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SSR primers of endangered Heritiera littoralis Hernandia nymphaeifolia and application thereof

A technology of lotus leaves and primer pairs, applied in the field of molecular genetics, to achieve the effect of protecting biodiversity and maintaining ecological balance

Active Publication Date: 2022-02-08
HAINAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

No SSR molecular markers have been used to design primers to study the genetic diversity of A.

Method used

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  • SSR primers of endangered Heritiera littoralis Hernandia nymphaeifolia and application thereof
  • SSR primers of endangered Heritiera littoralis Hernandia nymphaeifolia and application thereof
  • SSR primers of endangered Heritiera littoralis Hernandia nymphaeifolia and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Design and screening of embodiment 1 specific SSR primer

[0032] 1. Mining of SSR sites based on the spliced ​​sequence of the transcriptome of A. japonicus

[0033] (1) Sample detection, library construction and sequencing:

[0034] a) Extraction of total RNA: send it to the company, and the company will use a kit to extract it. Determine the concentration and purity of the sample. After the RNA quality inspection meets the requirements, perform library construction, etc.;

[0035] b) Isolation and fragmentation of mRNA: Isolation: Utilize the characteristic of polyA in the mature mRNA of eukaryotes, and use the property of Oligo, a magnetic bead that specifically binds to polyA, to achieve the purpose of enriching mRNA. Fragmentation: Use fragmentation buffer to fragment mRNA

[0036] c) Synthesis of the first strand of cDNA: Six-base random primer was used to form the first strand of cDNA using mRNA as a template.

[0037] d) Synthesis of the second strand of c...

Embodiment 2

[0065] Example 2 Analysis of Genetic Diversity of Palismania chinensis in Hainan Island

[0066] Genetic diversity indicators mainly include observed number of alleles (Na, Observed number of alleles), effective number of alleles (Ne, Effective number of alleles), Shannon's information index (I, Shannon's information index), expected heterozygosity ( He, Expected heterozygosity) and observed heterozygosity (Ho, Observed heterozygosity) and gene flow (Nm, Gene flow), Nei's standard genetic distance (GD, Genetic distance), genetic differentiation coefficient Fst, F fixed index, etc. The number of alleles refers to the number of allele types actually observed at the corresponding loci, reflecting the level of genetic diversity of the population. The effective allele number is the reciprocal of the homozygosity in the population. It is one of the indicators reflecting the genetic variation of the population. It also reflects the interaction between alleles. The closer the effectiv...

Embodiment 3

[0069] Example 3 Analysis of Genetic Differentiation of Pyrethia chinensis Population

[0070] In order to explore the level of genetic differentiation among the populations of P. basilicum, we used GeneAlEx software to conduct 8 populations (1 natural population in Boao, Qionghai, 5 natural populations in Tanmen, Qionghai, 1 natural population in the eastern suburb of Wenchang, 1 natural population in Wenchang Huiwen) genetic differentiation between two populations (Table 6) and Nei's population genetic distance (Table 7), and molecular analysis of variance (AMOVA) between populations was also carried out (Table 8)

[0071] Table 6 Genetic differentiation coefficients among populations

[0072]

[0073]

[0074] Table 7 Genetic identity and genetic distance

[0075]

[0076] Table 8 AMOVA analysis

[0077]

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Abstract

The invention belongs to the field of molecular genetics, and discloses application in the aspects of genetic diversity, genetic structure analysis and the like of Hainan Island Hernandia nymphaeifolia populations developed on the basis of SSR molecules, wherein genetic structure characteristics and genetic diversity of different populations of Hernandia nymphaeifolia are analyzed, gene exchange and genetic differentiation degrees among different populations are clarified, phylogenetic evolution and geographical differentiation of endangered Heritiera littoralis Hernandia nymphaeifolia populations are clarified, a foundation is laid for molecular biology and genetics research of the endangered Heritiera littoralis Hernandia nymphaeifolia, and a theoretical basis is provided for protection of genetic diversity of Hernandia nymphaeifolia, effective utilization of resources and the like. According to the invention, the SSR marker developed by adopting the transcriptome high-throughput technology is high in speed, high in efficiency and low in cost.

Description

technical field [0001] The invention belongs to the field of molecular genetics, and relates to an SSR primer, in particular to a specific SSR primer of the endangered semi-mangrove plant Aurantia chinensis. Background technique [0002] Hernandia nymphaeifolia (J.Presl) Kubitzki is an evergreen tree belonging to the genus Hernandia nymphaeifolia, which has certain medicinal value because of its lignin and alkaloid content. At present, there are few studies on the endangered Neptunia chinensis at home and abroad. Liu Liping, Wu Yinsheng, etc. conducted research on the components of the branches and seeds of the lotus leaf, and found that it contains a variety of anti-tumor and leukemia and other substances, verifying its important medicinal value. Li Jiawen et al. conducted a clustering analysis of the comparative evolution of the chrysanthemum and its close relatives, and classified it into the order Camphora. They also verified that the chloroplast genomes of the chrysant...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/686C12N15/11G16B40/00
CPCC12Q1/6895C12Q1/686G16B40/00C12Q2600/13C12Q2563/107C12Q2535/122C12Q2525/151Y02A50/30
Inventor 王勇倪靓谭佐莉张修含张世杰于靖
Owner HAINAN NORMAL UNIV
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