A molecular method for rapid identification of closely related species of Juniperus

Abstract

A molecular method for rapid identification of closely related species of Juniperus. The steps are: 1. Extract the total leaf DNA or seed endosperm DNA from the dried leaves or seeds of the Juniperus species; 2. From the DNA samples that have passed the test, Use the nuclear gene Chi1 primer for PCR amplification and sequencing; 3. Purification of the PCR amplification product; 4. Sequencing reaction of the purified product; 5. Further purification of the sequencing reaction product; 6. Data collection of the purified product using a genetic analyzer; Or use a genetic analyzer to sequence and collect data on the PCR product amplified by the nuclear gene Chi1; 7. Open the original sequence peak chart obtained by sequencing with software, and read the base data information for individuals with high sequencing quality; the original sequence ratio Corrected and corrected; identified the sequences of nuclear genes Chi1 of Juniperus juniper (JM), Juniper juniper square branch (JS) and Juniperus tibetans (JT); 8. Analysis and comparison of differences between sequences degree, for species identification; it has the characteristics of rapid and accurate identification.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a molecular method for quickly identifying close relatives of the genus Juniper. Background technique [0002] The rapid identification and identification of species is fundamental to conservation biology and biodiversity research. However, for a long time, people's identification and identification of plant species has mainly relied on the obvious external morphological characteristics between species, such as plant flowers, fruits, seeds, roots, stems and leaves. At the same time, it also relies on the practical experience and guidance of professional taxonomy personnel. However, in the current situation where traditional taxonomy professionals are sharply reduced, it is difficult to accurately identify and identify small differences in external morphological characteristics between species with taxonomic identification methods based on external morpholog...

AI Technical Summary

Problems solved by technology

[0004] However, for gymnosperm groups, due to the slow mutation rate of chloroplast genes, coupled with the long generation cycle of gymnosperms (from seed development to individual plant maturity to reproducing seeds, it takes about 15 years) and The large effective population size (number) resulted in the inability of the chloroplast DNA barcode rbcL+matK+trnH-psbA marker combination and nuclear ITS gene fragments to accurately distinguish gymnosperm relatives

This is especially true for Juniper species with a recent speciation history, resulting in substantial sharing of chloroplast DNA fragments and nuclear ITS gene fragment loci among Juniper species, making it difficult to use these standard DNA barcoding molecular marker combinations. Difficulty in accurately identifying and identifying closely related species of Juniperus

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