Selection and checking of the gene marker related to the high royal jelly quantity trait of apis mellifera lindauer
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Abstract:
In order to find out the gene markers related to the high jelly quantity trait in Western honey, genomic DNA from four kinds of Apis mellifera Lindauer were amplified with 12 random primers. A differential DNA fragment P 2 316 bp related to this trait was obtained from the polymorphic electrophoretic map of the PCR product with one of the primers.The results of Southern hybridization showed that this DNA fragment can only be the DNA of Apis mellifera Lindauer with high jelly quantity trait .It is concluded that the DNA fragment P 2 316 bp could be a specific gene marker of the high jelly quantity trait of Apis mellifera Lindauer.Keywords:
genomic DNA
Trait
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The established amplified fragmentlength polymorphism (AFLP) protocol was simplified and optimized for honey bee DNA (Apis mellifera L.). Compared to the original method, the following simplifications were made: (i) the digestion of DNA and ligation of the adapters are performed in one reaction vs. two, (ii) one restriction enzyme is used vs. two and (iii) amplification is accomplished in one reaction vs. two. PCR products are resolved in agarose-Synergel instead of polyacrylamide and are visualized by ethidium bromide staining rather than by autoradiography of labeled primers. Using the modified procedure for honey bee DNA, high reproducibility of the band patterns of PCR products and low sensitivity to the amplification conditions were seen. Analysis of honey bee DNA revealed considerable genetic variability within and between African and European bee samples. African- and European-specific fragments were found.
Ethidium bromide
Agarose
Agarose gel electrophoresis
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The experiment used royal jelly of Apis cerana and Apis mellifera as materials,extracted the DNA of royal jelly with modified phenol-chloroform method,the extracted DNA was analyzed by RAPD with 35 arbitrary primers.The results showed that: in the 35 primers that were tested,1 obtained the same bands.5 were polymorphous,a total of 32 bands were obtained,of which 24 were polymorphic.
Apis cerana
Royal Jelly
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Primer (cosmetics)
Molecular marker
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Sexing
genomic DNA
Cloning (programming)
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Cucumis melo ( C. melo L.) groups is an important plant with increasing economic value. The improvement of the varieties with great tastes like melon and resistance to insect and unfavorable weather conditions similar to Thai-melon has been needed. This research aimed to study genetic diversity and relationship between molecular markers and sweetness trait of C. melo L. by using Target Region Amplified Polymorphism Polymerase Chain Reaction (TRAP-PCR) technique. Specific primers which were designed based on the sequence of nine genes involving in metabolic pathway of sugar combining with twelve arbitrary primers were used in TRAP-PCR. It was found that 59 pairs of primers were able to amplify DNA generating total 379 amplicons. The polymorphic data was used to create dendrogram based on Jaccard’s similarity index through UPGMA. The results revealed the similarity index ranged from 0.69 to 0.95. Studied plants could be divided into 3 groups with a similarity coefficient of 0.8, corresponding to the sweetness (% brix). The 220 of polymorphic bands were detected with a percentage of 58.05. Analysis of the relationship between molecular markers and sweetness trait showed thatCM01 (X174) marker was found to correlate with the sweetness of 87.07 percent, resulting from the use of primers, Neutral invertase 1 with Sa17_800. The results of genetic diversity and developed DNA markers in this study will assist breeding processes and selection of sweetness trait in C. melo L. to be more efficient, more accurate and faster. Keywords: Cucumis melo L., TRAP-PCR, Genetic diversity, Molecular marker, sweetness
Cucumis
Sweetness
Molecular marker
Germ plasm
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The F1 population derived from the cross between Jingyu and Flavortop cultivars were used as materials for screening. The molecular markers closely linked to free stone gene (F). By RAPD finger printing technique and BSA(bulked segregate analysis) an amplified polymorphic DNA fragment, OPI07-1000, was screened from those plant populations. The distance of genetic linkage to the free stone gene (F) was 2.5 cM. The fragment was reclaimed, cloned and sequenced. The DNA marker was actually 1 054 bp. It is suggested that the sequence of the marker might be used as a basis for synthesizing the specific PCR primers and the probe for detecting free stone gene of peach fruit and molecular marker selection in peach breeding.
Bulked segregant analysis
Molecular marker
Genetic linkage
Genetic distance
Marker-Assisted Selection
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Sex-linked molecular markers are being obtained, which would be essential to be used in the screening of different sex of dioecious plants at the seedling stage. Furthermore, it is important in cloning the gene related to the sex. In this study the random amplified polymorphic DNA (RAPD) technique was employed with the objective to find markers linked to sex determination in Asparagus. A total of 100 primers were tested with the same PCR cycling procedure. A female-associated fragment with a length of about 867bp was generated with S12 primer. The fragment was cloned and sequenced, showing it is abundant in AT and contains 2 shorter open reading frames. In order to convert the RAPD marker into SCAR (sequence characterized amplified regions) marker, 24bp specific primers were constructed and used for PCR amplifying. The female-linked dominant SCAR marker was obtained, which would be efficient to identify the different sex of Asparagus officinalis L.
Asparagus
Primer (cosmetics)
Cloning (programming)
Molecular marker
Marker gene
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The DNA was extracted from mature Spinacia oleracea L.separately by the method of CTAB,then used as template after mixed by gender to be amplified with the optimized RAPD reaction condition.A total of 160 primers(10 bp) and 40 pairs of primers were screened,44 primers of which produced clear patterns.And the further study showed that the primer S1497 could generate a common DNA fragment in the male pool but absent in the female pool.The S1497 fragment was extracted from agarose gel and cloned into pUCm-T vector,and then transformed into E.coli JM109.It was sequenced after test.The sequencing result shows that this sequence is 1978 bp at full length.This means that the specific fragment could be used as a base to clone sex-linked gene of Spinacia oleracea L.
Spinacia
Primer (cosmetics)
Agarose
genomic DNA
clone (Java method)
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Honey bee, Apis mellifera L. (Hymenoptera: Apidae) DNA has alleles, at three polymorphic restriction sites, which were initially found almost exclusively in samples from eastern Europe. These markers were first detected with three labeled, cloned, probes on DNA blots. In this study, the polymorphic sites were made easier to analyze with the polymerase chain reaction (PCR). The amplification primers and conditions are described. The regions with the polymorphic sites were mapped within the sequences of the honey bee genome and were found within three separate linkage groups. The genomic locations of the defining restriction sites and PCR primers are given. With the PCR-based method of detection, additional samples of bees from Europe and Africa were examined. Within the eastern European group, the three markers were at high frequencies in Italian bees (subspecies Apis mellifera ligustica Spinola), two markers were at high frequencies and one marker was at a lower frequency in Carniolan bees (Apis mellifera carnica Pollman), and all three markers were absent or at low frequencies in Caucasian bees (Apis mellifera caucasica Gorbachev). The three markers were absent or at low frequencies in western European bees (Apis mellifera mellifera L and Apis mellifera iberica Goetze) and in South African bees (Apis mellifera scutellata Lepeletier).
Subspecies
genomic DNA
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Medicago sativa
Germ plasm
Parthenogenesis
Primer (cosmetics)
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