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Method for detecting phalaenopsis amabilis mutants

A phalaenopsis and mutant technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as inability to separate samples, poor experimental repeatability and stability, and save production costs. Effect

Active Publication Date: 2017-05-10
ENVIRONMENTAL HORTICULTURE RES INST OF GUANGDONG ACADEMY OF AGRI SCI
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Problems solved by technology

[0003] At present, RAPD detection methods are used for the detection of Phalaenopsis mutants. For example, in 2013, Xiao Wenfang et al. disclosed the RAPD detection methods for several common Phalaenopsis tissue culture mutants. Using 7 random primers, the 'Hongtong' Phalaenopsis and its flowers The nick-type mutants were distinguished, and the 'Sky Red' Phalaenopsis and its flower color chimeric mutants could be distinguished by using 6 primers, but the relevant primers could not be found from 50 random primers to distinguish the 'Hongtong' Phalaenopsis labialized mutant
Although the RAPD detection method is also suitable for the detection of mutants, it has insurmountable defects: (1) The experimental repeatability and stability are extremely poor; (2) Gel electrophoresis can only separate DNA fragments with large differences in length , but cannot separate those samples with the same or little difference in molecular weight

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  • Method for detecting phalaenopsis amabilis mutants
  • Method for detecting phalaenopsis amabilis mutants
  • Method for detecting phalaenopsis amabilis mutants

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Embodiment 1

[0018] (1) Extraction of total DNA: Take 0.2 g of the root tip tissues of four Phalaenopsis 'Mantianhong', 'Raining Radiance', V31 and 'Red Pearl' respectively, and follow the conventional extraction methods or conventional reagents for total DNA in this field The total DNA of four kinds of Phalaenopsis 'Mantianhong', 'Radiance', V31 and 'Red Pearl' were extracted from the box.

[0019] (2) PCR reaction: perform PCR with ten pairs of primers including SSR1, SSR2, SSR3, SSR4, SSR5, SSR6, SSR7, SSR8, SSR9 and SSR10.

[0020] The PCR reaction system was 6.0 µL of 2.5×Buffer V, 1.0 µL of upstream and downstream primers (5 µM), Taq enzyme (5U·µL -1 ) 0.1 µL, DNA 1.0 µL, ddH 2 O fill up to 15 µL;

[0021] The reaction program was 95 °C for 5 min, 35 cycles of 94 °C for 30 sec, 58 °C for 35 sec, 72 °C for 35 sec, and 60 °C for 30 min.

[0022] The sequences of ten pairs of primers for SSR1, SSR2, SSR3, SSR4, SSR5, SSR6, SSR7, SSR8, SSR9 and SSR10 are shown in Table 1.

[0023] Ta...

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Abstract

The invention belongs to the technical field of monitoring of plant varieties and in particular discloses a method for detecting phalaenopsis amabilis mutants. Four varieties including 'HongMantianHong', 'Guangmangsishe', V31 and 'Hongzhenzhu' and mutants thereof can be completely distinguished through utilizing combination of ten pairs of primers including SSR1, SSR2, SSR3, SSR4, SSR5, SSR6, SSR7, SSR8, SSR9 and SSR10; sequences of the ten pairs of primers including the SSR1, the R2, the SSR3, the SSR4, the SSR5, the SSR6, the SSR7, the SSR8, the SSR9 and the SSR10 are shown as SEQ: 1 to 20. The detection method disclosed by the invention can be used for screening the mutants, which do not meet growth requirements, at a bottle seedling period, so that the production of large-scale production is saved.

Description

technical field [0001] The invention relates to the technical field of plant variety monitoring, in particular to a method for detecting Phalaenopsis mutants. Background technique [0002] Phalaenopsis is a class of orchids native to the tropical regions of Asia. It has peculiar flower shapes and rich colors, and has extremely high ornamental value and economic value. The large-scale production of Phalaenopsis currently mainly propagates seedlings through tissue culture, which makes its production cost higher and the growth cycle longer. It takes about 18 months from the time when the seedlings are produced in a bottle to flowering and selling. Then, due to many mutagenic factors in the tissue culture process, Phalaenopsis will produce a certain amount of mutants in the tissue culture process, and the mutants do not meet the requirements of commercial production, which will increase the production cost and affect large-scale commercialization. Production is very bad. There...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/686C12Q1/6895C12Q2600/156C12Q2563/107C12Q2565/125
Inventor 肖文芳李佐吕复兵陈和明
Owner ENVIRONMENTAL HORTICULTURE RES INST OF GUANGDONG ACADEMY OF AGRI SCI
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