In order to screen microsatellites for conservation genetics studies of the species, a total of 23 microsatellite loci from Korean goral (Naemorhedus caudatus), including 15 previously developed loci and 8 new loci in this study, were tested. Eleven microsatellites were screened and subjected to cross-species amplification using a test panel of four Caprinae species, Japanese serows (Capricornis crispus), Chinese gorals (Naemorhedus goral), Northern chamois (Rupicapra rupicapra) and domestic goats (Capra hircus). In addition, all eleven microsatellites (SY3A, SY12A, SY12B, SY48, SY58, SY71, SY76, SY84, SY84B, SY112, and SY129) satisfied the criteria to be a core set of microsatellites. This core set of microsatellites and cross-species amplification of Korean goral microsatellites were found to be helpful for high-resolution studies for conservation and management of Korean goral and other endangered Caprinae species.
Animals ; Conservation of Natural Resources ; Genetic Variation ; Hybridization, Genetic/*genetics ; Microsatellite Repeats/*genetics ; Republic of Korea ; Ruminants/*genetics

Carrier State ; Gene Expression Profiling ; Hepatitis B ; genetics ; Humans ; Hybridization, Genetic ; Nucleic Acid Hybridization ; methods

We obtained intergenus somatic hybrid between Alhagi pseudalhagi and Astragalus cicer by using electroporation. Agrobacterium rhizogenes A4-transformed A. pseudalhagi protoplasts were treated with iodoacetamide so that they were unable to sustain divisions. A. cicer protoplasts were isolated from a methionine-resistant mutant and did not survive in the medium without plant growth regulator. The intergenus somatic hybrid cells were selected based on physiological complementation between the two parents. We optimized some parameters of electroporation, such as direct current impulse, alternating current impulse and the impulse number. We identified ten hybrid clones by morphological observation, checking the chromosome numbers, isoenzyme analysis and random amplified polymorphic DNA analysis, and obtained regenerated plantlets from three hybrid clones.
Astragalus Plant ; genetics ; physiology ; Electroporation ; Fabaceae ; genetics ; physiology ; Hybridization, Genetic ; Transformation, Genetic

In order to expand gene resources and improve Brassica napus cultivars, protoplasts isolated from hypocotyls of Brassica napus cv. Huayou No. 3 and Eruca sativa were fused by PEG-high Ca2+-high pH. Fusion frequency was up to 18.2% when fusion system contained 5 x 10(5) protoplasts/mL, and when PEG concentration of fusion agents were 35% and when fusion time was 25 min. Then the fused protoplasts were cultured by the method of thin liquid layer at the density of 1 x 10(5) protoplasts/mL in improved KM8p medium supplemented with 1.0 mg/L 2,4-D, 0.5 mg/L NAA, 0.5 mg/L 6-BA, 200 mg/L inositol, 300 mg/L protein hydrolysate, and the combinations of 0.1 mol/L sucrose and 0.2 mol/L glucose and 0.2 mol/L mannitol for osmotic regulator, the frequency of callus regeneration was up to 6.8%. When the micro-calli transferred to the proliferation medium that contained B5 salts, 0.087 mol/L sucrose, 0.2 mg/L 2,4-D, 0.5 mg/L NAA, 0.2 mg/L 6-BA and 0.5% Agar, pH 5.8, have grown up to 3-5 mm of diameter, the calli were transferred to the differentiation medium that contained MS salts, 0.087 mol/L sucrose, 0.1 mg/L IAA, 0.8 mg/L 6-BA, 0.8% Agar, pH5.8, the shoots were regenerated in 4 weeks and its frequency was up to 32.8%. Then 2-3 cm shoots were transferred to 1/2 MS medium with 0.5 mg/L IBA+0.2mg/L 6-BA, plantlets were obtained in 14 days and the plantlet frequency was up to 88%. When the protoplasts of Eruca sativa were treated with UV radiation for 2 minutes calli and plantlets have been regenerated, treated for 4 min only calli have been regenerated, and treated for more than 5 min calli have not been regenerated. The callus regeneration and callus proliferation and plant regeneration from symmetric fusion were more than from asymmetric fusion. 16 hybrid plantlets have been regenerated on 21 piece of hybrid calli identified by cytology method.
Brassica ; genetics ; Brassicaceae ; genetics ; Cell Fusion ; Hybrid Cells ; Hybridization, Genetic ; Protoplasts ; Regeneration ; Ultraviolet Rays

Chromosomal translocation is a kind of common chromosomal abnormality. The carriers with chromosomal translocation could have more chance of normal pregnancy with the help of fluorescence in situ hybridization(FISH). This is a review aimed at analyzing the meiosis types of the translocation chromosome. The strategy of preimplantation genetic diagnosis for the carriers with chromosomal translocation is also discussed.
Humans ; In Situ Hybridization, Fluorescence ; methods ; Meiosis ; genetics ; Preimplantation Diagnosis ; methods ; Translocation, Genetic ; genetics

OBJECTIVETo analyze the meiotic segregation results of male reciprocal chromosome translocation by fluorescence in situ hybridization (FISH).

METHODSMulti-color FISH using 3 combined probes located in any 3 chromosome segments on both sides of two breakpoints was performed on the de-condensed sperm head to analyze the sperm chromosomal contents and segregation patterns.

RESULTSFour male reciprocal translocation carriers were included in the study, with the karyotypes of 46, XY, t(2;18) (p16; q23); 46, XY, t(4;6) (q34;q21); 46, XY, t(8;13) (q23;q21) and 46, XY, t(4;5) (4q31;5q13), respectively. The results showed that 4 carriers had different proportions of various segregated spermatozoa. The spermatozoa of alternate, adjacent-1, adjacent-2, 3:1, non-disjunction in meiosis II, and 4:0 or diploidy accounted for 27.1%-49.4%, 26.9%-37.6%, 2.7%-15.7%, 8.6%-32.7%, 0.2%-1.9%, and 0.1%-0.4%, respectively.

CONCLUSIONFor each-reciprocal translocation carrier seems to have a particular meiotic segregation results, FISH analysis on sperm head should be done for each carrier in order to provide an accurate genetic counseling.

Chromosome Breakage ; Heterozygote ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Meiosis ; genetics ; Spermatozoa ; metabolism ; Translocation, Genetic ; genetics

Leymus racemosus had a high resistant capacity to wheat scab (Fusarum head blight). The transfer of scab resistant gene from L. racemosus to Triticum aestivum is of great significance for broadening the germplasm of wheat resistance. To obtain Triticum aestivum-Leymus racemosus translocation line with scab resistance, we irradiated the pollen of T. aestivum-L. racemosus disomic addition line DA7Lr by ⁶⁰Co-γ-rays 1 200 R (100 R/min) prior to pollinating to emasculation T. aestivum cv. Chinese Spring. One plant with one translocation chromosome was detected in the M1 by GISH. The plant with one translocation chromosome was self-pollinated, and at meiotic metaphase I its progenies with two translocation chromosomes were analyzed for chromosome pairing behavior in their pollen mother cells (PMCs). One rod bivalent was observed at meiotic metaphase I, indicating that the plant with two translocation chromosomes was one translocation homozygote. Sequential GISH-FISH analysis, using Oligo-pAs1-2 and Oligo-pSc119.2-2 as probe, translocation line was confirmed as T6DL·7LrS. The translocation line had higher resistance to wheat scab and feasibility to be used as a new source in wheat breeding resistant to scab disease.
Chromosomes, Plant ; Disease Resistance ; genetics ; In Situ Hybridization, Fluorescence ; Plant Breeding ; Plant Diseases ; genetics ; Poaceae ; genetics ; Pollen ; Translocation, Genetic ; Triticum ; genetics

OBJECTIVETo establish a detection method for HBV drug-resistant mutations related to lamivudine, adefovir and entecavir by optimization and assessment of reverse hybridization system.

METHOD26 degenerated probes covering 10 drug-resistant hotspots of 3 drugs were synthesized and immobilized on the same positively charged nylon membrane. PCR products labeled with digoxigenin were hybridized with corresponding probes. To improve the sensitivity and specificity, 4 reaction steps of reverse hybridization were optimized including the number of labeled digoxigenin, the energy intensity of UV cross-linking, hybridization and stringency wash conditions. To prove the feasibility, the specificity, sensitivity and accuracy of this system were assessed respectively.

RESULTSensitive and specific results are obtained by the optimization of the following 4 reaction steps: the primers labeled with 3 digoxigenin, energy intensity of UV cross-linking for 1500 x 0.1 mJ/cm², hybridization at 42 degrees C and stringency wash with 0.5 x SSC and 0.1% SDS solution at 44 degrees C for 30 min. In the assessment of system, the majority of probes have high specificity. The quantity of PCR product with a concentration of 10 ng/μl or above can be detected by this method. The concordant rate between reverse hybridization and direct sequencing is 93.9% in the clinical sample test.

CONCLUSIONThough the specificity of several probes needs to be improved further, it is a simple, rapid and sensitive method which can detect HBV resistant mutations related to lamivudine, adefovir and entecavir simultaneously. Due to the short distance between 180 and 181, likewise 202 and 204, the sequence of the same probe covers two codon positions, and hybridization will be interfered by each other. To avoid such interference, the possible solution is that probes are designed by arranging and combining various forms of two near codons.

DNA, Viral ; genetics ; Drug Resistance, Viral ; genetics ; Hepatitis B virus ; drug effects ; genetics ; Humans ; Hybridization, Genetic ; Mutation ; Nucleic Acid Hybridization ; methods ; Oligonucleotide Array Sequence Analysis ; Sensitivity and Specificity

OBJECTIVETo select superior mating combinations from different F1 generations of Dendrobium officinale.

METHODTwenty-six superior parents from four provenances were selected to make up 26 mating combinations. The agronomic traits of different F1 generation were measured, counted, and analyzed by DPS software.

RESULTThere were significant differences between different mating combinations among F1 generations. The variation among full-sib families was significantly higher than that among half-sib families, while the variation inside full-sib families was significantly lower than that inside half-sib families. Twelve agronomic traits were simplified into four principal components, of which the accumulative contribution rate was 84.127 3%. Twenty-six mating combinations have been divided into six groups, plants from the fifth and sixth group grew much better than the others.

CONCLUSIONThe selection of parents should be emphasized by using hybrid vigor. There were higher genetic gains and superior uniformity inside families when selection and breeding occurs in full-sib families than in half-sib families. Eight superior full-sib families (83 x 34, 66 x 9, 68 x 2, 91 x 69, 66 x 65, 69 x 91, 17 x 66, 66 x 17) have been selected which showed a good agronomic traits of seedlings.

Agriculture ; economics ; Cluster Analysis ; Dendrobium ; genetics ; growth & development ; Hybridization, Genetic ; Seedlings ; growth & development ; Software

The explants were obtained from stem segments of hybrid combination of Dendrobium officinale germplasms (Zhejiang Yandang Mountain x Yunnan Guangnan). The screened culture mediums for buds induction and multiplication, which were 1/2MS + IBA 1.0 mg x L(-1) +6-BA 1.0 mg x L(-1) and 1/2MS + IBA 1.5 mg x L(-1) +6-BA 0.5 mg x L(-1) respectively, were applied to 159 germplasms of D. officinale from Zhejiang, Yunnan, Guangxi, Hunan, etc. The medium for axillary buds induction had universality with 94.3% induction rate. During buds multiplication, there were significant differences in proliferation effect among germplasms, and three proliferation forms i.e. single bud, multiple buds and protocorm were differentiated from different germplasms. The results showed that different germplasms of D. officinale had specific requirements for culture medium. Therefore, developing special culture medium for breeding clones of D. officinale is urgent and has important application values.
Culture Media ; chemistry ; Dendrobium ; genetics ; growth & development ; physiology ; Hybridization, Genetic ; Regeneration