Method for in-vivo identification of flower bud differentiation period of monthly pollen Chinese rose

Abstract

The invention relates to a living body identification method for a flower bud differentiation period of China rose. The method for identifying the flower bud differentiation period of the China rose in vivo comprises the following steps: (1) determining the position of a mixed bud of an annual branch or an annual branch, and segmenting the mixed bud; the method comprises the following steps: dividing three sections from top to bottom according to morphology, namely an upper part, a middle part and a lower part, and selecting mixed buds at the upper part; (2) determining the growth period of the upper mixed buds according to the number of bud scales, the opening degree of the bud scales and the leaf extension condition of the upper mixed buds; and 3) determining the flower bud differentiation period of the flower bud according to the leaf extension number and the leaflet expansion degree of the flower bud. The technical problem to be solved is that the flower bud at the upper part of the annual branch or the annual branch is subjected to living body observation by naked eyes or a magnifying lens, the flower formation transformation and flower bud differentiation period can be distinguished, and the flower bud cannot be damaged.

Description

Technical Field

[0001] The invention belongs to the technical field of identifying the differentiation period of rose flower buds, and in particular relates to a method for identifying the differentiation period of rose flower buds in vivo. Background Art

[0002] Rose is a plant of the genus Rosa in the family Rosaceae. It is one of the top ten traditional famous flowers in my country and one of the most popular fresh cut flowers in the world. It is known as the "Queen of Flowers" and is widely cultivated and used around the world. It has extremely high economic and cultural value. 'Old Blush' rose (Rosa chinensis 'Old Blush') is an important ancient Chinese rose variety with a simple genetic background, continuous flowering and strong resistance. It is not only one of the important parents for the breeding of modern roses, but also has rich genomic data resources. It is currently a model material for international rose biology research and woody continuous flowering plant research.

[0003] Flowering and flower development are hot topics and difficulties in current research. One of the basic prerequisites for studying the flowering mechanism is to identify the period of flower bud differentiation. However, the rose 'Yueyuefen' is different from other roses. It has objective characteristics such as fast flower bud differentiation rate and small flower buds. Usually, it can only be identified by conventional paraffin section preparation technology, which is not only time-consuming and labor-intensive, but also damages the flower buds, affecting subsequent research. Summary of the invention

[0004] In view of this, the main purpose of the present invention is to provide a method for living identification of the flower bud differentiation period of 'Yueyuefen' rose. The technical problem to be solved is to use the naked eye or a magnifying glass to conduct living observation of the upper flower buds of annual branches or current year branches, so as to identify the differentiation period of flower buds without causing damage to the flower buds.

[0005] The purpose of the present invention and the technical problem to be solved are achieved by adopting the following technical solutions. The present invention proposes a method for identifying the flower bud differentiation period of 'Yueyuefen' rose in vivo, comprising the following steps: 1) Determine the position of the mixed buds of the one-year-old branches or the current year branches and divide them into three sections from the top to the bottom according to morphology, namely the top, middle and bottom, and select the mixed buds in the top; 2) According to the number of bud scales, the degree of bud scale opening, and the extension of leaves of the upper mixed buds, the growth period of the upper mixed buds can be determined; 3) Determine the differentiation period of the flower bud based on the number of leaves extending from the flower bud and the degree of expansion of the leaflets.

[0006] The purpose of the present invention and the technical problems to be solved can be further achieved by adopting the following technical measures.

[0007] Preferably, in the aforementioned method for in vivo identification of the flower bud differentiation period of 'Yueyuefen' rose, in step 1), the upper mixed buds are mixed buds in the upper 1 / 3 of the branch morphology.

[0008] Preferably, in the aforementioned method for in vivo identification of the flower bud differentiation period of 'Yueyuefen' rose, in step 1), the upper mixed buds are mixed buds at the upper 1 / 3 of the branch morphology and at the leaf axils.

[0009] Preferably, in the aforementioned method for in vivo identification of the flower bud differentiation period of 'Yueyuefen' rose, in step 1), the upper mixed buds are mixed buds at the morphological upper 1 / 3 section and leaf axils of annual branches or current year branches.

[0010] Preferably, in the aforementioned method for in vivo identification of the differentiation period of flower buds of 'Yueyuefen' rose, in step 2), the step of clarifying the growth period of the upper mixed buds according to the number of bud scales, the degree of opening of the bud scales, and the extension of the leaves of the upper mixed buds includes: when the number of bud scales is <5, it is determined that the upper mixed bud is in the undifferentiated stage; when the number of bud scales is ≥5, it is determined that the upper mixed bud has entered the flowering transformation and is in the reproductive growth stage.

[0011] Preferably, in the aforementioned method for in vivo identification of the differentiation period of flower buds of 'Yueyuefen' rose, in step 2), the step of clarifying the growth period of the upper mixed buds according to the number of bud scales, the degree of opening of the bud scales, and the extension of leaves of the upper mixed buds includes: when the number of bud scales <5 and the degree of opening of the bud scales <50%, it is determined that the upper mixed buds are in the undifferentiated stage; when the number of bud scales ≥5 and the degree of opening of the bud scales ≥50%, it is determined that the upper mixed buds have entered the flowering transformation and are in the reproductive growth stage.

[0012] Preferably, in the aforementioned method for in vivo identification of the differentiation period of flower buds of 'Yueyuefen' rose, in step 2), the step of clarifying the growth period of the upper mixed buds according to the number of bud scales, the degree of opening of the bud scales and the extension of leaves of the upper mixed buds includes: when the number of bud scales <5, the degree of opening of the bud scales <50%, and the leaves are "not extended", it is determined that the upper mixed buds are in the undifferentiated stage; when the number of bud scales ≥5, the degree of opening of the bud scales ≥50%, and the leaves are "extended", it is determined that the upper mixed buds have entered the flowering transformation and are in the reproductive growth stage.

[0013] Preferably, in the aforementioned method for in vivo identification of the flower bud differentiation period of 'Yueyuefen' rose, in step 2), the number of bud scales, the degree of opening of bud scales, and the extension of leaves of the upper mixed buds are observed with the naked eye or with the aid of a magnifying glass.

[0014] Preferably, in the aforementioned method for in vivo identification of the differentiation period of flower buds of the rose 'Yueyuefen', in step 3), the step of determining the differentiation period of the flower bud according to the number of leaves extending from the flower bud and the degree of expansion of the leaflets includes: when 1-2 leaves extend, determining that the flower bud is in the early to middle and late stages of flower bud differentiation; when more than 2 leaves extend, determining that the flower bud is in the late stage of flower bud differentiation.

[0015] Preferably, in the aforementioned method for living identification of the differentiation period of flower buds of 'Yueyuefen' rose, in step 3), the step of clarifying the differentiation period of the flower bud according to the number of leaves extending from the flower bud and the degree of expansion of the leaflets includes: when 1-2 leaves are extended and the degree of expansion of the leaflets is <50%, it is determined that the flower bud is in the early stage of flower bud differentiation, that is, the flowering transition period or the sepal primordium differentiation period; when 1-2 leaves are extended and the degree of expansion of the leaflets is ≥50%, it is determined that the flower bud is in the middle and late stages of flower bud differentiation, that is, the petal primordium differentiation period or the stamen primordium differentiation period; when more than 2 leaves are extended and the degree of expansion of the leaflets is ≥50%, it is determined that the flower bud is in the late stage of flower bud differentiation, that is, the pistil primordium differentiation period.

[0016] Preferably, in the aforementioned method for in vivo identification of the flower bud differentiation period of 'Yueyuefen' rose, in step 3), the number of leaves protruding from the upper flower buds and the degree of leaflet expansion are observed with the naked eye or with the aid of a magnifying glass.

[0017] Compared with the prior art, the method for in vivo identification of the flower bud differentiation period of 'Yueyuefen' rose described in the present invention has the following beneficial effects: 1. According to the present invention, flower buds in different developmental states can be quickly identified, and the internal flower bud differentiation period of the flower buds can be accurately determined based on their external developmental morphology; 2. The external development state of the flower bud of the present invention corresponds to the differentiation period of the flower bud inside the flower bud, and the differentiation period of the flower bud can be judged only by observing the external development state of the flower bud with the naked eye or with the help of a magnifying glass without using histological preparation techniques such as paraffin sections, so that the differentiation period of the flower bud can be quickly determined on site; 3. The present invention directly observes the flower bud buds without destroying the plants or taking flower bud samples, thereby realizing non-destructive identification of the flower bud differentiation period.

[0018] The above description is only an overview of the technical solution of the present invention. In order to more clearly understand the technical means of the present invention and implement it according to the contents of the specification, the preferred embodiments of the present invention are described in detail as follows. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 A schematic diagram of a branch according to an embodiment of the present invention; Figure 2 It is a statistical diagram of bud types at different sites of the present invention; Figure 3 This is a correlation analysis diagram between the various stages of flower bud differentiation and the number of bud scales according to an embodiment of the present invention; Figure 4 This is a correlation analysis diagram between the various stages of flower bud differentiation and the degree of bud scale opening in an embodiment of the present invention; Figure 5 This is a correlation analysis diagram between each stage of flower bud differentiation and the number of leaf extensions according to an embodiment of the present invention; Figure 6 This is a correlation analysis diagram between each stage of flower bud differentiation and the degree of leaflet expansion in an embodiment of the present invention; 1- current year branches; 2- one year old branches. DETAILED DESCRIPTION

[0020] In order to further explain the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the specific implementation method, structure, characteristics and effects of the method for in vivo identification of the flower bud differentiation period of the rose "Yueyuefen" proposed by the present invention are described in detail below in combination with the preferred embodiments. In the following description, different "one embodiment" or "embodiment" do not necessarily refer to the same embodiment. In addition, specific features, structures or characteristics in one or more embodiments can be combined in any suitable form.

[0021] Unless otherwise specified, the materials, reagents, etc. involved below are all commercially available products well known to those skilled in the art; unless otherwise specified, the methods described are all methods known in the art. Unless otherwise defined, the technical terms or scientific terms used should have the usual meanings understood by ordinary technicians in the field to which the present invention belongs. If no specific experimental steps or conditions are specified below, the operations or conditions of the conventional experimental steps described in the literature in the field can be followed.

[0022] Some embodiments of the present invention provide a method for in vivo identification of the flower bud differentiation period of the rose 'Yueyuefen', comprising the following steps: Step 1: Determine the branch type and select the one-year-old branches or current-year branches with the strongest flowering ability; the number of flowers on perennial branches (two-year-old branches and above) is relatively small, so only one-year-old branches and current-year branches are counted during the specific implementation; Step 2: Determine the position of the mixed buds on the branches and divide them into sections; divide the annual branches or current year branches of the 'Yueyuefen' rose into three sections from top to bottom according to morphology, namely the upper part, the middle part, and the lower part, and select the upper part that is most likely to develop into flower buds for observation; Step 3: Classify the selected upper mixed buds according to their five external appearance states. The five judgment indicators include: the number of bud scales, the degree of bud scale opening, the extension of leaves, the number of leaf extensions, and the expansion of leaflets. The number of bud scales and the number of leaf extensions need to be counted; the opening of bud scales is divided into two states: "not open" or "slightly open" when the bud scale opening degree is less than 50%, and "half open" or "fully open" when the bud scale opening degree is ≥50%; the extension of leaves is divided into two states: "not extended" and "extended"; the expansion of leaflets is divided into two states: "not extended" or "slightly extended" when the leaflet expansion degree is less than 50%, and "half extended" or "fully extended" when the leaflet expansion degree is ≥50%.

[0023] Step 4: First, comprehensively judge whether it has entered the flowering transition based on the number of bud scales, the degree of opening of bud scales, and the extension of leaves of the upper mixed buds. When the number of bud scales is less than 5, the degree of opening of bud scales is less than 50%, the bud scales are "not open" or "slightly open", and the leaves are "not extended", it is in an undifferentiated state, that is, vegetative growth; when the number of bud scales is ≥5, the degree of opening of bud scales is ≥50%, the bud scales are "half open" or "fully open", and the leaves are "extended", it is judged to have entered the flowering transition, that is, reproductive growth; Step 5: Then, comprehensively judge the differentiation period of flower buds according to the number of leaves extending from the flower buds and the degree of expansion of leaflets. When 1-2 leaves extend, the degree of expansion of leaflets is less than 50%, and the leaflets are "not expanded" or "slightly expanded", the flower bud is in the early stage of flower bud differentiation, that is, the flowering transition period or the sepal primordium differentiation period; when 1-2 leaves extend, the degree of expansion of leaflets is ≥50%, the leaflets are "half expanded" or "fully expanded", the inside of the flower bud is in the middle and late stages of flower bud differentiation, that is, the petal primordium differentiation period or the stamen primordium differentiation period; when more than 2 leaves extend, the degree of expansion of leaflets is ≥50%, the leaflets are "half expanded" or "fully expanded", the inside of the flower bud is in the late stage of flower bud differentiation, that is, the pistil primordium differentiation period.

[0024] In some preferred embodiments, the method comprises the following steps: 1) Determine the branch type: One-year-old branches and current-year branches are the most flowering-capable branch types. Select one-year-old branches or current-year branches from vigorous growing rose plants (see Figure 1 ).

[0025] 2) Find the upper buds of annual branches: Find the mixed buds located on the upper part of the selected branches, observe the morphological upper 1 / 3 of the branches, and find the mixed buds in the leaf axils.

[0026] 3) Identify whether the mixed buds have entered reproductive growth: carefully push aside the leaves, and carefully observe the number of bud scales, the degree of opening of bud scales, and whether they extend out of the leaves with the naked eye or with the help of a magnifying glass under natural light. If the number of bud scales is ≥5, and the bud scales are "half open" or "fully open", it can be identified that it has entered the reproductive growth stage and started flower bud differentiation.

[0027] 4) Simple judgment of flower bud differentiation process: when 1-2 leaves are extended, the degree of leaflet expansion is <50%, and the leaflets are "not expanded" or "slightly expanded", the flower bud is in the early stage of flower bud differentiation, that is, the flowering transition period or the sepal primordium differentiation period; when 1-2 leaves are extended, the degree of leaflet expansion is ≥50%, and the leaflets are "half expanded" or "fully expanded", the flower bud is in the middle and late stages of flower bud differentiation, that is, the petal primordium differentiation period or the stamen primordium differentiation period; when more than 2 leaves are extended, the degree of leaflet expansion is ≥50%, and the leaflets are "half expanded" or "fully expanded", the flower bud is in the late stage of flower bud differentiation, that is, the pistil primordium differentiation period.

[0028] The present invention will be further described below in conjunction with specific embodiments, but this should not be construed as limiting the scope of protection of the present invention. Some non-essential improvements and adjustments made to the present invention by technicians in this field based on the above-mentioned contents of the present invention still fall within the scope of protection of the present invention.

[0029] Example 1 In the spring of 2024, live flower buds of potted 'Yueyuefen' roses were sampled and identified at the Xiaotangshan base of the National Flower Engineering Technology Research Center, and a total of 240 were identified. All mixed buds on the annual branches of 'Yueyuefen' roses were selected every 5 days, with three biological replicates each time. The buds in the leaf axils were observed in turn under natural light, and the three indicators of bud length, bud width, and bud point spacing were measured with a vernier caliper. Five indicators were recorded: the number of bud scales, the opening state of bud scales, the extension of leaves, the number of leaves, and the expansion of leaflets.

[0030] Then, the buds on the branches were cut off with a single-sided blade, quickly stored in 50% FAA fixative, and the development period inside the buds was identified by paraffin sectioning. After identification, the undifferentiated period, flowering transition period, sepal primordium differentiation period, petal primordium differentiation period, stamen primordium differentiation period, and pistil primordium differentiation period can be distinguished. In the undifferentiated period, the growth cone was smooth and convex as a whole; in the flowering transition period, the growth cone gradually flattened; in the sepal primordium differentiation period, the two sides of the growth cone showed slight protrusions; in the petal primordium differentiation period, the inner side of the original sepal primordium protrusion continued to produce protrusions; in the stamen primordium differentiation period, several small protrusions were produced on the inner side of the sepal primordium and petal primordium; in the pistil primordium differentiation period, several protrusions were produced in the middle of the growth cone.

[0031] Based on the analysis of the apparent characteristics and internal anatomy of each bud, the conclusion is: (1) The upper part of the annual branch should be selected as the sampling site, as the upper part is the site where flower buds are most likely to form; (2) The five indicators of bud scale number, bud scale opening degree, leaf extension, leaf extension number, and leaflet expansion are important indicators for judging the flower bud differentiation process of 'Yueyuefen'; however, bud length, bud width, and bud point spacing are affected by individual differences of plants, and the values ​​fluctuate greatly, which are not indicative for judging the flower bud differentiation period; (3) When the number of bud scales is less than 5, the degree of opening of bud scales is less than 50%, the bud scales are "not open" or "slightly open", and the leaves are "not extended", the inside of the flower bud is in an undifferentiated state, that is, vegetative growth; when the number of bud scales is ≥5, the degree of opening of bud scales is ≥50%, the bud scales are "half open" or "fully open", and the leaves are "extended", the inside of the flower bud enters the flowering transition, that is, reproductive growth; (4) As the flower buds elongate and extend leaves, the progress of flower bud differentiation can be easily judged based on the number of leaves extended. When 1-2 leaves extend, the degree of leaflet expansion is less than 50%, and the leaflets are "not expanded" or "slightly expanded", the flower bud is in the early stage of flower bud differentiation, that is, the flowering transition period or the sepal primordium differentiation stage; when 1-2 leaves extend, the degree of leaflet expansion is ≥50%, and the leaflets are "half expanded" or "fully expanded", the flower bud is in the middle and late stages of flower bud differentiation, that is, the petal primordium differentiation period or the stamen primordium differentiation period; when more than 2 leaves extend, the degree of leaflet expansion is ≥50%, and the leaflets are "half expanded" or "fully expanded", the flower bud is in the late stage of flower bud differentiation, that is, the pistil primordium differentiation period.

[0032] The specific results are as follows.

[0033] (1) Based on the statistics of the development results of different sites at the top, middle and bottom of one-year-old branches ( Figure 2 ), the percentages of flower buds, leaf buds, and undeveloped buds were 80.27%, 15.65%, and 4.08%, respectively. Therefore, the upper part of the one-year-old branches should be selected as the sampling site.

[0034] (2) Statistical analysis of the correlation between flower bud differentiation period and bud scale number ( Figure 3 ), the average number of bud scales in the undifferentiated period is 1.98, the average number of bud scales in the flowering transition period is 5.00, the average number of bud scales in the sepal primordium differentiation period is 5.67, the average number of bud scales in the petal primordium differentiation period is 5.63, the average number of bud scales in the stamen primordium differentiation period is 5.78, and the average number of bud scales in the pistil primordium differentiation period is 5.86. Therefore, when the number of bud scales is <5, the flower bud is in an undifferentiated state, and when the number of bud scales is ≥5, the flower bud enters the flowering transition.

[0035] 3) Statistical analysis of the correlation between flower bud differentiation period and bud scale opening degree ( Figure 4 ), the opening degree of bud scales is low in the undifferentiated period, with an average opening degree of 23%, and the opening degree of bud scales is less than 50%, which is a "not open" or "slightly open" state; the opening degree of bud scales is 65% in the flowering transition period, 83% in the sepal primordium differentiation period, 88% in the petal primordium differentiation period, 95% in the stamen primordium differentiation period, and 100% in the pistil primordium differentiation period. After entering the flowering transition period, the opening degree of bud scales is high, and the opening degree of bud scales is ≥50%, which is a "half open" or "fully open" state. Therefore, when the opening degree of bud scales is less than 50%, the bud scales are "not open" or "slightly open", the flower bud is in the undifferentiated period, and when the opening degree of bud scales is ≥50%, the bud scales are in a "half open" or "fully open" state, the flower bud enters the flowering transition.

[0036] 4) Statistical analysis of the correlation between flower bud differentiation period and leaf outgrowth number ( Figure 5 ), the average number of leaves in the undifferentiated period is 0.19, the average number of leaves in the flowering transition period is 1.13, the average number of leaves in the sepal primordium differentiation period is 1.22, the average number of leaves in the petal primordium differentiation period is 1.55, the average number of leaves in the stamen primordium differentiation period is 1.69, and the average number of leaves in the pistil primordium differentiation period is 2.75. Therefore, when 1-2 leaves are extended, the inside of the flower bud is in the early to middle and late stages of flower bud differentiation (flowering transition period to stamen primordium differentiation period); when more than 2 leaves are extended, the inside of the flower bud is in the late stage of flower bud differentiation (pistil primordium differentiation period).

[0037] 5) Statistical analysis of the correlation between flower bud differentiation period and leaflet expansion degree ( Figure 6 ), after entering the flowering transition, the average leaflet expansion degree is as follows: 38% in the flowering transition period, 40% in the sepal primordium differentiation period, the leaflet expansion degree is <50%, which is a "not expanded" or "slightly expanded" state; 83% in the petal primordium differentiation period, 93% in the stamen primordium differentiation period, and 100% in the pistil primordium differentiation period, the leaflet expansion degree is high, and the leaflet expansion degree is ≥50%, which is a "semi-expanded" or "fully expanded" state. Therefore, when the leaflet expansion degree is <50%, the leaflet is "not expanded" or "slightly expanded", the inside of the flower bud is in the early stage of flower bud differentiation, that is, the flowering transition period or the sepal primordium differentiation stage; when the leaflet expansion degree is ≥50%, the leaflet is "semi-expanded" or "fully expanded", the inside of the flower bud is in the middle and late to late stages of flower bud differentiation (petal primordium differentiation period to pistil primordium differentiation period).

[0038] Example 2 In the summer of 2024, according to the summary method of Example 1, multiple upper buds of the current year's branches of the rose 'Yueyuefen' were selected for identification and verified by paraffin sections. A total of 185 buds were observed and identified.

[0039] The results showed that the percentage of buds in the undifferentiated period was 100% when the number of bud scales was <5, the degree of bud scale opening was <50%, the bud scales were "not open" or "slightly open", and the leaves were "not extended"; The number of bud scales was ≥5, the degree of bud scale opening was ≥50%, the bud scales were "half-opened" or "fully-opened", and the leaves were "extended", the buds that entered the flowering transformation accounted for 84.31%; 1-2 leaves were extended, the degree of leaflet expansion was less than 50%, and the leaflets were "not expanded" or "slightly expanded". 42.31% of the flower buds were in the flowering transition period, and 57.69% were in the sepal primordium differentiation period. 1-2 leaves were extended, the degree of leaflet expansion was ≥50%, and the flower buds with leaflets "half-expanded" or "fully expanded" were in the stage of sepal primordium differentiation at 3.57%, in the stage of petal primordium differentiation at 50%, and in the stage of stamen primordium differentiation at 46.43%; Among the flower buds with more than two leaves extended and leaflets expanded ≥50%, and leaflets "half-expanded" or "fully-expanded", 4.29% were in the stage of stamen primordium differentiation, and 95.71% were in the stage of pistil primordium differentiation.

[0040] The above results confirm that the flower bud differentiation period meets the judgment criteria, which once again proves the reliability of the technology of the present invention.

[0041] Therefore, the present invention can be used to efficiently complete the identification of the flower bud differentiation process of the rose 'Yueyuefen' in a living state with non-destructive sampling.

[0042] Example 3 In the autumn of 2024, according to the summary method of Example 1, multiple upper buds of the current year's branches of the rose 'Yueyuefen' were selected for identification and verified by paraffin sections. A total of 136 buds were observed and identified.

[0043] The results showed that the percentage of buds in the undifferentiated period was 100% when the number of bud scales was <5, the degree of bud scale opening was <50%, the bud scales were "not open" or "slightly open", and the leaves were "not extended"; The number of bud scales was ≥5, the degree of bud scale opening was ≥50%, the bud scales were "half-opened" or "fully-opened", and the leaves were "extended", the buds that entered the flowering transformation accounted for 76.96%; 1-2 leaves were extended, the degree of leaflet expansion was less than 50%, and the leaflets were "not expanded" or "slightly expanded". 57.78% of the flower buds were in the flowering transition period, and 42.22% were in the sepal primordium differentiation period; 1-2 leaves were extended, the degree of leaflet expansion was ≥50%, and the flower buds with leaflets "half-expanded" or "fully expanded" were in the stage of petal primordium differentiation at 52.63%, and in the stage of stamen primordium differentiation at 47.37%; For flower buds with more than 2 leaves extended and leaflets expanded ≥50%, and leaflets "half-expanded" or "fully expanded", 100% of them are in the stage of pistil primordium differentiation.

[0044] The above results confirm that the flower bud differentiation period meets the judgment criteria, which once again proves the reliability of the technology of the present invention.

[0045] Therefore, the present invention can be used to efficiently complete the identification of the flower bud differentiation process of the rose 'Yueyuefen' in a living state with non-destructive sampling.

[0046] In the specification of the present invention, a lot of specific details are described. However, it is understood that embodiments of the present invention can be practiced without these specific details. In some embodiments, well-known methods, structures and techniques are not shown in detail so as not to obscure the understanding of this specification.

[0047] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

[0048] In addition, various embodiments of the present invention may be arbitrarily combined, and as long as they do not violate the concept of the present invention, they should also be regarded as the contents disclosed by the present invention.

[0049] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Any simple modification, equivalent change and modification made to the above embodiment according to the technical essence of the present invention still falls within the scope of the technical solution of the present invention.

Claims

1. A method for identifying the flower bud differentiation period of 'Yueyuefen' rose in vivo, characterized in that: The following steps are involved: 1) Determine the position of the mixed buds of the annual branches or current year branches and divide them into sections; According to morphology, it is divided into three sections from the upper part to the lower part, namely the upper part, the middle part and the lower part, and the mixed buds in the upper part are selected; 2) According to the number of bud scales, the degree of bud scale opening, and the extension of leaves of the upper mixed buds, the growth period of the upper mixed buds can be determined; 3) Determine the differentiation period of the flower bud based on the number of leaves extending from the flower bud and the degree of expansion of the leaflets.

2. The method for living identification of the flower bud differentiation period of the rose 'Yueyuefen' as claimed in claim 1, characterized in that: In step 1), the upper mixed buds are mixed buds in the upper 1 / 3 section of the branch morphology.

3. The method for living identification of the flower bud differentiation period of 'Yueyuefen' rose as claimed in claim 2, characterized in that: In step 1), the upper mixed buds are mixed buds at the upper 1 / 3 of the branch morphology and at the leaf axils.

4. The method for living identification of the flower bud differentiation period of 'Yueyuefen' rose as claimed in claim 3, characterized in that: In step 1), the upper mixed buds are mixed buds at the morphological upper 1 / 3 section and leaf axils of annual branches or current year branches.

5. The method for living identification of the flower bud differentiation period of 'Yueyuefen' rose as claimed in claim 1, characterized in that: In step 2), the step of clarifying the growth period of the upper mixed bud according to the number of bud scales, the degree of opening of the bud scales, and the extension of the leaves of the upper mixed buds includes: when the number of bud scales is <5, it is determined that the upper mixed bud is in the undifferentiated stage; when the number of bud scales is ≥5, it is determined that the upper mixed bud has entered the flowering transformation and is in the reproductive growth stage.

6. The method for living identification of the flower bud differentiation period of the rose 'Yueyuefen' as claimed in claim 5, characterized in that: In step 2), the step of clarifying the growth period of the upper mixed buds according to the number of bud scales, the degree of opening of the bud scales, and the extension of leaves of the upper mixed buds includes: when the number of bud scales is <5 and the degree of opening of the bud scales is <50%, it is determined that the upper mixed buds are in the undifferentiated stage; when the number of bud scales is ≥5 and the degree of opening of the bud scales is ≥50%, it is determined that the upper mixed buds have entered the flowering transformation and are in the reproductive growth stage.

7. The method for living identification of the flower bud differentiation period of 'Yueyuefen' rose as claimed in claim 6, characterized in that: In step 2), the step of clarifying the growth period of the upper mixed bud according to the number of bud scales, the degree of opening of the bud scales, and the extension of the leaves of the upper mixed buds includes: when the number of bud scales is <5, the degree of opening of the bud scales is <50%, and the leaves are "not extended", it is determined that the upper mixed bud is in the undifferentiated stage; when the number of bud scales is ≥5, the degree of opening of the bud scales is ≥50%, and the leaves are "extended", it is determined that the upper mixed bud has entered the flowering transformation and is in the reproductive growth stage.

8. The method for living identification of the flower bud differentiation period of the rose 'Yueyuefen' as claimed in claim 1, characterized in that: In step 2), the number of bud scales, the degree of opening of bud scales, and the extension of leaves of the upper mixed buds are observed with the naked eye or with the aid of a magnifying glass.

9. The method for living identification of the flower bud differentiation period of the rose 'Yueyuefen' as claimed in claim 1, characterized in that: In step 3), the step of determining the differentiation period of the flower bud according to the number of leaves extending from the flower bud and the degree of expansion of the leaflets includes: when 1-2 leaves are extended, it is determined that the flower bud is in the early to middle and late stages of flower bud differentiation; when more than 2 leaves are extended, it is determined that the flower bud is in the late stage of flower bud differentiation.

10. The method for identifying the flower bud differentiation period of the rose 'Yueyuefen' in vivo as claimed in claim 9, characterized in that: In step 3), the step of determining the differentiation period of the flower bud according to the number of leaves extending from the flower bud and the degree of expansion of the leaflets comprises: when 1-2 leaves are extended and the degree of expansion of the leaflets is less than 50%, it is determined that the flower bud is in the early stage of flower bud differentiation; when 1-2 leaves are extended and the degree of expansion of the leaflets is ≥50%, it is determined that the flower bud is in the middle and late stages of flower bud differentiation; when more than 2 leaves are extended and the degree of expansion of the leaflets is ≥50%, it is determined that the flower bud is in the late stage of flower bud differentiation; the number of leaves extending from the upper flower bud and the degree of expansion of the leaflets are observed with the naked eye or with the aid of a magnifying glass.

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