桉树浸提液水体染色行为研究

为探求桉树水体染色行为的机制与原理,于广西高峰林场界牌分场采集尾叶桉桉树叶、桉树树干皮、桉树枝皮、桉树枯落物进行不同条件处理,即不同p H值、响应时间和不同金属阳离子,观察对比桉树各部位浸提物水体吸光度的变化情况,利用双因素方差分析法获得导致“黑水”现象的主要因素。结果表明,p H值和离子类型因素显著影响桉树各个器官浸提物水体吸光度,其中碱性条件下吸光度更高。Fe^(2+)条件下桉树各个器官浸提物水体吸光度最大,其次为Fe^(3+)条件,Na^(+)、Mg^(2+)、Ca^(2+)条件下桉树各个器官浸提物水体吸光度相对较低。说明碱性土壤及土壤铁离子高的环境下更容易出现桉树水体染色现象,即“黑水”现象。

Doubling Efficiency of Maize Haploids Treated with Different Methods

Haploid seedlings were inducted from different maize materials.At 2-3-leaf stage,maize haploids were treated with 0.06% colchicine and 2.0% dimethyl sulfoxide（DMSO） by dipping root,dripping heart leaf and acupuncturing growing point,respectively.The doubling rate and mortality rate in different treatments were analyzed by variance analysis and multiple comparisons.The result showed that growing point acupuncturing method exhibited the highest doubling efficiency with an average doubling rate of 23%,seed-setting rate of 21.4%,and mortality rate of 16.3%.Composed with other two chemical doubling methods,growing point acupuncturing method significantly improved the doubling rate of maize haploids with a lower application dose of colchicine.This study laid the foundation for industrial application of haploid breeding techniques.

Morphological Difference and Karyotype of Pelteobagrus fulvidraco in Dongting Lake Water System

[Objective] The research aimed to study the morphological characteristics and karyotype of Pelteobagrus fulvidraco in Dongting Lake water system.[Method] By using the conventional biological morphometry,PHA and colchicine injection method in vivo,the morphological characteristics and karyotype of P.fulvidraco in Yuanshui River and Lishui River of Dongting Lake were analyzed.[Result] In three ratio traits including standard length/head length,standard length/caudal peduncle depth,head length/snout length,P.fulvidraco of Yuanshui River and Lishui River had significant differences（P0.05）.However,the number and karyotype of their chromosomes were same.The chromosome number was 2n = 52,and the karyotype formula was 20M＋12SM＋10ST＋10T.The number of chromosome arm was 84.[Conclusion] The research result had certain theoretical guidance significance for the protection and utilization of wild P.resource in Dongting Lake water system.

Study on the Polyploid Induction of Lycium ruthenicum Murr.

With apical leaves of Lycium ruthenicum Murr as experimental material, the polyploids of L. ruthenicum were induced with colchicine solution, and total four polyploidy plants were identified by chromosome courts. The results showed that tetraploid plants of L. ruthenicum were successfully induced with 300 mg/L of colchicine solution after 7 and 21 d of induction, with 400 mg/L of colchicine solution after 14 days of induction, and with 500 mg/L of colchicine solution after 7 days of induction, respectively.

QTLs Mapping for Salinity Tolerance at Seedling Stage or Rice(Oryza sativa L.) Using Chromosome Segment Substitution Lines

In this study, a population of chromosome segment substitution lines （CSSLs） derived from the cross between 9311 （indica） and Nipponbare （japonica） was employed to map the quantitative trait loci （QTLs） for salt tolerance under the salt stress simulated with 0.5% NaCI, using survival rate as the index. The data were analyzed by QTL IciMapping v3.1, and the results showed that one QTL （QSsr3） related to salt tolerance was located in the vicinity of the marker RM1350 on chromosome 3, into a genetic interval of 113.2-132.8 cM, with a contribution rate of 17.75%. The additive effect was 10.9, indicating that the QTL derived from the parent Nipponbare improved the salt tolerance of rice at seedling stage. This study will provide a theoretical basis for the selection of salt tolerant rice germplasm.

Mapping QTLs Associated with Sheath Blight Resistance Using Chromosome Segment Substitution Lines of Rice(Oryza sativa L.)

In this study, a population of 119 chromosome segment substitution lines （CSSLs） derived from backcross between indica 9311 and japonica Nipponbare was employed to map quantitative trait loci （QTL） associated with sheath blight resis-tance in rice with toothpick inoculation method. A total of three sheath blight resis-tance-associated QTLs （qsb8-1, qsb8-2 and qsb8-3） were identified, which were lo-cated on adjacent molecular markers RM3262, RM5485 and RM3496 of chromo-some 8; the genetic interval was 81.7cM-91.7cM, 91.7cM-108.1cM and 108.1cM-119.6cM, respectively. The additive effect of qsb8-2 was negative, indicating that sheath blight resistance of susceptible parent harboring qsb8-2 fragment was en-hanced; additive effects of qsb8-1 and qsb8-3 were positive, indicating that sheath blight resistance of susceptible parent harboring qsb8-1 and qsb8-3 fragments was reduced.

Observation of Mitosis and Meiosis in Rice Cells by Simple Squash Method

[Objective] The aim of this study was to establish a feasible squashing technique for chromosome and obtain data of rice chromosome. [Method] With the materials of rice root tips and anther, the specimen was prepared by the modified squash method, and microscopic observation of mitosis and meiosis in rice cells was also carried out. [ Result] Mitosis in rice cells included interphase, prophase, metaphase, anaphase and telophase. Chromosome in metaphase shortened to the minmum, which was a good time for observing and investigating chromosome. However, meiosis in rice cells included meiosis Ⅰ and meiosis Ⅱ. Chromosome replication appeared in meiosis Ⅰ, while cell division only appeared in meiosis Ⅱ. [ Conclusion] The modified squashing technique for rice chromosome can obtain accurate data of rice chromosome, which provides evidence for genetic breeding.

Characterization of a S-locus-related Receptor-like Kinase Cluster in Rice Chromosome 4

We have identified 14 S _locus glycoprotein (SLG)_related protein kinase genes in a 323 kb contig of rice (Oryza sativa L.) chromosome 4 and we detected the transcription pattern of this gene cluster by reverse transcription_polymerase reaction (RT_PCR). RT_PCR results revealed that nine putative genes were transcribed in rice and these genes had the different expression patterns: two genes are expressed predominantly in reproductive tissues while the other seven genes are expressed in both reproductive and vegetative tissues. Analysis of the predicted amino acid sequences demonstrated that the extracellular receptor domains are highly homologous to SLG of Brassica, whereas the cytoplasmic kinase domains contain conserved amino acids present in serine/threonine kinases.

Construction of a DNA library from chromosome 4 of rice(Oryza sativa)by microdissection

A simple method to create a chromosome-specific DNA library of rice, including microdissection, amplification,characterization and cloning, is described. Rice chromosome 4 from a metaphase cell has been isolated and amplified by the Linker Adapter PCR (LA-PCR). The PCR products were labeled as probes with DIG-11-dUTP using the random priming method.Southern blot analysis with rice genomic DNA and specific RFLP markers demonstrated that the PCR products were derived from rice chromosome 4. A large library comprising over 100,000recombinant plasmid microclones from rice chromosome 4was constructed. Colony hybridization showed that 58% of the clones contained single or low-copy sequences and 42%contained repetitive sequences. The size of inserts generated by PCR ranged from 140bp to 500bp.This method will facilitate cloning of the specific chromosome DNA markers and important genes of rice.

Genetic algorithms and aquifer parameter identification

In order to identify aquifer parameter,authors develops an improved combinatorial method called best chromosome clone plus younger generation chromosome prepotency genetic algorithm (BCC-YGCP-GA), based on a decimal system simple genetic algorithm (SGA). The paper takes unsteady state flows in a two dimensional, inhomogeneous, confined aquifer for a ideal model, and utilizes SGA and BCC-YGCP-GA coupled to finite element method for identifying aquifer hydraulic conductivity K 1 ,K 2 ,K 3 and storage S 1 ,S 2 ,S 3 , respectively. It is shown from the result that GSA does not reach convergence with 100 generations, whereas convergence rate of BCC-YGCD-GA is very fast. Objective function value calculated by BCC-YGCD-GA is 0 001 29 with 100 generations, and hydraulic conductivity and storage of three zones are almost equal to the "true" values of ideal model.

Intestinal alkaline phosphatase in the colonic mucosa of children with inflammatory bowel disease

AIM： To investigate intestinal alkaline phosphatase （iAP） in the intestinal mucosa of children with inflammatory bowel disease （IBD）. METHODS： Colonic biopsy samples were taken from 15 newly diagnosed IBD patients and from 10 healthy controls. In IBD patients, specimens were obtainedboth from inflamed and non-inflamed areas. The lAP mRNA and protein expression was determined by reverse transcription-polymerase chain reaction and Western blotting analysis, respectively. Tissue localiza- tion of lAP and Toll-like receptor （TLR） 4 was investi- gated by immunofluorescent staining. RESULTS： The lAP protein level in the inflamed muco- sa of children with Crohn＇s disease （CD） and ulcerative colitis （UC） was significantly decreased when compared with controls （both P 〈 0.05）. Similarly, we found a significantly decreased level of lAP protein in the in- flamed mucosa in CD compared with non-inflamed mucosa in CD （P 〈 0.05）. In addition, the iAP protein level in inflamed colonic mucosa in patients with UC was decreased compared with non-inflamed mucosa in patients with CD （P 〈 0.05）. lAP protein levels in the non-inflamed mucosa of patients with CD were similar to controls, lAP mRNA expression in inflamed colonic mucosa of children with CD and UC was not significant- ly different from that in non-inflamed colonic mucosa with CD. Expression of lAP mRNA in patients with non- inflamed mucosa and in controls were similar. Co-local- ization of lAP with TLR4 showed intense staining with a dotted-like pattern, lAP was present in the inflamed and non-inflamed mucosa of patients with CD, UC, and in control biopsy specimens, irrespective of whether it was present in the terminal ileum or in the colon. However, the fluorescent signal of TLR4 was more pro- nounced in the colon compared with the terminal ileum in all groups studied. CONCLUSION： Lower than normal lAP protein levels in inflamed mucosa of IBD patients may indicate a role for lAP in inflammatory lesions in IBD. Based on our results, administration of exogenous lAP enzyme to pa- tients with the active form of IBD may be a therapeutic option.

Cytogenetic Analysis of Three Species of Astyanax Genus （Pisces, Characidae, Incertae sedis） from Freshwaters of Upper Paraguay Basin, Mato Grosso State, Brazil

Cytogenetic studies have shown that the genus Astyanax has extensive karyotypic variability among species and many are the works developed about this group in different river basins in Brazil. The Tangara da Serra area （Mato Grosso State, Brazil）, which contains tributaries of both Amazon and Upper Paraguay River basins is, therefore, a natural watershed. Thus, in this work, the karyotypes of three species of Astyanax from the Sepotuba River＇s tributary streams were analyzed by using conventional methods of chromosome analysis. Astyanax marionae presented 2n =48（12m＋ 10sm＋ 12st＋ 14a） and NF （fundamental number） = 82 with evident heterochromatin in several chromosomal regions. Astyanax cf. asuncionensis presented 2 n = 50 （20m ＋ 12sm ＋ 6st ＋ 12a）, NF = 94 chromosomes and with evident heterochromatin in few chromosomes and Ag-NOR in one chromosome pair. Astyanax cp. scabripinnis presented 2 n = 50 （12m ＋ 10sm ＋ 10st ＋ 18a）, NF = 82 with evident heterochromatin at most of the chromosomes in pericentromeric region and a strongly marked block on a telocentric chromosome pair. The Ag-NORs were observed near the heterochromatic region. The karyotypic differences found, probably occurred by chromosomal rearrangements, resulting in a heterogeneous model of chromosomal evolution who probably involved both Robertsonian chromosomal rearrangements to A. marionae, as not Robertsonian to other Astyanax species. This indicates that although these species share the same hydrographic basin （Upper Paraguay） are isolated in different microbasins and the absence in the gene flow may have allowed the establishment of independent rearrangements in each population.

Polyploidy Induction of Pteroceltis tatarinowii Maxim.

[Objective] This study was conducted to obtain tetraploid Pteroceltis tatar# nowii Maxim. with excellent ornamental traits. [Method] The stem apex growing points of Pteroceltis tatarinowii Maxim. were treated with different concentrations of colchicine solution for different hours to figure out a proper method and obtain poly- ploids. [Result] The most effective induction was obtained by treatment with 0.6%- 0.8% colchicine for 72 h with 34.2% mutation rate. Flow cytometry and chromosome observation of the stem apex growing point of P. tatarinowii Maxim. proved that the tetraploid plants were successfully obtained with chromosome number 2n=4x=36. [Conclusion] The result not only fills the blank of polyploid breeding of P. tatarinowii, but also provides an effective way to broaden the methods of cultivation of fast- growing, high-quality, disease-resilience, new varieties of Pteroceltis.

DECOMPOSITION OF BLACK LIQUOR BY ULTRASOUND PROCESS

Ultrasound (US)-induced cavitation is an effective way in oxidizing organic contaminants in wastewater either as the independent operation unit or in combination with other oxidation methods. In this paper, black liquor and filtrate after acidifying and settling were sonicated. The effect of working parameters on ultrasonic degradation of black liquor, such as different combination methods, frequency, power supply, initial concentration, pH, duration time, ionic strength and catalyst were studied. The results were as follows: (1) At the conditions of 40kHz, 100W, 4h, pH at 6 and temperature 30?℃, utilizing US/US-H2O2/US-Fenton, weak-orange filtrate turned to colloid with the increase of time and little sediment produced after settling. Especially filtrate came to be milk white collides and upper water approached to nearly achromatic by US-Fenton. Compared with the US, US-H2O2/US-Fenton COD (Chemical oxidation demand) removal ratio can increase 15%, 30% respectively. Because of more hydroxyl radicals produced in the reaction process; (2) At the condition of 100W and 4h, the degradation efficiency of black liquor was better at 40kHz over at 20kHz. Moreover black liquor can be biodegraded easily. Those based on that the big molecule of contaminants in aqueous solution can be changed into the little molecule with ultrasound (3) At the condition of 40kHz and 4h, the COD removal ratio of black liquor was more higher at 60W than at 80W, while the removal ratio of COD at 60W was nearly close to the ratio at 100W; (4) The initial concentration of black liquor influenced the effect of sonochemical degradation; (5) The variation of pH had no effect on degradation; (6) The longer the duration time, the greater the removal ratio of COD. But this ratio increased slowly after 4h; (7) Adding 0.2g/L NaCl to change the ionic strength of the black liquor, the COD removal ratio can increase 10%; (8) The degradation rates increased by the coexistent catalysts of TiO2, Co2+ and Ag+.

Molecular mapping for seedling cold tolerance QTLs in rice

The quantitative trait loci (QTLs) for cold tolerance relative characters were identified with microsatellite markers.Ten QTLs located on chromosome 1, 3, 4, 5, 6, 8, 9, 11(two)and 12 were detected for seedling height at different low temperature. Only 2 of these were detected at the same locus at four environments, 1 was significant at three environments, 6 were significant at two environments and 1 was significant at one environment. Seven QTLs located on chromosome 1(two), 2(two),5,6,8 were found for low temperature chlorosis resistance and five QTLs located on chromosome 3, 4, 7, 8, 11 resistant to chilling injury. The amount of variation explained by individual QTL ranged from 4.85%to 49.34%. There was no linkage relationship among the three characters, which indicates seedling cold tolerance is a complex character and is controlled by different QTLs.

Study on the extra X chromosome＇s effect on the hormone level of Klinefelter＇s syndrome

Klinefelter＇s syndrome is an inherited （genetic） disorder found only in men caused by at least one extra X chromosome in a cell. Does the extra X chromosome have any effect on the hormone level of Klinefelter＇s Syndrome？ In this paper, 25 subjects with Klinefelter＇s syndrome, 30 infertile subjects and 36 normal men without Klinefelter＇s syndrome were compared each other in endocrinology profile and cytogenetics. Subjects with Klinefelter＇s syndrome were identified by the karyotypes 47, XXY or 47, XXY/46XY, and positive of the X-chromatins （Barr bodies）. Hormone analysis of subjects with Klinefelter＇s syndrome showed that the testosterone （T） values were lower than those of the normal subjects, while the FSH and LH values were higher than those of the normal people; in the infertile experiment subjects without Klinefelter＇s Syndrome, the karyotypes are 46, XY, with negative of the X-chromatins. The testosterone （T） values of these subjects were also lower than those of the normal people, but the FSH and LH values were within the normal range. These results indicated that endocrinological test on infertile subjects can be used to determine whether a cytogenetic analysis is necessary, and hence exclude non- Klinefelter＇s syndrome. The mechanism of the occurrence of this difference, its clinical applications and the relationship among the karyotypes, the endocrinological test and the severity of the phenotype are discussed. Lyon＇s hypothesis stating that only one of the two X-chromosomes is genetically active in female cells, but our study concluded that the extra X chromosome do have effect on the hormone level of Klinefelter＇s Syndrome.

Development and characterization of synthetic amphiploids of Oryza sativa and Oryza latifolia

Oryza sativa and Oryza latifolia belong to the AA and CCDD genomes of Oryza, respectively. In this study, amphiploids were obtained from the tube seedlings of O. sativa × O. latifolia F1 hybrids by treatment with colchicine, an agent for chromosome doubling. Subse- quently, amphiploids were investigated using the methods of morphology, genomic in situ hybridization, and molec- ular markers. Amphiploids were characterized by a shorter plant height, larger diameter of stem, longer and wider leaves, darker leaf color, decreased spikelets per panicle and panicle length, and larger spikelets and anthers than the original F1 hybrid. Based on the mitotic metaphase chro- mosome number of the investigated root tips, the somatic chromosome number of the amphiploid is 2n = 72. Additionally, the amphiploid is an allohexaploid, and its genomic constitution is AACCDD by genomic in situ hybridization analysis. Finally, the amphiploids were identified to be true using 37 polymorphic markers at the DNA level.

Genotoxicity biomarkers in aquatic bioindicators

Pollution of the aquatic environment is an ever-growing problem, as waters are the ultimate sink for the large number of xenobiotics from multiple sources. DNA damaging agents have a significant ecological relevance since they are implicated in many pathological processes and exert effects beyond that of individual being active through following generations. A large number of methods have been applied to evaluate genotoxic damage in different aquatic species. Comet assay, as method for de- tecting DNA alterations, and micronucleus test, as an index of chromosomal damage are the most widely applied and validated methods in field studies. These methods were applied in different vertebrate and invertebrate aquatic species, but only mollusk and fish species have been employed in routine biomonitoring programs. Mussels, due to their widely geographical distribution and the suitability for caging represent the bioindicator of choice in field studies. Mytilus species is the most used marine mussel. The use of fish is limited to specific geographic areas. The present review mainly focuses on the application of comet assay and micronucleus test in mussels. A number of biomonitoring studies in mussels, using comet assay or micronucleus test, revealed exposure to different classes of genotoxic compounds with a good discrimination power. The different evidence from the two as- says, reflects different biological mechanisms for the two genetic endpoints, DNA damage and chromosomal damage, suggesting their combined application in the field. Different endogenous and exogenous factors have been shown to modulate the genotoxic responses in mussels, acting as confounding factors in environmental monitoring. The use of standardized protocol for caging, sampling and genotoxity evaluation is critical in biomonitoring studies. The use of a multimarker approach coupling genotoxicity biomarkers with physiological and biochemical factors allows to have a complete picture of the environmental pollution [Current Zoology 60 （2）： 273-284, 2014].

Review:Whole genome amplification in preimplantation genetic diagnosis

Preimplantation genetic diagnosis (PGD) refers to a procedure for genetically analyzing embryos prior to implantation,improving the chance of conception for patients at high risk of transmitting specific inherited disorders.This method has been widely used for a large number of genetic disorders since the first successful application in the early 1990s.Polymerase chain reaction (PCR) and fluorescent in situ hybridization (FISH) are the two main methods in PGD,but there are some inevitable shortcomings limiting the scope of genetic diagnosis.Fortunately,different whole genome amplification (WGA) techniques have been developed to overcome these problems.Sufficient DNA can be amplified and multiple tasks which need abundant DNA can be performed.Moreover,WGA products can be analyzed as a template for multi-loci and multi-gene during the subsequent DNA analysis.In this review,we will focus on the currently available WGA techniques and their applications,as well as the new technical trends from WGA products.