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Method for obtaining, breeding and storing peanut interspecies hybridization variety, and identifying molecular cytology

A preservation method and peanut seed technology, applied in the fields of botanical equipment and methods, horticultural methods, plant regeneration, etc., can solve the problems of poor chromatin heterogeneity, infertility, inability to continue generations, etc., and overcome incompatibility. Sex, the effect of enriching the genetic basis

Active Publication Date: 2013-12-18
HENAN ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0005] Due to the incompatibility of interspecific hybridization, it is difficult to obtain well-developed and viable seeds (ovules) by hybridization among many species of the genus Peanut, and the interspecific hybrid F 1 Highly sterile and infertile, unable to continue for generations; at the same time, the chromatin heterogeneity between peanut and wild species is poor, which brings difficulties to the identification of interspecific hybrids

Method used

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  • Method for obtaining, breeding and storing peanut interspecies hybridization variety, and identifying molecular cytology
  • Method for obtaining, breeding and storing peanut interspecies hybridization variety, and identifying molecular cytology
  • Method for obtaining, breeding and storing peanut interspecies hybridization variety, and identifying molecular cytology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1. Peanut Interspecific Hybrid F 1 method of obtaining

[0049] The cultivated species Baisha 1016 (preserved by the Institute of Economic Crops, Henan Academy of Agricultural Sciences) was used as the female parent, and the wild species A. macedoi F 1 ,Specific steps are as follows:

[0050] (1) Artificial hybridization

[0051] In Baisha 1016 with A. macedoi At the beginning of flowering period, from 16:00 to 18:00 in the afternoon every day, select the calyx micro-cracked flower buds of the female parent, gently poke the petals with tweezers, remove the stamens, and mark the nodes where the stamens are removed; the next day Collect the male parent flower in the morning, take its pollen, apply the pollen on the stigma of the female parent plant where the stamen flower has been removed with tweezers; after routine fertilization, watering, fix the branches after the fruit needles are exposed, harvest after 60-80 days, and observe the hybridization Pods ...

Embodiment 2

[0060] Example 2. Peanut Interspecific Hybrid F 1 The breeding and preservation method, comprises the following steps:

[0061] (1) Take F from the culture medium under the ultra-clean bench 1 For seedlings, cut the seedlings into 2-3 sections, each section retaining at least one leaf, and place the terminal bud section of the seedlings in a solid rooting medium composed of MS, 0.8 mg / L NAA (naphthaleneacetic acid) and 3% sucrose , cultivated for 15-30 days, until a complete plant is obtained;

[0062] (2) Place the other sections of the seedlings in a solid medium consisting of MS, 0.1 mg / L NAA, 0.4 mg / L 6-BA (6-benzyl adenine) and 3% sucrose, and light at a constant temperature of 25°C Induce bud differentiation in the culture room for 15-30 days, then inoculate into solid rooting medium composed of MS, 0.8 mg / L NAA and 3% sucrose, and culture for 15-30 days until a complete plant is formed;

[0063] By repeating the above two steps, the hybrid F can be propagated and p...

Embodiment 3

[0064] Example 3. Peanut distant hybrid F 1 The molecular cytological identification method, the specific steps are as follows:

[0065] (1) Take the female parent Baisha 1016 and the male parent A. macedoi and hybrid F 1 The leaves of the plants were extracted from the female parent, male parent and hybrid F by CTAB method. 1 DNA of the plant;

[0066] (2) Use Baisha 1016 and A. macedoi The specific marker E66 was used for PCR reaction, followed by polyacrylamide gel electrophoresis analysis, the steps are as follows:

[0067] Perform PCR reaction first: PCR reaction volume is 10 μl, including template genomic DNA 0.9 μl (25 ng / ml), SSR primer (R) 0.2 μl (10 μM), SSR primer (F) 0.2 μl (10 μM), 10 ×buffer 1μl, dNTP (2.5 mM) 0.9 μl, MgCl 2 (25 mM) 0.8 μl, Taqase 0.1 μl (0.5U), ddH2 O 5.9 μl. Mix the above ingredients and centrifuge, add a drop of paraffin oil to cover to prevent water from evaporating;

[0068] Then carry out PCR amplification: the reaction is carri...

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Abstract

The invention relates to a method for obtaining, breeding and storing a peanut interspecies hybridization variety F1, and identifying molecular cytology. The method comprises the following steps of: taking cultispecies white sand 1016 as a female parent and taking a wild seed A.macedoi as a male parent to carry out artificial hybridization; carrying out isolated culture on an ovule and a young embryo to obtain the F1; in a breeding process, placing a seedling terminal bud region into a solid rooting culture medium and culturing to obtain a whole set of a plant; in an identifying process, respectively extracting gene groups DNA (Deoxyribonucleic Acid) on the female parent, the male parent and the hybrid F1; utilizing specific markers E66 of the female parent and the male parent to carry out PCR (Polymerase Chain Reaction) reaction; and carrying out gel electrophoresis analysis to identify the reality of the hybrid F1. According to the method disclosed by the invention, great breakthroughs on the aspects of developing and utilizing a peanut wild species gene resource are obtained and the incompatibility of the interspecies hybridization is overcome, so that the hybrid F1 is bred and stored for a long period and a convenience condition is provided for transferring and utilizing good genes of the peanut wild species; the specific makers are combined with a molecular cytology technology to successfully identify the hybrid F1; and the method has the important active effects on a technology of enriching a heredity basis of peanut varieties and optimizing breeding.

Description

technical field [0001] The invention relates to peanut breeding technology, in particular to peanut interspecific hybrid variety F 1 The methods of acquisition, reproduction preservation and molecular cytology identification. Background technique [0002] The genetic diversity of cultivated peanut has been decreasing during the domestication process, and the available genetic resources are limited. Especially in recent years, the intensive breeding and wide application of varieties have made the genetic basis of peanut varieties more and more narrow, and the production is under the stress of various diseases and insect pests and adversity (drought, waterlogging, salinity, low temperature), resulting in the quality of peanuts. decrease and decrease in production. How to improve the resistance and yield of peanuts to pests and adversity has become the main goal of peanut breeding work. [0003] The genus Arachis includes about 80 species, which are annual or perennial speci...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H1/02A01H4/00C12Q1/68
Inventor 张新友杜培徐静秦利黄冰艳董文召汤丰收
Owner HENAN ACAD OF AGRI SCI
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