2001 Vol. 22, No. 1
2001, 22(1): 1-8.
In the present paper,the cladistics method is used to analyze the phylogenetic relationship of 16 genera of Tinginae (Hemiptera:Heteroptera:Tingidae) from northern China.33 characters with 88 states were chosen based on the morphological comparison.The character matrix was formed through ingroup and outgroup analysis and computed using the computer programs MacClad (version 3.0.1),AutoDecay (version 3.0),PAUP (version 3.1.1) and Hennig 86 (version 1.5),and the same most parsimonious tree and Nelson consensus cladogram were obtained from both computation to explain the phylogenetic relationship among the genera (L=118,CI=0.454,RI=0.529).We categorized these genera involved in the analysis phylogenetically into 6 groups according to the results.They are Agramma group,Leptoypha group,Dictyla group,Catoplatus group,Physatocheila group and Derephysia group.The phylogenetic relationship among the involved genera is presented as(((((Derephysia group=(Lasiacantha,(Acalypta,(Galeatus,(Dictyonota,(Derephysia,Sphaerista))))),Physatocheila group =((Elasmotropis,Tingis),((Oncochila,Cochlochila),Physatocheila))),Catoplatus group=Catoplatus),Dictyla group=(Monosteira,Dictyla)),Leptoypha group=Leptoypha),Agramma group=Agramma).These groups will not be regarded as higher classific taxa until more genera and data are available and further analysis are carried out.
Electrophoretic Analysis of Isozymes and Discussion about Species Differentiation in Three Species of Genus Gymnocypris
2001, 22(1): 9-19.
By using the method of electrophoresis,three isozymes (lactate dehydrogenase,malate dehydrogenase and esterase) of three species of genus Gymnocypris were described and analyzed from North Tibet in this paper.The results showed that all three isozymes presented interspecific difference and distinct differentiation among individuals in the same population,and there was no electrophorectic difference between males and females.Analysis of relationships among three naked carps indicated a high degree of similarity between G.selincuoensis and G.cuoensis,whereas low degree between G.selincuoensis and G.namensis.Furthermore,three isozymes presented expression of null alleles,and the duplicate genes of LDH-A,LDH-B,s-MDH-A and m-MDH-B also expressed in some individuals.Compared to other tetraploid fishes,three naked carps retained more functional duplicate genes and null alleles.This suggests fishes of genus Gymnocypris are at the early stage of evolution after polyploidization than that of fishes of Catostomidae,it directly related to the later time of schizothoracine fishes originate as well as severe environment.
2001, 22(1): 20-26.
In this study,we had developed a fast and easy method to extract total DNA from migratory locusts.We could extract about 50 μg and 100 μg DNA from single male and female individual respectively.The extracted total DNA was about 45 kb in length with OD/OD between 1.5 and 2.2,which could satisfy the requirements of RAPD and PCR on DNA template.In order to obtain high-molecular-weight DNA from migratory locusts and repeatable RAPD results,we used autoclaved tips to collect precipitated DNA after being extracted with phenol/chloroform instead of collecting by centrifugation.We tested experimental conditions including different PCR amplifiers,tubes and concentrations of template DNA,Taq DNA polymerase,dNTP and primer that might affect RAPD results.Within some limits,the concentrations of template DNA,Taq DNA polymerase,dNTP and primer did not greatly affect RAPD profiles.The optimized reaction conditions of RAPD for three Chinese locust subspecies in 25 μL reaction volume were as follows:20 ng template DNA,0.1 mol/L dNTP,0.2 μmol/L primer,1 U Taq polymerase.Some advice was also given,especially all individuals of all populations should be amplified with every primer at the same reaction conditions.With the optimized reaction conditions for RAPD,we analyzed the genetic relationship among three locust subspecies in China.The genetic distances generated by RAPDDIST between populations belonging to the same subspecies were less than those to different subspecies.In the UPGMA Phenogram of three locust subspecies,the two populations of L.m.manilensis and L.m.tibetensis were clustered together with a bootstrap of 100%,respectively.The Bameng population of L.m.migratoria was clustered with two populations of L.m.manilensis with a bootstrap of 66%,meaning that the Bameng population of L.m.migratoria could also clustered with L.m.tibetensis.In the UPGMA Phenogram of all individuals of three subspecies,all individuals of one subspecies were clustered together to form a branch,so three subspecies were significantly different in DNA polymorphism.Thus,RAPD method could be used to separate the three subspecies.There was a gene flow between the populations of L.m.tibetensis and L.m.manilensis.In order to get the true relationships among the three subspecies,more samples and primers should be used in the analysis on the three locust subspecies with RAPD,and other molecular techniques should be applied to test the results of RAPD.
Phylogenetic Relationships of Chinese Brown Frogs Inferred from Mitochondrial DNA Sequence of 12 S Ribosome RNA Gene
2001, 22(1): 27-32.
Aligned 393 base pairs of the mitochondrial 12S rRNA gene from 6 species of Chinese brown frogs,and Fejervarya limnocharis,Pelophylax nigromaculata and P.plancyi selected as outgroup were sequenced.Pairwise comparisons showed that the site variability between ingroup and outgroup members ranged from 7.3% to 23.1%.The variability among ingroup members ranged from 0.0% to 9.2%.Phylogenetic analyses based on these DNA sequence data by the neighbor-joining and maximum parsimony methods showed:1)These six species of Chinese brown frogs grouped together and formed a monophyletic clade (BPs value >90%).2)The six species of ingroup were divided two sister groups,I.e.Rana chensinensis,R.amurensis,and R.huanrenensis clustered together (BPs value >94%),and R.omeimontis,R.chaochiaoensis and R.zhenhaiensis formed another cluster (BPs value >50%).3)R.chaochiaoensis was more closed to R.zhenhaiensis than R.omeimontis.4)The Yuzhong population of R.chensinensis related to R.huanrenensis while the Mudanjiang population of R.chensinensis was close to R.amurensis.This indicates that the differentiation between the two populations of R.chensinensis is likely to be at species level.So R.kukunoris including the Yuzhong population of R.chensinensis and R.dybowskii including the Mudanjiang population of R.chensinensis may be revived as two valid species.
2001, 22(1): 33-40.
The acquired immunodeficiency syndrome caused by HIV-1 is spreading all over the world.The slow progress in AIDS therapy and vaccine partially imputes to be lack of appropriate animal models used to study AIDS pathogenesis and to evaluate vaccines.To find an animal model is an imperative task in HIV research.Tree shrew is widely used in biomedical research,and is susceptible to many medically important viruses.Whether tree shrew (Tupaia belangeri) can be infected by HIV-1 is a valuable approach.In present study,wild tree shrews from Yunnan Province were maintained in laboratory animal facility for more than 2 weeks before sacrifice.Tree shrew spleen and peripheral blood lymphocytes and monocytes/macrophages and human peripheral blood lymphocytes and monocytes were separately infected by 5 HIV-1 strains (HIV-1[ⅢB],HIV-1[JR-FL],HIV-1[Ada-M],HIV-1[Ba-L],HIV-1[SF162]),which use different coreceptors after the cells were activated by allogeneic lymphocytes,PHA and IL-2 for 72 h.The HIV-1 infected cells were then cultured in vitro for 15 days.The proliferation rates and viability of HIV-1 infected human immunocytes obviously dropped on day 15,but those of tree shrew immunocytes did not despite either infected or not infected by HIV-1.HIV-1 particles in the infected culture supernatant and proviral DNA in the cells were respectively detected by RT-PCR using primer sk145/431 and by PCR using primer sk68/69 on days 1,3,5,7,9,12,15 after infection with HIV-1.In addition,the expression of HIV-1 specific antigens on the tree shrew and human immunocytes after infection were detected by flow cytometry,for which an AIDS patient plasma was used as the first antibody,and FITC-conjugated sheep anti-human-IgG as the second antibody.HIV-1 RNA and proviral DNA were respectively found in the culture supernatant of the HIV-1 infected human immunocytes and in the infected cells.Using flow cytometry,HIV-1 specific antigens were also measurable on the surface of these cells.However,neither HIV-1 RNA in the supernatant from the infected HIV-1 tree shrew immunocytes nor the proviral DNA from the infected tree shrew immunocytes could be detected.The HIV-1 specific antigens were not demonstrated on the surface of the HIV-1 infected tree shrew immunocytes.Taken together,these experimental results suggested that the tree shrew immunocytes can not be in vitro infected by these HIV-1 strains.It seems to be caused by the structure differences of the HIV-1 receptors (CD4) and coreceptors (CCR5 or CXCR4) between human and tree shrew immunocytes.
The Recruitment of Brown Adipose Tissue and Expression of Uncoupling Protein Gene in Brandt's Vole during Cold Exposure
2001, 22(1): 41-45.
Brandts voles (Microtus brandti) were randomly divided into seven groups and exposed to cold temperature［12L∶12D,(4±2)℃］ for 12 hours,1,3 ,7,14,21 and 28 days respectively;the control group was kept in warm place ［12L∶12D,(25±2)℃］.Compared with the control,the weight of BAT and the total contents of DNA in BAT decreased during cold exposure from 12 hours to 3 days,but increased significantly from 7 to 28 days.Cold exposure also induced the increase of protein contents of BAT.In molecular level,the contents of UCP mRNA in BAT increased significantly and reached peak at 21 days.The results suggested that cold exposure could induce the recruitment of BAT cell and the expression of UCP gene,resulting in the increase of adaptive thermogenesis in Brandts voles.
The characteristic of feeding sites for Crested Ibis (Nipponia nippon) in winter was analyzed using plot methods based on radio-telemetry study.Three types of feeding sites for Crested Ibis in winter were recorded:paddy field,river shoal and reservoir.Among them,paddy field is the main feeding site for adults,river shoal for juveniles.Analysis of variance and factor analysis show that the main factors influencing the feeding site selection of Crested Ibis are the elevation,the degree of width,the area of feeding sites,the disturbance of human activities,the coverage of vegetation and the softness of soil.Restoration and protection of paddy fields in winter are important for the conservation of Crested Ibis.
2001, 22(1): 51-57.
To further understand the mechanism of sexual difference of vocalization and the relation among the vocal behavior,vocal control nuclei and testosterone level in the serum,sound spectrogram,volume of three vocal control nuclei (HVC high vocal-control center,RA robutus archistriatalis nucleus and Area X),weight of testis and testosterone levels in the serum were investigated in the songbird (Lonchura striata swinhoei) from post-hatching 5 days to 120 days.The results were as follows:1) Both the male and female produced several kinds of calls before post-hatch 45 days and there was no sexual difference among these calls between the male and female.Male birds began to sing only after post-hatching 45 days.However,they could not sing the same complex and stable songs as the adult males until post-hatch 120 days.Female birds could not produce any song as the male.2) HVC,RA and Area X in the female were 6,4,and 2 times less than the volumes of the corresponding nuclei in the male,respectively.HVC,RA and Area X developed in different ways.Their rapid increasing period was not parallel to the time when the male birds learned singing,indicating that the development of the vocal control nuclei was not completely subjected to the vocal behavior.3) Only the testis developed very well or the testosterone in the serum reached adequately high level,male birds could produce quite complex and stable songs,suggesting that testosterone level in the serum could be one of the important factors in the mature song production.
A Study on the Ant Communities on West Slope at Different Elevation of the Gaoligongshan Mountain Nature Reserve in Yunnan,China
2001, 22(1): 58-63.
The ant communities and their species diversity with altitudinal zonation on west slope of the Gaoligongshan Mountain Nature Reserve were studied for the first time.Along with the increasing of altitude,number of dominant species increases at the north and north-middle sections of the reserve,but decreases at the south section.While the altitude increasing,percentage of dominant species decreases at the north and north-middle sections,but increases at the south section.It is inferred that along with the increasing of altitude the species number decreases.This study also suggested that the density of individuals decrease while the altitude increasing.At the north and north-middle sections the predominant index decreases while altitude increasing,but the index increases at middle-south and south sections.Along with the altitude increasing,the species diversity index increases at north section,without regularity at north-middle section,and decreases at middle-south and south sections.At north,north-middle and middle-south sections,evenness index increases while altitude increasing,but the index decreases at south section.Similarity coefficient between the ant communities on west slope at all the four sections is almost all between 0.00-0.25,with only one coefficient surpasses this range.We think the basic rules of the species diversity of ant communities on west slope under the state of virgin vegetation should be that along with the altitude increasing,number of dominant species decreases,percentage of dominant species increases,species number decreases,individual density decreases,predominant index increases,species diversity index decreases,and evenness index decreases.The unusual situation observed in the practical investigation might be caused by the destroying of vegetation on the foot of the mountain and half way up the mountain.
Effects of Lipid Nutrition on Hepatopancreatic Ultrastructure of Eriocheir sinensis Larvae (Crustacea:Decapoda)
2001, 22(1): 64-68.
From March to July of 1998 Eriocheir sinensis larvae were cultured by feeding with rotifers treated with lipid enrichment 50 DE-G at early zoeal developmental stages and with Artemia nauplii at postlarval developmental stages.The hepatopancreatic ultrastructure of the larvae was observed through transmission electronic microscopy.The results show:1) the hepatopancreatic length increase with the larval development,and the cells of hepatopancreas consist of four types,which are embryo cell (E cell),resorptive cell (R cell),fibrillar cell (F cell) and secretory cell (B cell) respectively;2) E cells are stem cells that can be differentiated into other three types of cells;3) B cells have 1 to 2 large vacuoles and numerous rough endoplasmic reticulum (Rer);4) R cells store some nutrients such as lipid droplets for the larval development and have some vacuoles,large quantity of smooth endoplasmic reticulum (Ser) and free ribosomes,and many mitochondria with high electron density matrix and plenty of ridges can be observed in R cells;5) F cells have plenty of zymogen granules near the apical cells;6) compared with hepatopancreatic ultrastructure of the larvae fed with rotifers cultured by bakers yeast,significant difference can be observed in R cells.After enriched by lipid nutrients,R cells have more lipid droplets,and structural changes of mitochondria can be seen.The results indicate that ultrastructural changes in R cells of hepatopancreas could reflect the nutritional status of the larvae.
2001, 22(1): 69-73.
The zygote doesnt begin to cleave until the time 52 hours after it is spawned.The pattern of cleavage is superficial cleavage.The blastula is formed when the total numbers of the blastomeres add up to 256.In the late stage of the blastula,the presumptive endoderm cells and the other cells invaginate in the yolk and form gastrulation.Mandible rudiment and the antenna rudiment form in the bridge like cells group.The stomodeum forms in the middle of the ventral plate.With the forming of the antennula,the egg nauplius stage is identified.
2001, 22(1): 74-77.
The present paper studies the intrinsic and peripheral connecting of rostrolateral area of the anterior dorsal ventricular ridge (ADVR)in the lizard,Gekko gecko by means of anterograde and retrograde transport of HRP.The results show:1) rostrolateral area of the ADVR has a circuit pathway between its central core and superficial cell plate;2)the periventricular zone of ADVR has a widespread connections between its rostrolateral and caudal area;3)the circuit pathway between the central core of rostrolateral area of ADVR and pallial thickening is the connection hub between the two visual pathways.
2001, 22(1): 77-82.
The cell nucleus as the controller of all genetic and physiological activities within the cell,is the most prominent marker of eukaryotic cells.The formation of the cell nucleus is the key event during the origin of eukaryotic cells.The first appearance of the primitive cell nucleus indicates the emergence of the first eukaryotic cell.The research on the origin of the cell nucleus not only enriches the modern cell biology and evolutionary biology,but might also even influence the further development of the molecular cell biology by stimulating cell biologists to consider the significance of the viewpoint of evolution to cell researches.However,for a long time,on the origin of the cell nucleus there were only several reckless and assumptions and a few earnest efforts which were unsuccessful or only indirectly related to the formation of the nucleus.One of the main reasons for this situation seems to be the lack of a practical way for the study.Through searching for a long time we found a way.The essential point of the way is to consider the comprehension of the primitive nucleus as the key link for understanding the whole process of and early evolution of the cell nucleus.We have already known that the prokaryotic ancestor of eukaryotes must be one kind of ancient archaea (Li,1999).The primitive cell nucleus should be an intermediate link between the archaeal nucleoid and the typical cell nucleus.In order to obtain some features of the original nucleus,We d better study the most primitive protists we can find today and investigate all aspects of their cell nucleus thoroughly.Then,combined with the present knowledge on archaea we would be able to propose a hypothetical model for the primitive cell nucleus,and arrange various possible experiments to examine it from various aspects in order to test,to modify,to improve it,or replace it with a new one.Along this way,we would finally obtain a convincing model of the primitive cell nucleus.Then,combined the model with the present knowledge on archaea,protists and eukaryotic cells we could establish an all-sided hypothesis on the origin and early evolution of the cell nucleus for further examinations.Further along this way we would closer and closer approach the real evolutionary process.This is a realistic way.Along this way we have already negated the dinoflagellate nucleus model and the related hypothesis and established the diplomonad nucleus model for the primitive cell nucleus and proposed a rather complete theory on the origin and early revolution of the cell nucleus,including the origins of nuclear envelope,eukaryotic chromosomes and nucleolus (Li,1999).
Intermediate-Type Filament-LaminaNuclear Matrix of Vegetative Cell and Resting Cyst in Pseudourostyla cristata
2001, 22(1): 83-84.