A method for identification of distant hybrid offspring of peony and peony

An identification method and a distant hybridization technology are applied in the identification field of the distant hybrid progeny of peony and peony, and can solve the problems of scarcity, low hybrid germination rate, long hybridization time and the like.

Inactive Publication Date: 2011-11-30
BEIJING FORESTRY UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] There are not many studies on intergroup hybrid kinship in China. The main reason is that there are not many domestic intergroup hybrid resources. As we all know, intergroup hybrids originated in Japan and developed in the United States. They are expensive and scarce. The lack of domestic scientific research materials in this area
However, there are not many studies on the identification of hybrids, which may be due to the fact that distant hybridization still faces many problems such as hybridization incompatibility of parents, sterility after pollination, too long hybridization time, poor flowering of parents, and incompatibility between parents. The genetic relationship is not clear, the germination rate of the hybrid is low, or it can germinate but the fertility of the F1 generation is low, etc.
Affected by these objective conditions, my country's research work on Paeonia intergroup hybrids, a miracle in the Paeonia breeding circle, is limited.

Method used

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  • A method for identification of distant hybrid offspring of peony and peony
  • A method for identification of distant hybrid offspring of peony and peony
  • A method for identification of distant hybrid offspring of peony and peony

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1 Hybrid Seedling Selection and Morphological Character Investigation

[0029] Method for obtaining hybrid seedlings: pick the flower buds of the male parent back during the dew stage, pick off the anthers indoors, and dry them in the shade for 24 hours in a dry place without direct sunlight. Gently tap the anthers until the pollen is completely scattered, separate the pollen and anthers with a sieve, collect the sieved pollen in a clean vial, label it and store it in a -20°C refrigerator for later use. Select a robust plant as the female parent, use tweezers or bare hands to pinch off or tear off the petals and stamens in the female parent's flower bud stage or dew stage, cover the flower head with a sulfuric acid paper bag and squeeze the bag tightly with a paper clip. After 3-4 days of female parent bagging, pollination begins when the stigma begins to secrete mucus. Pollination was carried out before 11:00 or after 17:00, repeated for three consecutive d...

Embodiment 2

[0039] Embodiment 2 Amplification and sequencing of chloroplast genes

[0040] After the young leaves of 21 hybrid offspring and 2 parents in Example 1 were dried on silica gel, the total DNA was extracted using the Plant Genome Extraction Kit of Tiangen Biotechnology Company, and the operation was performed according to the kit instructions.

[0041] Two chloroplast gene fragments petB-petD and rps16-trnQ were selected for sequencing. The amplification primers of petB-petD are shown in SEQ ID No.1 and 2; the amplification primers of rps16-trnQ are shown in SEQ ID No.3 and 4.

[0042] The polymerase chain reaction (PCR) was completed on the My CyclerTM Thermal Cycler type PCR instrument of BIO-RAD Company. The amplification system is a 25 μl system: template 20-30ng, 10×PCR buffer 2.5 μl, dNTP 80mmol / L, primer 5umol / L, Taq enzyme 1U (Beijing Zexing Biotechnology Co., Ltd.) plus double distilled water to 25ul. The amplification procedure is:

[0043] Pre-denaturation at 94°C...

Embodiment 3

[0056] Amplification and sequencing of embodiment 3SSR markers

[0057] From the 14 pairs of primers screened by Yuan Junhui (his doctoral thesis: The origin of peony and Yan'an peony 2009.12), 10 pairs of primers with high polymorphism and which can be used for experimental samples were selected for amplification (Table 3) (SEQ ID No.5~SEQ ID No.24).

[0058] Table 3 SSR primers

[0059]

[0060] The 5' end of the upstream primer of each pair of primers was fluorescently labeled, wherein Pdel05, Pdel20, Pdel29b, and Pdel33 were labeled with FAM, Pdel02-2, Pdel07, Pdel22, and Pdel35 were labeled with HEX, and Jx02-2 and Jx17 were labeled with ROX.

[0061] The polymerase chain reaction (PCR) was completed on the My CyclerTM Thermal Cycler type PCR instrument of BIO-RAD Company. The amplification system is a 20 μl system: template 20-30ng, PCR Mix (Beijing Biotech Biotechnology Company) 10 μl, 5 μmol / L primers 1 μl each, and double distilled water 20 μl.

[0062] The ampl...

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Abstract

The invention relates to a method for identifying plant hybrid progeny, in particular to a method for identifying distant hybrid progeny of peony and peony. The identification method provided by the invention comprises: identification and screening of hybrid seedling early form, identification of female parent by chloroplast DNA, identification of male parent by SSR molecular marker. Through the combination of morphology and molecules, identify hybrid offspring, identify true and false hybrids, study and summarize the relationship between true hybrid seedlings and parents, determine the heritable external morphological indicators, and obtain a reasonable hybrid at the molecular genetic level The genetic distance between the species and the parent, the degree of kinship, and finally a reasonable identification standard system is established on the basis of morphological and molecular genetics, so that the later selection process can be carried out effectively.

Description

technical field [0001] The invention relates to a method for identifying plant hybrid progeny, in particular to a method for identifying distant hybrid progeny of peony and peony. Background technique [0002] The genus Paeonia includes about 35 species, which are currently divided into three groups, namely, the peony group (sect. Moutan DC), the peony group (sect. Paeonia) and the North American peony group (sect. Onaepia Lindl). "King of flowers" peony and "flower phase" peony are traditional famous flowers with a long history of cultivation in China. They are known as "two unique flowers" and their beauty has always been respected by people. Because the two have great differences in leaf type, flower type, flowering period and growth habit, although they are all Paeoniaceae Paeoniaceae plants, the woody peony belongs to the peony group, and the herbaceous peony belongs to the Paeoniae group. For a long period of time in history, people generally believed that the hybridi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 成仿云张娜娜袁军辉韩丽晓
Owner BEIJING FORESTRY UNIVERSITY
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