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1.
Oleic acid is incorporated into an insoluble fraction left over after lipid extraction in Scenedesmus acutus. This incorporation is extremely sensitive to the chloroacetamide herbicide, metazachlor (I50= ca 20 nM). Therefore, factors influencing the incorporation of radioactivity from oleic acid into this non-lipid fraction were investigated. S. acutus cells were cultivated under various conditions with or without inhibitors and [14C]-oleic acid was supplied to the algae; the lipids were extracted and the radioactivity incorporated in the remaining fraction monitored. The inhibition seemed specific for chloroacetamides and related classes since it was also observed with alachlor, dimethenamid and mefenacet (an oxyacetamide). In contrast, it could not be found with diuron, oryzalin, nor could it be observed with a non-herbicidal metazachlor derivative or iodoacetamide. Incorporation of oleic acid into that fraction required meta-bolically active cells and was stimulated by light. Other fatty acids (16:0, 18:2, and 18:3) were also incorporated into the non-lipid fraction but their incorporation was not inhibited by metazachlor. Among other components, the fraction contains proteins. However, a possible specific effect of chloroacetamides on the binding of oleic acid to proteins or on the in vitro activity of lipid transfer proteins could not be detected. Not much is known yet about mechanism and chemistry of oleic acid incorporation but this finding opens a new path for investigations towards the primary target of these herbicides.  相似文献   

2.
Identification of units within species worthy of separate management consideration is an important area within conservation. Mitochondrial DNA (mtDNA) surveys can potentially contribute to this by identifying phylogenetic and population structure below the species level. The American crocodile (Crocodylus acutus) is broadly distributed throughout the Neotropics. Its numbers have been reduced severely with the species threatened throughout much of its distribution. In Colombia, the release of individuals from commercial captive populations has emerged as a possible conservation strategy that could contribute to species recovery. However, no studies have addressed levels of genetic differentiation or diversity within C. acutus in Colombia, thus complicating conservation and management decisions. Here, sequence variation was studied in mtDNA cytochrome b and cytochrome oxidase I gene sequences in three Colombian captive populations of C. acutus. Two distinct lineages were identified: C. acutus‐I, corresponding to haplotypes from Colombia and closely related Central American haplotypes; and C. acutus‐II, corresponding to all remaining haplotypes from Colombia. Comparison with findings from other studies indicates the presence of a single “northern” lineage (corresponding to C. acutus‐I) distributed from North America (southern Florida), through Central America and into northern South America. The absence of C. acutus‐II haplotypes from North and Central America indicates that the C. acutus‐II lineage probably represents a separate South American lineage. There appears to be sufficient divergence between lineages to suggest that they could represent two distinct evolutionary units. We suggest that this differentiation needs to be recognized for conservation purposes because it clearly contributes to the overall genetic diversity of the species. All Colombian captive populations included in this study contained a mixture of representatives of both lineages. As such, we recommend against the use of captive‐bred individuals for conservation strategies until further genetic information is available.  相似文献   

3.
ABSTRACT

Biomphalaria alexandrina snails have been used as bioindicators for freshwater qaulity and the effects of some herbicides such as butralin, glyphosate-isopropylammonium and pendimethalin). In the present study the effect of these three herbicides on snail biochemistry was examined. The results indicated that the herbicides increased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the haemolymph of B. alexandrina snails and significantly decreased total protein and albumin content. Light microscopical examinations of haemocytes monolayers of B. alexandrina snails showed three different cell types (small cells, granulocytes and hyalinocytes). All three herbicides caused abnormalities in cell shapes. Flow cytometric analysis of haemocytes from B. alexandrina demonstrated that circulating haemocyte populations could be divided into two main subtypes differing in their granularity (granulocytes or hyalinocytes) and size (large and small cells). In addition, the flow cytometric analysis showed that the total number of dead haemocytes in the haemolymph was significantly increased in treated groups compared to the control group. Phagocytosis in groups treated with the herbicides was highly significantly increased compared to the control indicating a very strong response of the treated snails. The results of the alkaline comet assay of DNA damage demonstrated that these herbicides have a genotoxic effect.  相似文献   

4.
Displacement of native plant species by non‐indigenous congeners may affect associated faunal assemblages. In endangered salt marshes of south‐east Australia, the non‐indigenous rush Juncus acutus is currently displacing the native rush Juncus kraussii, which is a dominant habitat‐forming species along the upper border of coastal salt marshes. We sampled insect assemblages on multiple plants of these congeneric rushes in coastal salt marshes in Sydney, New South Wales, Australia, and compared the abundance, richness, diversity, composition and trophic structure between: (i) J. acutus and J. kraussii at invaded locations; and (ii) J. kraussii at locations either invaded or not invaded by J. acutus. Although J. acutus supported a diverse suite of insects, species richness and diversity were significantly greater on the native J. kraussii. Moreover, insect assemblages associated with J. kraussii at sites invaded by J. acutus were significantly different from, and more variable than, those on J. kraussii at non‐invaded sites. The trophic structure of the insect assemblages was also different, including the abundance and richness of predators and herbivores, suggesting that J. acutus may be altering consumer interactions, and may be spreading in part because of a reduction in herbivory. This strongly suggests that J. acutus is not playing a functionally similar role to J. kraussii with respect to the plant‐associated insect species assemblages. Consequently, at sites where this non‐indigenous species successfully displaces the native congener, this may have important ecological consequences for community composition and functioning of these endangered coastal salt marshes.  相似文献   

5.
ABSTRACT. The dinoflagellate Tintinnophagus acutus n. g., n. sp., an ectoparasite of the ciliate Tintinnopsis cylindrica Daday, superficially resembles Duboscquodinium collini Grassé, a parasite of Eutintinnus fraknoii Daday. Dinospores of T. acutus are small transparent cells having a sharply pointed episome, conspicuous eyespot, posteriorly positioned nucleus with condensed chromosomes, and rigid form that may be supported by delicate thecal plates. Dinospores attach to the host via a feeding tube, losing their flagella, sulcus, and girdle to become spherical or ovoid cells. The trophont of T. acutus feeds on the host for several days, increasing dramatically in size before undergoing sporogenesis. Successive generations of daughter sporocytes are encompassed in an outer membrane or cyst wall, a feature not evident in trophonts. Tintinnophagus acutus differs from D. collini in host species, absence of a second membrane surrounding pre‐sporogenic stages, and failure to differentiate into a gonocyte and a trophocyte at the first sporogenic division. Phylogenetic analyses based on small subunit (SSU) ribosomal DNA (rDNA) sequences placed T. acutus and D. collini in the class Dinophyceae, with T. acutus aligned loosely with Pfiesteria piscicida and related species, including Amyloodinium ocellatum, a parasite of fish, and Paulsenella vonstoschii, a parasite of diatoms. Dubosquodinium collini nested in a clade composed of several Scrippsiella species and Peridinium polonicum. Tree construction using longer rDNA sequences (i.e. SSU through partial large subunit) strengthened the placement of T. acutus and D. collini within the Dinophyceae.  相似文献   

6.
The parasite contamination make difficult laboratory cultivation of green algae and decrease significantly the production of microalgal biomass during industrial cultivation. In the present study the influence of the endogenous abscisic acid content (determinate by gas chromatography) on the host-parasite relationship in different ontogenetic phases of the host Scenedesmus as well as resistance induction after treatment of synchronous algal culture with ABA were studied. Synchronization of algae was carried out by alteration of light and dark periods. The age groups under study were: autospores (at the beginning of the light period), growing cells, mature cells (belonging to in the end of the light period) and mature cells, starting to release autospore at the beginning of dark phases. The higher levels of endogenous ABA during the algal autospore formation as well as exogenous ABA supply of (10−5 M) inhibited the infection process in the pathosystem green microalga Scenedesmus acutus and unicellular fungal parasite Phlyctidium scenedesmi. The treatment with fluridone 10−7 M (an inhibitor of ABA biosynthesis) increased the host susceptibility during all ontogenetic phases. The susceptibility of S. acutus to the chytridial infection depended on the endogenous ABA level during different ontogenetic stages of the alga.  相似文献   

7.
Many herbicides of different chemical structure inhibit photosynthetic electron flow by interrupting the photosyn‐thetic electron flow by interrupting the photosynthetic electron transport chain between the primary acceptor (QA) and the secondary acceptor (QB) of photosystem 2 (PS2). Thermoluminescence (TL) originates from PS2, and the bands of the glow curve can be related to the charge recombination between positively charged donors and negatively charged acceptors. The glow curve of TL is strongly influenced by addition of PS2 herbicides. The herbicide treatment shifts the peak position and activation energy of the TL band related to QA, suggesting that herbicide binding affects the midpoint redox potential not only of Q B but also that of QA. On the basis of the band shift the herbicides of various chemical structures can be classified into different “thermodynamical” groups which relfect the differences in the binding properties of these herbicides. As a new approach TL seems to be a useful technique in studying the mechanism and site of action of herbicides that inhibit electron transport of PS2.  相似文献   

8.
An in vitro embryo culture protocol was developed for Scirpus acutus Muhl. A maximum of 85.6% of germination was obtained when isolated embryos were cultured in vitro, a result similar to those reported in the literature with traditional dormancy breaking treatments. In vitro seedling development was optimal in half-strength Murashige and Skoog (1962) liquid medium. An average of 3–4 shoots were produced from the initial seedlings. Clusters of plantlets were successfully acclimatized and transferred to soil. These results corroborate the findings of previous studies stating that seed dormancy in Scirpus is caused by the seed/fruit coats. In vitro embryo culture thus allows for the production of Scirpus acutus Muhl. seedlings that can be transferred to natural or artificial wetlands.  相似文献   

9.
Different approaches to the study of life cycle strategies of Antarctic copepods are described in an attempt to shed new light on our present knowledge. To date, most studies were carried out on abundance, horizontal and vertical distribution and stage composition during different seasons and in various regions. Hence, the seasonal pictures had to be compiled from different years and sampling regions. The physiological method includes measurements on e.g. egg production, feeding, respiration and excretion rates, C:N and O:N ratios, lipid and protein contents. However, both physiological and biochemical data are still rare. Results of field observations are given in this paper for investigations conducted within the last 15 years in the eastern Weddell Sea, while data of physiological parameters are based on a broader geographical region. In the eastern Weddell Sea, eight copepod species account for about 95% of copepod abundance and for more than 80% of copepod biomass. Within the calanoids, the small species Microcalanus pygmaeus dominates by numbers with 66%, while the large species Calanoides acutus and Calanus propinquus comprise together 52% of the biomass. Species abundance is lowest in winter and highest in summer/autumn, however, seasonal changes in the abundance of M. pygmaeus are small and this species occurs in similar quantities throughout the year. All copepod species show a distinct seasonal vertical distribution pattern and they occur in upper water layers in summer, in contrast to the other seasons. However, the depth layers of maximum concentration differ between species. The ontogenetic vertical migration is most pronounced in C. acutus and relatively weak in C. propinquus. The age structure also shows seasonal differences with the youngest population observed in summer for C. acutus, C. propinquus, Ctenocalanus citer or autumn for Metridia gerlachei, whereas the M. pygmaeus population is oldest during summer. The youngest copepodite stage and the males are not always present in C. acutus and C. propinquus. In contrast, all developmental stages and both sexes occur throughout the year in M. gerlachei, M. pygmaeus and C. citer. Gonad maturation in the dominant calanoid species proceeds well before the onset of phytoplankton production in the eastern Weddell Sea. However, the highest portion of females with ripe gonads and hence highest egg production rates coincide with the productive period in spring and summer. In autumn, ovaries of the three larger species C. acutus, C. propinquus and M. gerlacheiare all spent. In contrast, the percentage of ripe females of the two smaller species, C. citer and M. pygmaeus, stays high in autumn. Egg production rates are highly variable within one region and species. Many copepods accumulate large depots of lipid, mainly wax esters. In contrast, five species (C. propinquus, C. simillimus, Euchirella rostromagna, Stephos longipes and Paralabidocera antarctica) almost exclusively synthesise triacylglycerols and not wax esters. The lipid content exhibits distinct seasonal patterns, and is highest in autumn. A seasonal difference is also obvious in metabolic activities with lowest rates during the dark season. The adaptation to the pronounced seasonality in the Southern Ocean differs greatly between copepod species, and most Antarctic copepods stay active during the dark season. Calanoides acutus seems to be the only true diapause species. Calculations of summer developmental rates and winter mortality rates of the large species C. acutus and C. propinquus suggest that both species have a 1-year life cycle with few females overwintering and probably spawning a second time. In contrast, a 2-year life cycle is more likely in R. gigas. However, life cycle durations of all species studied are still uncertain and regional differences are very probable.  相似文献   

10.
Lipid content, fatty acid composition, and feeding activity of the dominant Antarctic copepods, Calanoides acutus, Calanus propinquus, and Metridia gerlachei, were studied at a quasi-permanent station in the eastern Weddell Sea in December 2003. During 3 weeks of the spring phytoplankton development, total lipid levels of females and copepodite stages V (CVs) of C. acutus were almost doubled. Meanwhile, only a slight increase in total lipid content occurred in M. gerlachei, and no clear trend was observed in lipids of C. propinquus females. The pronounced increase of lipids in C. acutus was due to an accumulation of wax esters. The proportion of wax esters in the lipids of M. gerlachei was clearly lower, while triacylglycerols played a more important role. In C. propinquus, triacylglycerols were the only neutral lipid class. There were no pronounced changes in the feeding activity of M. gerlachei, whereas the feeding activity of C. acutus had rapidly increased with the development of the phytoplankton bloom in December, which explains its rapid lipid accumulation. The combination of gut content and fatty acid trophic marker analyses showed that C. acutus was feeding predominantly on diatoms. The typical diatom fatty acid marker, 16:1(n-7), slightly decreased and the tracer for flagellates, 18:4(n-3), increased in females and CVs of C. acutus. This shift indicates the time, when the significance of flagellates started to increase. The three copepod species exhibited different patterns of lipid accumulation in relation to their trophic niches and different duration of their active phases. The investigations filled a crucial data gap in the seasonal lipid dynamics of dominant calanoid copepods in the Weddell Sea in December and support earlier hypotheses on their energetic adaptations and life cycle strategies.  相似文献   

11.
Summary Three corn (Zea mays L.) lines resistant to imidazolinone herbicides were developed by in vitro selection and plant regeneration. For all three lines, resistance is inherited as a single semidominant allele. The resistance alleles from resistant lines XA17, XI12, and QJ22 have been crossed into the inbred line B73, and in each case homozygotes are tolerant of commercial use rates of imidazolinone herbicides. All resistant selections have herbicide-resistant forms of acetohydroxyacid synthase (AHAS), the known site of action of imidazolinone herbicides. The herbicide-resistant phenotypes displayed at the whole plant level correlate directly with herbicide insensitivity of the AHAS activities of the selections. The AHAS activities from all three selections have normal feedback regulation by valine and leucine, and plants containing the mutations display a normal phenotype.  相似文献   

12.
Rapid and widespread evolution of multiple herbicide resistance in global weed species endowed by increased capacity to metabolize (degrade) herbicides (metabolic resistance) is a great threat to herbicide sustainability and global food production. Metabolic resistance in the economically damaging crop weed species Lolium rigidum is well known but a molecular understanding has been lacking. We purified a metabolic resistant (R) subset from a field evolved R L. rigidum population. The R, the herbicide susceptible (S) and derived F2 populations were used for candidate herbicide resistance gene discovery by RNA sequencing. A P450 gene CYP81A10v7 was identified with higher expression in R vs. S plants. Transgenic rice overexpressing this Lolium CYP81A10v7 gene became highly resistant to acetyl-coenzyme A carboxylase- and acetolactate synthase-inhibiting herbicides (diclofop-methyl, tralkoxydim, chlorsulfuron) and moderately resistant to hydroxyphenylpyruvate dioxygenase-inhibiting herbicide (mesotrione), photosystem II-inhibiting herbicides (atrazine and chlorotoluron) and the tubulin-inhibiting herbicide trifluralin. This wide cross-resistance profile to many dissimilar herbicides in CYP81A10v7 transgenic rice generally reflects what is evident in the R L. rigidum. This report clearly showed that a single P450 gene in a cross-pollinated weed species L. rigidum confers resistance to herbicides of at least five modes of action across seven herbicide chemistries.  相似文献   

13.
Ethylene and Ethane Production from Sulfur Dioxide-injured Plants   总被引:8,自引:4,他引:4       下载免费PDF全文
After alfalfa (Medicago sativa) seedlings were exposed to approximately 0.7 microliter per liter SO2 for 8 hours, elevated ethylene and ethane production was observed. Ethylene production peaked about 6 hours and returned to control levels by about 24 hours following the fumigation, while ethane production peaked about 36 hours and was still above control levels 48 hours after the fumigation. Light had an opposite effect upon the production of the two gases: ethane production rates were higher from plants held in light, whereas ethylene production rates were higher from those held in the dark. Peak ethylene and ethane production rates from SO2-treated plants were about 10 and 4 to 5 times greater, respectively, than those of the control plants. Ethylene appeared to be formed primarily from stressed yet viable leaves and ethane from visibly damaged leaves. The different time courses and light requirements for ethylene and ethane production suggest that these two gases were formed via different mechanisms. Light appears to have a dual role. It enhances SO2-induced cellular damage and plays a role for repairs.  相似文献   

14.
The mode of action of paraquat (1,1-dimethyl-4,4-bipyridinium) and the mechanism of resistance to it were studied in leaves of atrazine/paraquat co-resistant (R) and susceptible (S) biotypes of horseweed (Conyza canadensis) collected from Hungarian vineyards. The application of 0·5 mol m?3 paraquat by spraying onto the surface of the leaves of intact plants in the light rapidly led to typical symptoms of paraquat action in the initial period in both biotypes, i.e. inhibition of CO2 fixation, suppression of variable chlorophyll fluorescence (Fv), decrease of oxygen evolution and stimulation of ethane production. The inhibitory effect of paraquat in the S plants was irreversible, whereas it was transient in the R plants and those plants recovered gradually afterwards. The R plants recovered from the inhibitory effect of paraquat only in the light, and an increase in light intensity was found to have a pronounced effect on the recovery of Fv. The mechanism of resistance to paraquat in C. canadensis is discussed.  相似文献   

15.
The present study investigates the effect of anthocyanin (ACN), phenolic acid (PA) fractions, and their combination (ACNs:PAs) from wild blueberry powder (Vaccinum angustifolium) on the speed of endothelial cell migration, gene expression, and protein levels of RAC1 and RHOA associated with acute exposure to different concentrations of ACNs and PAs. Time-lapse videos were analyzed and endothelial cell speed was calculated. Treatment with ACNs at 60 μg/mL inhibited endothelial cell migration rate ( P ≤ 0.05) while treatment with PAs at 0.002 μg/mL ( P ≤ 0.0001), 60 μg/mL ( P ≤ 0.0001), and 120 μg/mL ( P ≤ 0.01) significantly increased endothelial cell migration rate compared with control. Moreover, exposure of HUVECs to ACNs:PAs at 8:8 μg/mL ( P ≤ 0.05) and 60:60 μg/mL increased ( P ≤ 0.001) endothelial cell migration. Gene expression of RAC1 and RHOA significantly increased 2 hours after exposure with all treatments. No effect of the above fractions was observed on the protein levels of RAC1 and RHOA. Findings suggest that endothelial cell migration is differentially modulated based on the type of blueberry extract (ACN or PA fraction) and is concentration-dependent. Future studies should determine the mechanism of the differential action of the above fractions on endothelial cell migration.  相似文献   

16.
Summary Chlorsulfuron and/or imazaquin resistant mutants of Chlamydomonas reinhardtii strain CW15 have been obtained and shown to have actolactate synthase (ALS) with altered sensitivity to one or both of these herbicides. Herbicide resistance in the three mutants described is allelic, and resistance appears to result from a dominant or semidominant mutation in a single, nuclear gene. Imazaquin and chlorsulfuron resistant ALS from imazaquin and chlorsulfuron resistant mutants, together with single-gene Mendelian inheritance of these phenotypes, suggests that ALS is the sole site of action of the two herbicides in Chlamydomonas. A high degree of cross resistance between the two herbicides was found in only one mutant. This mutant (IM-13) was selected for resistance to imazaquin and has a high level of in vitro resistance to both imazaquin (270-fold increased I50) and chlorsulfuron (900-fold increased I50). In another mutant selected for resistance to imazaquin (IMR-2), hyper-sensitivity to chlorsulfuron was found. A mutant selected for resistance to chlorsulfuron (CSR-5), had a substantial degree of resistance of chlorsulfuron (80-fold increased I50), but not to imazaquin (7-fold increased I50).  相似文献   

17.
The gene of tryptophan 7-halogenase was isolated from the Pseudomonas aureofaciens ACN strain producing pyrrolnitrin, a chlorocontaining antibiotic, and sequenced. A high homology degree (over 95%) was established for the genes and the corresponding halogenases from P. aureofaciens ACN and P. fluorescens BL915. The tryptophan 7-halogenase gene was amplified by PCR, and the corresponding enzyme was expressed in Escherichia coli cells using the pBSII SK+ vector.  相似文献   

18.
Heat shock proteins (hsp) are a highly conserved group of proteins that are synthesized as a response to different forms of stress (heat, toxic chemicals, diseases, non-physiological pH changes). Because of their high sensitivity to changes in the environment, these proteins were suggested as possible early biomarkers of exposure in ecotoxicological studies. The purpose of the present study was to check the suitability of hsp32 and hsp70 as biomarkers of in vitro exposure to environmentally relevant carcinogens: polycyclic aromatic hydrocarbons (PAHs), their nitro-derivates, aromatic amines, acrylonitrile (ACN) and the mixture of organic compounds adsorbed onto ambient airborne particles (extractable organic matter, EOM).The expression of hsp32 and hsp70 was studied in human diploid lung fibroblasts (HEL cells) and human monocytic leukaemia cells (THP-1 cells) incubated in vitro with different concentrations of dibenzo[a,l]pyrene (DB[a,l]P), 1-nitropyrene, (NP), 4-aminobiphenyl (ABP), ACN and EOM for different periods of time. The incubation of cells with DB[a,l]P, NP, ABP and EOM did not result in increased levels of hsp32 or hsp70, either in dose- or time-dependent manner. ACN induced the expression of hsp32 as well as hsp70 in HEL and THP-1 cells, which probably reflects its ability to induce oxidative stress. We conclude that hsp32 and hsp70 are not suitable biomarkers of an early exposure to PAHs, their nitro-derivates, aromatic amines or EOM under the conditions used.  相似文献   

19.
Open-water, marginal-ice and in-ice zones were sampled in the Weddell Sea during November and December, 1993 in an effort to examine the influence of the early spring bloom on the diet and population structure of the three biomass dominant copepods: Metridia gerlachei, Calanus propinquus, and Calanoides acutus. The abundance of all three species in the upper 200 m was highest at stations in the open water, but individually, each species displayed a unique trend. M. gerlachei, which showed the least variability, was significantly more abundant in open water than in the marginal-ice zone. The abundance of Calanus propinquus was higher in open water than in the marginal-ice zone or in the ice. Calanoides acutus displayed the highest variability, with significant differences between all three ice-cover zones. Diet analysis revealed no significant differences in the number of food items within each ice-cover zone and diatoms were the most numerous item identified in the guts of all three species. However, M. gerlachei and Calanus propinquus also contained metazoan material, while Calanoides acutus did not. There were dramatic differences in the age composition of the species between the zones. Early copepodite stages of all three species predominated at the ice edge and in open water. Numbers of M. gerlachei adult females were roughly equivalent in all three zones while Calanoides acutus and Calanus propinquus adult females composed a higher fraction of the total population within the ice. These results compare well with life-history data compiled by other authors and reinforce the importance of the ice edge to bloom-dependent Antarctic zooplankton. Accepted: 5 April 1999  相似文献   

20.
Drits  A. V.  Pasternak  A. F.  Kosobokova  K. N. 《Hydrobiologia》1994,(1):201-207
Ingestion, respiration and excretion rates as well as lipid and protein body content of the dominant Antarctic copepod Calanoides acutus (CIV to adult females) were studied during the period covering the end of phytoplankton bloom (February) to the beginning of transition to overwintering (March-April). Daily rations measured with gut fluorescence varied from 2.2 to 2.7% in surface C. acutus. Weight-specific respiration and excretion rates in deep C. acutus decreased by a factor of 11 and 3.5–3.8 compared to their surface counterparts. High lipid (up to 455 µg ind–1) and protein (198 µg ind–l) content was observed in surface C. acutus CV in February; a month later the animals with similar lipid and protein content were found in the depth (500–1000 m layer). Their lipid reserves were enough to overwinter and probably to ascend, molt and reproduce. At the same time some of the deep CVs had much lower protein and lipid content and could survive only for 4–5 months. Our own and literature data led to the conclusion that females of C. acutus reach adulthood at the age of more than one year while development of males could be completed in one year.  相似文献   

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