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A kind of method for breeding high-yield Wenguan fruit line

A high-yielding technology for sorbifolium sorbifolium, applied in the fields of botany equipment and methods, biochemical equipment and methods, and microbial measurement/inspection, can solve the problem of the lack of methods for identifying self-fertile, high-yielding and superior plants of sorbifolium sorbifolium New varieties breeding technology and other issues

Active Publication Date: 2021-04-13
INST OF BOTANY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] So far, there is no method for identifying self-fertile, high-yielding and superior plants of X. sorbifolium and its new variety breeding technology

Method used

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  • A kind of method for breeding high-yield Wenguan fruit line
  • A kind of method for breeding high-yield Wenguan fruit line
  • A kind of method for breeding high-yield Wenguan fruit line

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1, the acquisition of X. sorbifolium genomic DNA

[0064] The genomic DNA of the sample was extracted by the CTAB method, and the specific operation was as follows:

[0065] 1) Preheat the CTAB extract in a 65°C water bath;

[0066] 2) Grind the sample quickly in liquid nitrogen, transfer the powdered material into a 2mL centrifuge tube, add preheated CTAB extract (3-5ml of extract per gram of sample), and keep warm at 65°C for 30-60min, every Gently invert and mix for 10 minutes;

[0067] 3) Centrifuge at 11000rpm for 5min, take the supernatant and transfer it to a new centrifuge tube;

[0068] 4) Add an equal volume of phenol / chloroform (1:1, volume ratio), mix thoroughly, centrifuge at 11,000 rpm for 10 min, and transfer the supernatant to a new centrifuge tube;

[0069] 5) Add an equal volume of chloroform, mix thoroughly, centrifuge at 11000rpm for 10min, and transfer the supernatant to a new centrifuge tube;

[0070] 6) Repeat steps 4) and 5);

[0...

Embodiment 2

[0076] Embodiment 2, SSR primer screening

[0077] (1) PCR reaction system:

[0078] SSR (total 20 μL): ddH 2 7.2 μL of O, 10 μL of MIX, 0.3 μL of forward primer F, 0.3 μL of reaction primer R, 2 μL of DNA template (genomic DNA of X. sorbifolium obtained in Example 1), and 0.2 μL of Taq.

[0079] (2) The PCR reaction adopts the following cycle parameters:

[0080] SSR PCR amplification program: pre-denaturation at 94°C for 5 minutes; denaturation at 94°C for 30 s, annealing at 54°C for 35 s, extension at 72°C for 40 s, a total of 35 cycles; final extension at 72°C for 3 min.

[0081] (3) Primer information:

[0082] Table 1 Screening primer information

[0083]

[0084]

[0085] (4) The PCR product was added with loading buffer, denatured at 94° C. for 10 min, then analyzed by vertical electrophoresis on a 6% denaturing polyacrylamide gel, and observed after silver staining. Partial picture of primer screening in denaturing polyacrylamide gel electrophoresis (PAGE) ...

Embodiment 3

[0086] Embodiment 3, detection by capillary electrophoresis

[0087] Using the 19 pairs of SSR primers (Table 1) with clear bands, high polymorphism and good repeatability screened in Example 2 above, fluorescent primers were synthesized, and the fragment size was detected by capillary electrophoresis analysis.

[0088] 1. PCR amplification

[0089] (1) PCR reaction system:

[0090] SSR fluorescent primer system (total 20 μL): ddH 2 O 14.8 μL, dNTP 0.4 μL, Buffer 2 μL, forward primer F 0.3 μL (20 μM), reverse primer R 0.3 μL (20 μM), DNA template 2 μL, Taq 0.2 μL.

[0091] (2) The PCR reaction adopts the following cycle parameters:

[0092] SSR PCR amplification program: pre-denaturation at 94°C for 5 min; denaturation at 94°C for 30 s, annealing at 54°C (annealing temperature fluctuates around 54°C) for 35 s, extension at 72°C for 40 s, a total of 35 cycles; final extension at 72°C for 3 min.

[0093] 2. Capillary electrophoresis analysis

[0094] After mixing the formam...

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Abstract

The invention discloses a method for breeding high-yield sorbifolium sorbifolium strains. The method comprises: collecting open-pollinated seeds of high-yield Xanthocarpus sorbifolium, after germination, using SSR molecular markers to identify the male parent, screening for a self-compatibility rate of more than 60%, an ovule fertility rate of more than 90%, and a single plant seed yield Compared with the control tree under the same conditions, the mother plant is more than 30% higher; take the sorbifolium sorbifolium container sapling as the rootstock, graft the selected single plant scion, identify the male parent of the seeds, and confirm that it is self-fertilization. Carry out the second generation grafting and paternal identification. If the self-fertilization is confirmed again, the mother plant is determined to be the final single plant; the scion is taken, and the 3-5-year-old X. radix radix seedlings are used as the rootstock to graft and multiply to obtain a self-fertile high-yield X. sorbifolia strain. The invention adopts means such as SSR molecular markers, container seedling cultivation, grafting and the like to carry out self-fertile high-yield improved variety cultivation of X. sorbifolium, so as to solve the shortage problem of high-yield improved variety of X. sorbifolium.

Description

technical field [0001] The invention belongs to the field of new plant variety cultivation, and relates to a method for breeding high-yield sorbiola sorbifolium strains, in particular to a method for breeding self-fertile high-yield radix sorbifolia strains. Background technique [0002] Xingguan fruit is a precious woody oil-bearing plant unique to northern my country. Its seeds have high oil content and good oil quality. It is listed as healthy edible oil by the State Grain Administration. However, the fruit setting rate of X. sorbifolium is very low, which seriously limits its development and utilization. Since the 1950s, my country has always attached great importance to the breeding of high-yield varieties of sorbifolium, and some scientific research institutes have also done a lot of work to cultivate some local varieties. However, these improved varieties have not been widely recognized and promoted in many years of production practice, because they do not exhibit st...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68A01H1/02A01H1/04A01G2/30A01G17/00
CPCA01H1/02A01H1/04A01G17/005A01G18/00A01G22/00
Inventor 周庆源
Owner INST OF BOTANY CHINESE ACAD OF SCI
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