Objective

2

To 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

.

Method

2

Single 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.

Result

2

There 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.

Conclusion

2

The 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.