Molecular detection method for prunusmume flower bud dormancy state

A dormant state, flower bud technology, applied in the field of molecular biology, can solve problems such as ambiguity

Active Publication Date: 2017-11-14
BEIJING FORESTRY UNIVERSITY
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  • Abstract
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Problems solved by technology

As far as plum blossoms are concerned, the physiological mechanism of dormancy has been studied in depth, but at the molecular level, there are still many unclear mechanisms in the process of bud dormancy, especially the release of flower bud dormancy, such as the determination of genes that play a regulatory role in the process of dormancy release, and the release of low temperature. The mechanism of dormancy, how to regulate photoperiod, etc.

Method used

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  • Molecular detection method for prunusmume flower bud dormancy state
  • Molecular detection method for prunusmume flower bud dormancy state
  • Molecular detection method for prunusmume flower bud dormancy state

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Experimental program
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Effect test

Embodiment 1

[0029] Materials: Starting from October 15th, every 7 days, the plum blossom branches of the Jiufeng "Green Calyx" variety were collected, 10 branches and about 100 flower buds were collected each time, and they were randomly divided into 3 groups, and each group was counted separately The number of flower buds on the branch.

[0030] Cultivation under artificial conditions: Insert the bottom of the raw paper strips of the year into water, and put them into an artificial climate chamber for cultivation. The artificial conditions are set at 25°C for 16 hours during the day, 16°C for 8 hours at night, and the humidity is 70%. The bottom was subtracted every 2 days, and the germination of flower buds was observed every day. After 10 days of cultivation, the number and germination rate of flower buds in each group were recorded respectively.

[0031] Germination rate statistics: According to the germination rate and the time required for germination, determine the dormant state of...

Embodiment 2

[0045] Flower bud samples were collected on November 1st and 8th, and RNA extraction, cDNA reverse transcription, and qPCR quantitative analysis were performed on them. The expression level of No. 1 is only 2% (less than 10%) higher than that of No. 1, which shows that the flower buds of No. 1 and No. 8 are in the quasi-dormant stage. In order to verify the results, the branches of No. 1 and No. 8 were hydroponically cultured, and it was found that the germination rate was only 0%, which also met the definition of quasi-dormant proposed by Lang.

[0046] Flower bud samples were collected on December 10th and 18th, and RNA extraction, cDNA reverse transcription, and qPCR quantitative analysis were performed on them. It was found that the expression levels of the Pm011905 gene were very high on the 10th and 18th, and the expression level on the 18th was good. 50% (more than 10%) higher than the expression level of No. 10, it shows that the flower buds of No. 10 and No. 18 are in...

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Abstract

The invention relates to the field of molecular biology and in particular discloses application of Pm011905 genes (SEQ ID NO.1) in an aspect of monitoring a prunusmume flower dormancy state and a molecular detection method for a prunusmume flower bud dormancy state. The method comprises the following steps: (1) collecting flower buds of to-be-detected prunusmume flower twice, respectively extracting RNA of the flower bud and performing inverse transcription to obtain cDNA, and by taking the cDNA as a template, carrying out a fluorescent quantitative PCR (Polymerase Chain Reaction) on Pm011905 genes by utilizing primers shown as SEQ ID NO.2-3; and (2) comparing the expression quantity of the Pm011905 genes of the flower buds collected twice, and judging the dormancy stage of the prunusmume flower buds. The method disclosed by the invention first discovers that the dormancy state of the flower buds can be monitored or judged by virtue of the change of the expression quantity of the Pm011905 genes of the flower buds. The invention provides a simple, convenient, rapid and accurate method capable of detecting the prunusmume flower bud dormancy state on a molecular level.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a method for detecting the dormant state of plum flower buds by molecular markers. Background technique [0002] Dormancy is a beneficial biological characteristic, and it is a biological adaptation to environmental conditions and seasonal changes obtained by higher plants through long-term evolution. The induction, maintenance, and release of flower bud dormancy are a complex and continuous biological process, which is the result of the interaction between plant internal regulation and external environmental factors. According to the inducement of dormancy, Lang (1987) called the temporary cessation of visible growth of plant structures containing meristems as dormancy, and divided them into para-dormancy, endo-dormancy and ecological dormancy ( eco-dormancy) and divided the whole process of dormancy into five stages: dormancy-like, dormancy-like-endormancy, endormancy, endorma...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q1/6895C12Q2531/113C12Q2563/107
Inventor 张启翔张智勇程堂仁王佳郑唐春卓孝康
Owner BEIJING FORESTRY UNIVERSITY
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