Cytogenetic characterization of Agrobacterium rhizogenes transformed root lines of Rauvolfia serpentina
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Agrobacterium rhizogenes contains Ri plasmid,the rolBgenes is on the T-DNA of Ri plasmid,rolB genes can form a lot of hairy root through the wound infection of Agrobacterium rhizogenes.By the transformation of hairy root can get regenerate plants which show many stabilizes in the phenotypic variation of the genetic,so it has a broad application prospect in plant genetic improvement.The biological characteristics of Agrobacterium rhizogenes and the structure and function of Ri plasmid were summarized,as well as rolBgene loci and characteristics,Agrobacterium rhizogenes transformation mechanism and the methods of the hairy root induction,the influence factors of inducing hairy root,identifying and hairy plantlet regeneration after the eradication of bacteria and so on.In addition,the application in plant sciences was reviewed.
Plantlet
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In order to increase the efficiency of the Magiscan metaphase location and karyotyping system, its software and mode of operation have been changed. In the new multiple‐cell karyotyping method, interactions by the operator are only required for relocation and counting of metaphases, but not for karyotyping. Metaphases are located and their coordinates recorded automatically as before. The first metaphase in the list is relocated, displayed on the screen, and counted by the operator. It is then karyotyped automatically while the operator relocates and counts the next metaphase in the list. This procedure continues until an appropriate number of metaphases have been counted and karyotyped. Finally a composite karyotype is printed out. Each karyotype is represented by a column of 23 chromosome pairs (1–22 and XX or XY) and all columns are lined up next to each other. Most chromosomes are correctly classified into the composite karyotype. Minor structural abnormalities are detected by comparing pairs of homologues. Overlapped, close touching, and grossly abnormal chromosomes are often misclassified or rejected and shown beneath the classified chromosomes. A trained cytotechnician can easily detect even small chromosome abnormalities on the composite karyotype. A clinical feasibility study indicates that the procedure can be used for routine cytogenetic analysis.
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Chromosome number
Polyploid
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Akin to the situation in humans, dogs are frequently affected by tumors of the prostate. The malignancies share many similarities between both species, for example, median age at the onset of the disease and metastatic behavior. In human prostatic tumor samples, investigations of prepared metaphase spreads showed a variety of chromosomal aberrations, with trisomies of chromosomes 7, 8, and 17 as the leading cytogenetic abnormalities. In this article we present one case of a canine adenocarcinoma of the prostate, including clinical examination and establishment of a cell line from a tumor sample obtained from the affected 10-year-old male Briard. Searching for similarities between both species in respect to chromosomal changes within the tumor samples, we investigated prepared metaphases of the canine cell line cytogenetically. These investigations presented a highly rearranged karyotype showing a large biarmed marker consisting of material from chromosomes 1 and 2 in addition to centromeric fusions between dog chromosomes 1 and 5 that both could be identified in every metaphase investigated, while centric fusions of chromosomes 4 and 5 occurred in up to 50% of the metaphases. The cell line grew very well and showed evidence of being spontaneously immortalized when it crossed the 20th passage.
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Taking the diploid and autotetraploid mini-watermelon as meterial,the diploid and autotetraploid miniwatermelons' karyotype were analyzed using traditional tabletting method.The results showed that the diploid miniwatermelon karyotype formula was 2n=2x=22=22mwith the index of the As.k(karyotypic asymmetry)of 58.90%,with 11pairs metacentric chromosomes.The karyotype belonged to 1A,with ultimately symmetry karyotype.In addition,the tetraploid mini-watermelon karyotype formula was 2n=4x=44=44m,As.k was 56.45%,same as diploid.The results proved that tetraploid mini-watermelon resulted in the chromosome doubling of diploid.
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Giemsa C-banding patterns and karyotype of chromosomes were analyzed in the root-tip cells of diploid Thinopyron elongatum. A modified seed germinating method was developed for obtaining the C-banding patterns in diploid Thinopyron elonggatum. The results of The C-banding analysis showed the significant differences among the seven pairs of chromosomes in diploid Thinopyron elonggatum. The intensive C-bands were stained steadily on the intercalary, terminal, subcentromeric and centromeric regions of Chromosomes. We found that there are three pairs of metacentric chromosomes and four pairs of submetacentric chromosomes in diploid Thinopyron elonggatum. The karyotype formula is 2n=2x=14=6m+8sm. Present results provided a basic cytological data and will be useful for the further studies in Thinopyron elongatum.
Giemsa stain
G banding
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A formula is proposed for estimating statistically the degree of somatic chromosome association during metaphase. Standard chi-square tests can then be used to determine the presence or absence of somatic association between homologous chromosomes in selected species. The method requires that the chromosomes be morphologically distinguishable and that their circular arrangement on the metaphase plate has not been distorted by the squash used during their preparation.
Association (psychology)
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OBJECTIVE To induce hairy roots of Gynostemm apentaphyllum by Agrobacterium rhizogenes strains. METHODS Hairy roots were induced by the co-culture method of explants and Agrobacterium rhizogenes strains. Effects of different Agrobacterium rhizogenes strains, explants, pre(co)-culture time, Bacterial concentration, infecting time, As concentration and antibiotic medium on the transformation frequency were studied. RESULTS The highest induction frequency was obtained form leaf 2 days co-cultivation, which were induced by Agrobacterium rhizogenes OD 600 0. 8 for 10 min, 100 micromol/L As and MS + 300 mg/L Cab. CONCLUSION Hairy roots were induced by co-cultivation and the optimum induced condition were determined.
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Objective To establish the genetic transformation system of Phytolacca americana L. by Agrobacterium rhizogenes mediated and analysis of hairy root strains biomass for the base of Phytolacca americana L. research using genetic engineering technology. Methods Activated Agrobacterium rhizogenes R1601,transforming leaves and stems of Phytolacca americana L. by using different concentrations of Agrobacterium rhizogenes R1601; Hairy roots identified by PCR; randomly selected hairy roots to culture,identified the biomass differences of different strains. Results Phytolacca americana L. is sensitive by Agrobacterium rhizogenes R1601 infected; Four concentrations can induce hairy roots of Phytolacca americana L. highly efficient,the genetic transformation efficiency of stems and leaves can reach 100 % if A600 is 1. 0; PCR identification showed the T-DNA sequences had integrated into the hairy roots of Phytolacca americana L.; The biomass analysis showed the biomass of different hairy root strains was significantly different. Conclusion The genetic transformation system highly efficient of Phytolacca americana L. by Agrobacterium rhizogenes has been established,the biomass of different hairy root strains was significantly different.
Hairy Root Culture
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