A method for treating winter jujube scion using ubiquitin and its application in grafting

Abstract

The present invention belongs to the technical field of fruit tree grafting, and specifically relates to a method for treating winter jujube scions using ubiquitin and its application in grafting. The method for treating winter jujube scions comprises: (1) preparing a ubiquitin sodium chloride solution; (2) soaking treatment; (3) irradiation treatment. The present invention uses ultraviolet light irradiation treatment on the scions soaked in ubiquitin, so that ubiquitin fully penetrates into the cells and tissues of the scions to play a role in regulating gene transcription and expression. Grafting is to graft the treated winter jujube scions onto rootstocks, and the rootstocks include winter jujube trees, sour jujube trees, and golden silk jujube trees. Using soaked and irradiated winter jujube scions for grafting can effectively prevent the methylation of lysine in the histone of the winter jujube scions, significantly reduce the frequency of the "reversion" phenomenon of the scions, and enable the excellent traits of the scions to be expressed stably and long-term, resulting in large fruit yield, excellent quality, and high economic value.

Description

Technical Field

[0001] The invention belongs to the technical field of fruit tree grafting, and particularly relates to a method for processing winter jujube scions using ubiquitin and an application thereof in grafting. Background Art

[0002] Grafting is a commonly used asexual reproduction method for winter jujube trees. It can not only maintain the good disease resistance, adaptability and vitality of the rootstock, but also help shorten the childhood of the jujube tree and achieve stable and high yields as soon as possible.

[0003] The young shoots of winter jujube will have gene mutations, which are also called bud mutations. Some mutants have one or several traits that are better than the mother plant. When these mutants are grafted onto other jujube trees as scions, their excellent traits can be expressed in the short term, but after 3-5 years, they will be inhibited, unstable or even unable to express due to the methylation of histone lysine in the scion, and will revert to the traits before the mutation. This phenomenon of poor expression of mutant traits is the "atavism" phenomenon of the scion.

[0004] Zhandong No. 2 is an excellent bud mutation variety of ordinary winter jujube. The difference in its characteristics from ordinary winter jujube is that it has short internodes, large fruit, high sugar content and crispy flesh. Using the branches of Zhandong No. 2 winter jujube to make scions and graft them onto winter jujube, golden silk jujube or sour jujube trees, a large number of winter jujubes with the above excellent traits can be produced in a short period of time. However, in long-term planting, the "reversion" phenomenon of Zhandong No. 2 winter jujube grafted plants has a high incidence and obvious manifestations. The survey shows that the probability of "reversion" of Zhandong No. 2 scions grafted through branches within five years after grafting is 35%-55%, and the excellent traits of short internodes, large fruit, high sugar content and crispy flesh are lost. The sugar content of the fruit drops from no less than 28% to 18%-23%, seriously affecting the fruit yield and quality.

[0005] Ubiquitin is a type of protein with a relatively low molecular weight, with only 76 amino acid residues and a highly conserved sequence. The most widely used application of ubiquitin in the prior art is to regulate protein degradation. In addition to regulating protein degradation and removing erroneous proteins, ubiquitin also has a regulatory effect on cell growth cycle, DNA replication, chromosome structure, gene expression and transcription, and signal transduction. However, there is little research on the application of ubiquitin in plant cultivation and reproduction, especially in the field of grafting. Summary of the invention

[0006] In view of the problem in the prior art that the winter jujube scion is prone to "reversion" during long-term cultivation after grafting, it is difficult to maintain excellent traits for a long time, and the fruit yield and quality are affected, the present invention provides a method for treating winter jujube scion using ubiquitin and applies it in grafting. The present invention uses ultraviolet light irradiation to treat the scion soaked in ubiquitin and then grafted, which can effectively prevent the methylation of lysine in the histone of the winter jujube scion, significantly reduce the frequency of the "reversion" phenomenon of the scion, and enable the excellent traits of the scion to be expressed stably for a long time.

[0007] In a first aspect, the present invention provides a method for treating winter jujube scion using ubiquitin, comprising the following steps:

[0008] (1) preparing ubiquitin sodium chloride solution: dissolving ubiquitin in a sodium chloride solution to prepare a ubiquitin sodium chloride solution;

[0009] (2) soaking treatment: soaking the winter jujube scion in a ubiquitin sodium chloride solution to obtain a soaked winter jujube scion;

[0010] (3) Irradiation treatment: The soaked winter jujube scion is irradiated with ultraviolet light, and other light sources are avoided during the irradiation process to obtain the soaked and irradiated winter jujube scion. The use of ultraviolet light irradiation can promote the germination of the winter jujube scion while promoting the effect of ubiquitin.

[0011] Furthermore, in step (1), ubiquitin is dissolved in a sodium chloride solution with a mass concentration of 1% to prepare a ubiquitin sodium chloride solution with a ubiquitin mass concentration of 0.1%-0.5%.

[0012] Preferably, the winter jujube scion in step (2) is a scion made by cutting branches of the winter jujube tree of the Zhandong No. 2 variety.

[0013] Furthermore, in step (2), the ubiquitin sodium chloride solution is placed in an ultrasonic generator, and the ultrasonic generator is started and then the winter jujube scion is immersed. During the immersion process, the ultrasonic generator is turned on to effectively promote the ubiquitin in the ubiquitin sodium chloride solution to fully penetrate into the cells and tissues of the winter jujube scion, and the ultrasonic generator includes an ultrasonic cleaning machine.

[0014] Furthermore, the frequency of the ultrasonic generator is 28-40KHz, the water temperature during immersion is 30-38°C, and the immersion time is 30-60min.

[0015] Furthermore, in step (3), the soaked winter jujube scion is placed in a light incubator and irradiated with ultraviolet light for 24-36 hours, and the ultraviolet light irradiance is 80-100uw / cm 2 , the ambient temperature during the irradiation process is 5-10℃.

[0016] Furthermore, after step (3) is completed, the soaked and irradiated winter jujube scions are placed in a light-proof device for storage, and the light-proof device includes a black light-proof bag.

[0017] In a second aspect, the present invention provides an application of the above-mentioned winter jujube scion processing method in grafting, wherein the winter jujube scion that has been soaked and irradiated is grafted onto a rootstock, and the rootstock includes a winter jujube tree, a sour jujube tree, and a golden thread jujube tree; the "reversion" rate of the winter jujube scion that has been soaked and irradiated is ≤5% after grafting for 5 years.

[0018] Furthermore, the soaked and irradiated winter jujube scion is immediately wrapped with a black plastic film after grafting, and the black plastic film is removed 3-5 days after grafting.

[0019] Further, after removing the black plastic film, an orange plastic bag is immediately used to wrap the soaked and irradiated winter jujube scion, and the bag is removed after 10-15 days of wrapping; the orange plastic bag is made of low-density polyethylene, with a thickness of 2-4 threads and a size of 3-5cm*8-12cm; the color of the orange plastic bag is expressed in RGB as R=250-255, G=120-130, and B=0-40. Using an orange plastic bag to wrap the soaked and irradiated winter jujube scion can allow the soaked and irradiated winter jujube scion to fully receive the orange light that promotes the germination and nutritional growth of the jujube tree, while filtering out other light, especially effectively filtering out the purple light that inhibits the growth of new buds, which can improve the survival rate of the scion and help stabilize the excellent traits of Zhandong No. 2.

[0020] After grafting, wrapping the soaked and irradiated winter jujube scions with black plastic film and orange plastic bags successively can improve the survival rate of the scions and help prolong the action time of ubiquitin, thereby further stabilizing the excellent traits of Zhandong No. 2. At the same time, wrapping the grafted scions with black plastic film first and then with orange plastic bags can increase the sensitivity of the grafted scions to orange light, which is more conducive to promoting the germination of new buds on the scions.

[0021] The beneficial effects of the present invention are:

[0022] The present invention uses ultraviolet irradiation treatment on the scion soaked in ubiquitin, so that ubiquitin can fully penetrate into the cells and tissues of the scion to play a role in regulating gene transcription and expression. Using the soaked and irradiated winter jujube scion for grafting can effectively prevent the methylation of lysine in the histone of the winter jujube scion, significantly reduce the frequency of the "reversion" phenomenon of the scion after many years of grafting, and enable the excellent traits of the scion to be expressed stably for a long time. The "reversion" rate of the scion after five years of grafting using the soaked and irradiated winter jujube scion is ≤5%, and the winter jujube yield is large, the quality is excellent, and the economic value is high. DETAILED DESCRIPTION

[0023] In order to enable those skilled in the art to better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0024] Example 1

[0025] A method for treating winter jujube scion using ubiquitin comprises the following steps:

[0026] (1) preparing a ubiquitin sodium chloride solution: dissolving ubiquitin in a sodium chloride solution having a mass concentration of 1% to prepare a ubiquitin sodium chloride solution having a mass concentration of 0.3%;

[0027] (2) Soaking treatment: The sodium chloride solution is placed in an ultrasonic cleaning machine, the ultrasonic frequency is set to 28 kHz, the water temperature is controlled to 30° C. After starting the ultrasonic cleaning machine, the scion made by cutting the branches of the winter jujube tree of the Zhandong No. 2 variety is placed in the ultrasonic cleaning machine and soaked for 30 minutes to obtain the soaked winter jujube scion;

[0028] (3) Irradiation treatment: The soaked winter jujube scion was placed in a light incubator and irradiated with ultraviolet light for 24 hours at an irradiance of 80uw / cm 2 During the irradiation process, avoid exposure to other light sources and control the ambient temperature to 5-10°C to obtain the winter jujube scion after soaking and irradiation.

[0029] Example 2

[0030] A method for treating winter jujube scion using ubiquitin comprises the following steps:

[0031] (1) preparing a ubiquitin sodium chloride solution: dissolving ubiquitin in a sodium chloride solution having a mass concentration of 1% to prepare a ubiquitin sodium chloride solution having a mass concentration of 0.1%;

[0032] (2) Soaking treatment: The sodium chloride solution is placed in an ultrasonic cleaning machine, the ultrasonic frequency is set to 36 kHz, the water temperature is controlled to 34° C. After starting the ultrasonic cleaning machine, the scion made by cutting the branches of the winter jujube tree of the Zhandong No. 2 variety is placed in the ultrasonic cleaning machine and soaked for 60 minutes to obtain the soaked winter jujube scion;

[0033] (3) Irradiation treatment: The soaked winter jujube scion was placed in a light incubator and irradiated with ultraviolet light for 30 hours at an irradiance of 90uw / cm 2 During the irradiation process, avoid exposure to other light sources and control the ambient temperature to 5-10°C to obtain the winter jujube scion after soaking and irradiation.

[0034] Example 3

[0035] A method for treating winter jujube scion using ubiquitin comprises the following steps:

[0036] (1) preparing a ubiquitin sodium chloride solution: dissolving ubiquitin in a sodium chloride solution having a mass concentration of 1% to prepare a ubiquitin sodium chloride solution having a mass concentration of 0.5%;

[0037] (2) Soaking treatment: The sodium chloride solution is placed in an ultrasonic cleaning machine, the ultrasonic frequency is set to 40 kHz, the water temperature is controlled to 38° C. After starting the ultrasonic cleaning machine, the scion made by cutting the branches of the winter jujube tree of the Zhandong No. 2 variety is placed in the ultrasonic cleaning machine and soaked for 50 minutes to obtain the soaked winter jujube scion;

[0038] (3) Irradiation treatment: The soaked winter jujube scion was placed in a light incubator and irradiated with ultraviolet light for 36 hours at an irradiance of 100 uw / cm 2 During the irradiation process, avoid exposure to other light sources and control the ambient temperature to 5-10°C to obtain the winter jujube scion after soaking and irradiation.

[0039] Example 4

[0040] The winter jujube trees were used as rootstocks, and the soaked and irradiated winter jujube scions obtained in Example 1 were used to graft 216 winter jujube trees in a jujube garden of 3 mu. After grafting, the soaked and irradiated winter jujube scions were immediately wrapped with a transparent plastic film. The transparent plastic film was removed 3 days after grafting.

[0041] After 5 years of grafting, the appearance of the scion, the yield, size, sugar content and brittleness of the grafted winter jujube trees were counted, and the incidence of the "reversion" phenomenon and the survival rate of the scion of the grafted winter jujube trees were calculated. It was calculated that the "reversion" rate of the grafted winter jujube trees in Example 4 was 3.24%, and the scion survival rate was 90.28%.

[0042] As a comparison, untreated winter jujube scions of the Zhandong No. 2 variety were directly grafted onto winter jujube rootstocks. Five years after grafting, the appearance, yield, size, sugar content and brittleness of the grafted winter jujube trees were counted, and the incidence of "reversion" and the survival rate of the grafted winter jujube trees were calculated. The calculation showed that the "reversion" rate of the grafted winter jujube trees was 48.61%, and the scion survival rate was 92.13%.

[0043] Example 5

[0044] The 216 jujube trees in a jujube garden of 3 mu were grafted using the soaked and irradiated winter jujube scions obtained in Example 1. The soaked and irradiated winter jujube scions were immediately wrapped with a black plastic film after grafting. The black plastic film was removed 4 days after grafting, and the irradiated winter jujube scions were immediately wrapped with an orange plastic bag, and the bag was removed after 12 days of wrapping; the orange plastic bag was made of low-density polyethylene, with a thickness of 3 threads and a size of 4 cm×10 cm; the color of the orange plastic bag was expressed in RGB color mode as R=250, G=125, and B=20.

[0045] After 5 years of grafting, the appearance of the scion, the yield, size, sugar content and brittleness of the grafted winter jujube trees were counted, and the incidence of the "reversion" phenomenon and the survival rate of the scion of the grafted winter jujube trees were calculated. According to statistics, the grafted winter jujube trees of Example 5 did not have the "reversion" phenomenon, and the scion survival rate was 98.15%.

[0046] As a comparison, untreated scions were directly grafted onto sour jujube rootstocks. Five years after grafting, the appearance, yield, size, sugar content and brittleness of the grafted winter jujube trees were counted, and the incidence of "reversion" and the survival rate of the scions were calculated. The calculation showed that the "reversion" rate of the grafted winter jujube trees was 37.96%, and the scion survival rate was 91.67%.

[0047] Example 6

[0048] The winter jujube tree was used as the rootstock, and the soaked and irradiated winter jujube scion obtained in Example 2 was used to graft 216 winter jujube trees in a jujube garden of 3 mu. The soaked and irradiated winter jujube scion was immediately wrapped with a black plastic film after grafting. The black plastic film was removed 5 days after grafting, and the soaked and irradiated winter jujube scion was immediately wrapped with an orange plastic bag, and the bag was removed after 15 days of wrapping; the orange plastic bag was made of low-density polyethylene, with a thickness of 4 threads and a size of 5 cm×12 cm; the color of the orange plastic bag was expressed in RGB color mode as R=255, G=130, and B=40.

[0049] After 5 years of grafting, the appearance of the scion, the yield, size, sugar content and brittleness of the grafted winter jujube trees were counted, and the incidence of the "reversion" phenomenon of the grafted winter jujube trees was calculated. The calculation showed that the "reversion" rate of the grafted winter jujube trees in Example 6 was 3.70%.

[0050] Example 7

[0051] The winter jujube tree was used as the rootstock, and the soaked and irradiated winter jujube scion obtained in Example 3 was used to graft 216 winter jujube trees in a jujube garden of 3 mu. The soaked and irradiated winter jujube scion was immediately wrapped with a black plastic film after grafting. The black plastic film was removed 3 days after grafting, and the soaked and irradiated winter jujube scion was immediately wrapped with an orange plastic bag, and the bag was removed after 10 days of wrapping; the orange plastic bag was made of low-density polyethylene, with a thickness of 4 threads and a size of 3 cm×8 cm; the color of the orange plastic bag was expressed in RGB color mode as R=250, G=120, and B=20.

[0052] After 5 years of grafting, the appearance of the scion, the yield, size, sugar content and brittleness of the grafted winter jujube trees were counted, and the incidence of the "reversion" phenomenon of the grafted winter jujube trees was calculated. It was calculated that the "reversion" rate of the grafted winter jujube trees in Example 7 was 2.78%.

[0053] The present invention has been described in detail by way of preferred embodiments, but the present invention is not limited thereto. Without departing from the spirit and essence of the present invention, a person of ordinary skill in the art may make various equivalent modifications or substitutions to the embodiments of the present invention, and such modifications or substitutions shall be within the scope of the present invention. Any person of ordinary skill in the art may easily conceive of changes or substitutions within the technical scope disclosed by the present invention, and such changes or substitutions shall be within the scope of protection of the present invention.

Claims

1. Application of a method for treating winter jujube scion using ubiquitin in grafting, It is characterized in that The winter jujube scion processing method comprises the following steps: (1) Dissolving ubiquitin in a sodium chloride solution to prepare a ubiquitin sodium chloride solution; (2) soaking the winter jujube scion in a ubiquitin sodium chloride solution to obtain a soaked winter jujube scion; (3) irradiating the soaked winter jujube scion with ultraviolet light, avoiding exposure to other light sources during the irradiation process, and obtaining the soaked and irradiated winter jujube scion; The application is to graft the soaked and irradiated winter jujube scion onto the rootstock, which includes winter jujube tree, sour jujube tree and golden silk jujube tree; The "reversion" rate of winter jujube scion grafted after soaking and irradiation is ≤5% 5 years later.

2. The use according to claim 1, It is characterized in that Step (1) dissolving ubiquitin in a sodium chloride solution with a mass concentration of 1% to prepare a ubiquitin sodium chloride solution with a mass concentration of 0.1%-0.5%.

3. The use according to claim 1, It is characterized in that The winter jujube scion in step (2) is a scion made by cutting branches of the winter jujube tree of the Zhandong No. 2 variety.

4. The use according to claim 1, It is characterized in that In step (2), the ubiquitin sodium chloride solution is placed in an ultrasonic generator, and the winter jujube scion is placed in the ultrasonic generator for immersion after the ultrasonic generator is started.

5. The use according to claim 4, It is characterized in that The frequency of the ultrasonic generator is 28-40KHz, the temperature during immersion is 30-38°C, and the immersion time is 30-60min.

6. The use according to claim 1, It is characterized in that Step (3) placing the soaked winter jujube scion in a light incubator and irradiating it with ultraviolet light for 24-36 hours, with an ultraviolet light irradiance of 80-100uw / cm 2 , the ambient temperature during the irradiation process is 5-10℃.

7. The use according to claim 1, It is characterized in that After step (3) is completed, the soaked and irradiated winter jujube scions are placed in a light-proof device for storage.

8. The use according to claim 1, It is characterized in that Immediately after grafting, wrap the soaked and irradiated winter jujube scion with black plastic film, and remove the black plastic film 3-5 days after grafting.

9. The use according to claim 8, It is characterized in that After removing the black plastic film, immediately use orange plastic bags to wrap the soaked and irradiated winter jujube scions, and remove the bags after wrapping for 10-15 days; the orange plastic bags are made of low-density polyethylene, with a thickness of 2-4 threads and a size of 3-5cm*8-12cm; the color of the orange plastic bags is expressed in RGB color mode as R=250-255, G=120-130, B=0-40.