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A molecular detection method for the dormancy state of plum flower buds

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

Active Publication Date: 2020-09-22
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • A molecular detection method for the dormancy state of plum flower buds
  • A molecular detection method for the dormancy state of plum flower buds
  • A molecular detection method for the dormancy state of plum flower buds

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0030] Cultivation under artificial conditions: Insert the bottom of the raw paper into the water and put it in an artificial climate room 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 cut off every 2 days, and the bud germination was observed daily. After 10 days of cultivation, the number of buds germinated and the germination rate of each group were recorded.

[0031] Germination rate statistics: According to the germination rate and the time required for germination, determine the dormant state of flower buds. If the germination rate is 0, the flower bud is in the dormant stage; i...

Embodiment 2

[0045] The flower bud samples were collected on November 1 and 8 respectively, and RNA extraction, cDNA reverse transcription, and qPCR quantitative analysis were performed on them. It was found that the expression of Pm011905 gene in the samples of No. 1 and No. 8 was very low, and that of No. 8. The expression level of No. 1 is only 2% (less than 10%) higher than that of No. 1, indicating that the flower buds of No. 1 and No. 8 are in the dormant-like stage. In order to verify the results, the shoots of No. 1 and No. 8 were subjected to hydroponic culture, and it was found that the germination rate was only 0%, which also met the definition of dormancy-like dormancy proposed by Lang.

[0046] The flower bud samples were collected on December 10th and 18th respectively, and RNA extraction, cDNA reverse transcription, and qPCR quantitative analysis were performed on them. It was found that the expression levels of Pm011905 genes on the 10th and 18th were very high, and the express...

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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 blossom buds with molecular markers. Background technique [0002] Dormancy is a beneficial biological characteristic, and a biological adaptability to environmental conditions and seasonal changes obtained by higher plants through long-term evolution. The induction, maintenance and release of flower bud dormancy is 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 it into para-dormancy, endo-dormancy, and ecological dormancy. eco-dormancy), and the whole process of dormancy is divided into five stages: class dormancy, class dormancy-internal dormancy, internal dormanc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12Q1/6851
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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