Abstract
Chrysanthemummorifolium Ramat. ‘Daboju’ is a C. morifolium cultivar with important ornamental and medicinal values, and is often used in the treatment of colds, blurred vision, dizziness, and itchy skin. As the morphological characteristics of C. morifolium ‘Daboju’ are very similar to those of other C. morifolium cultivars, they are often confused in practice. However, the medicinal value and practical use of C. morifolium depends on using the correct rapid and accurate identification of C. morifolium ‘Daboju’ and its differentiation from other, morphologically similar C. × morifolium cultivars. Twenty-one polymorphic start codon-targeted (SCoT) primers were amplified in 21 distinct C. morifolium cultivars. One cultivar-specific DNA marker was developed with the aim of the rapid and accurate identification of C. morifolium ‘Daboju’ and its differentiation from other, similar C. morifolium cultivars. Twenty-one polymorphic start codon-targeted (SCoT) primers were amplified in 21 distinct C. morifolium cultivars. One cultivar-specific 385-bp amplicon (named SCoT36-385), amplified only in C. morifolium ‘Daboju’ (and in all samples of this cultivar), was identified, cloned, and sequenced. Subsequently, a sequence-characterized amplified region (SCAR) marker (named DBJF/DBJR), generating a 360-bp amplicon, was developed from SCoT36-385 and tested for amplification in all 21 C. morifolium cultivars, ten C. morifolium ‘Daboju’ populations, and different simulated adulterations of ‘Daboju’ with other cultivars. The primers amplified the specific 360-bp-long DNA fragment in all the tested C. morifolium ‘Daboju’ samples but failed in the absence of ‘Daboju’. The detection limit of the SCAR primer pair (DBJF/DBJR) was 100 pg of DNA extracted from C. morifolium ‘Daboju’. Hence, this SCAR marker has a very high detection sensitivity, and can be used for accurate and rapid identification of C. morifolium ‘Daboju’. It can play an important role in ensuring the quality of medicinal preparations and protecting C. morifolium ‘Daboju’ germplasm resources in breeding programs and in identifying lines generated from this cultivar.
Highlights
Chrysanthemum, Chrysanthemum morifolium Ramat., belongs to the family Asteraceae, and it has a long history of cultivation and use as a popular ornamental and herbal medicine use and cultivation in China, South Korea, Thailand, and Japan [1,2]
We developed a useful sequence-characterized amplified region (SCAR) marker for ‘Daboju’ identification via the use of start codon-targeted (SCoT) primers, which are PCR-based gene-targeted markers, in which the SCoT primers are based on conserved regions flanking the translation start codon of genes [38]
The results showed that a clear, specific amplicon of bp was generated from the ‘Daboju’ DNA samples by the primer pair, but no amplicons were detected from DNA samples from any of the other 20 chrysanthemum cultivars
Summary
Chrysanthemum, Chrysanthemum morifolium Ramat., belongs to the family Asteraceae, and it has a long history of cultivation and use as a popular ornamental and herbal medicine use and cultivation in China, South Korea, Thailand, and Japan [1,2]. As a traditional medicinal plant, C. morifolium flowers have significant pharmacological effects for treating headaches, dizziness, sore carbuncles, swelling, poison, and cold or wind heat [3]. As growing conditions and processing methods vary between production areas, different regions produce different medicinal preparations. ‘Chuju,’ ‘Gongju,’ ‘Hangju,’ or ‘Huaiju’ types, based on origin and processing methods, are legal Chinese medicinal materials, according to the 2015 edition of the Pharmacopoeia of the People’s Republic of China [3]. C. morifolium preparations are slightly cold in effect, with a sweet and bitter taste, with effects on wind, clearing heat, liver problems (ping mingmu tea), and for detoxification issues. Famous Chinese herbal medicine books, such as the ’Chinese Medicine Dictionary’ and ’Chinese Materia Medica,’ recorded that the ‘Boju’ type was the highest quality of medicinal chrysanthemum in China, and was mainly produced in Bozhou City, An’hui Province. In 2014, the Ministry of Agriculture of the People’s Republic of China officially approved the registration and awarded the protection of agricultural products from ‘Boju’ under geographical indication [3]
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