scholarly journals Sex Determination During Inflorescence Bud Differentiation in Monoecious Pistacia chinensis Bunge

Forests ◽  
2019 ◽  
Vol 10 (3) ◽  
pp. 202 ◽  
Author(s):  
Qian Bai ◽  
Chenyi Zhu ◽  
Xia Lei ◽  
Tao Cao ◽  
Shuchai Su ◽  
...  
Keyword(s):  
Sex Determination ◽  
Early Stage ◽  
Sex Differentiation ◽  
Banding Patterns ◽  
Vegetative Period ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Sex Determination Mechanisms

Pistacia chinensis Bunge is widely acknowledged to be dioecious, but rare monoecious individuals have been found. However, the origin of monoecism and the sex differentiation of different sex types remain intriguing questions. Here, sex expressions were explored by identification of sex-associated DNA markers, determination of the sex stability after grafting, and histological characterization of inflorescence bud development using anatomical analysis. The results showed that (1) although polymorphisms among individuals existed, the banding patterns of Polymerase Chain Reaction (PCR) products for different sex types on the same monoecious tree were consistent; (2) the sex expressions of grafted trees were not consistent with those of scions, indicating that monoecism probably did not originate from a stable bud mutation; and (3) both males and females underwent a bisexual period, then the stamen primordia in female buds degenerated into the second round tepals, while the pistil primordia in male buds gradually disappeared. During the sex differentiation phase, female buds were spindle-shaped, while the male buds were full teardrop-shaped, and male buds were bigger than female buds. Taken together, no sex-associated DNA marker was found, sex expressions were unstable after grafting, and the alternative sex organs appeared in the early stage of sex differentiation, suggesting that sex determination occurred during floral development instead of the early vegetative period. These results indicated that the sex expressions may be affected by environmental factors, increasing the understanding of sex determination mechanisms in P. chinensis and other species.

scholarly journals Sex determination in gibbons of genus Nomascus using non-invasive method

2016 ◽  
Vol 85 (4) ◽  
pp. 363-366 ◽  
Author(s):  
Petra Bolechová ◽  
Kateřina Ječmínková ◽  
Michal Hradec ◽  
Tomáš Kott ◽  
Jana Doležalová
Keyword(s):  
Sex Determination ◽  
Direct Examination ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Non Invasive ◽  
Faecal Samples ◽  
Invasive Method ◽  
Polymerase Chain ◽  
Template Dna

Gibbons of the genus Nomascus have a strong sexual dimorphism and dichromatism. As they mature, both sexes develop sex-specific pelage colour. In combination with physical similarities in the genitalia with both sexes, there are problems with determining the sex of young individuals compared to other genus of gibbons. This is a pilot study applying a multiplex polymerase chain reactions based on a non-invasive method for sex determination of gibbons. The study was conducted on 22 faecal samples from gibbons of the genus Nomascus. The animals were monitored by staff so that the samples were identified correctly and each sample was collected immediately after the defecation. Results confirmed the sex in all adult and juvenile animals with known sex; and 2 females and 5 males in juveniles were determined with unknown sex. The results of direct examination completely corresponded with the PCR results. The PCR reaction with template DNA isolated from faecal material required BSA usage, however, we observed the occurrence of nonspecific fragments. This did not affect the reliability of our results and we confirmed the usability of this method for this genus.

scholarly journals First Isolation and Molecular Typing of Pathogenic and Intermediate Leptospira Species from Urine of Symptomatic Dogs

2021 ◽  
Vol 8 (12) ◽  
pp. 304
Author(s):  
Ivana Piredda ◽  
Loris Bertoldi ◽  
Giuseppe Benvenuto ◽  
Bruna Palmas ◽  
Aureliana Pedditzi ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Zoonotic Risk ◽  
Leptospira Spp ◽  
Blood And Urine Samples ◽  
Healthy Dogs ◽  
Polymerase Chain ◽  
Sequence Types ◽  
Leptospira Species

Aim of this study was to evaluate, the presence and diversity of Leptospira spp. in blood and urine samples collected from 175 owned-dogs from Sardinia, Italy. After determination of leptospiral infection by microscopic agglutination test (MAT), urine from MAT-positive dogs were examined by real-time polymerase chain reaction (lipL32 rt-PCR) and then isolated by culture. In order to characterize obtained serovars, positive cultures were then subjected to 16S rRNA and secY sequencing, phylogenetic analysis and Multilocus Sequence Typing (MLST). Results showed that seven dogs (4%; 95% CI: 0–55) had Leptospira DNAs in their urine and five strains were isolated from urine cultures. The three different sequence types (ST17, ST198 and ST24) belonging to Leptospira interrogans genomospecies identified by MLST analyses in this study, confirmed that the leptospiral infection was widespread in Sardinian dogs. We also reported the first characterization of a new Leptospira spp. isolated from urine of one dog living in the study area. Whole genome sequencing and phylogenetic analysis, confirmed that this genospecies was closely related to Leptospira hovindhougenii, an intermediate Leptospira spp. with unknown pathogenicity previously isolated from a rat in Denmark. Further studies are required to clarify whether healthy dogs that shed leptospires in their urine could represent a zoonotic risk for humans in this region.

scholarly journals Characterization of a New Alloantigen (SH) on the Human Neutrophil Fcγreceptor IIIb

Blood ◽  
1997 ◽  
Vol 89 (3) ◽  
pp. 1027-1034 ◽  
Author(s):  
Juergen Bux ◽  
Ernst-Ludwig Stein ◽  
Philippe Bierling ◽  
Patricia Fromont ◽  
Mary Clay ◽  
...  
Keyword(s):  
Nucleotide Sequence Analysis ◽  
Polymorphism Analysis ◽  
Mutational Event ◽  
Coding Region ◽  
Specific Pcr ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Antigen Frequency ◽  
Neonatal Neutropenia

Abstract Polymorphic structures of the neutrophil Fcγreceptor IIIb (FcγRIIIb) result in alloantibody formation that causes alloimmune neonatal neutropenia and transfusion reactions. Alloantigens located on FcγRIIIb include the antigens NA1 and NA2. In four cases of alloimmune neonatal neutropenia, granulocyte-specific alloantibodies directed against a thus far unknown antigen were detected by granulocyte agglutination and immunofluorescence tests in the maternal sera. By the use of the monoclonal antibody–specific immobilization of granulocyte antigens (MAIGA) assay, the new antigen, termed SH, was located on the FcγRIIIb. Nucleotide sequence analysis of the FcγRIIIb coding region from a SH(+) individual showed a single-base C→A mutation at position 266, which results in an Ala78Asp amino acid substitution. A family study confirmed that this nucleotide difference is inherited, and corresponds to the SH phenotype. Serologic typing of 309 randomly selected individuals showed an antigen frequency of 5% in the white population. The same frequency was found by genotyping, for which a technique based on polymerase chain reaction (PCR) using sequence-specific primers (PCR-SSP) was developed. Typing of all SH(+) individuals for NA1 and NA2, and PCR-restriction fragment length polymorphism analysis of the NA-specific PCR products from five SH(+) individuals using the SH-specific endonuclease SfaN I showed that SH antigen is very probably the result of an additional mutational event in the NA2 form of the FcγRIIIB gene. Immunochemical studies also demonstrated that the SH determinants reside on the 65- to 80-kD NA2 isoform of the FcγRIIIb. Our findings show the existence of an additional polymorphism of the FcγRIIIb, which can result in alloantibody formation causing alloimmune neonatal neutropenia.

Rapid sex determination of chick embryos using the polymerase chain reaction1

1995 ◽  
Vol 6 (2) ◽  
pp. 119-130 ◽  
Author(s):  
James N. Petitte ◽  
A. Elizabeth Kegelmeyer
Keyword(s):  
Sex Determination ◽  
Chick Embryos ◽  
Polymerase Chain

Using polymerase chain reaction for rapid sex-determination of a case of sirenomielia

1998 ◽  
Vol 40 (3) ◽  
pp. 282-285
Author(s):  
Yuko Jin ◽  
Akihiko Endo ◽  
Masami Shimada ◽  
Michiyoshi Minato ◽  
Masaaki Takada ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Sex Determination ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals PENENTUAN JENIS KELAMIN BURUNG KENARI (Serinus canaria) BERDASARKAN GEN Chromodomain Helicase DNA-Binding 1 (CHD1)

2020 ◽  
Vol 7 (1) ◽  
Author(s):  
Afif Muhammad Akhrom ◽  
Indarjulianto Soedarmanto ◽  
Yanuartono Yanuartono ◽  
Trini Susmiati ◽  
Alfarisa Nururrozi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Sex Determination ◽  
Mature Female ◽  
Mature Male ◽  
Chain Reaction ◽  
Blood Samples ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Female Canaries

Phenotype determination of sex in young canaries is very low in accuracy. This study aimed to develop a genotypic sexing method in canaries. This study used 12 canaries consisting of 3 mature males, 3 mature females and 6 one-month-old canaries. Phenotypic sexing by cloacal observation was done on all birds, continued by genotypic sexing to identification CHD1 gene using polymerase chain reaction (PCR). The PCR used blood samples for mature canaries, and feather for mature and one-month-old canaries. The results of phenotypic observations showed that all mature male canaries had prominent and pointed cloaca forms, all mature females had flat and wide, whereas all one-month-old birds had a flat cloaca. The result of PCR showed a single band (500 bp) for mature male and double bands (500 bp and 300 bp) for mature female canaries. The PCR results of one-month-old canaries showed that there were one male and five females. Based on this study, it was concluded that genotypic sexing using the PCR method is effective in the sex determination of canaries.Keywords: canary, CHD1, genotype, PCR, sexing ABSTRAKPenentuan jenis kelamin burung kenari muda secara fenotip akurasinya sangat rendah. Penelitian ini bertujuan untuk menentukan jenis kelamin burung kenari secara genotip. Penelitian ini menggunakan 12 ekor burung kenari, terdiri dari 6 ekor dewasa (3 jantan, 3 betina) serta 6 ekor umur 1 bulan. Semua burung ditentukan jenis kelaminnya dengan mengamati kloaka dan identifikasi gen CHD1 menggunakan teknik polymerase chain reaction (PCR). Sampel DNA berasal dari darah dan bulu untuk burung dewasa serta bulu untuk burung umur 1 bulan. Pengamatan fenotip menunjukkan bahwa burung kenari dewasa jantan mempunyai bentuk kloaka menonjol dan runcing, dewasa betina berbentuk datar dan lebar, sedangkan semua burung umur 1 bulan mempunya bentuk kloaka datar. Hasil identifikasi gen CHD1 diperoleh adanya 1 pita gen sekitar 500 bp dari sampel darah dan bulu semua burung kenari dewasa jantan, dan 2 pita gen sekitar 500 bp dan 300 bp dari sampel semua burung kenari betina dewasa. Hasil PCR pada sampel burung umur 1 bulan menunjukkan bahwa 1 ekor jantan dan 5 ekor betina. Berdasarkan penelitian ini dapat disimpulkan bahwa penentuan jenis kelamin secara genotip menggunakan gen CHD1 dapat dilakukan pada burung kenari.

scholarly journals SEX DETERMINATION OF BALI STARLING (Leucopsar rothschildi) USING MOLECULAR SEXING

2015 ◽  
Vol 2 (1) ◽  
pp. 114
Author(s):  
Putu Indra Pramana Wirastika ◽  
Ignatius Pramana Yuda ◽  
Felicia Zahida
Keyword(s):  
Dna Binding ◽  
Sex Determination ◽  
Gel Electrophoresis ◽  
Morphological Characters ◽  
Molecular Sexing ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Pcr Products ◽  
Conventional Methods

<p>Bali Starling (Leucopsar rothschildi) are monomorphic at the age of nestling. For the conservation of bird it important is to determine its sex at the earlier stage. Conventional methods have limitations. This study applied PCR-based molecular sexing to answer this issue. This study aimed to obtain the most effective molecular primers to identify the sex of Bali starling. The most common used combination of P2/P8, 2550F/ 2718R and 1237L/1272H primers, which amplify CHD1 gene (Chromo-helicase-DNA-binding) were evaluated. DNA samples were obtained from secondary wing feathers of young Bali Starling. Separation in agarose gel electrophoresis of PCR products showed that the three primers were successfully amplified the samples with different degrees of success, that was 90% (P2/P8), 86.7% (2550F/2718R), and 73.3% (1237L/1272H), respectively. However, only the combination of P2/P4 and 2550F/2718R primers was able to sex Bali Starling based on observation of PCR products on agarose gel. The sizes of the genes were slightly different with those reported on previous studies. Most of the results of molecular sexing were in accordance with the sex based on morphological characters.</p><p><br /><strong>Keywords</strong> : Bali starling, Leucopsar rothschildi, molecular sexing, CHD gene</p>

scholarly journals Characterization of clone-specific rearrangement T-cell receptor gamma- chain genes in lymphomas and leukemias by the polymerase chain reaction and DNA sequencing

Blood ◽  
1994 ◽  
Vol 84 (2) ◽  
pp. 574-581 ◽  
Author(s):  
M Kneba ◽  
I Bolz ◽  
B Linke ◽  
J Bertram ◽  
D Rothaupt ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Polymerase Chain Reaction ◽  
T Cell ◽  
Dna Sequencing ◽  
Cell Lines ◽  
Chain Reaction ◽  
Gamma Chain ◽  
Pcr Products ◽  
Polymerase Chain

The structures of rearranged gamma-chain T-cell antigen receptor (TCR) genes were analyzed in 5 cases of T-cell acute lymphoblastic leukemia (T-ALL), in 15 cases of peripheral T-cell non-Hodgkin's lymphoma (T- NHL), in 1 case with large granular CD8 lymphocytosis, 1 case with CD8 lymphocytosis after autologous bone marrow transplantation for Hodgkin's disease, and in 2 cases with nonneoplastic diseases. Rearranged V-J TCR gamma-gene segments were amplified by the polymerase chain reaction (PCR). Because most of the biopsy tissue or bone marrow samples contained significant amounts of admixed nonmalignant T-cells, direct DNA sequencing of the PCR products yielded mixed sequence data because of coamplification of clonal together with polyclonal TCR gamma V-N-J junctions. Reliable data could only be obtained by cloning the V gamma-J gamma PCR products and sequencing several (4 to 10) randomly chosen clones. In the polyclonal samples, all PCR-derived clones differed in their specific V-N-J junctions, as expected. In the two T- cell lines and in most of the T-cell malignancies, monoclonal PCR products could be identified by the demonstration of clonally restricted V-N-J junctions. In most cases, this information yielded the desired clone-specific sequence and showed a background population of polyclonal TCR gamma cells in each specimen, except for those that were obtained from the T-ALL samples, the cell lines, or the NHL samples with high tumor cell fraction. The results obtained by PCR-directed sequencing were confirmed by temperature-gradient gel electrophoresis (TGGE) that showed distinct DNA bands only with the PCR products containing predominant (ie, monoclonal) TCR gamma V-N-J junctions. By combined sequence and TGGE analysis, it was found that PCR/TGGE is able to distinguish between monoclonal and polyclonal TCR gamma-PCR products. This finding prompted us to complete the analysis of the TCR gamma locus in the samples by PCR/TGGE using primer mixes which covered all possible V gamma and J gamma recombinations. Monoclonality was shown with all mixes by PCR/TGGE in 21 of 24 (87%) of the lymphoproliferations. In summary, the present study shows that the combination of amplifying TCR gamma V-N-J junctions by PCR with the identification of clonal PCR products by TGGE and DNA sequencing is a reliable method for the characterization of clonal TCR gamma sequences.

scholarly journals Molecular analysis of glycophorin C deficiency in human erythrocytes

Blood ◽  
1993 ◽  
Vol 81 (10) ◽  
pp. 2799-2803
Author(s):  
R Winardi ◽  
M Reid ◽  
J Conboy ◽  
N Mohandas
Keyword(s):  
Mechanical Stability ◽  
Mutant Gene ◽  
Erythrocyte Shape ◽  
Normal Individuals ◽  
Pcr Products ◽  
Protein Isoform ◽  
Polymerase Chain ◽  
Exon 4 ◽  
Glycophorin C

Human erythrocyte glycophorin C plays a functionally important role in maintaining erythrocyte shape and regulating membrane mechanical stability. We report here the characterization of the glycophorins C and D deficiency in erythrocytes of the Leach phenotype. Glycophorin C gene is encoded by 4 exons. Amplification of reticulocyte cDNA from Leach phenotype and normal individuals generated a 140-bp fragment when using primers spanning exons 1 and 2. However, no polymerase chain reaction (PCR) products were detected in the Leach phenotype using primers flanking either exons 1 and 3 or exons 1 and 4, suggesting that the 3' end of the mRNA was missing or altered. Exon 4 also appeared to be missing from Leach genomic DNA, based on both Southern hybridization and PCR. These results indicate that an absence of glycophorin C and glycophorin D in erythrocytes from these Leach phenotype individuals is a consequence of a deletion or marked alteration of exon 3 and exon 4 of their glycophorin C gene. Surprisingly, the mutant gene encodes an mRNA stable enough to be detected in circulating reticulocytes. Although this mRNA could encode an N-terminal fragment of glycophorin C, these protein isoform(s) would not be expressed in the membrane because they lack the transmembrane and cytoplasmic domains.

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