A Multiplex PCR assay for Discriminating Charlock from Rapeseed: Implications for Seed Testing

Author: Seyed Hossein Jamali, Leila Sadeghi and Maryam Najafian

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Abstract

So far, a number of specific markers have been introduced for genome identification and phylogenic studies in Brassicaceae family. We introduced a new application for these markers i.e. identifying charlock as a noxious weed in seed samples of rapeseed. A multiplex PCR assay was designed based on 625, 190 and 325 bp fragments of Brassica Transparent Testa Glabra 1 (TTG1) gene, 5S ribosomal DNA, and cruciferin gene as internal control. The resulting presence/absence amplification pattern could differentiate Sinapis arvensis genome (SarSar) from that of Brassica napus (AACC). As an alternative to chemical tests, this assay can complement visual identification wherein charlock seeds are discerned based on seed morphology by seed analysts. However, the discriminating characteristics are completely hidden once working samples are taken from film-coated seeds. Moreover, interaction of chemicals used in film coating might result in misclassification. Therefore, the introduced molecular assay

Keywords

charlock, rapeseed, noxious weed, seed testing

Conclusion

So far, a number of specific markers have been introduced for genome identification and phylogenic studies in Brassicaceae family. We introduced a new application for these markers i.e. identifying charlock as a noxious weed in seed samples of rapeseed. A multiplex PCR assay was designed based on 625, 190 and 325 bp fragments of Brassica Transparent Testa Glabra 1 (TTG1) gene, 5S ribosomal DNA, and cruciferin gene as internal control. The resulting presence/absence amplification pattern could differentiate Sinapis arvensis genome (SarSar) from that of Brassica napus (AACC). As an alternative to chemical tests, this assay can complement visual identification wherein charlock seeds are discerned based on seed morphology by seed analysts. However, the discriminating characteristics are completely hidden once working samples are taken from film-coated seeds. Moreover, interaction of chemicals used in film coating might result in misclassification. Therefore, the introduced molecular assay

References

In spite of being a valuable source of resistance to diseases and pod shattering (Snowdon et al., 2000; Liu et al., 2014) and recently novel type of cytoplasmic male sterility (Liu et al., 2015), charlock or wild mustard (Sinapis arvensis L.) is a notorious weed of Brassica crops. Moreover, when it comes to determining other seeds in rapeseed (Brassica napus L.) seed samples, it is also considered as a noxious weed along with other seeds like wild radish (Raphanus raphanistrum) and turnip weed (Rapistrum rugosum). Maximum number of restricted seeds in a noxious weed exam varies between countries as defined in national seed standards. For instance, Iran's seed standard tolerate up to seven weed seed (including charlock) in a 100g sample of rapeseed. There is also much more stringent standard by restricting up to two seeds in a half pound sample of certified rapeseed bag as mandated by Oregon Seed Certification Service, USA. Although rapeseed and charlock could be roughly distinguished f

How to cite this article

Seyed Hossein Jamali, Leila Sadeghi and Maryam Najafian (2017). A Multiplex PCR assay for Discriminating Charlock from Rapeseed: Implications for Seed Testing , Biological Forum – An International Journal 9(2): 87-91.