Abstract
BackgroundAt present, the distinctness, uniformity, and stability (DUS) testing of flue-cured tobacco (Nicotiana tabacum L.) depends on field morphological identification, which is problematic in that it is labor intensive, time-consuming, and susceptible to environmental impacts. In order to improve the efficiency and accuracy of tobacco DUS testing, the development of a molecular marker-based method for genetic diversity identification is urgently needed.ResultsIn total, 91 simple sequence repeats (SSR) markers with clear and polymorphic amplification bands were obtained with polymorphism information content, Nei index, and Shannon information index values of 0.3603, 0.4040, and 0.7228, respectively. Clustering analysis showed that the 33 study varieties, which are standard varieties for flue-cured tobacco DUS testing, could all be distinguished from one another. Further analysis showed that a minimum of 25 markers were required to identify the genetic diversity of these varieties. Following the principle of two markers per linkage group, 48 pairs of SSR markers were selected. Correlation analysis showed that the genetic relationships revealed by the 48 SSR markers were consistent with those found using the 91 SSR markers.ConclusionsThe genetic fingerprints of the 33 standard varieties of flue-cured tobacco were constructed using 48 SSR markers, and an SSR marker-based identification technique for new tobacco varieties was developed. This study provides a reliable technological approach for determining the novelty of new tobacco varieties and offers a solid technical basis for the accreditation and protection of new tobacco varieties.
Highlights
At present, the distinctness, uniformity, and stability (DUS) testing of flue-cured tobacco (Nicotiana tabacum L.) depends on field morphological identification, which is problematic in that it is labor intensive, timeconsuming, and susceptible to environmental impacts
Thereafter, China developed and released the first domestic tobacco DUS testing standard, the Guidelines for the Conduct of Tests for Distinctness, Uniformity, Stability – FlueCured Tobacco (Nicotiana tabacum L.; YC/T 369–2010) [5], which was based on the General Directives for the Conduct of Tests of Distinctness, Uniformity, Stability for New Varieties of Plants (GB/T 19557.1–2004) [6] and the tobacco testing guidelines of the UPOV [4]
We addressed the lack of molecular marker-based technologies for estimating the distinctness, uniformity, and stability of flue-cured tobacco varieties by carrying out a population genetics study and constructing simple sequence repeats (SSR) fingerprints of 33 standard flue-cured tobacco varieties that are commonly used in DUS testing [5]
Summary
The distinctness, uniformity, and stability (DUS) testing of flue-cured tobacco (Nicotiana tabacum L.) depends on field morphological identification, which is problematic in that it is labor intensive, timeconsuming, and susceptible to environmental impacts. Distinctness, uniformity, and stability (DUS) are three technical and scientific criteria for the protection of new plant varieties [2, 3]. Using the UPOV DUS testing guidelines as an example, China developed a series of crop DUS testing guidelines and promoted the use of these guidelines in the protection of new plant varieties [3]. Thereafter, China developed and released the first domestic tobacco DUS testing standard, the Guidelines for the Conduct of Tests for Distinctness, Uniformity, Stability – FlueCured Tobacco (Nicotiana tabacum L.; YC/T 369–2010) [5], which was based on the General Directives for the Conduct of Tests of Distinctness, Uniformity, Stability for New Varieties of Plants (GB/T 19557.1–2004) [6] and the tobacco testing guidelines of the UPOV [4]
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