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1.
Summary Somaclonal variation, which is a welcome source of genetic variation for crop breeding, is unwanted when direct regenerants have to be used in tissue culture mass propagation (eg. in many forest trees), or in the regeneration of genetically transformed plants. Random amplified polymorphic DNA (RAPD) was used to analyse somatic embryos and plants regenerated from embryogenic cell lines in Norway spruce, Picea abies (L.) Karst. RAPD facilitated the identification of clones, as material from the same cell lines shared identical patterns of amplified fragments, whereas regenerants from different cell lines were easily distinguishable by their respective patterns. For comparisons with explant donor genotypes, cell lines were initiated from cotyledons. Some of the seedlings that had parts of their cotyledons removed were grown on as control plants. Somatic embryos regenerated from cotyledon cell lines showed no aberrations in RAPD banding patterns with respect to donor plants. We conclude that gross somaclonal variation is absent in our plant regeneration system.Abbreviations ESM
embryogenic suspensor mass
- RAPD
random amplified polymorphic DNA
- RFLP
restriction fragment length polymorphism
- (2,4-dichlorophenoxy)acetic acid
2,4-D
- 1-naphthaleneacetic acid
NAA 相似文献
2.
Genetic variation within and between two cultivars of red clover (Trifolium pratense L.): Comparisons of morphological,isozyme, and RAPD markers 总被引：2，自引：0，他引：2
Summary Morphological, isozyme and random amplified polymorphic DNA (RAPD) markers were used to estimate genetic variation within and between cultivars of red clover (Trifolium pratense L.), an important temperate forage legume. Two cultivars of red clover, Essi from Europe and Ottawa from Canada, were evaluated. Six monogenic morphological characters were observed for 80 plants from each of these two cultivars. All six morphological loci were polymorphic in the cultivar Essi whereas only four loci were polymorphic in the cultivar Ottawa. Forty plants from each cultivar were assayed for isozyme markers. A total of 21 enzyme-coding loci with 43 alleles was detected using twelve enzyme systems. Thirteen and nine of these loci were polymorphic in Essi and Ottawa, respectively. The mean number of alleles per locus was 1.81 in Essi and 1.67 in Ottawa. Seventeen random 10-mer primers were screened for RAPD markers. Nine primers which gave clear and consistent amplified products were used to assay 20 individuals from each cultivar. Each primer gave from 7 to 20 amplified bands with an average of 14.8 bands per primer. One hundred and eight of 116 putative loci were polymorphic in Essi and 90 of 98 loci were polymorphic in Ottawa. High within-cultivar variation was observed in both cultivars using both isozyme and RAPD markers. This high polymorphism makes these markers useful for germplasm characterization and genetic studies in red clover. 相似文献
3.
V. Orbović M. Ćalović Z. Viloria B. Nielsen F. G. Gmitter Jr. W. S. Castle J. W. Grosser 《Euphytica》2008,161(3):329-335
Five populations of lemon plants [Citrus limon (L.) Burm] obtained from undeveloped ovules through different tissue culture procedures were examined for the presence of
somaclonal and irradiation-induced genetic variation. Tested groups were: (1) nucellar seedlings; (2) organogenic, regenerated
via adventitious buds from nucellar seedling internodes; (3) embryogenic population, regenerated from non-irradiated nucellar
callus via somatic embryogenesis; (4) embryogenic population, regenerated from irradiated nucellar callus via somatic embryogenesis;
and (5) protoplast-derived, regenerated via somatic embryogenesis. Genomic DNA samples from 360 plants (72 from each group)
were screened for polymorphism among RAPD fingerprints amplified by 10 decamer primers. Among all tested plants, genetic variation
was detected only within the group of plants recovered from irradiated embryogenic calli. Out of 72 plants from that group,
three had RAPD fingerprints different from the rest of the population, and fourth plant was found to be cytochimeric, consisting
of diploid and tetraploid cells as revealed by flow cytometry. In all other populations of regenerated plants, we did not
come across any plants with changed ploidy level. 相似文献
4.
Four years' old micropropagated plants regenerated by enhanced axillary branching from shoot buds of a single genotype of
Robinia pseudoacacia were characterized by RAPDs. Random amplified polymorphic DNA analysis was carried outusing 19 random 10-mer DNA primers
and 286 RAPD bands were examined which showed 30% polymorphism. Similarity indices ranged from 0.86 to 0.96 among different
plants based on RAPD data. The UPGMA dendrogram was constructed based on similarity indices which showed clustering of different
plants into subgroups based on similarity values. Our results suggest that somaclonal DNA sequence variations are present
even when organized cultures such as shoot buds were used as explant for micro-propagation.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
5.
Summary DNA polymorphism among five Asparagus officinalis L. cultivars-Imperial, Snow, Steline, UC-157 and Larac, as detected by random amplified polymorphic DNA (RAPD), is reported. Thirty one decamer primers were tested. and twenty six of them yielded amplification products. Fourteen primers gave products with at least one polymorphic DNA fragment. Among a total of 119 amplified fragments 33 were polymorphic. These RAPD markers enabled the identification of asparagus cultivars. Unique markers for cultivars were: Snow-bands 475 bp, 772 bp, 412 bp, 935 bp and 820 bp amplified by primers D5, OPA-07, OPA-09, OPA-10 and OPA-18, respectively. Steline-bands 645 bp, 680 bp and 997 bp amplified by primers A32, OPA-03 and OPA-09, respectively. A band 903 bp, amplitied by primer OPA-12, is a marker for Imperial, and a band 420 bp, amplified by primer D52, is a marker for Larac. Cultivar UC-157 could be identified by a combination of shared polymorphic bands. The pairwise marker difference between cultivars ranged from 0.08 to 0.17. A phenogram of the genetic relationship based on RAPD fits with the known origin of the cultivars. 相似文献
6.
Summary Thirty-five rice (Oryza sativa L.) varieties, including 18 japonica, 5 javanica and 12 indica subspecies and 12 lettuce (Lactuca sativa L.) varieties were identified taxonomically, using PCR with originally designed 21 RAPD (Random Amplified Polymorphic DNA) primers and 8 sequence-specific primers, used for amplifying four specific DNA fragments. Use of these primers revealed polymorphisms among varieties in rice and lettuce and facilitates DNA fingerprinting. Dendrograms of both species based on polymorphisms were constructed and genetical relationships were established. In rice, half the number of amplified bands were polymorphic and almost all varieties differentiated. However, differentiation of minor genetic alterations among somaclonal variants or mutants and their mother varieties was not feasible. In L. sativa, 47% of the amplified fragments were polymorphic and all 12 varieties were differentiated. Some of the PCR fragments were variety or type specific, which could be used for indicators for type-selection. The dendrogram obtained showed differentiated clusters of crisphead, leaf and butterhead type, findings in good accord with the classification based on the genetic background. 相似文献
7.
Somatic hybrids were obtained from electrofused protoplasts derived from embryogenic suspension cultures of tetraploid cotton (G. hirsutum L. cv. Coker 201) and embryogenic callus of diploid wild cotton G. davidsonii. The regenerants were initially identified as hybrids by RAPD (random amplified polymorphic DNA) analysis. Subsequently, observation on chromosome counting, morphology and SSR (simple sequence repeat) confirmed the hybrid status. Cytological investigation of the metaphase root-tip cells of the regenerated plants revealed there were 74 to 84 chromosomes in the plants, close to the expected 78 chromosomes. SSR analysis revealed the regenerated plants contained specific genomic fragments from both fusion partners, further confirmed their hybridity. The morphology of the plants was intermediate between the two fusion partners. The regenerants were difficult to develop into mature plants because their roots browned and they wilted from the stem apex before forming 3 to 5 true leaves. The hybrid plants were transferred to soil by grafting in vitro onto rootstocks. 相似文献
8.
Amit Kaushik Navinder Saini Sunita Jain Poonam Rana R.K. Singh Rajinder K. Jain 《Euphytica》2003,134(2):231-238
Segregation for salinity tolerance and ISSR markers based molecular polymorphism were investigated in a F3 plant population raised via single-seed descent method from a cross between salt-tolerant indica rice variety CSR10 and salt-susceptible
premium traditional Basmati rice variety Taraori Basmati HBC19. A total of 130 F3plants were evaluated individually for salinity tolerance on 1–9 scale on the basis of seedling growth parameters; the average
score ranged between 1.7 to 8.3. Frequency distribution curve obtained using the salinity tolerance data of F3 population and a chi-square analysis, showed a good fit to a normal distribution. Eleven plants each in the category of salt-tolerant
and salt-susceptible were selected from the segregating F3 population for ISSR marker analysis. A total of 149 bands (4–11 bands per primer) ranging from 200 to 3530 bp were scored
for the two rice varieties and the selected CSR10 × HBC19 segregating F3 plants using 26 ISSR primers. Of these, 89 were monomorphic and 60 were polymorphic. Of the 60 polymorphic bands,36 and 20
bands were specific to CSR10 andHBC19 respectively. The remaining four bands were amplified using UBC primers 810,848, 853
and 886 and present in only some of the CSR10 × HBC19 F3 plants. Notably, ISSR primers with dinucleotide repeat motif and 5'-anchored end amplified more number of bands (7.0 bands/primer)
compared to3'-anchored dinucleotide primers (5.4bands/primer), but 3'-anchored dinucleotide primers revealed higher level
of polymorphism (2.6 polymorphic bands/primer) compared to 5'-anchoreddinucleotide primers (1.43 polymorphic bands/ primer).
While distribution of majority of the polymorphic bands were more or less in the expected ratios in salt-tolerant and/or salt-sensitive
F3segregating plants, but some of the bands amplified using UBC ISSR primers 823, 825,826, 849, 853, 864, 866 and 884 showed
highly skewed distribution. Such polymorphic bands stand greater chances of having a linkage with the genes/ QTLs for salinity
tolerance and shall be the target for further studies.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
9.
Riccardo Aversano Salvatore Savarese Jose Maria De Nova Luigi Frusciante Maria Punzo Domenico Carputo 《Euphytica》2009,165(2):353-361
In this work we detected the extent of variability at nuclear and cytoplasmic DNA level of regenerated plants belonging to
Solanum genotypes with a different genetic background and somatic chromosome number. As for the nuclear characterization, a total
of 66 (18.5%) polymorphic bands were scored using 13 ISSR primers on 45 randomly selected regenerants. Our results show that
the regenerants obtained from clone cmm 1T and, at lower level, those from cph 1C are unstable under in vitro conditions or
rather more prone to in vitro-induced stress leading to somaclonal variation than the other genotypes used. Two types of changes
were observed: disappearance of parental ISSR fragments, termed “loss”; appearance of novel ISSR fragments, termed “gain”.
The most frequent event occurring in the regenerants was the loss of fragments (41 bands). Regenerated plants were analyzed
with seven plastid universal primers to determine the cytoplasmic composition at chloroplast level. All cpDNA primer pairs
tested produced amplicons of the same size in all genotypes analyzed and no polymorphic fragments were observed with any universal
primers used. Our results show that under in vitro culture conditions genotype affects the integrity of the genome. In addition,
the absence of polymorphism at plastid level confirms the greater genetic stability of cytoplasmic DNA. 相似文献
10.
“薄皮”ד垂枝”板栗RAPD遗传连锁图构建 总被引：1，自引：1，他引：0
使用20组共400条RAPD随机引物,在"薄皮"和"垂枝"两个板栗亲本无性系及其5个杂交后代进行筛选,共有187条引物获得了清晰、重复性好的多态性条带,占供试引物的46.75%.应用筛选出的多态性引物在两亲本及其60个杂交后代中进行扩增,获得的多态性条带在后代中的分离比用卡方测验(α=0.05)分析,结果共有143个标记符合1:1测交分离比,其中来源于母本"薄皮"板栗的有88个,来源于父本"垂枝"的有55个.采用作图软件Mapmaker和回交群体模型分别对符合测交分离比的标记进行亲本特异的标记分群和连锁分析,设定LOD值为3和最大重组值0为0.50作为可信统计度与最大连锁标记数量之间的最佳组合,其中母本的标记定位了10个连锁组,父本的标记定位了7个连锁组.母本"薄皮'板栗遗传连锁图共包含88个标记,覆盖了板栗基因组总长约823.1 cM(Kosambi,以下同),占基因组的66.7%;父本"垂枝"板栗遗传连锁图共包含55个标记,覆盖了板栗基因组总长约720.8 cM,占基因组的41.7%.母本"薄皮"板栗遗传连锁图上标记间的遗传距离为1.6～26.5 cM,平均9.4 cM,连锁组的大小从15.5 cM到166.3 cM,平均为82.3 cM;父本"垂枝"板栗遗传连锁图上标记间的遗传距离为2.1～39.9 cM,平均13.1 cM,连锁组的大小从53.5 cM到139.6 cM,平均为103.0 cM. 相似文献
11.
基于RAPD标记的芥蓝种质资源遗传多样性分析 总被引：1，自引：1，他引：0
调查了44份芥蓝种质的植物学性状,并利用RAPD分子标记分析了其遗传多样性。结果从150条随机引物中筛选出20个引物,20个引物共扩增出177条谱带,其中多态谱带105条,平均每个引物扩增出8.9条谱带和5.3条多态性谱,多态性比率为59.32%。基于RAPD标记,利用NTSYS-pc2.11构建了聚类树状图谱,遗传距离为0.70时,44份芥蓝资源可聚成六大类群。芥蓝种质存在着一定的遗传多样性,但原产华南而且主要产区也在华南,遗传多样性要小于芸薹属其他蔬菜。 相似文献
12.
13.
Giuseppe Mandolino Andrea Carboni Manuela Bagatta V.M. Cristiana Moliterni Paolo Ranalli 《Euphytica》2002,126(2):211-218
DNA from female and male hemp (Cannabis sativa L.) plants belonging to nine different varieties were screened with180 RAPD primers in a search for sex-associated DNA markers.
About 1500bands were produced in total, nine primers were found yielding one or two DNA bands amplified in all nine male DNA
bulks and absent in all female DNA bulks. These putatively male-associated markers were then scored in three different F1progenies,
deriving from a cross between a common male parent and three different female plants. The sex of the progeny was accurately
scored on the basis of the floral phenotype, and the presence of the nine male-associated markers was verified by RAPD analysis.
In all three progenies examined, all the male plants showed the DNA markers previously identified by bulk segregant analysis
(BSA) on the hemp varieties, while all the female plants lacked them. The fact that the association between these markers
and the staminate phenotype is found when examining male plants of distantly related varieties, and that such linkage is never
broken when different progenies are examined, strongly supports the hypothesis that the markers found are physically located
on the Y chromosome, in a region excluded from recombination during meiosis. Another marker was shown to be present in the
male parent, in all the male plants of each progeny, and in 50% of the female progenies, while it was absent in the female
parent; the possible occurrence of markers deriving from multiple amplification sites of the genome is discussed.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
14.
Genetic variation among five elite winter barley cultivars (H. vulgare L.) currently grown in Bulgaria was assessed at the molecular level using restriction fragment length polymorphism (RFLP)
and randomly amplified polymorphic DNA (RAPD) markers. The present study sampled RFLPs in four well characterized multigene
families in barley: the seed storage protein loci; the 18S, 5.8S and 26S ribosomal DNA loci; the loci coding for 5S ribosomal
RNA and the loci coding subunit α of ATP-A complex in the mitochondrial genome. RFLPs were detected in three out of five investigated
chromosomal loci in the barley cultivars studied. RAPD assay using arbitrary 10-base primers was applied to generate amplified
length polymorphic markers in barley. Overall a total of 15 polymorphic phenotypes were found among the studied barley cultivars
by using 11 out of 25 tested primers. All RAPDs were considered as dominant genetic markers except for two, where PCR and
Southern blot
analysis indicated the presence of codominant amplification products. Five RAPD polymorphisms in F1 and F2 progenies of the cross between Alpha and Obzor were inherited in Mendelian fashion. The determined values for the genetic
variation proved a high genetic similarity among the tested cultivars. Genetic similarity (GS) calculated from RFLP and RAPD data ranged from 0.888 to 0.997 with a mean GS – 0.933.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
15.
This paper describes the relative efficiency of three marker systems, RAPD, ISSR, and AFLP, in terms of fingerprinting 14
rice genotypes consisting of seven temperatejaponica rice cultivars, three indica near-isogenic lines, three indica introgression lines, and one breeding line of japonica type adapted to high-altitude areas of the tropics with cold tolerance genes. Fourteen RAPD, 21 ISSR, and 8 AFLP primers
could produce 970 loci, with the highest average number of loci (92.5) generated by AFLP. Although polymorphic bands in the
genotypes were detected by all marker assays, the AFLP assay discriminated the genotypes effectively with a robust discriminating
power (0.99), followed by ISSR (0.76) and RAPD (0.61). While significant polymorphism was detected among the genotypes of
japonica and indica through analysis of molecular variance (AMOVA), relatively low polymorphism was detected within the genotypes of japonica rice cultivars. The correlation coefficients of similarity were significant for the three marker systems used, but only the
AFLP assay effectively differentiated all tested rice lines. Fingerprinting of backcross-derived resistant progenies using
ISSR and AFLP markers easily detected progenies having a maximum rate of recovery for the recurrent parent genome and suggested
that our fingerprinting approach adopting the ‘undefined-element-amplifying’ DNA marker system is suitable for incorporating
useful alleles from the indica donor genome into the genome of temperate japonica rice cultivars with the least impact of deleterious linkage drag. 相似文献
16.
17.
Aromatic rices are preferred by the consumers all over the world due to its flavour and palatability. Although a large number
of these collections are available, little systematic analysis of genetic diversity has been carried out. With the objective
of identification and classification of aromatic rice genotypes, RAPD profiling was employed using 58 random decamer primers.
Most of these primers (96.5%) detected polymorphism among the genotypes. Of the 465 amplified bands, 314 were polymorphic.
Cluster analysis based on Jaccard's similarity coefficient using UPGMA grouped all the traditional tall, photosensitive, low
yielding, long grained ‘basmati’ aromatics together. The short grained aromatic cultivars, formed a different cluster with
high level of average similarity among themselves. The dendrogram based on 58 primers was highly similar to that based on
10 and 15 primers with matrix correlation (r) of 0.88 and 0.91, respectively. This suggested that a set of 10 primers can
be employed for an initial assessment of genetic diversity in a large number of collections. All the rice genotypes included
in the study could be distinguished from each other at the level of 19 to 186 polymorphic bands between individuals in pair
wise comparison over all the 58 primers. Probability of identical profiles by chance suggested that about 1041 genotypes can be unambiguously differentiated by RAPD fingerprints obtained by 58 primers. A diagrammatic mode of presentation
of DNA fingerprints of the aromatic rices based on 10 of the informative primers was developed.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
18.
Identification and DNA polymorphism of the S‐locus receptor kinasegene (SRK) was analysed by pollen tube tests, polymerase chain reaction‐cleaved amplified polymorphic sequence (PCR‐CAPS) and nucleotide sequencing. SRK‐specific primers that can distinguish class and class II S haplotypes amplified single DNA fragments of 900‐1050 bp. The DNA fragments of 22 inbred lines amplified with a class SRK‐specific primer pair determined seven types with HinfI and EcoRII. In addition, the DNA fragments of 17 inbred lines amplified with a class II SRK‐specific primer pair determined three types with Hinf1. Nucleotide sequencing of the DNA fragments amplified from 10S haplotypes showed that exons of the 3′‐end in SRK are highly conserved, and that there is much variation of the introns, which produced polymorphism of the band pattern in PCR‐CAPS profiles. The S haplotypes of the plants were determined by restriction analysis of PCR products and agreed with results based on pollen tube growth tests. The PCR‐CAPS analysis using specific primer pairs of SRK is considered to be useful for S allele identification in breeding programmes. 相似文献
19.
20.
Summary The genetic similarities of eight closely related rye cultivars were estimated using two molecular marking techniques: restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD). Cultivars were evaluated for variation by 11 random cDNA and genomic clones used in combination with four restriction enzymes and 40 decamer primers. A total of 53 polymorphic RFLP fragments and 94 polymorphic RAPD fragments were observed. Based on the presence/absence of fragments, two genetic similarity matrices were calculated which were then used in cluster analysis. Differences between pair of cultivars were observed in RFLP and RAPD dendrograms. RFLP analysis produced estimates of genetic relationships more in accordance with the partially known pedigree of the cultivars than did RAPD analysis. The use of bulk samples of DNA in these analyses affected the sensitivity of RAPD assays more strongly. Dendrograms which took into account all fragments produced, either by RFLP or RAPD, reflected better the relationships between cultivars than did dendrograms based on only one type of marker. This reflects the importance of the number of markers used in determining the genetic relationships between genotypes. 相似文献