Management of Vitis and Brassica Germplasm

We discuss a series of studies in our units employing molecular genetic markers in collection management, primarily for identity and diversity assessment and partitioning of genetic variation. Isozymes, random amplified polymorphic DNAs (RAPDs), and simple sequence repeat DNAs (SSRs) have been used for these purposes. We analyzed a range of Brassica oleracea accessions at six isozyme loci. Unique isozyme profiles (or fingerprints) were found for 40% of the individual genotypes within accessions. While isozymes were extremely valuable for partitioning genetic variability between and among subspecies, they failed to identify accessions and subspecies. Furthermore, relationships found did not correspond to those predicted by taxonomy. In a study of three species of Chinese vegetable brassicas using 112 RAPD markers, we were able to unambiguously distinguish all 52 accessions studied, despite some intra-accession variability. In addition, cluster analysis correctly grouped all individuals of the same species, but below that rank, taxonomic groupings occasionally broke down. RAPD profiles were found that unambiguously distinguished the three Brassica species from one another, but, for subspecies, no such profiles were found. In another RAPD study of B. oleracea subsp. capitata (cabbage), a closely related set of cultivars were not distinguishable, although more distantly related cultivars were. We had disappointing results with a RAPD study of Vitis accessions. DNA was extracted from the leaves of 23 greenhouse-grown and 52 field-grown vines. Twelve of the 23 greenhouse vines were rooted cuttings collected from 12 of the field-grown vines. Unfortunately, the RAPD profiles of all vines grown in the same location (whether greenhouse or vineyard) were more similar to one another than were profiles from the same clone grown in the two different locations. We are studying whether this result is due to physiological differences in plants growing under different conditions, to differences between PCR reagent lots, to pathogen infestation, or to DNA sample contamination. In a study of 23 accessions representing 15 Vitis species and three species hybrids, we used six different SSR markers to identify individual genotypes. We were able to unambiguously distinguish all genotypes, except two that were identical at all six loci. Review of planting records revealed that the two genotypes were probably the same grape clone. SSR results were not congruent with known taxonomic relationships or geographic origin of genotypes. The SSR polymorphisms found in even this small subset of the Vitis collection in principle make possible the identification of more than 130 trillion different genotypes. This high level of polymorphism, however, makes our particular SSR loci of limited use for identification of species and for the determination of genetic relationships. Molecular genetic markers offer a powerful, efficient approach to assessing questions of identity, relationship, and diversity in germplasm collections, but markers need to be selected based on their suitability for the particular task.

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Molecular genetic markers for thyroid FNAB

Nuklearmedizin ◽

10.1055/s-0037-1616610 ◽

2015 ◽

Vol 54 (03) ◽

pp. 94-100 ◽

Cited By ~ 4

Author(s):

P. B. Musholt ◽

T. J. Musholt

Keyword(s):

Genetic Markers ◽

Thyroid Nodules ◽

Molecular Genetic ◽

Genetic Alterations ◽

Diagnostic Tools ◽

Ultrasound Screening ◽

Thyroid Malignancy ◽

Thyroid Carcinomas ◽

Molecular Genetic Markers ◽

The Impact

SummaryAim: Thyroid nodules > 1 cm are observed in about 12% of unselected adult employees aged 18–65 years screened by ultrasound scan (40). While intensive ultrasound screening leads to early detection of thyroid diseases, the determination of benign or malignant behaviour remains uncertain and may trigger anxieties in many patients and their physicians. A considerable number of thyroid resections are consecutively performed due to suspicion of malignancy in the detected nodes. Fine needle aspiration biopsy (FNAB) has been recommended for the assessment of thyroid nodules to facilitate detection of thyroid carcinomas but also to rule out malignancy and thereby avoid unnecessary thyroid resections. However, cytology results are dependent on experience of the respective cytologist and unfortunately inconclusive in many cases. Methods: Molecular genetic markers are already used nowadays to enhance sensitivity and specificity of FNAB cytology in some centers in Germany. The most clinically relevant molecular genetic markers as pre-operative diagnostic tools and the clinical implications for the intraoperative and postoperative management were reviewed. Results: Molecular genetic markers predominantly focus on the preoperative detection of thyroid malignancies rather than the exclusion of thyroid carcinomas. While some centers routinely assess FNABs, other centers concentrate on FNABs with cytology results of follicular neoplasia or suspicion of thyroid carcinoma. Predominantly mutations of BRAF, RET/PTC, RAS, and PAX8/PPARγ or expression of miRNAs are analyzed. However, only the detection of BRAF mutations predicts the presence of (papillary) thyroid malignancy with almost 98% probability, indicating necessity of oncologic thyroid resections irrespective of the cytology result. Other genetic alterations are associated with thyroid malignancy with varying frequency and achieve less impact on the clinical management. Conclusion: Molecular genetic analysis of FNABs is increasingly performed in Germany. Standardization, quality controls, and validation of various methods need to be implemented in the near future to be able to compare the results. With increasing knowledge about the impact of genetic alterations on the prognosis of thyroid carcinomas, recommendations have to be defined that may lead to individually optimized treatment strategies.

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