-1-a (0.800–1.000) and Ldh-b (0.487–0.750) in A. sinensis demonstrated increased frequency from north to south. Such tendency suggests that the allele frequency in these three loci may be correlated with the species’ geographic distributions. A. sinensis showed higher genetic diversity than A. peregrina as indicated by higher mean number of alleles per locus (A = 1.9–2.3 in A. sinensis and 1.7–2.2 in A. peregrina), percentage of polymorphic loci (56.3%–68.8% in A. sinensis and 43.8%–56.3% in A. peregrina), and the observed heterozygosities (H _o = 0.072–0.096 in A. sinensis and 0.70–0.107 in A. peregrina). The observed heterozygosities of the six populations were all noticeably lower than the Hardy-Weinberg expectations, mostly due to heterozygote deficiency in the populations of both species. The overall mean F _ST were small (F _ST = 0.045, P > 0.05 in A. sinensis populations and 0.087, P > 0.05 in A. peregrina populations). Nei’s genetic identity (I) estimates indicate low intraspecific (>0.95) but higher interspecific (0.377–0.447) genetic diversity. The cluster analysis based on modified Roger’s genetic distance (D) showed that the two species were divided into two branches. Both species are of limited dispersal capacity and a moderate geographical barrier might significantly restrict the gene exchange among populations, resulting in accumulation of local genetic differentiations. The A. sinensis populations used in this study were separated from each other by 155.2 to 271.4 km and the A. peregrina populations were separated from each other by 78.8 to 174.9 km with observable physical barriers. The allozyme data showed only minimal genetic differentiation at population level, most likely as a result of gene exchange. It is reasoned that natural factors and human agricultural activities might have facilitated migration and dispersal for the two species. __________ Translated from Acta Ecologica Sinica, 2005, 25(10): 2574–2481 [译自: 生态学报]     

Functional androdioecy in the rare endemic tree Tapiscia sinensis

Tapiscia sinensis, a rare endemic woody plant with both male and hermaphrodite individuals, is distributed in southern China. Whether T. sinensis is functionally androdioecious is unknown. In this study, we compare the male fitness between male and hermaphrodite individuals and perform pollination experiments in different habitats, identify the ability of actual siring of pollen from hermaphrodites and males under natural pollination, and discuss the evolution and maintenance of androdioecy in T. sinensis. Research suggests that flowers and fruits grow synchronously on hermaphrodite plants of T. sinensis from April to June. The males of T. sinensis had more than twice the genetic contribution of hermaphrodites through their male function and the fruit set from male pollination and cross‐pollination was the highest in all of the treatments, whereas that from self‐pollination was the lowest. Additionally, paternity analysis showed that the hermaphroditic pollen could result in siring success under natural pollination. The results showed that T. sinensis is a functionally androdioecious tree, that male individuals might evolve from a hermaphroditic ancestor and that the synchronous growth of flowers and fruit in hermaphrodites might facilitate the evolution and maintenance of androdioecy in T. sinensis.     

Root endophytic Chaetomium cupreum promotes plant growth and detoxifies aluminum in Miscanthus sinensis Andersson growing at the acidic mine site

Miscanthus sinensis Andersson grows naturally at the Hitachi mine. The root‐zone soil was acidic and contained high concentrations of Cu, Pb, Zn and exchangeable Al. Adventitious roots accumulated high concentrations of Al and Fe, but not other heavy metals. The purpose of this study was to elucidate the mechanism of tolerance of Al in M. sinensis, focusing on its chemical interaction with root endophytes. We isolated Chaetomium cupreum, which produced siderophores, from adventitious roots of M. sinensis via CAS assay. In inoculation tests, C. cupreum promoted M. sinensis seedling growth and increased Al and Fe uptake in the roots, although C. cupreum did not stimulate M. sinensis to produce Al detoxicants, such as citric and malic acids. Observation of the pattern of Al localization in the roots clarified that C. cupreum reduced Al toxicity in M. sinensis via compartmentalizing Al into fungal mycelia surrounding the roots and creating a less toxic Al‐localization pattern, allocating Al to the epidermis, endodermis and stele of roots. In conclusion, our results indicated that C. cupreum increases Al tolerance in M. sinensis growing at the acidic mine site.     

Juvenile Chinese mitten crabs (<Emphasis Type="Italic">Eriocheir sinensis</Emphasis>) in the Thames estuary: distribution,movement and possible interactions with the native crab <Emphasis Type="Italic">Carcinus maenas</Emphasis>

The Chinese mitten crab, Eriocheir sinensis is a successful invader. Whilst non-breeding adult E. sinensis have been associated with the destruction of riverbanks, little is known about the ecology of this species in its invaded areas. This is especially true of the juveniles which are a key migratory stage. Intertidal surveys along the Thames estuary indicated an increase in the population since the 1990s. Juvenile E. sinensis were abundant in the sampled upper tidal region of the Thames, refuging under boulders in the intertidal at low tide. Seasonal differences in sampled populations were observed, with a significantly lower abundance of crabs found during winter compared to summer. Mark-recaptures indicated movement in the intertidal occurring during high tide, with an influx of new crabs evident after a single tidal cycle. Endogenous rhythms were also apparent, with peaks in activity occurring corresponding with night-time high tide conditions. E. sinensis juveniles can successfully exclude similar sized native Carcinus maenas from shelters in the laboratory, regardless of which species originally inhabited the shelter. This may have implications for native estuarine Carcinus populations, which can rely on such intertidal shelters for refuges.     

Studies on the Genetic Control of a Nitrogenase Component in Leguminous Root Nodules

An attempt was made to determine whether Rhizobium or the host legume carries genetic information required for synthesis of the enzyme complex nitrogenase. No rhizobial mutants were found which could reduce N₂. Anaerobically-grown rhizobial cells contained neither component of nitrogenase. Rhizobium strain 32H1 was used to produce root nodules on Vigna sinensis and Phaseolus aureus. Other V. sinensis plants were inoculated with Rhizobium strain 29C2. The iron-molybdenum component of nitrogenase (fraction I) from these three combinations was purified by DEAE-cellulose chromatography and preparative electrophoresis. The final sample formed a single band after analytical electrophoresis on polyacrylamide gel. No differences in mobility were observed between the three nitrogenase samples during short periods of analytical electrophoresis. Longer periods of electrophoresis produced degradative products with similar electrophoretic mobilities from each nitrogenase sample. Amino acid analysis of fraction I samples from the three host-symbiont combinations suggested that the protein from the V. sinensis-Rhizobium 29C2 combinations was different from the other two samples. These results are interpreted as preliminary evidence that Rhizobium carries the genetic information for fraction I of nitrogenase.