US 11,053,555 B2
Method for identifying genetic relatedness of liriodendron hybrids
Jinhui Chen, Nanjing (CN); Jisen Shi, Nanjing (CN); Xiaofei Long, Nanjing (CN); and Tielong Cheng, Nanjing (CN)
Assigned to NANJING FORESTRY UNIVERSITY, Nanjing (CN)
Appl. No. 17/42,937
Filed by NANJING FORESTRY UNIVERSITY, Nanjing (CN)
PCT Filed Oct. 31, 2018, PCT No. PCT/CN2018/113082
§ 371(c)(1), (2) Date Sep. 29, 2020,
PCT Pub. No. WO2019/223245, PCT Pub. Date Nov. 28, 2019.
Claims priority of application No. 201810498902.0 (CN), filed on May 23, 2018.
Prior Publication US 2021/0032708 A1, Feb. 4, 2021
Int. Cl. C12P 19/34 (2006.01); C12Q 1/6895 (2018.01); C12Q 1/686 (2018.01)
CPC C12Q 1/6895 (2013.01) [C12Q 1/686 (2013.01); C12Q 2600/13 (2013.01)] 7 Claims
 
1. A method for identifying genetic relatedness of Liriodendron hybrids, comprising the following steps:
1) extracting DNA from leaves of tree cross offspring to obtain an extracted DNA;
2) performing a first PCR detection and then a second PCR detection on the extracted DNA, wherein in the first PCR detection, performing a first PCR amplification with primer pair 18.2 on the extracted DNA, and then
identifying a female parent with a 400 bp specifically-amplified product as Liriodendron chinense, and identifying a female parent with a 382 bp specifically-amplified product as Liriodendron tulipifera; and
wherein in the second PCR detection, performing a second PCR amplification on the female parent identified as Liriodendron chinense with primer pair 700, performing a third PCR amplification on the female parent identified as Liriodendron tulipifera with the primer pair 700 to obtain a 700 bp specifically-amplified product, identifying a female parent with the 700 bp specifically-amplified product as a Liriodendron chinense tree originating from a western provenance, and identifying a female parent with a 242 bp specifically-amplified product as a Liriodendron chinense tree originating from an eastern provenance;
wherein, a sequence of a first upstream primer of the primer pair 18.2 is shown in SEQ ID NO. 1, and a sequence of a first downstream primer of the primer pair 18.2 is shown in SEQ ID NO. 2; a sequence of a second upstream primer of the primer pair 700 is shown in SEQ ID NO. 3, and a sequence of a second downstream primer of the primer pair 700 is shown in SEQ ID NO. 4;
wherein, the western provenance is a Monan provenance of Guizhou province, and the eastern provenance is a Lushan provenance of Jiangxi province.