Molecular method for quickly identifying closely related species of juniperus

Abstract

The invention discloses a molecular method for quickly identifying closely related species of juniperus. The molecular method comprises the following steps: firstly, extracting leaf total DNA (Deoxyribose Nucleic Acid) or seed endosperm DNA from dry leaves or seeds of juniperus species; secondly, performing PCR (Polymerase Chain Reaction) amplification and sequencing on DNA samples which are qualified by detection through a karyogene Chil primer; thirdly, purifying a PCR amplified product; fourthly, performing a sequencing reaction on the purified product; fifthly, further purifying the sequencing reaction product; sixthly, performing data collection on a purified product by using a genetic analyzer, or performing the sequencing and the data acquisition on the PCR product amplified by the karyogene Chil through the genetic analyzer; seventhly, opening an original sequence peak chart obtained by sequencing through software, reading basic group data information of individuals with high sequencing quality, comparing and correcting an original sequence and identifying the sequences of sabina convallium (JM), a sabina chinensis (JS), juniperus tibetica (JT) and karyogene (Chil) as the closely related species of the juniperus; eighthly, analyzing and comparing the difference degree among the sequences and identifying the species. The molecular method has the characteristics of quickness and accuracy for identification.

Description

Technical field [0001] The invention belongs to the technical field of molecular biology, and particularly relates to a molecular method for quickly identifying related species of Juniper. Background technique [0002] The rapid identification and identification of species is the basis of conservation biology and biodiversity research. However, for a long time, people's identification and recognition of plant species has mainly relied on the obvious external morphological features 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 classification personnel. However, in the current situation where there is a sharp decline in traditional taxonomy professionals, taxonomic identification methods based on the external morphological characteristics of species are difficult to accurately identify and identify the small differences in external morphological characteristi...

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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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