Limits...
Molecular cloning and development of RAPD-SCAR markers for Dimocarpus longan variety authentication.

Yang L, Fu S, Khan MA, Zeng W, Fu J - Springerplus (2013)

Bottom Line: The specific bands with expected sizes were amplified in five D. longan samples but not in others.To identify and characterize the difference between D. longan and D. confinis, PCR amplifications were performed again.Therefore, our study provides an effective and precise PCR-based diagnostic method and markers to identify D. longan species.

View Article: PubMed Central - PubMed

Affiliation: Research Center for Preclinical Medicine, Luzhou Medical College, Luzhou, Sichuan 646000 China.

ABSTRACT
As an edible fruit and source of traditional medicine, D. longan is grown in most areas of Southern China. Identification of D. longan cultivars by using molecular markers is important genetically. In this study, we cloned fragments from improved randomly amplified polymorphic DNA (RAPD), and developed stably diagnostic sequence-characterized amplified region (SCAR) markers. The specific RAPD bands of D. longan cultivars from Guangxi, with size ranging from 500 bp to 900 bp were gel-purified, cloned and sequenced. Four clones named LY2-1, LY4-7, LY4-8 and LY5-2 were identified. In order to investigate whether the fragments were specific for the species, four pairs of SCAR primers were then designed. PCR amplifications were conducted to analyze 18 samples including different D. longan cultivars and other species. The specific bands with expected sizes were amplified in five D. longan samples but not in others. To identify and characterize the difference between D. longan and D. confinis, PCR amplifications were performed again. The specific bands with expected sizes were found in D. longan but not in D. confinis by SCAR markers LY2-1, LY4-7 and LY5-2, respectively. These results showed that our developed SCAR markers could be very useful as a specific D. longan variety authentication. Therefore, our study provides an effective and precise PCR-based diagnostic method and markers to identify D. longan species.

No MeSH data available.


Related in: MedlinePlus

Analysis of the PCR amplicons of a SCAR marker LY 4–8. The sample origins and orders indicate on the Figure 4. Lane “M” indicates the DNA molecular weight marker DL600.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3795202&req=5

Fig6: Analysis of the PCR amplicons of a SCAR marker LY 4–8. The sample origins and orders indicate on the Figure 4. Lane “M” indicates the DNA molecular weight marker DL600.

Mentions: To generate stable longan-specific diagnostic SCAR markers from RAPD markers, four pairs of primers (LY2-1 L and LY2-1R; LY4-7 L and LY4-7R; LY4-8 L and LY4-8R; LY5-2 L and LY5-2R) (Table 1) were designed and synthesized based on cloned sequences in Figure 2. The designed SCAR primer pairs were then used to amplify the genomic DNA from 18 of collected DNA samples to test the amplification species-specificity. PCR results indicated that the PCR products with expected size were observed only in five D. longan samples by SCAR marker LY2-1 (Figure 4), SCAR marker LY4-7 (Figure 5), SCAR marker LY4-8 (Figure 6), and SCAR marker LY5-2 (Figure 7), without any amplification in other species we tested (Figures 4,5,6,7), which indicated that all four SCAR markers are longan-specific. The lack of this specific amplicon in the other species indicated the efficacy of these marker in distinguishing the longan group from the others.Figure 4


Molecular cloning and development of RAPD-SCAR markers for Dimocarpus longan variety authentication.

Yang L, Fu S, Khan MA, Zeng W, Fu J - Springerplus (2013)

Analysis of the PCR amplicons of a SCAR marker LY 4–8. The sample origins and orders indicate on the Figure 4. Lane “M” indicates the DNA molecular weight marker DL600.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3795202&req=5

Fig6: Analysis of the PCR amplicons of a SCAR marker LY 4–8. The sample origins and orders indicate on the Figure 4. Lane “M” indicates the DNA molecular weight marker DL600.
Mentions: To generate stable longan-specific diagnostic SCAR markers from RAPD markers, four pairs of primers (LY2-1 L and LY2-1R; LY4-7 L and LY4-7R; LY4-8 L and LY4-8R; LY5-2 L and LY5-2R) (Table 1) were designed and synthesized based on cloned sequences in Figure 2. The designed SCAR primer pairs were then used to amplify the genomic DNA from 18 of collected DNA samples to test the amplification species-specificity. PCR results indicated that the PCR products with expected size were observed only in five D. longan samples by SCAR marker LY2-1 (Figure 4), SCAR marker LY4-7 (Figure 5), SCAR marker LY4-8 (Figure 6), and SCAR marker LY5-2 (Figure 7), without any amplification in other species we tested (Figures 4,5,6,7), which indicated that all four SCAR markers are longan-specific. The lack of this specific amplicon in the other species indicated the efficacy of these marker in distinguishing the longan group from the others.Figure 4

Bottom Line: The specific bands with expected sizes were amplified in five D. longan samples but not in others.To identify and characterize the difference between D. longan and D. confinis, PCR amplifications were performed again.Therefore, our study provides an effective and precise PCR-based diagnostic method and markers to identify D. longan species.

View Article: PubMed Central - PubMed

Affiliation: Research Center for Preclinical Medicine, Luzhou Medical College, Luzhou, Sichuan 646000 China.

ABSTRACT
As an edible fruit and source of traditional medicine, D. longan is grown in most areas of Southern China. Identification of D. longan cultivars by using molecular markers is important genetically. In this study, we cloned fragments from improved randomly amplified polymorphic DNA (RAPD), and developed stably diagnostic sequence-characterized amplified region (SCAR) markers. The specific RAPD bands of D. longan cultivars from Guangxi, with size ranging from 500 bp to 900 bp were gel-purified, cloned and sequenced. Four clones named LY2-1, LY4-7, LY4-8 and LY5-2 were identified. In order to investigate whether the fragments were specific for the species, four pairs of SCAR primers were then designed. PCR amplifications were conducted to analyze 18 samples including different D. longan cultivars and other species. The specific bands with expected sizes were amplified in five D. longan samples but not in others. To identify and characterize the difference between D. longan and D. confinis, PCR amplifications were performed again. The specific bands with expected sizes were found in D. longan but not in D. confinis by SCAR markers LY2-1, LY4-7 and LY5-2, respectively. These results showed that our developed SCAR markers could be very useful as a specific D. longan variety authentication. Therefore, our study provides an effective and precise PCR-based diagnostic method and markers to identify D. longan species.

No MeSH data available.


Related in: MedlinePlus