Method for screening fine progeny strain polyploidy plant of Chinese gooseberry Hort 16A

A Chinese kiwifruit, hort16a technology, applied in the field of plant genetics and breeding, can solve the problems of uncertainty, heavy workload, and many influencing factors, and achieve the effect of significant progress, easy access, and significant substantive characteristics.

Inactive Publication Date: 2016-12-07
SOUTHWEST FORESTRY UNIVERSITY
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Problems solved by technology

For plants such as kiwifruit, which have a large number of chromosomes and large phenotypic changes, making ploidy identification difficult, the former is cumbersome to operate, with many influencing factors and heavy workload, and it is very difficult to obtain a clear number of chromosome sheets; the latter screening The result is that the polyploid plant physiology is changed by the environment, usually only as a reference, not deterministic

Method used

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  • Method for screening fine progeny strain polyploidy plant of Chinese gooseberry Hort 16A
  • Method for screening fine progeny strain polyploidy plant of Chinese gooseberry Hort 16A
  • Method for screening fine progeny strain polyploidy plant of Chinese gooseberry Hort 16A

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Embodiment Construction

[0037] (1) Extract the total RNA from the leaves of Actinidia sinensis tissue cultured seedlings using the Plant RNA Rapid Extraction Kit. Under the conditions of 1.0% agarose gel, 110V voltage, and electrophoresis for 25 minutes, the quality of the extracted total RNA was detected to observe whether the bands were clear, whether there was degradation, and whether there was DNA contamination.

[0038] (2) Prepare the reaction solution to remove genomic DNA (see Table 4), then prepare the reaction system (the mixture is prepared on ice, see Table 5), reverse transcribe the RNA into cDNA, and use the reverse transcription product for PCR amplification:

[0039] Table 4 Removal of Genomic DNA Reaction System

[0040]

[0041] Gently blow and mix with a pipette, inactivate DNase I at 37°C for 30 min; at 65°C for 10 min.

[0042] Table 5 Reaction system for reverse transcription of RNA into cDNA

[0043]

[0044] The setting of reverse transcription reaction conditions: 25°...

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Abstract

The invention discloses a method for screening a fine progeny strain polyploidy plant of Chinese gooseberry Hort 16A, and relates to the screening of a fine strain polyploidy of the plant. The method comprises the following steps: extracting total RNA of a leaf of a tissue culture seedling, reversely transcribing RNA into cDNA, selecting 1 to 2 differential genes having an up-regulation trend in transcriptomes of diploid and polyploidy plants, taking a housekeeping gene Elongation factor EC as a reference gene, fluorescently quantifying a PCR (Polymerase Chain Reaction), drawing an amplification curve and a solubility curve for the differential genes, selecting the differential genes, calculating a relative transcript level, and taking the tissue culture seedling, whose differential genes are more than twice those of the diploid, as the tissue culture seedling of the polyploidy. The theory and the method for screening the polyploidy variant plant are innovated, and the efficient feasible method is provided for screening the polyploidy plant of the Chinese gooseberry.

Description

technical field [0001] The invention belongs to plant genetics and breeding, and relates to a screening method for polyploids of excellent strains of plants. Background technique [0002] Kiwifruit is a deciduous vine of Actinidiaceae, native to my country. Following Hayward, New Zealand systematically bred the cultivar 'Hort 16A' of Actinidia sinensis from the Actinidia sinensis group in the 1990s. The best variety has applied for international patents. In 2010, the School of Forestry of Southwest Forestry University selected a batch of high-yielding and excellent lines of 'Hort 16A' descendants from the seedlings of 'Hort 16A' open pollination. This line has high fruit-setting rate, strong growth potential, unique fruit flavor and long storage period. It is a promising strain with independent intellectual property rights. [0003] Polyploids generally have the characteristics of thick branches, thick leaves, large fruits, high yield, strong stress resistance and strong ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q1/6895C12Q2600/166C12Q2531/113C12Q2545/101C12Q2563/107
Inventor 张汉尧刘小珍李升星张先昂刘艳艳陈海
Owner SOUTHWEST FORESTRY UNIVERSITY
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