A Rapid PCR-RFLP Method for Assessing Heterozygosity of Murraya paniculata Germplasm
10.13422/j.cnki.syfjx.20231512
- VernacularTitle:千里香杂合度PCR-RFLP快速评估方法的建立
- Author:
Bocheng WANG
1
;
Ziyuan CHEN
2
;
Zhongyi HUA
2
;
Hui TIAN
1
;
Wenbo XIE
3
;
Yuan YUAN
2
Author Information
1. Guangxi University of Chinese Medicine， Nanning 530200， China
2. National Resource Center for Chinese Materia Medica， China Academy of Chinese Medical Sciences， Beijing 100700， China
3. China Resources Sanjiu Medical & Pharmaceutical Co. Ltd.， Shenzhen 518029， China
- Publication Type:Journal Article
- Keywords:
Murraya paniculata;
polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP）;
single nucleotide polymorphisms （SNPs）;
heterozygosity;
whole genome resequencing
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2024;30(4):29-34
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo establish a rapid method for evaluating the heterozygosity of Murraya paniculata germplasm materials and provide as a foundation for developing germplasm breeding and innovation measures for M. paniculata. MethodSingle nucleotide polymorphisms （SNPs） were screened from the genome resequencing data of 65 plants of M. paniculata. A self-written script was used to transform 20 SNPs into restriction fragment length polymorphism （RFLP） markers. Polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） was employed to detect the 20 RFLP markers in 12 M. paniculata germplasm accessions， and the heterozygosity of M. paniculata germplasm accessions was calculated based on the number of enzyme-cutting bands at the 20 RFLP marker sites. Plink was used to calculate the whole genome heterozygosity of 12 M. paniculata germplasm accessions， and the results obtained with different methods were compared. ResultThere was no significant difference in the heterozygosity calculated by the PCR-RFLP method and the genome resequencing method. The PCR-RFLP and genome resequencing methods identified 8 and 9 germplasm accessions， respectively， with a heterozygosity level less than 30%. Seven germplasm accessions with heterozygosity less than 30.00% were calculated by both methods. ConclusionThe PCR-RFLP method established in this study for evaluating the heterozygosity of M. paniculata germplasm demonstrates the precision of 87.5% and the accuracy of 77.8%. This method serves as a reference for developing heterozygosity evaluation methods in other medicinal plant germplasm resources.
