A method for promoting oil palm callus proliferation and somatic embryo induction by using baicalin

Abstract

The application discloses a method for promoting oil palm callus proliferation and somatic embryo induction by using baicalin. The tender yellow leaf of oil palm is used as a tissue culture explant, and Y3 medium is used as a basic medium, and baicalin is added to promote the induction and proliferation of oil palm callus and somatic embryo generation. In the callus induction and proliferation stage, 0.5 mg / L of baicalin can replace part of the effect of 2,4-D or dicamba, reduce the dosage of 2,4-D or dicamba, reduce the risk of subsequent regeneration deformity, shorten the callus induction time by 20 days, and increase the callus proliferation rate by 48.19%; in the somatic embryo induction stage, 0.3 mg / L of baicalin can completely replace the plant hormone 6-BA, and the somatic embryo induction time is shortened by 31 days, and the somatic embryo induction rate is increased by 5%. Therefore, the addition of baicalin in the medium can improve the efficiency of oil palm callus proliferation and somatic embryo induction, shorten the induction time, and has a good popularization and application prospect in the field of oil palm tissue culture and rapid propagation.

Description

Technical Field

[0001] This invention relates to the field of plant tissue culture technology, and in particular to a method for promoting callus proliferation and somatic embryo induction in oil palm using baicalin. Background Technology

[0002] Oil palm, known as the "King of Oils," produces approximately 80 million tons of palm oil globally, with China importing about 6 million tons annually, making it a crucial guarantee for my country's edible oil supply security. However, the development of my country's oil palm industry is constrained by a shortage of high-quality seedlings.

[0003] In recent years, the Coconut Research Institute of the Chinese Academy of Tropical Agricultural Sciences has cultivated a new oil palm variety, "Hot Oil No. 1," through introduction and selection. This variety produces oval or elliptical fruits with a smooth, orange-red exocarp and a thick mesocarp. After three years of planting in Hainan, over 80% of the plants bear fruit annually, producing 10-15 bunches per plant, with each bunch weighing 10-15 kg. The oil yield per bunch is 22.9%-29.3%, and the palm oil contains 46.0%-46.7% unsaturated fatty acids. In the eighth year after planting, the yield per bunch reaches 860.3 kg / mu (approximately 0.067 hectares), equivalent to 239.2 kg of oil, making it a high-yielding variety. Tissue culture technology, as an important method for rapid asexual propagation, can achieve clonal propagation of superior individual plants while maintaining the genetic characteristics of the maternal parent, which is of great significance for the promotion and application of superior varieties such as "Hot Oil No. 1." However, existing oil palm tissue culture systems mainly rely on high concentrations of herbicide-based plant hormones (such as 2,4-D or dicamba), which often leads to long callus formation cycles, low proliferation efficiency, low somatic embryogenesis rates, poor reproducibility, and excessively long seedling development cycles (usually 24-30 months). Therefore, reducing the risk of synthetic hormones, improving somatic embryo induction rates, and shortening the culture cycle are key technologies for constructing a rapid propagation system for oil palm tissue culture seedlings. Breakthroughs in this technology can provide technical solutions for the asexual propagation of superior oil palm varieties, achieving a leapfrog innovation in the rapid asexual propagation of oil palm seedlings, and providing strong technical support for the high-quality expansion and increased production of my country's oil palm industry. Summary of the Invention

[0004] To address the aforementioned deficiencies in the prior art, this invention proposes a method for promoting callus proliferation and somatic embryo induction in oil palm using baicalin, thereby resolving the problems mentioned in the background art.

[0005] To achieve the above objectives, the present invention provides the following technical solution:

[0006] A method for promoting callus proliferation and somatic embryo induction in oil palm using baicalin includes the following steps:

[0007] (1) Callus induction: Take the tender yellow leaves of mature oil palm as explants, cut them into small sections, soak them in sterile water and dry them, and inoculate them into the callus induction medium. Oil palm callus tissue was obtained in 95-110 days.

[0008] The composition and formulation of the callus induction medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.1~0.5 mg / L of baicalin, 4~6 g / L of plant gel, 30~40 g / L of sucrose, and any one of 5~10 mg / L of dicamba or 5~10 mg / L of 2,4-D.

[0009] (2) Callus proliferation culture: The oil palm callus obtained in step (1) is inoculated into the callus proliferation culture medium, and the proliferated oil palm callus is obtained in 25-30 days;

[0010] The composition and formulation of the callus proliferation medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.1~0.5 mg / L of baicalin, 30~40 g / L of sucrose, and any one of 5~10 mg / L of dicamba or 5~10 mg / L of 2,4-D.

[0011] (3) Somatic embryo induction culture: The oil palm callus proliferation tissue obtained in step (2) was transferred to the somatic embryo induction culture medium. After 3 months of culture, it was observed whether white spherical somatic embryos appeared on the surface. Oil palm somatic embryos were obtained in 94-150 days.

[0012] The composition and formulation of the somatic embryo induction medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.1~0.5 mg / L of baicalin, 6~10 g / L of plant gel, and 15~20 g / L of sucrose.

[0013] Preferably, the Y3 culture medium formula (per liter) is: calcium chloride dihydrate 29.4 g / L, potassium chloride 149.6 g / L, ammonium chloride 53.6 g / L, potassium nitrate 202.4 g / L, sodium dihydrogen phosphate 27.56 g / L, boric acid 0.3092 g / L, magnesium sulfate heptahydrate 24.8 g / L, manganese sulfate 0.848 g / L, potassium iodide 0.832 g / L, copper sulfate 0.0248 g / L, and zinc sulfate 0.7 g / L. 2 g / L, nickel chloride 0.0024 g / L, cobalt chloride 0.024 g / L, sodium molybdate dihydrate 0.024 g / L, disodium EDTA 3.724 g / L, ferrous sulfate heptahydrate 1.5 g / L, thiamine hydrochloride 0.1012 g / L, pyridoxine hydrochloride 0.1028 g / L, nicotinic acid 0.1 g / L, L-asparagine 8.8 g / L, L-arginine 10 g / L, L-glutamine 10 g / L.

[0014] Preferably, the preparation method for baicalin (English name: Baicalin; CAS No.: 21967-41-9) is as follows: Weigh 25 mg of baicalin powder, add 5 mL of dimethyl sulfoxide (DMSO), and shake or sonicate for 10-15 min until completely dissolved to prepare a stock solution with a concentration of 5 g / L. After sterilizing the stock solution by filtration through a 0.22 μm microporous membrane, dispense 0.5-1 mL into sterile centrifuge tubes and store at -4°C protected from light for later use. When using, take an appropriate amount of the stock solution and add it to sterilized culture medium cooled to 50-60°C, mix well, and then dispense.

[0015] Preferably, the method for obtaining explants in step (1) includes the following steps: select “Hot Oil No. 1” oil palm that has been flowering and fruiting normally for more than 3 years, remove the outer 2-3 leaves, take a stem segment 30-60cm above the growth point, wipe it with 75% alcohol by volume; then peel off 2-3 layers of leaf sheaths to expose the inner leaves, thus obtaining the tender yellow leaves of mature oil palm.

[0016] Preferably, the width of the segment in step (1) is 0.5~1cm, and the soaking time is 15~20min.

[0017] Preferably, the callus induction medium in step (1) is cultured in the dark at a temperature of 27℃~28℃ and a humidity of 55%~65%, and subcultured once every 30 days.

[0018] Preferably, the pH of the callus induction medium in step (1) is adjusted to 5.80~5.95 before sterilization.

[0019] Preferably, the callus proliferation medium in step (2) is cultured in the dark at a temperature of 27℃~28℃ and a humidity of 55%~65%, and subcultured once every 15 days.

[0020] Preferably, the pH of the callus proliferation culture medium in step (2) is adjusted to 5.80~5.85 before sterilization.

[0021] Preferably, the culture conditions of the somatic embryo induction medium in step (3) are dark culture at a temperature of 27℃~28℃ and a humidity of 55%~65%, and subculture once every 30 days.

[0022] Preferably, the pH of the embryo induction culture medium in step (3) is adjusted to 5.85~5.90 before sterilization.

[0023] The method involved in this invention has applications in improving the callus proliferation rate or somatic embryo induction rate of oil palm and shortening the induction cycle.

[0024] Compared with the prior art, the beneficial effects of the present invention are:

[0025] (1) This invention is the first to use baicalin in the callus induction, callus proliferation, and somatic embryo induction stages of young yellow leaves of oil palm. In the callus induction and callus proliferation stages, baicalin replaces part of 2,4-D or dicamba, reducing the risk of subsequent regeneration malformation and effectively increasing the callus proliferation rate and shortening the callus induction time of young yellow leaves; in the somatic embryo induction stage, baicalin can completely replace the plant hormone 6-BA, effectively increasing the somatic embryo induction rate and shortening the somatic embryo induction time.

[0026] (2) Adding 0.5 mg / L baicalin during the callus induction and callus proliferation stages can partially replace the efficacy of 2,4-D or dicamba, reduce the dosage of 2,4-D or dicamba, shorten the callus induction time by 20 days, and increase the callus proliferation rate by 48.19%. During the somatic embryo induction stage, baicalin can completely replace the plant hormone 6-BA, while increasing the somatic embryo induction rate by 5% and shortening the induction time by 31 days. Attached Figure Description

[0027] Figure 1 The study investigated the effect of adding baicalin on the proliferation of oil palm callus; where A represents the effect of the commonly used oil palm callus proliferation medium (10 mg / L dicamba + 0 mg / L baicalin, control group) after 30 days of inoculation; and B represents the effect of the callus proliferation medium prepared in this invention (5 mg / L dicamba + 0.3 mg / L baicalin, treatment group) after 30 days of inoculation.

[0028] Figure 2 The effect of adding baicalin on the induction of somatic embryos in oil palm was investigated. A represents the effect of the commonly used oil palm somatic embryo induction medium (5 mg / L 6-BA + 0 mg / L baicalin, control group) after 150 days of inoculation; B represents the effect of the somatic embryo induction medium prepared in this invention (0 mg / L 6-BA + 0.3 mg / L baicalin, treatment group) after 150 days of inoculation. Detailed Implementation

[0029] To enable those skilled in the art to better understand the technical content of the present invention, the technical solution of the present invention will be further described in detail below with reference to specific embodiments.

[0030] Unless otherwise specified, the experimental methods used in the embodiments of this invention are conventional methods. In the following embodiments and comparative examples, unless otherwise specified, all raw materials can be obtained commercially or prepared by conventional methods.

[0031] Example 1

[0032] A method for promoting callus proliferation and somatic embryo induction in oil palm using baicalin includes the following steps:

[0033] (1) Callus induction: Take the young yellow leaves of mature oil palm as explants, cut them into small segments with a width of 0.5 cm, soak them in sterile water for 15 min and then dry them. Inoculate them into the callus induction medium. The culture conditions are 27℃ and 55% humidity in the dark. Subculture once every 30 days, and oil palm callus tissue is obtained after 95 days.

[0034] The method for obtaining the explants includes the following steps: Select "Hot Oil No. 1" oil palm that has been flowering and fruiting normally for 5 years, remove the outer 3 leaves, take a stem segment 30m above the growth point, wipe it with 75% alcohol by volume; then peel off 3 layers of leaf sheaths to expose the inner leaves, thus obtaining the tender yellow leaves of mature oil palm.

[0035] The composition and formulation of the callus induction medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.5 mg / L baicalin, 4 g / L plant gel, 30 g / L sucrose, and 5 mg / L 2,4-D; the pH of the callus induction medium is adjusted to 5.95 before sterilization.

[0036] (2) Callus proliferation culture: The oil palm callus obtained in step (1) was inoculated into the callus proliferation culture medium. The culture conditions were 27°C and 55% humidity in the dark. The callus was subcultured once every 15 days and collected after 30 days.

[0037] The composition and formulation of the callus proliferation medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.5 mg / L of baicalin, 30 g / L of sucrose, and 5 mg / L of 2,4-D; the pH of the callus proliferation medium is adjusted to 5.85 before sterilization.

[0038] (3) Somatic embryo induction culture: The oil palm callus proliferation tissue obtained in step (2) was transferred to the somatic embryo induction culture medium. The culture conditions were 27℃ and 55% in the dark. The culture was subcultured once every 30 days, and oil palm somatic embryos were obtained in 94 days.

[0039] The composition and formulation of the somatic embryo induction medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.3 mg / L baicalin, 6 g / L plant gel, and 15 g / L sucrose. The pH of the somatic embryo induction medium is adjusted to 5.90 before sterilization.

[0040] Example 2

[0041] A method for promoting callus proliferation and somatic embryo induction in oil palm using baicalin includes the following steps:

[0042] (1) Callus induction: Take the young yellow leaves of mature oil palm as explants, cut them into small segments with a width of 1 cm, soak them in sterile water for 20 min and then dry them. Inoculate them into the callus induction medium. The culture conditions are dark culture at a temperature of 28℃ and a humidity of 65%. Subculture once every 30 days, and oil palm callus tissue is obtained in 110 days.

[0043] The method for obtaining the explants includes the following steps: Select "Hot Oil No. 1" oil palm that has been flowering and fruiting normally for 3 years, remove the outer 2 leaves, take a stem segment 60cm above the growth point, wipe it with 75% alcohol by volume; then peel off 2 layers of leaf sheaths to expose the inner leaves, thus obtaining the tender yellow leaves of mature oil palm.

[0044] The composition and formulation of the callus induction medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.1 mg / L baicalin, 6 g / L plant gel, 40 g / L sucrose, and 5 mg / L 2,4-D; the pH of the callus induction medium is adjusted to 5.80 before sterilization.

[0045] (2) Callus proliferation culture: The oil palm callus obtained in step (1) was inoculated into the callus proliferation culture medium. The culture conditions were 28℃ and 65% in the dark. The callus was subcultured once every 15 days and collected after 30 days.

[0046] The composition and formulation of the callus proliferation medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.1 mg / L of baicalin, 40 g / L of sucrose, and 5 mg / L of 2,4-D; the pH of the callus proliferation medium is adjusted to 5.80 before sterilization.

[0047] (3) Somatic embryo induction culture: The oil palm callus proliferation tissue obtained in step (2) was transferred to the somatic embryo induction culture medium. The culture conditions were 28℃ and 65% in the dark. The culture was subcultured once every 30 days, and oil palm somatic embryos were obtained after 150 days.

[0048] The composition and formulation of the somatic embryo induction medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.1 mg / L baicalin, 10 g / L plant gel, and 20 g / L sucrose. The pH of the somatic embryo induction medium is adjusted to 5.85 before sterilization.

[0049] Example 3

[0050] A method for promoting callus proliferation and somatic embryo induction in oil palm using baicalin includes the following steps:

[0051] (1) Callus induction: Take the young yellow leaves of mature oil palm as explants, cut them into small segments with a width of 0.5 cm, soak them in sterile water for 15 min and then dry them. Inoculate them into the callus induction medium. The culture conditions are dark culture at a temperature of 27℃ and a humidity of 55%. Subculture once every 30 days, and oil palm callus tissue is obtained after 100 days.

[0052] The method for obtaining the explants includes the following steps: Select "Hot Oil No. 1" oil palm that has been flowering and fruiting normally for 4 years, remove the outer 2 leaves, take a stem segment 30cm above the growth point, wipe it with 75% alcohol by volume; then peel off 2 layers of leaf sheaths to expose the inner leaves, thus obtaining the tender yellow leaves of mature oil palm.

[0053] The composition and formulation of the callus induction medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.3 mg / L of baicalin, 4 g / L of plant gel, 30 g / L of sucrose, and 5 mg / L of dicamba; the pH of the callus induction medium is adjusted to 5.95 before sterilization.

[0054] (2) Callus proliferation culture: The oil palm callus obtained in step (1) was inoculated into the callus proliferation culture medium. The culture conditions were 27°C and 55% humidity in the dark. The callus was subcultured once every 15 days and collected after 30 days.

[0055] The composition and formulation of the callus proliferation medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.3 mg / L of baicalin, 30 g / L of sucrose, and 5 mg / L of dicamba; the pH of the callus proliferation medium is adjusted to 5.85 before sterilization.

[0056] (3) Somatic embryo induction culture: The oil palm callus proliferation tissue obtained in step (2) was transferred to the somatic embryo induction culture medium. The culture conditions were 27℃ and 55% in the dark. The culture was subcultured once every 30 days, and oil palm somatic embryos were obtained after 100 days.

[0057] The composition and formulation of the somatic embryo induction medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.5 mg / L baicalin, 6 g / L plant gel, and 15 g / L sucrose. The pH of the somatic embryo induction medium is adjusted to 5.90 before sterilization.

[0058] Example 4

[0059] A method for promoting callus proliferation and somatic embryo induction in oil palm using baicalin includes the following steps:

[0060] (1) Callus induction: Take the young yellow leaves of mature oil palm as explants, cut them into small segments with a width of 1cm, soak them in sterile water for 20min and then dry them. Inoculate them into the callus induction medium. The culture conditions are dark culture at a temperature of 28℃ and a humidity of 65%. Subculture once every 30 days, and oil palm callus tissue is obtained in 105 days.

[0061] The method for obtaining the explants includes the following steps: Select “Hot Oil No. 1” oil palm that has been flowering and fruiting normally for 4 years, remove the outer 3 leaves, take a stem segment 60cm above the growth point, wipe it with 75% alcohol by volume; then peel off 3 layers of leaf sheaths to expose the inner leaves, thus obtaining the tender yellow leaves of mature oil palm.

[0062] The composition and formulation of the callus induction medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.1 mg / L baicalin, 6 g / L plant gel, 40 g / L sucrose, and 5 mg / L dicamba; the pH of the callus induction medium is adjusted to 5.80 before sterilization.

[0063] (2) Callus proliferation culture: The oil palm callus tissue obtained in step (1) was inoculated into the callus proliferation culture medium. The culture conditions were 28℃ and 65% in the dark. The culture was subcultured once every 15 days, and the oil palm callus proliferation tissue was obtained after 30 days.

[0064] The composition and formulation of the callus proliferation medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.1 mg / L of baicalin, 40 g / L of sucrose, and 5 mg / L of dicamba; the pH of the callus proliferation medium is adjusted to 5.80 before sterilization.

[0065] (3) Somatic embryo induction culture: The oil palm callus proliferation tissue obtained in step (2) was transferred to the somatic embryo induction culture medium. The culture conditions were 28℃ and 65% in the dark. The culture was subcultured once every 30 days, and oil palm somatic embryos were obtained after 150 days.

[0066] The composition and formulation of the somatic embryo induction medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.1 mg / L baicalin, 10 g / L plant gel, and 20 g / L sucrose. The pH of the somatic embryo induction medium is adjusted to 5.85 before sterilization.

[0067] Example 1: Callus Induction Rate

[0068] The following experiment was conducted: callus rate (%) = number of explants induced to produce callus / total number of explants × 100%, as detailed in Table 1. 2,4-D (10 mg / L) and dicamba (10 mg / L) were used as control groups. Hormone concentrations were gradually decreased while 0–0.5 mg / L baicalin was added. The callus induction rate and initial callus formation time of young yellow leaves of oil palm were statistically analyzed. As the concentration of 2,4-D or dicamba decreased, the callus induction rate decreased, and the induction time increased accordingly. When the hormone concentration was below a certain threshold, the callus induction rate was too low and had no practical application value. Adding baicalin while decreasing the hormone concentration significantly shortened the callus induction period. Therefore, within a suitable hormone concentration range, adding baicalin to the callus induction medium can effectively shorten the callus induction time.

[0069] Table 1 Effects of baicalin on callus induction in oil palm

[0070]

[0071] Note: The callus induction rate was statistically analyzed 150 days after inoculation.

[0072] Example 2: Callus proliferation rate

[0073] The experiment was set up as follows: callus proliferation rate (%) = ((callus weight after 30 days - inoculated callus weight) / inoculated callus weight) × 100%, details are shown in Table 2 and... Figure 1 As shown, 2,4-D (10 mg / L) and dicamba (10 mg / L) were used as control groups. The hormone concentrations were gradually decreased while 0–0.5 mg / L baicalin was added. Callus induced from young yellow leaves of oil palm was weighed before inoculation and 30 days after inoculation. The callus proliferation rate decreased with decreasing 2,4-D or dicamba concentrations, but excessively high concentrations of dicamba inhibited callus proliferation. Adding baicalin while decreasing the hormone concentration significantly increased the oil palm callus proliferation rate, increasing it by 48.19% compared to media with higher concentrations or without baicalin. Therefore, within a suitable hormone concentration range, adding baicalin to the callus proliferation medium can effectively improve the callus proliferation rate.

[0074] Table 2. Effects of baicalin on callus proliferation in oil palm.

[0075]

[0076] Experimental Example 3: Somatic Embryo Induction Rate

[0077] The experiment was set up as follows: Somatic embryo induction rate (%) = number of calluses induced into somatic embryos / total number of calluses × 100%, as detailed in Table 3 and... Figure 2As shown, a somatic embryo induction medium supplemented with 6-BA (5 mg / L) was used as the control group, and a somatic embryo induction medium supplemented with baicalin (0.1 mg / L~0.5 mg / L) prepared in this invention was used as the experimental group. Both groups were inoculated with proliferated oil palm callus for somatic embryo induction. It was found that adding baicalin to the control group (sophore induction rate 7%) inhibited somatic embryo induction (2%~4%); adding baicalin (0.3 mg / L) to the somatic embryo induction medium without the plant hormone 6-BA significantly increased the somatic embryo induction rate (12%) and shortened the induction period.

[0078] Table 3 Effects of baicalin on the induction of oil palm somatic embryos

[0079]

[0080] Note: The statistical time for somatic embryo induction rate is 150 days after inoculation.

[0081] In summary, in this invention, baicalin mainly enhances callus proliferation and somatic embryo induction efficiency in young yellow leaves, and shortens the induction time. As a flavonoid compound, baicalin acts as an antioxidant, scavenging excess reactive oxygen species generated by cells under external stress (high concentrations of auxin, trauma, etc.) and maintaining normal cell growth. Furthermore, it may regulate the transport and metabolism of auxin, and integrate auxin with other plant hormones, ROS, and transcriptional regulatory signaling pathways, ultimately regulating callus proliferation and differentiation.

[0082] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for promoting callus proliferation and somatic embryo induction in oil palm using baicalin, characterized in that, Includes the following steps: (1) Callus induction: Take the tender yellow leaves of mature oil palm as explants, cut them into small sections, soak them in sterile water and dry them, and inoculate them into the callus induction medium. Oil palm callus tissue was obtained in 95-110 days. The composition and formulation of the callus induction medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.3~0.5 mg / L of baicalin, 4~6 g / L of plant gel, 30~40 g / L of sucrose, and either 5 mg / L of dicamba or 5 mg / L of 2,4-D. (2) Callus proliferation culture: The oil palm callus tissue obtained in step (1) is inoculated into the callus proliferation culture medium, and oil palm callus proliferation tissue is obtained in 25-30 days; The composition and formulation of the callus proliferation medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.3~0.5 mg / L of baicalin, 30~40 g / L of sucrose, and either 5 mg / L of dicamba or 5 mg / L of 2,4-D. (3) Somatic embryo induction culture: The oil palm callus proliferation tissue obtained in step (2) was transferred to the somatic embryo induction culture medium, and oil palm somatic embryos were obtained in 94-150 days; The composition and formulation of the somatic embryo induction medium are as follows: Y3 medium is used as the basal medium, and each liter of basal medium contains 0.3 mg / L of baicalin, 6-10 g / L of plant gel, and 15-20 g / L of sucrose.

2. The method according to claim 1, characterized in that, The method for obtaining explants in step (1) includes the following steps: Select oil palms that have flowered and fruited normally for more than 3 years, remove the outer 2-3 leaves, take a stem segment 30-60cm above the growth point, wipe it with 75% alcohol by volume; then peel off 2-3 layers of leaf sheaths to expose the inner leaves, thus obtaining the tender yellow leaves of mature oil palms.

3. The method according to claim 1, characterized in that, The width of the segment in step (1) is 0.5~1cm, and the soaking time is 15~20min.

4. The method according to claim 1, characterized in that, The callus induction medium in step (1) is cultured in the dark at a temperature of 27℃~28℃ and a humidity of 55%~65%, and subcultured once every 30 days.

5. The method according to claim 1, characterized in that, Before sterilization, the pH of the callus induction medium in step (1) is adjusted to 5.80~5.

95.

6. The method according to claim 1, characterized in that, The callus proliferation medium in step (2) is cultured in the dark at a temperature of 27℃~28℃ and a humidity of 55%~65%, and subcultured once every 15 days.

7. The method according to claim 1, characterized in that, Before sterilization, the pH of the callus proliferation medium in step (2) is adjusted to 5.80~5.

85.

8. The method according to claim 1, characterized in that, The culture conditions for the embryo induction medium in step (3) are: dark culture at a temperature of 27℃~28℃ and a humidity of 55%~65%, with subculture every 30 days.

9. The method according to claim 1, characterized in that, Before sterilization, the pH of the embryo induction medium in step (3) is adjusted to 5.85~5.

90.

10. The application of the method according to any one of claims 1 to 9 in the following aspects: (1) Improve the callus proliferation rate or somatic embryo induction rate of oil palm; (2) Shorten the callus induction period of oil palm.

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