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Zhi-li Suo, Wen-ying Li, Juan Yao, Hui-jin Zhang, Zhi-ming Zhang, and Di-xuan Zhao

Tree peony cultivars are usually classified according to flower characteristics (flower form and flower color) which are commonly affected by environmental influences and developmental levels. Judgment of flower forms may also depend on the observer. Precise and rapid cultivar identification methods are also required to manage cultivar collections as well as tree peony breeding programs. The objective of this paper is to analyze the discriminatory ability of leaf morphology and Intersimple sequence repeat (ISSR) marker systems for tree peony cultivars. As a result, although there exist large variations of leaf morphology of tree peony cultivars, the morphological characteristics of biternately compound leaves 3, 4, and 5 from the base of a shoot at the middle part of a plant are relatively stable with smaller variations within cultivars (2.7% to 27.1%, 16.8% on average) and with larger differentiations among cultivars (72.9% to 97.3%, 83.2% on average). Statistical and principal components analyses indicate that 12 leaf morphological characteristics are valuable for cultivar classification. ISSR markers present a precisely discriminatory power in tree peony cultivar classification without environmental influences. The cultivars with multiple flower forms, which makes it difficult to make judgment by means of a flower-form-based classification system, have been significantly characterized using leaf morphology or ISSR markers.

Open access

Xiuli Lv, Yuan Guan, Jian Wang, Yanwei Zhou, Qunlu Liu, and Zequn Yu

To reveal the genetic diversity and genetic relationships of China’s Bergenia germplasm, 28 Bergenia accessions from different regions in China were analyzed by 24 intersimple sequence repeat (ISSR) markers. The results showed that 318 sites were amplified in all germplasm, including 307 polymorphic sites, and the percentage of polymorphic sites was 96.54%. Cluster analysis showed that the 28 accessions were divided into three categories, with a similarity coefficient of 0.5475. Bergenia purpurascens was clustered into one category; B. scopulosa was clustered into one category; and B. tianquaninsis, B. emeiensis, B. stracheyi, and B. crassifolia were clustered into one category. The results of the cluster analysis indicated that the 28 accessions were not completely classified by origin. Using the ISSR marker technique to analyze the phylogenetic relationship of Bergenia germplasm is helpful for identifying valuable resources and providing a theoretical basis for the selection of breeding parents.

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Kirk W. Pomper, Sheri B. Crabtree, Shawn P. Brown, Snake C. Jones, Tera M. Bonney, and Desmond R. Layne

The pawpaw [Asimina triloba (L.) Dunal.] is a tree fruit native to many areas of the southeastern and mid-western United States. Kentucky State University (KSU) is designated as a satellite repository for Asimina for the U.S. Department of Agriculture (USDA), National Plant Germplasm System (NPGS). An assessment of the level of genetic diversity in cultivated pawpaw would assist in development of the future germplasm repository collection strategies for cultivar improvement. The objectives of this study were to identify intersimple sequence repeat (ISSR) markers that segregate in a simple Mendelian fashion and to use these markers to assess genetic diversity in 19 pawpaw cultivars. Leaf samples from the 34 progeny of controlled crosses (1-7-1 × 2-54 and reciprocal) and the parents were collected, DNA was extracted, and subjected to the ISSR methodology using the University of British Columbia microsatellite primer set #9. Seven primers yielded 11 Mendelian markers with either a 3:1 or 1:1 ratio that was confirmed by chi-square analysis. Analysis of genetic diversity using 10 of the ISSR markers from 19 pawpaw cultivars revealed a moderate to high level of genetic diversity, with a percent polymorphic loci P = 80 and an expected heterozygosity He = 0.358. These diversity values are higher than those reported for cultivated pawpaw using isozyme or randomly amplified polymorphic DNA (RAPD) markers, indicating that the ISSR marker methodolgy has a higher level of discrimination in evaluating genetic diversity in pawpaw and/or pawpaw has greater levels of genetic diversity than previously found.

Free access

Daniel Potter, Fangyou Gao, Giovanna Aiello, Charles Leslie, and Gale McGranahan

The utility of intersimple sequence repeat (ISSR) markers for identification of English or Persian walnut (Juglans regia L.) cultivars was explored. Four cultivars were screened with 47 ISSR primers; eight of these primers, which generated reproducible and informative data, were selected for further study. Two individuals from each of 48 cultivars, including many currently important in the California walnut industry as well as accessions from Europe and Asia, were then examined with the eight ISSR primers. Polymerase chain reaction (PCR) products were separated on agarose gels and stained with ethidium bromide. Fifty-four bands were scored as present or absent in each cultivar; of these, 31 (57%) were polymorphic among the 48 cultivars. Combined data from the eight ISSR primers provided a unique fingerprint for each of the cultivars tested. Fifteen of the cultivars could be distinguished from all others with just one primer, 31 with a minimum of two primers, and two required three primers. Pairwise genetic distances between the cultivars were calculated and a dendrogram was generated using the neighbor-joining algorithm. Some of the groupings in the dendrogram corresponded to groups which, based on known pedigrees, are genealogically closely related. Others included accessions from diverse genetic and/or geographic origins. These results can be attributed to a combination of the limitations of the ISSR method for inferring genetic relationships, on the one hand, and the complex history of walnut cultivar development involving extensive exchange and breeding of germplasm from different geographic regions, on the other.

Free access

Kai-Ge Zhao, Ming-Qin Zhou, Long-Qing Chen, Donglin Zhang, and Gituru Wahiti Robert

repeat marker and RAPD amplifications. Intersimple sequence repeat marker amplification was performed in a 25-μL reaction volume, containing 40 ng of template DNA, 1× PCR buffer (MBI Fermentas, Lithuania), 2 m m MgCl 2 , 0.2 m m each of dNTP, 0.2 μ m

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Jian-Feng Geng, Cheng-Song Zhu, Xiao-Wei Zhang, Yan Cheng, Yuan-Ming Zhang, and Xi-Lin Hou

Brassica rapa L. ssp. chinensis (L.) Hanelt, known as nonheading chinese cabbage in China, is an important vegetable in eastern Asia and its genetic improvement requires a genetic linkage map. The first genetic linkage map of nonheading chinese cabbage using 112 doubled haploid lines derived from a released F1 hybrid cultivar Shulü between two lines SW-3 and Su-124 was constructed in this paper. One hundred thirty-eight molecular markers were mapped into 14 linkage groups. Among these markers, there were 77 sequence-related amplified polymorphism markers, 27 simple sequence repeat markers, 21 random amplification polymorphic DNA markers, and 13 intersimple sequence repeat markers. Chi-square tests showed that 54 markers are distorted from Mendelian segregation ratios, and the direction of the distortion is mainly toward the maternal parent SW-3. The distortion affects not only the estimation of genetic distance, but also the order of distorted markers on a same linkage group. Given a specific marker order, the authors proposed a multipoint approach to correct the linkage map in an unbiased manner in an F2 population while considering distorted, dominant, and missing markers. A new method was used to correct the linkage map in the doubled haploid population mentioned earlier considering new, distorted, and missing markers. The total length of the corrected linkage map was 1923.75 cM, with an average marker spacing of 15.52 cM. The map will facilitate selective breeding and mapping of quantitative trait loci.

Full access

Yi-Lun Liao, Wen-Shin Lin, and Shu-Yun Chen

variation among all accessions tested by intersimple sequence repeat markers was shown in a previous study ( Xi et al., 2016 ). The authors also reported that the proportion of triglyceride in the accessions test could be a useful indicator for quality

Free access

Todd A. Burnes, Robert A. Blanchette, Jason A. Smith, and James J. Luby

in labeling or propagation may have occurred and it was wrongly designated as immune. Recently, fingerprinting techniques using random amplified polymorphic DNA and intersimple sequence repeat markers have become available to determine relatedness of

Free access

Zhi-li Suo, Xiao-qing Zhao, Jian-peng Zhao, Xiao-chong Zhao, and Fu-fei Chen

.X. 2005 Application of leaf morphology and intersimple sequence repeat markers in classification of tree peony (Paeoniaceae) cultivars HortScience 40 329 334 Wang, L.Y. 1998 Chinese tree peony (English ed.) China For

Free access

Xinyi Zhang, Li Liao, Zhiyong Wang, Changjun Bai, and Jianxiu Liu

), amplified fragment length polymorphisms (AFLPs), and simple sequence repeats (SSRs) ( Kalia et al., 2011 ). Intersimple sequence repeat markers can be amplified simply and directly without knowledge of the flanking sequences, thus enabling easier development