Celery cultivars (Apium graveolens var. dulce) in North America have a narrow genetic base. Twenty-two celery, one celeriac and one annual cultivar were screened for polymorphic RAPD (Random Amplified Polymorphic DNA) markers with 28 arbitrary 10-mer primers. Among the total 231 bands obtained, 28 (12%) of the bands were polymorphic among the 24 accessions screened, but only 18 (7.8%) were polymorphic within the 22 celery cultivars. These markers are sufficient to distinguish each of the cultivars used. The average number of marker differences is 6.2 between two celery cultivars, 13.5 between the celeriac and the remaining cultivars, and 16.5 between the annual and the other cultivars. The relationship among the celery cultivars disclosed from this study is basically consistent with that observed using total protein and isozyme markers. RAPD technology provides a new alternative for cultivar identification in celery.
Xiaofeng Yang and Carlos F. Quiros
Dror Sharon, Avital Adato, Samir Mhameed, Uri Lavi, Jossi Hillel, Maria Gomolka, Conny Epplen and Jorg Thomas Epplen
Plant genomes contain polymorphic repetitive sequences that can be used as DNA markers. Minisatellites (16 to 64 bp per repeat) and simple-sequence repeats (2 to 6 bp per repeat) are the most polymorphic markers found in plant and animal genomes. In this study, the hybridizations between genomic DNA and variable number of tandem repeat probes were examined in Arabidopsis thaliana L. (Heynn), onion (Allium cepa L.), tomato (Lycopersicon esculentum L.), wheat (Triticum aestivum L.), avocado (Persea americana Mill.), litchi (Chinensis Sonn.), mango (Mangifera indica L.), and Carica species. Some of the probes detected polymorphic sequences in all the species, but others were useful only for one or two species. None of the probes gave clear band patterns in either onion or wheat. The in-gel hybridization method was similar to Southern blot hybridization using the simple-sequence repeat probes.
François Luro, Frédéric Laigret, Joseph-Marie Bové and Patrick Ollitrault
We used three short repetitive nucleotide sequences [(GTG)5, (TCC)5, and (GACA)4] either as radiolabeled probes for hybridization with restricted citrus DNA or as single primers in polymerase chain reaction amplification experiments with total genomic DNA. We tested the ability of the sequences to discriminate between seedlings of zygotic or nucellar origin in the progeny of a Volkamer lemon (Citrus volkameriana Ten. & Pasq.) tree. The genetic variability within two species [Citrus sinensis (L.) Osbeck (sweet oranges) and Citrus reticulata Blanco and relatives (mandarins)] also was evaluated. DNA amplified fingerprinting with single primers was the more successful technique for discriminating between nucellar and zygotic seedlings. Although we were not able to distinguish among 10 cultivars of C. sinensis, all 10 C. reticulata cultivars tested were distinguishable. However, it still is difficult to identify the putative parents of a hybrid plant when the two parental genomes are closely related.
N. Nikoloudakis, G. Banilas, F. Gazis, P. Hatzopoulos and J. Metzidakis
Random amplified polymorphic DNA (RAPD) markers were used to study the genetic diversity and to discriminate among 33 Greek olive (Olea europaea L.) cultivars. Three feral forms from Crete and five foreign cultivars recently introduced into Greece were also included. Nineteen primers were selected which produced 64 reproducible polymorphic bands in the 41 olive genotypes studied, with an average of 3.4 informative markers per primer. The RAPD markers resulted in 135 distinct electrophoretic patterns, with an average of 7.1 patterns per primer. Based on either unique or combined patterns, all genotypes could be identified. Genetic similarities between genotypes were estimated using the Dice similarity index and these indicated that a high degree of diversity exists within the Greek olive germplasm. Using the unweighted pair-group method (UPGMA) most cultivars were clustered into two main groups according to their fruit size or commercial use (table or olive oil). However, poor correlation was detected between clustering of cultivars and their principal area of cultivation. RAPD marker data were subjected to nonmetric multidimentional scaling (NMDS) which produced results similar to those of the UPGMA analysis. The results presented here contribute to a comprehensive understanding of cultivated Greek olive germplasm and provide information that could be important for cultural purposes and breeding programs.
Chemda Degani, Jiusheng Deng, Avigdor Beiles, Ruth El-Batsri, Moshe Goren and Shmuel Gazit
There is widespread confusion and uncertainty concerning the identity of lychee cultivars: the same cultivar may be known under different names and different cultivars may appear under the same name. In the present study, the potential of intersimple sequence repeat (ISSR) for the identification of 66 lychee cultivars and accessions and a determination of their genetic relationships was evaluated, using 32 primers containing different simple sequence repeat motifs. Of the 194 bands produced, 124 (64%) were polymorphic. A set of six ISSR primers was sufficient to distinguish all cultivars and accessions. Thus, cultivars which are morphologically very similar and have identical isozyme profiles can be distinguished by ISSR analysis. However, seven pairs of accessions, each considered to be the same cultivar, were found to be identical by ISSR analysis. Nei and Li band-sharing distances and Nei genetic distances were calculated among the cultivars and two similarity dendrograms were generated using the neighbor-joining algorithm. Results showed that the ISSR technique is a valuable tool for identification of lychee cultivars and analysis of their genetic relationships.
Young-ju Kim and David H. Byrne
Isozyme analysis has been used for cultivar identification, but little has been done with the genus Rosa. One hundred and sixty rose accessions (species, cultivars, and hybrids) were characterized for isozyme phenotypes using starch gel electrophoresis. Six enzyme systems were stained on three electrode buffer systems. ACP, MDH, and 6PGD were run on morpholine citrate (pH 6.1) and histidine (pH 5.7), PGI and PGM were run on histidine (pH 5.7) and lithium borate (pH 8.3), and SKDH was run on morpholine citrate (PH 6.1) and lithium borate (PH 8.3). The most variable isozymes were MDH and 6PGD. MDH and 6PGD revealed 10 and 9 bands, respectively. This study showed that isozyme variability exists in roses and can be useful in their classification into species groups.
Arthur Q. Villordon and Don R. LaBonte
Clonal propagation assures the maintenance of genetic purity of a sweetpotato variety. The existence of foundation seed programs further contributes to the conservation of favorable genetic constitution in a commercial cultivar. However, the improvement of current maintenance procedures is necessary as shown by the occurrence of mutations and the decline of certain commercial varieties. Information on the nature and extent of changes in sweetpotato would therefore be useful in this regard.
`Jewel' clones obtained from eight state foundation seed programs were subjected to yield tests and a RAPD-based assay. Differences in nearly all yield grades were detected during the 1991, 1992, and 1993 seasons. The yield of U.S. No. 1 grade roots varied from 27% to 46%. The quality factors measured also varied: % alcohol insoluble solids varied by 13%, while sucrose ranged from 9.6% to 19%. Total DNA was extracted from each clone and assayed against 40 primers. All primers produced amplified fragments. A total of 110 reproducible bands was generated by 38 primers. Putative polymorphic markers were scored in 21 (18.58%) of these bands based on the presence or absence of amplified products. The results suggest an underlying cause for the variability observed in phenotypic traits within sweetpotato clones.
J. Baral and P.W. Bosland
Domesticated chile (Capsicum annuum L. var. annuum) is a widely cultivated spice and vegetable crop. It originated in the Western Hemisphere, but spread rapidly throughout the globe after the voyage of Columbus. However, very little is known about the genetic diversity of chile in Asia and especially in Nepal. Thus, research was conducted to document morphological as well as molecular characterization of C. annuum var. annuum landraces collected from Nepal. Genetic diversity in C. annuum var. annuum landraces from Nepal was investigated using randomly amplified polymorphic DNA (RAPD) markers and compared with that of C. annuum var. annuum landraces from the center of diversity, Mexico. RAPD marker based cluster analysis of C. annuum var. annuum clearly separated each accession. All accessions of C. annuum var. annuum from Nepal grouped into a single cluster at a similarity index value of 0.80, whereas, accessions from Mexico grouped into eight different clusters at the same similarity level indicating greater genetic diversity in Mexican accessions. RAPD analysis indicated that the Nepalese chile population went through an additional evolutionary bottleneck or founder effect probably due to intercontinental migrations. Some Nepalese accessions had unique RAPD markers suggesting that additional sources of genetic variation are available in Nepalese germplasm.
Jit B. Baral and Paul W. Bosland
The species status of two morphologically closely related species, Capsicum frutescens L. and C. chinense Jacq., was investigated using typological, phylogenetic, and biological species concepts. Diagnostic morphological differences, randomly amplified polymorphic DNA (RAPD) marker-based cluster analysis, and hybrid analyses were used to delimit C. frutescens and C. chinense. In many cases two morphological characters, calyx constriction and flower position, can separate accessions of C. frutescens from C. chinense. The RAPD-based analysis clearly separated accessions of C. frutescens and C. chinense into two distinct groups. The average genetic similarity within C. frutescens and C. chinense accessions was 0.85 and 0.8, respectively, whereas the average genetic similarity between C. frutescens and C. chinense accessions was only 0.38. The progenies obtained from C. frutescens and C. chinense hybridization had reduced fertility. Based on these evidences, C. frutescens and C. chinense represent two morphologically diagnosable, phylogenetically distinct, and reproductively isolated species.
G. Acquaah, T.G. Isleib and A.E. Ferguson
The future of molecular markers in common bean (Phaseolus vulgaris L.) breeding and genetics lies in the discovery of more useful markers than now available. One-dimensional SDS/PAGE analysis revealed four phaseolin types, “S,” “T,” “C,” and “H,” in proportions of 23.5%, 49.2%, 24.8%, and 2.5%, respectively. Molecular heterogeneity of phaseolin subunits was not apparent. On the basis of the phaseolin types and seed size, ≈75% of the landraces from Malawi probably were introduced from the Andean primary center of common bean domestication. The remaining 25% were small-seeded and probably originated from the Meso-American center of domestication of common bean. In Malawi, some amount of hybridization has occurred between genotypes from the two centers of domestication.