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1.
Osipova ES Koveza OV Troitskiĭ AV Dolgikh IuI Shamina ZB Gostimskiĭ SA 《Genetika》2003,39(12):1664-1672
RAPD (Random Amplified Polymorphic DNA) and ISSR (Inter Simple Sequence Repeats) markers were used to analyse the genetic divergence between the regenerated plants derived from callus cultures and the original maize line A188. Analysis of polymorphism by using 38 RAPD- and 10 ISSR-oligonucleotide primers showed that the differences between eight examined somaclones and the original line ranged from 6.5 to 23%. As confirmed using new primers, the regenerants derived from callus cultures grouped into two clusters according to their origin. The regenerants isolated from calluses grown for eight months differed from one another and the original line to a larger extent than the regenerants obtained from two-month callus cultures. In some somaclones, molecular marking of the regenerants revealed specific RAPD and ISSR fragments that were absent in other somaclones or the original maize line. On the basis of six specific fragments (five RAPD and one ISSR), SCAR (Sequence Characterized Amplified Region) markers were developed. Specific polymorphism revealed with random primers was completely confirmed using five SCAR markers. Polymorphism of one SCAR marker differed from that revealed with random primers. Five SCAR fragments were inherited as simple dominant traits. One SCAR fragment displayed codominant inheritance. 相似文献
2.
Analysis of Specific RAPD and ISSR Fragments in Maize (Zea mays L.) Somaclones and Development of SCAR Markers on Their Basis 总被引:1,自引:0,他引:1
E. S. Osipova O. V. Koveza A. V. Troitskij Yu. I. Dolgikh Z. B. Shamina S. A. Gostimskij 《Russian Journal of Genetics》2003,39(12):1412-1419
RAPD (Random Amplified Polymorphic DNA) and ISSR (Inter Simple Sequence Repeats) markers were used to analyse the genetic divergence between the regenerated plants derived from callus cultures and the original maize line A188. Analysis of polymorphism by using 38 RAPD- and 10 ISSR-oligonucleotide primers showed that the differences between eight examined somaclones and the original line ranged from 6.5 to 23%. As confirmed using new primers, the regenerants derived from callus cultures grouped into two clusters according to their origin. The regenerants isolated from calluses grown for eight months differed from one another and the original line to a larger extent than the regenerants obtained from two-month callus cultures. In some somaclones, molecular marking of the regenerants revealed specific RAPD and ISSR fragments that were absent in other somaclones or the original maize line. On the basis of six specific fragments (five RAPD and one ISSR), SCAR (Sequence Characterized Amplified Region) markers were developed. Specific polymorphism revealed with random primers was completely confirmed using five SCAR markers. Polymorphism of one SCAR marker differed from that revealed with random primers. Five SCAR fragments were inherited as simple dominant traits. One SCAR fragment displayed codominant inheritance. 相似文献
3.
Sheila Jazmín Reyes-Zambrano Carlos Alberto Lecona-Guzmán Federico Antonio Gutiérrez-Miceli Víctor Manuel Ruiz-Valdiviezo Arely Anayansi Vargas-Díaz 《Phyton》2021,90(6):1727-1739
Agave americana L. callus were exposed to different concentrations of ethyl methanesulphonate (EMS) 0, 15, 30, 45 and 60 mM and to different times of exposure (2 and 4 h). The viability and capacity of shoot formation were shown to be affected when the callus were exposed to high concentrations (30–60 mM). Only the callus exposed to 15 mM EMS presented shoot formation; the exposure time of two hours produced the largest quantity of shoots regenerated per callus (21 shoots/callus). In order to generate somaclonal variants resistant to Fusarium oxysporum, a selection pressure was applied through of a culture filtrate (CF) of 100 ppm of the fungus. This was made in callus obtained in the treatment with 15 mM EMS during 2 h of exposure. The CF caused oxidation and necrosis in 71.25% of the callus; however, they were capable of generating shoots (3.5 shoots/callus). Molecular markers type RAPD, ISSR and DAMD were used to evaluate the genetic variation arising from the mutations caused by EMS on control plants and 16-month-old somaclonal variants. The polymorphic information content (PIC) for each one of the initiating groups was: 0.28 (DAMD), 0.09 (ISSR) and 0.14 (RAPD). DAMD revealed a greater percentage of polymorphism than RAPD and ISSR. Polymorphic bands were detected in the somaclonal variants. This indicated that the EMS caused genetic variation in the regenerated plants conferring resistance to them against Fusarium oxysporum. 相似文献
4.
利用RAPD和ISSR分子标记分析怀地黄种质遗传多样性 总被引:42,自引:0,他引:42
用RAPD与ISSR技术对怀地黄的8个品种和2个脱毒品系进行了种质遗传多样性分析。分别从80条RAPD引物和44条ISSR引物中筛选出适合怀地黄种质分析的17条RAPD引物和10条ISSR引物,用于RAPD和ISSR分析。17条RAPD引物共扩增出177条带, 多态性位点数为109; 多态性位点比率为61.58%;平均多样性指数(I)为0.3135;每个位点的有效等位基因数(Ne)是1.3641; 10条ISSR引物共扩增出110条带. 多态性位点数为79; 多态性位点比率为71.58%;平均多样性指数(I)为0.3577;每个位点的有效等位基因数(Ne)是1.4037。 基于扩增条带数据库建立了各自的Jaccard遗传相关系数矩阵,构建了相似的分子树状图,将10个供试材料分为2类:一类群含组培85.5、大田85.5、组培9302、大田9302、金状元和金白6个材料;另一类群含北京1号、大红袍、地黄9104和野生地黄4个材料。两种分子标记的分析结果呈极显著正相关(r=0.649)。结果表明,RAPD与ISSR标记适合于怀地黄种质遗传多样性分析,ISSR标记技术是一种多态性和重复性优于RAPD技术的实用技术。 相似文献
5.
Identification of RAPD markers linked to the gene PM 1 for resistance to powdery mildew in wheat 总被引:17,自引:0,他引:17
X. Y. Hu H. W. Ohm I. Dweikat 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1997,94(6-7):832-840
Powdery mildew caused by Blumeria graminis DC. f. sp. triticiém. Marchal is an important disease of wheat (Triticum aestivum L. em Thell). We report here the identification of three random amplified polymorphic DNA (RAPD) markers closely linked to
a gene for resistance to B. graminis in wheat. RAPD-PCR (polymerase chain reaction) analysis was conducted using bulked segregant analysis of closely related
lines developed from a segregating F5 family. The F5 family was derived from a cross between the susceptible cultivar Clark and the resistant line Zhengzhou 871124. Genetic analysis
indicated that resistance of Zhengzhou 871124 to powdery mildew is conferred by the gene Pm1. After performing RAPD-PCR analysis with 1300 arbitrary 10-mer primers and agarose-gel electrophoresis, two RAPD markers,
UBC320420 and UBC638550, were identified to be co-segregating with the disease resistance. No recombinants were observed between either of the RAPD
markers and the gene for resistance to powdery mildew after analysis of 244 F2 plants. The third RAPD marker, OPF12650, was identified with denaturing gradient-gel electrophoresis (DGGE), and was determined to be 5.4±1.9 cM from the resistance
gene. UBC320420 and UBC638550 were present in wheat powdery mildew differential lines carrying the gene Pm1, suggesting linkage between these markers and the Pm1 resistance gene. Co-segregation between Pm1 and the two markers UBC320420 and UBC638550 was confirmed in a segregating population derived from a cross with CI14114, the wheat differential line carrying Pm1. The method of deriving closely related lines from inbred families that are segregating for a trait of interest should find
wide application in the identification of DNA markers linked to important plant genes. The RAPD marker UBC638550 was converted to a sequence tagged site (STS). RAPD markers tightly linked to target genes may facilitate selection and enable
gene pyramiding for powdery mildew resistance in wheat breeding programs.
Received: 10 December 1995 / Accepted: 13 September 1996 相似文献
6.
Meetul Kumar Gyan P. Mishra Raghwendra Singh Jitendra Kumar Pradeep K. Naik Shashi Bala Singh 《Physiology and Molecular Biology of Plants》2009,15(3):225-236
The phylogenetic relationships of 36 locally grown Prunus armeniaca genotypes which are collected from nine sampling sites from two valleys viz. Nubra (9,600 ft) and Leh (11,500 ft) of trans-Himalayan region were analyzed using 31 PCR markers (20 RAPDs and 11 ISSRs). This is the first report of molecular genetic diversity studies in apricot from this region of the world. RAPD analysis yielded 139 fragments, of which 136 were polymorphic, with an average of 6.8 polymorphic fragments per primer. ISSR analysis produced 58 bands, of which 56 were polymorphic, with an average of 5.09 polymorphic fragments per primer. The primers based on (CT)n produced maximum number of bands (nine) while, (AT)n and many other motifs gave no amplification. RAPD markers were found more efficient with regards to polymorphism detection, as they detected 97.84 % as compared to 96.5 % for ISSR markers. Clustering of genotypes within groups was not similar when RAPD and ISSR derived dendrogram were compared, whereas the pattern of clustering of the genotypes remained more or less the same in RAPD and combined data of RAPD + ISSR. The results of PCA analysis were comparable to the cluster analysis. These analyses, allowed us to identify the groups corresponding to the two apricot collection sites.Key words: Prunus armeniaca, Apricot, Genetic Diversity, RAPD, ISSR, AMOVA 相似文献
7.
Randomly amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) markers were applied to assess the genetic
stability of micropropagated olive (Olea europaea L. cv. Maurino) plants regenerated by axillary buds. Initial olive explants, isolated from one donor tree, were multiplied
on Murashige and Skoog medium for 12 repeated subcultures. A total of 40 RAPD and 10 ISSR markers resulted in 301 distinct
and reproducible band classes showing homogeneous RAPD and ISSR patterns. The amplification products revealed genetic stability
among the micropropagated plants and between them and the donor plant. The results demonstrate the genetic stability of nine
year old mature micropropagated olive plants cultured in field, and corroborated the fact that axillary multiplication is
the safest mode for multiplication of true to type plants. 相似文献
8.
Tui Ray Indrajit Dutta Prasenjit Saha Sampa Das S.C. Roy 《Plant Cell, Tissue and Organ Culture》2006,85(1):11-21
An efficient micropropagation protocol produced large number of plants of the three elite banana (Musa spp.) cultivars Robusta (AAA), Giant Governor (AAA) and Martaman (AAB) from shoot tip meristem. The genetic relationships
and fidelity among the cultivars and micropropagated plants as assessed by random amplified polymorphic DNA (RAPD) and inter-simple
sequence repeat (ISSR) markers, revealed three somaclonal variants from Robusta and three from Giant Governor. A total of
5330 RAPD and 2741 ISSR fragments were generated with 21 RAPD and 12 ISSR primers in micropropagated plants. The percentage
of polymorphic loci by RAPD and ISSR were found to be 1.75, 5.08 in Robusta and 0.83, 5.0 in Giant Governor respectively.
Among the two marker systems used, ISSR fingerprinting detected more polymorphism than RAPD in Robusta and Giant Governor
with most of the primers showing similar fingerprinting profile, whereas Martaman revealed complete genetic stability. 相似文献
9.
10.
Codonopsis lanceolata Benth. et Hook. f., commonly known as bonnet bellflower, is a high-valued herb medicine and vegetable. In this study, a large number of plants were regenerated via organogenesis from immature seed-derived calli in C. lanceolata by a simple and efficient method. Compared with the mother donor plant, the regenerated plants did not exhibit visible phenotypic variations in six major morphological traits examined at the stage of one-season-maturity under field conditions. To gain insight into the genomic stability of these regenerated plants, 63 individuals were randomly tagged among a population of more than 2,000 regenerants, and were compared with the single mother donor plant by two molecular markers, the inter-simple sequence repeats (ISSR) and randomly amplified polymorphic DNA (RAPD). Apparent genomic variation was detected in the 63 regenerants, whereas preexisting heterozygosiy in the donor plant was deemed minimal by testing 30 seedlings germinated from selfed seeds of the same donor plant. The percentages of polymorphic bands (PPB) in the ISSR and RAPD analysis were respectively 15.7 and 24.9% for the 63 regenerated plants. Cluster analysis indicates that the genetic similarity values calculated on the basis of RAPD and ISSR data among the 64 plants (63 regenerated and one donor) were respectively 0.894 and 0.933, which allow classification of the plants into distinct groups. Nineteen randomly isolated bands underlying the changed RAPD or ISSR patterns were sequenced, and three of them showed significant homology to known-function genes. Detailed pairwise sequence comparison at one locus between the donor plant and a regenerant revealed that insertion of two short (24 and 19 bp) stretches of nucleotides in the regenerated plant relative to the donor plant occurred in an apparently stochastic manner.Electronic Supplementary Material Supplementary material is available for this article at 相似文献
11.
A comparative analysis of genetic diversity in blackgram genotypes using RAPD and ISSR markers 总被引:16,自引:0,他引:16
Souframanien J Gopalakrishna T 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》2004,109(8):1687-1693
Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were used to study the DNA polymorphism in elite blackgram genotypes. A total of 25 random and 16 ISSR primers were used. Amplification of genomic DNA of the 18 genotypes, using RAPD analysis, yielded 104 fragments that could be scored, of which 44 were polymorphic, with an average of 1.8 polymorphic fragments per primer. Number of amplified fragments with random primers ranged from two (OPA-13) to nine (OPK-4) and varied in size from 200 bp to 2,500 bp. Percentage polymorphism ranged from 16.6% (OPK-7) to a maximum of 66.6% (OPE-5, OPH-2, and OPK-8), with an average of 42.7%. The 16 ISSR primers used in the study produced 101 bands across 18 genotypes, of which 55 were polymorphic. The number of amplified bands varied from two (ISSR 858) to ten (ISSR 810), with a size range of 200–2,200 bp. The average numbers of bands per primer and polymorphic bands per primer were 6.3 and 3.4, respectively. Percentage polymorphism ranged from 25% (ISSR 885) to 100% (ISSR 858), with an average percentage polymorphism of 57.5% across all the genotypes. The 3-anchored primers based on poly(GA) and poly(AG) motifs produced high average polymorphisms of 54.98% and 58.32%, respectively. ISSR markers were more efficient than the RAPD assay, as they detected 57.4% polymorphic DNA markers in Vigna mungo as compared to 42.7% for RAPD markers. The Mantel test between the two Jaccards similarity matrices gave r =0.32, showing low correlation between RAPD- and ISSR-based similarities. Clustering of genotypes within groups was not similar when RAPD and ISSR derived dendrogram were compared, whereas the pattern of clustering of the genotypes remained more or less the same in ISSR and combined data of RAPD and ISSR. 相似文献
12.
Applicability of inter-simple sequence repeat polymorphisms in wheat for use as DNA markers in comparison to RFLP and RAPD markers 总被引:43,自引:0,他引:43
T. Nagaoka Y. Ogihara 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1997,94(5):597-602
Inter-simple sequence repeat polymorphic DNA (ISSR) was evaluated for its applicability as a genetic marker system in wheat.
PCR was carried out with primers that annealed to simple sequence repeats. The resultant products were subjected to agarose-gel
electrophoresis, and the banding patterns were compared among six wheat accessions containing diploid, tetraploid, and hexaploid
members. Out of 100 examined, 33 primers produced distinguishable as well as polymorphic bands in each of the six accessions.
Although most of the primers that gave distinct bands (30 primers out of 33) contained dinucleotide repeats, each of the primers
with tri-, tetra-, and penta-nucleotide motifs also yielded discrete bands. Primers based on (AC)n repeats gave the most polymorphic bands. In total, 224 polymorphic bands were found in the comparison between Einkorn wheats
whereas, on the average, 120 polymorphic bands were detected between common wheats. ISSR primers produced several times more
information than RAPD markers. The extent of band polymorphism was similar to that of RFLP markers, and greater than that
of RAPDs. The genetic relationships of wheat accessions estimated by the polymorphism of ISSR markers were identical with
those inferred by RFLP and RAPD markers, indicating the reliability of ISSR markers for estimation of genotypes. These polymorphic
bands are potential candidates as novel markers for use in linkage-map construction in wheat. The characteristic features
of ISSR markers, i.e. polymorphism, generation of information and ease of handling, suggest their applicability to the analysis
of genotypes as well as to the construction of PCR-based genome maps of wheats.
Received: 15 September 1996 / Accepted: 25 October 1996 相似文献
13.
Occurrence of chromosomal variations and plant regeneration from long-term-cultured citrus callus 总被引:4,自引:0,他引:4
Summary Embryogenic callus of Anliucheng sweet orange (Citrus sinensis Osbeck) is theoretically diploid. However, significant chromosomal variations occurred when the calluses were subcultured
and preserved for a long time. Cytological observation revealed a variety of mitotic irregularities underlying the occurrence
of chromosomal variations. Despite the ubiquitous existence of chromosomal variations, long-term-cultured calluses were still
capable of producing somatic embryos and plants. Interestingly, chromosomal variants were selected against when somatic embryos
and plants regenerated from the embryogenic callus. Randomly amplified polymorphic DNA (RAPD) analysis was also carried out
to detect DNA sequence variation in regenerated plants derived from the embryogenic callus. No difference in banding patterns
was detected. It was clear that the plant regeneration from long-term-cultured callus was inclined to select against somaclonal
variations. 相似文献
14.
To control the genetic quality during the whole process of tissue culture of the traditional Chinese medicinal plant, Saussurea involucrate Kar. et Kir., DNA polymorphisms and genetic variations were investigated using randomly amplified polymorphic DNA (RAPD)
and inter-simple sequence repeats (ISSR) markers. The genetic stability/variation in tissue-cultured products, including three
calli, three adventitious shoots, regenerated plantlets and 2 year-old regenerated plantlets cultivated in the planting base
in Tianshan Mountain, were assessed compared with 1 year-old and 2 year-old seedlings cultivated in the same planting base
using aseptic seedlings as reference. Apparent genetic variation was detected in the 11 type of plant materials. The percentages
of polymorphic bands in the RAPD and ISSR analysis were, respectively, 35% and 33%. Cluster analysis indicated that the genetic
similarity values calculated on the basis of RAPD and ISSR data among the 11 type of plant materials were respectively ranged
from 0.823 to 0.995 with a mean of 0.878 and 0.825 to 0.974 with a mean of 0.885, which classified the samples into three
groups. The similarity coefficient also revealed that differences among three calli were not remarkable by both RAPD and ISSR
analysis, and only chemical components and growth properties needed consideration in the screening of callus used for the
next redifferentiation studies. But there are remarkable differences among three adventitious shoots analyzed by ISSR markers.
Therefore, RAPD and ISSR markers are efficient tools in genetic variation assessment and quality control in plant tissue culture
process. 相似文献
15.
Identification of DNA markers linked to the male sex in dioecious hemp (Cannabis sativa L.) 总被引:16,自引:0,他引:16
G. Mandolino A. Carboni S. Forapani V. Faeti P. Ranalli 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,98(1):86-92
A 400-bp RAPD marker generated by a primer of random decamer sequence has been found associated with the male sex phenotype
in 14 dioecious cultivars and accessions of hemp (Cannabis sativa L.). The primer OPA8 generates a set of bands, most of which polymorphic among all the individual plants tested, and 1 of
which, named OPA8400, present in all male plants and absent in female plants. A screening of 167 plants belonging to different genotypes for the
association of the OPA8400 marker with the sex phenotype revealed that only in 3 cases was the 400-bp band was present in plants phenotypically female;
on the contrary, in male plants the band was never missing, while in monoecious plants it was never present. Despite this
sex-specific association, the sequences corresponding to OPA8400 were present in both staminate and carpellate plants, as revealed by Southern blotting and hybridization with the cloned
RAPD band. The RAPD marker was sequenced, and specific primers were constructed. These primers generated, on the same genotypes
used for RAPD analysis, a SCAR marker 390 bp in length and male-specific. This SCAR is suitable for a precise, early and rapid
identification of male plants during breeding programs of dioecious and monoecious hemp.
Received: 16 January 1998 / Accepted: 30 April 1998 相似文献
16.
H. P. Gajera R. S. Tomar S. V. Patel R. R. Viradia B. A. Golakiya 《Journal of plant biochemistry and biotechnology.》2011,20(2):217-223
The genetic variability and relationships among 20 Mangifera indica genotypes representing 15 endangered and 5 cultivars, obtained from Indian Gir forest region, were analyzed using 10 random amplified polymorphic DNA (RAPD) and 21 inter simple sequence repeat (ISSR) markers. RAPD markers were more efficient than the ISSR assay with regards to polymorphism detection. Also, the average numbers of polymorphic loci per primer, average polymorphic information content (PIC) and primer index (PI) values were more for RAPD than for ISSR. But, total number of genotype specific marker loci, Nei’s genetic diversity (h), Shannon’s information index (I), total heterozygosity (Ht), average heterozygosity (Hs) and mean coefficient of gene differentiation (Gst) were more for ISSR as compared to RAPD markers. The regression test between the two Nei’s genetic diversity indexes showed low regression between RAPD and ISSR based similarities but maximum for RAPD and RAPD + ISSR based similarities. The pattern of clustering of genotypes within groups was not similar when RAPD and ISSR derived dendrogram were compared. Thus, both the markers were equally important for genetic diversity analysis in M. indica. 相似文献
17.
Bueno MA Agundez MD Gomez A Carrascosa MJ Manzanera JA 《International journal of plant sciences》2000,161(3):363-367
In vitro-induced cork oak (Quercus suber L.) embryos from anther cultures proved to be of haploid origin both by enzyme and RAPD gene marker analysis. The problem considered was to ascertain if embryo cultures originated either from a single haploid cell, from a microspore, or from multiple haploid cells. Therefore, a heterozygotic gene was searched for in the parent tree. The gene coding for shikimate dehydrogenase (SKDH1) proved to be heterozygous in the parental tree, and subsequently, these allozymes were screened for the embryos induced in anther cultures from the same tree. Only haploid embryos were found, confirming the microspore origin. Different genotypes were not identified inside each anther by isozyme analysis, probably because of selective pressure for one embryo early in development, but both parental SKDH1 alleles were found in the embryos of different anthers. The banding patterns detected by RAPD markers permitted the identification of multiple microspore origins inside each anther. 相似文献
18.
Molecular variation and fingerprinting of Leucadendron cultivars (Proteaceae) by ISSR markers 总被引:4,自引:0,他引:4
BACKGROUND AND AIMS: There are more than 80 species of Leucadendron and most are used as cut flowers. Currently, more than 100 cultivars are used by industry and many of them are interspecific hybrids. The origin of most cultivars is unclear and their genetic diversity and relationships have not been studied. This investigation was carried out to evaluate the genetic variation and relationships among 30 Leucadendron cultivars. METHODS: ISSR markers were applied to determine the genetic variation and to discriminate Leucadendron cultivars. Sixty-four ISSR primers were screened and 25 primers were selected for their ability to produce clear and reproducible patterns of multiple bands. KEY RESULTS: A total of 584 bands of 305-2400 bp were amplified, of which 97 % were polymorphic. A dendrogram generated using the Unweighted Pair Group Method with Arithmetic Average based on a distance measure of total character difference showed that the Leucadendron cultivars clustered into two main groups. Twenty-four of the 30 cultivars can be unequivocally differentiated, but identical profiles were observed for three cultivar pairs, 'Katie's Blush' and 'Silvan Red', 'Highlights' and 'Maui Sunset', and 'Yellow Crest' and 'Yellow Devil'. CONCLUSIONS: ISSR profiling is a powerful method for the identification and molecular classification of Leucadendron cultivars. A fingerprinting key was generated based on the banding patterns produced using two ISSR primers (UBC856 and UBC857). In addition cultivar-specific ISSR bands were obtained for 17 of the 30 Leucadendron cultivars tested. 相似文献
19.
S. V. Hajiyeva Z. I. Akparov N. A. Hasanov Z. P. Mustafayeva E. S. Hajiyev A. T. Mammadov V. I. Izzatullayeva S. M. Babayeva S. S. Sharifova A. M. Mammadov M. A. Abbasov 《Russian Journal of Genetics》2018,54(2):188-197
The article presents the results of a study of genetic polymorphism for the first time carried out on pomegranate varieties and forms of Azerbaijan origin using molecular markers. In total, 102 PCR fragments were identified, of which 80 were polymorphic. The high level of polymorphism (75.5%) and the rich genetic diversity were identified among the studied pomegranate collection. As a result of data analysis and on the basis of the values of the basic parameters (PIC, EMR, MI, RP, MRP) determining informativeness of markers, all 14 ISSR primers were suitable for genotyping pomegranate accessions. The most effective markers (UBC808, UBC811, UBC834, and UBC840) were identified among the set of primers tested. A dendrogram was constructed on the basis of the data obtained, which made it possible to group genotypes into 16 major clusters. The genetic similarity index ranged from 0.032 to 0.94. The study of the genetic relationship of different pomegranate varieties confirms the effectiveness of the ISSR method, which makes it possible to determine the level of genetic diversity, as well as to establish the relationship among the studied pomegranate accessions. 相似文献
20.
Occurrence of genetic variants during micropropagation is occasionally encountered when the cultures are maintained in vitro for long period. Therefore, the micropropagated multiple shoots of Vanilla planifolia Andrews developed from axillary bud explants established 10 years ago were used to determine somaclonal variation using random amplified polymorphic DNA (RAPD) and intersimple sequence repeats markers (ISSR). One thousand micro-plants were established in soil of which 95 plantlets (consisting of four phenotypes) along with the mother plant were subjected to genetic analyses using RAPD and ISSR markers. Out of the 45 RAPD and 20 ISSR primers screened, 30 RAPD and 7 ISSR primers showed 317 clear, distinct and reproducible band classes resulting in a total of 30 115 bands. However, no difference was observed in banding patterns of any of the samples for a particular primer, indicating the absence of variation among the micropropagated plants. Our results allow us to conclude that the micropropagation protocol that we have used for in vitro proliferation of vanilla plantlets for the last 10 years might be applicable for the production of clonal plants over a considerable period of time. 相似文献