Tissue culture method for promoting germination of zingiber zingiberensis

Abstract

The invention relates to the technical field of plant tissue culture. The invention provides a tissue culture method for promoting germination of zingiber officinale roscoe, which comprises the following steps of: sterilizing zingiber officinale roscoe tubers serving as explants, sequentially inoculating the explants to an induction culture medium, a proliferation culture medium and a rooting culture medium for culture, and then domesticating, hardening seedlings and transplanting to obtain finished zingiber officinale roscoe seedlings. According to the technical scheme, the germination rate, the multiplication coefficient and the rooting rate of the zingiber officinale roscoe tubers can be remarkably increased by culturing the zingiber officinale roscoe tubers through the optimized and blended culture media. Besides, the growth vigor and disease resistance of the zingiber officinale kimura et zingiber officinale seedlings can be further improved through reasonable application of leaf fertilizer and bactericide after transplanting, the transplanting survival rate is guaranteed, and reliable technical guarantee is provided for rapid propagation and large-scale planting of the zingiber officinale kimura et zingiber officinale kimura et zingiber et zingiber et zingiber.

Description

Technical Field

[0001] This invention relates to the field of plant tissue culture technology, and in particular to a tissue culture method for promoting the germination of ginger truncatum. Background Technology

[0002] Zingiber officinale is a perennial herbaceous plant belonging to the genus Zingiber in the family Zingiberaceae. It has high medicinal and ornamental value. Its tubers contain a variety of active ingredients and are widely used in traditional Chinese medicine. Additionally, its vibrant green leaves and unique flowers also make it suitable for cultivation as an ornamental plant.

[0003] However, the natural propagation rate of ginger is relatively slow. Conventional division propagation methods not only have a low propagation coefficient but are also greatly limited by seasonal and environmental conditions, making it difficult to meet the large market demand for ginger seedlings. Plant tissue culture technology, as an effective means of rapid plant propagation, has advantages such as fast propagation speed, no seasonal limitations, and the ability to maintain the superior characteristics of varieties, and has been widely used in the propagation of various plants.

[0004] Currently, research on tissue culture of *Gynostemma pentaphyllum* is limited, and existing tissue culture methods suffer from low tuber germination rates, slow proliferation rates, and poor rooting effects, severely hindering the large-scale propagation and industrial development of *Gynostemma pentaphyllum*. Therefore, developing a tissue culture method that can effectively promote the germination of *Gynostemma pentaphyllum* tubers is of significant practical importance. Summary of the Invention

[0005] The purpose of this invention is to provide a tissue culture method for promoting the germination of ginger, which can significantly improve the germination rate, proliferation coefficient and rooting rate of ginger tubers, providing a reliable technical guarantee for the rapid propagation and large-scale cultivation of ginger.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0007] This invention provides a tissue culture method for promoting the germination of ginger truncatum, comprising the following steps:

[0008] (1) The truncated ginger rhizomes were used as explants, and were cleaned and disinfected to obtain sterile explants;

[0009] (2) Sterile explants were sequentially inoculated into induction medium and proliferation medium to obtain adventitious bud sterile seedlings;

[0010] (3) After inoculating the aseptic seedlings of adventitious buds into the rooting medium and culturing for 15-20 days, the seedlings are acclimatized and hardened to obtain truncated ginger seedlings;

[0011] (4) When the truncated ginger seedlings have grown 3-5 strong roots, transplant them into a mixed substrate with a volume ratio of vermiculite: peat moss: raw red soil = (1-1.5):(1-1.8):2 to obtain the finished truncated ginger seedlings.

[0012] Preferably, step (1) involves washing and soaking with dish soap for 5-15 minutes.

[0013] Preferably, in step (1), alcohol and mercuric chloride are used sequentially for disinfection; the volume concentration of the alcohol is 70-80%, and the alcohol disinfection time is 10-20s; the mass concentration of the mercuric chloride is 0.08-0.12%, and the mercuric chloride disinfection time is 10-15min.

[0014] Preferably, the induction medium in step (2) is: MS + 0.8-1.5 mg / L zeatin + 0.2-0.4 mg / L NAA + 40-60 mg / L activated carbon + 0.8-1.2 g / L PPA + 20-30 g / L sucrose + 4-6 g / L agar; pH value is 5.7-5.9.

[0015] Preferably, the shoots are induced and cultured in an induction medium until they reach a length of 2-4 cm.

[0016] Preferably, the proliferation medium in step (2) is: MS + 0.8-1.8 mg / L zeatin + 0.1-0.3 mg / L NAA + 20-30 g / L sucrose + 4-6 g / L agar; pH value is 5.7-5.9.

[0017] Preferably, the adventitious shoots are cultured in a proliferation medium until they reach a length of 4-6 cm.

[0018] Preferably, the rooting medium in step (3) is: MS + 0.1-0.2 mg / L IBA + 0.1-0.3 mg / L IAA + 20-30 g / L sucrose + 4-6 g / L agar; pH value is 5.7-5.9.

[0019] Preferably, the cultivation temperature in steps (2) and (3) is 23-28℃, the light intensity is 1200-1500 Lux, and the light duration is 8-12h / d.

[0020] As a preferred option, the acclimatization and hardening of seedlings in step (3) is as follows: after acclimatizing the seedlings for 4-5 days with the cover off, move them to natural light and harden them at 24-26℃ for 5-7 days.

[0021] Preferably, the method further includes applying foliar fertilizer and / or pesticide after culturing for 20-30 days in step (4), wherein the foliar fertilizer is a 0.2-0.3% potassium dihydrogen phosphate solution and the pesticide is a 60-70% methyl thiophanate wettable powder.

[0022] Preferably, the application frequency is 5-7 days / time, and the number of applications is 2-3 times.

[0023] By adopting the above technical solution, the present invention has the following beneficial effects:

[0024] 1. The technical solution of this invention effectively reduces the contamination rate of explants and improves the success rate of tissue culture by strictly disinfecting the truncated ginger rhizome explants.

[0025] 2. The technical solution of this invention optimizes the formulation of induction medium, proliferation medium and rooting medium. By rationally adding different types and concentrations of plant growth regulators, the germination rate, proliferation coefficient and rooting rate of truncated ginger tubers are significantly improved.

[0026] 3. The truncated ginger seedlings cultivated by the technical solution of this invention are robust and have well-developed root systems. After transplanting, the seedlings' growth vigor and disease resistance are further improved by the reasonable application of foliar fertilizer and fungicide, resulting in a high transplant survival rate. This provides a reliable technical guarantee for the rapid propagation and large-scale planting of truncated ginger. Attached Figure Description

[0027] Figure 1 The seedlings of ginger obtained by the tissue culture method described in Example 1 of this invention are truncated ginger seedlings. Detailed Implementation

[0028] This invention provides a tissue culture method for promoting the germination of ginger truncatum, comprising the following steps:

[0029] (1) The truncated ginger rhizomes were used as explants, and were cleaned and disinfected to obtain sterile explants;

[0030] (2) Sterile explants were sequentially inoculated into induction medium and proliferation medium to obtain adventitious bud sterile seedlings;

[0031] (3) After inoculating the aseptic seedlings of adventitious buds into the rooting medium and culturing for 15-20 days, the seedlings are acclimatized and hardened to obtain truncated ginger seedlings;

[0032] (4) When the truncated ginger seedlings have grown 3-5 strong roots, transplant them into a mixed substrate with a volume ratio of vermiculite: peat moss: raw red soil = (1-1.5):(1-1.8):2 to obtain the finished truncated ginger seedlings.

[0033] In this invention, healthy, disease-free, and vigorous current-year truncated ginger rhizomes are selected as explants. They are washed and soaked with dish soap, and then rinsed with clean water. The volume concentration of the dish soap is 0.5-2%, more preferably 0.8-1.5%, and even more preferably 1%. The soaking time is preferably 5-15 minutes, more preferably 8-12 minutes, and even more preferably 10 minutes.

[0034] In this invention, the cleaned explants are disinfected sequentially with alcohol and then with mercuric chloride to obtain sterile explants. The volume concentration of the alcohol used in this invention is preferably 70-80%, more preferably 72-78%, and even more preferably 75%; the alcohol disinfection time is preferably 10-20 seconds, more preferably 12-18 seconds, and even more preferably 15 seconds; the mass concentration of the mercuric chloride is preferably 0.08-0.12%, more preferably 0.08-0.11%, and even more preferably 0.1%; the mercuric chloride disinfection time is preferably 10-15 minutes, more preferably 12-14 minutes, and even more preferably 13 minutes. Preferably, after both alcohol and mercuric chloride disinfection, the explants are rinsed with sterile water, and the number of rinses is preferably 3-5 times.

[0035] In this invention, sterile explants are sequentially inoculated into induction medium and proliferation medium to obtain aseptic seedlings with adventitious buds. The preferred formulation of the induction medium is MS + 0.8-1.5 mg / L zeatin + 0.2-0.4 mg / L NAA + 40-60 mg / L activated charcoal + 0.8-1.2 g / L PPA + 20-30 g / L sucrose + 4-6 g / L agar. A more preferred formulation is MS + 0.9-1.2 mg / L zeatin + 0.25-0.35 mg / L NAA + 45-55 mg / L activated charcoal + 0.9-1.1 g / L PPA + 22-28 g / L sucrose + 4.5-5.5 g / L agar. An even more preferred formulation is MS + 1 mg / L zeatin + 0.3 mg / L NAA + 50 mg / L activated charcoal + 1 g / L PPA + 25 g / L sucrose + 5 g / L agar. The pH value of the induction medium described in this invention is preferably 5.7-5.9, and more preferably 5.8. Preferably, the shoots are induced and cultured in the induction medium until they reach a length of 2-4 cm.

[0036] In this invention, the preferred formulation of the proliferation medium is MS + 0.8-1.8 mg / L zeatin + 0.1-0.3 mg / L NAA + 20-30 g / L sucrose + 4-6 g / L agar; more preferably, MS + 1-1.6 mg / L zeatin + 0.15-0.25 mg / L NAA + 22-28 g / L sucrose + 4.5-5.5 g / L agar; even more preferably, MS + 1.5 mg / L zeatin + 0.2 mg / L NAA + 25 g / L sucrose + 5 g / L agar. The preferred pH of the proliferation medium in this invention is 5.7-5.9, more preferably 5.8. In this invention, the adventitious shoots are preferably cultured in the proliferation medium until they reach a length of 4-6 cm.

[0037] In this invention, aseptic seedlings with adventitious buds are inoculated into a rooting medium for culture. The preferred formulation of the rooting medium is MS + 0.1-0.2 mg / L IBA + 0.1-0.3 mg / L IAA + 20-30 g / L sucrose + 4-6 g / L agar; more preferably, MS + 0.12-0.18 mg / L IBA + 0.15-0.25 mg / L IAA + 22-28 g / L sucrose + 4.5-5.5 g / L agar; even more preferably, MS + 0.15 mg / L IBA + 0.2 mg / L IAA + 25 g / L sucrose + 5 g / L agar. The preferred pH of the rooting medium in this invention is 5.7-5.9, more preferably 5.8. The preferred culture time in the rooting medium in this invention is 15-20 days, more preferably 16-19 days, and even more preferably 18 days.

[0038] In this invention, the cultivation temperatures in steps (2) and (3) are each preferably 23-28°C, more preferably 24-27°C, and even more preferably 26°C; the light intensity for cultivation is each preferably 1200-1500 Lux, more preferably 1300-1450 Lux, and even more preferably 1400 Lux; and the light duration for cultivation is each preferably 8-12 h / d, more preferably 9-11 h / d, and even more preferably 10 h / d.

[0039] In this invention, after the rooting medium is cultured, the seedlings are acclimatized and hardened off. The preferred method for acclimatizing and hardening off the seedlings is to uncover them and harden them off for 4-5 days, then move them to natural light and harden them off at 24-26℃ for 5-7 days.

[0040] In this invention, when the truncated ginger seedlings have grown 3-5 robust roots, they are transplanted into a mixed substrate for cultivation. The components of the mixed substrate preferably include vermiculite, peat moss, and raw red soil. The volume ratio of vermiculite, peat moss, and raw red soil is preferably (1-1.5):(1-1.8):2, further preferably (1.1-1.4):(1.3-1.7):2, and even more preferably 1.2:1.5:2.

[0041] In this invention, it is preferred to apply foliar fertilizer and / or pesticide after culturing in a mixed substrate for 20-30 days. The foliar fertilizer is preferably a 0.2-0.3% potassium dihydrogen phosphate solution, more preferably a 0.22-0.28% potassium dihydrogen phosphate solution, and even more preferably a 0.25% potassium dihydrogen phosphate solution. The pesticide is preferably a 60-70% methyl thiophanate wettable powder, more preferably a 62-68% methyl thiophanate wettable powder, and even more preferably a 65% methyl thiophanate wettable powder. The methyl thiophanate wettable powder in this invention is preferably an 800-1000 times dilution, more preferably an 850-950 times dilution, and even more preferably a 900 times dilution.

[0042] In this invention, when potassium dihydrogen phosphate solution is used as a foliar fertilizer, it can quickly provide plants with the two important elements of phosphorus and potassium, enhance the plant's resistance to stress, and promote photosynthetic processes.

[0043] In this invention, the use of methyl thiophanate wettable powder can effectively reduce the risk of microbial contamination, create a sterile or low-bacterial environment for the germination and subsequent growth of ginger rhizomes, ensure the healthy development of tissue culture seedlings, and improve the success rate of tissue culture.

[0044] In this invention, the application frequency is preferably 5-7 days / time, more preferably 6 days / time; the number of applications is 2-3 times, more preferably 2 times.

[0045] In this invention, the spraying time is preferably before 9 a.m. or after 4 p.m., and the spraying should be evenly applied to both sides of the leaves.

[0046] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0047] Example 1

[0048] A tissue culture method for promoting the germination of ginger buds, comprising the following steps:

[0049] Healthy, disease-free, and vigorous current-year ginger rhizomes were selected as explants. They were washed and soaked in 1% (v / v) detergent for 10 minutes and then rinsed with clean water. They were then disinfected with 75% (v / v) alcohol for 15 seconds and rinsed three times with sterile water. Finally, they were disinfected with 0.1% (w / w) mercuric chloride for 13 minutes and rinsed three times with sterile water to obtain sterile explants.

[0050] Sterile explants were inoculated onto an induction medium and cultured at 26°C with a light intensity of 1400 Lux and a photoperiod of 10 h / d until the shoot length reached 4 cm. Then, they were inoculated onto a proliferation medium and cultured at 26°C with a light intensity of 1400 Lux and a photoperiod of 10 h / d until the adventitious shoot length reached 5 cm, thus obtaining sterile adventitious shoot seedlings. The induction medium consisted of MS + 1 mg / L zeatin + 0.3 mg / L NAA + 50 mg / L activated carbon + 1 g / L PPA + 25 g / L sucrose + 5 g / L agar, with a pH of 5.8. The proliferation medium consisted of MS + 1.5 mg / L zeatin + 0.2 mg / L NAA + 25 g / L sucrose + 5 g / L agar, with a pH of 5.8.

[0051] Aseptic seedlings with adventitious buds were inoculated into rooting medium and cultured at 26°C with a light intensity of 1400 Lux and a light duration of 10 h / d for 18 days. The rooting medium consisted of MS + 0.15 mg / L IBA + 0.2 mg / L IAA + 25 g / L sucrose + 5 g / L agar, with a pH of 5.8.

[0052] After the rooting culture was completed, the seedlings were hardened off for 5 days after uncovering, and then moved to natural light and hardened off at 24℃ for 5 days to obtain truncated ginger seedlings;

[0053] When the truncated ginger seedlings have grown 3-5 strong roots, transplant them into a mixed substrate with a volume ratio of vermiculite, peat moss, and raw red soil of 1.2:1.5:2. After 30 days of cultivation, spray with 0.25% potassium dihydrogen phosphate solution and 900 times dilution of 65% methyl thiophanate wettable powder. It is best to spray evenly on both sides of the leaves before 9 am or after 4 pm, once every 6 days, for a total of 2 sprays.

[0054] Example 2

[0055] A tissue culture method for promoting the germination of ginger buds, comprising the following steps:

[0056] Healthy, disease-free, and vigorous current-year ginger rhizomes were selected as explants. They were washed and soaked in 0.5% (v / v) detergent for 15 minutes and then rinsed with clean water. They were then disinfected with 70% (v / v) alcohol for 18 seconds and rinsed 4 times with sterile water. Finally, they were disinfected with 0.12% (w / w) mercuric chloride for 15 minutes and rinsed 3 times with sterile water to obtain sterile explants.

[0057] Sterile explants were inoculated into induction medium and cultured at 26°C with a light intensity of 1200 Lux and a photoperiod of 11 h / d until the shoot length reached 3 cm. Then, they were inoculated into proliferation medium and cultured at 24°C with a light intensity of 1300 Lux and a photoperiod of 12 h / d until the adventitious shoot length reached 5 cm, obtaining sterile adventitious shoot seedlings. The induction medium consisted of MS + 1.2 mg / L zeatin + 0.2 mg / L NAA + 40 mg / L activated carbon + 0.8 g / L PPA + 30 g / L sucrose + 4 g / L agar, with a pH of 5.7. The proliferation medium consisted of MS + 1 mg / L zeatin + 0.3 mg / L NAA + 20 g / L sucrose + 4 g / L agar, with a pH of 5.8.

[0058] Aseptic seedlings with adventitious buds were inoculated into rooting medium and cultured at 25°C with a light intensity of 1200 Lux and a light duration of 12 h / d for 18 days. The rooting medium consisted of MS + 0.1 mg / L IBA + 0.3 mg / L IAA + 30 g / L sucrose + 4 g / L agar, with a pH of 5.7.

[0059] After the rooting culture was completed, the seedlings were hardened off for 4 days after uncovering and then moved to natural light. They were hardened off at 26℃ for 7 days to obtain truncated ginger seedlings.

[0060] When the truncated ginger seedlings have grown 3-5 strong roots, transplant them into a mixed substrate with a volume ratio of vermiculite, peat moss, and raw red soil of 1:1.8:2. After 30 days of cultivation, spray with 0.2% potassium dihydrogen phosphate solution and 1000 times dilution of 70% methyl thiophanate wettable powder. It is best to spray evenly on both sides of the leaves before 9 am or after 4 pm, once every 6 days, for a total of 2 sprays.

[0061] Example 3

[0062] A tissue culture method for promoting the germination of ginger buds, comprising the following steps:

[0063] Healthy, disease-free, and vigorous current-year ginger rhizomes were selected as explants. They were washed and soaked in 1.5% (v / v) detergent for 5 minutes and then rinsed with clean water. They were then disinfected with 80% (v / v) alcohol for 10 seconds and rinsed three times with sterile water. Finally, they were disinfected with 0.08% (w / w) mercuric chloride for 10 minutes and rinsed four times with sterile water to obtain sterile explants.

[0064] Sterile explants were inoculated into induction medium and cultured at 23°C with a light intensity of 1300 Lux and a photoperiod of 11 h / d until the shoot length reached 4 cm. Then, they were inoculated into proliferation medium and cultured at 27°C with a light intensity of 1400 Lux and a photoperiod of 9 h / d until the adventitious shoot length reached 6 cm, obtaining sterile adventitious shoot seedlings. The induction medium consisted of MS + 0.8 mg / L zeatin + 0.4 mg / L NAA + 60 mg / L activated carbon + 1.2 g / L PPA + 20 g / L sucrose + 6 g / L agar, with a pH of 5.7. The proliferation medium consisted of MS + 1.8 mg / L zeatin + 0.1 mg / L NAA + 30 g / L sucrose + 6 g / L agar, with a pH of 5.8.

[0065] Aseptic seedlings with adventitious buds were inoculated into rooting medium and cultured at 26°C with a light intensity of 1500 Lux and a light duration of 8 h / d for 16 days. The rooting medium consisted of MS + 0.2 mg / L IBA + 0.1 mg / L IAA + 20 g / L sucrose + 6 g / L agar, with a pH of 5.8.

[0066] After the rooting culture was completed, the seedlings were hardened off for 5 days after uncovering and then moved to natural light. They were hardened off at 25℃ for 7 days to obtain truncated ginger seedlings.

[0067] When the truncated ginger seedlings have grown 3-5 strong roots, transplant them into a mixed substrate with a volume ratio of vermiculite, peat moss, and raw red soil of 1.5:1:2. After 20 days of cultivation, spray with 0.3% potassium dihydrogen phosphate solution and 800 times dilution of 60% methyl thiophanate wettable powder. It is best to spray evenly on both sides of the leaves before 9 am or after 4 pm, once every 5 days, for a total of 3 sprays.

[0068] Comparative Example 1

[0069] Similar to Example 1, except that "then disinfect with 75% alcohol for 15 seconds and rinse with sterile water 3 times; then disinfect with 0.1% mercuric chloride for 13 minutes and rinse with sterile water 3 times" is replaced with "disinfect with 75% alcohol for 5 minutes and rinse with sterile water 3 times", and "wash and soak with 1% detergent for 10 minutes" is replaced with "wash and soak with clean water for 10 minutes".

[0070] Comparative Example 2

[0071] Similar to Example 1, except that "the composition of the induction medium is MS + 1 mg / L zeatin + 0.3 mg / L NAA + 50 mg / L activated carbon + 1 g / L PPA + 25 g / L sucrose + 5 g / L agar" is replaced with "the composition of the induction medium is MS + 1 mg / L zeatin + 0.3 mg / L NAA + 25 g / L sucrose + 5 g / L agar".

[0072] Comparative Example 3

[0073] Similar to Example 1, except that "the composition of the proliferation medium is MS + 1.5 mg / L zeatin + 0.2 mg / L NAA + 25 g / L sucrose + 5 g / L agar" is replaced with "the composition of the proliferation medium is MS + 0.2 mg / L NAA + 25 g / L sucrose + 5 g / L agar".

[0074] Comparative Example 4

[0075] Similar to Example 1, except that "the composition of the rooting medium is MS + 0.15 mg / L IBA + 0.2 mg / L IAA + 25 g / L sucrose + 5 g / L agar" is replaced with "the composition of the rooting medium is MS + 0.2 mg / L IAA + 25 g / L sucrose + 5 g / L agar".

[0076] Comparative Example 5

[0077] Similar to Example 1, except that “a mixed matrix with a volume ratio of vermiculite, peat moss and red laterite of 1.2:1.5:2” is replaced with “a mixed matrix with a volume ratio of vermiculite and perlite of 1:1”.

[0078] Comparative Example 6

[0079] Similar to Example 1, only the following was removed: "After 20 days of cultivation, spray with 0.3% potassium dihydrogen phosphate solution and 60% methyl thiophanate wettable powder at a dilution of 800 times. It is best to spray evenly on both sides of the leaves before 9 am or after 4 pm, once every 5 days, for a total of 3 sprays".

[0080] Experimental Example 1 (one)

[0082] Based on Example 1 and Comparative Examples 1-6, 20 explants of the same growth condition and size were randomly selected from each group for tissue culture. After induction culture to a shoot length of 4 cm, the number of induced shoots was counted, and the appearance of mycelia, fungal patches, or other signs of microbial growth on the surface of the explants was observed to determine whether the explants were contaminated. The induction rate and contamination rate were calculated. After obtaining aseptic seedlings with adventitious shoots through culture, the proliferation coefficient was observed and calculated.

[0083] Induction rate (%) = Number of explants that induced shoots / Number of inoculated explants × 100%

[0084] Contamination rate (%) = Number of contaminated explants / Number of inoculated explants × 100%

[0085] Proliferation coefficient = Total number of induced shoots / Number of explants inoculated

[0086] Table 1. Induction rate, contamination rate, and proliferation coefficient of different treatment groups.

[0087] Group Induction rate / % Pollution rate / % Proliferation coefficient Example 1 95 0 3.75 Comparative Example 1 75 35 2.90 Comparative Example 2 60 0 2.00 Comparative Example 3 90 5 3.50 Comparative Example 4 95 0 3.70 Comparative Example 5 90 0 3.70 Comparative Example 6 95 0 3.60

[0088] As can be seen from Table 1, the tissue culture method described in this invention has the best induction rate and proliferation coefficient for Ginger scutellaria, with a contamination rate of basically 0. (two)

[0090] Based on Example 1 and Comparative Examples 1-6, 20 explants of the same growth condition and size were randomly selected from each group for tissue culture. After transplanting to a mixed substrate and culturing for 30 days, the number of surviving seedlings was counted and the survival rate was calculated.

[0091] Transplant survival rate (%) = (Number of surviving seedlings / Number of transplanted seedlings) × 100%

[0092] Table 2. Survival rate of transplanted seedlings in different treatment groups.

[0093]

[0094]

[0095] As can be seen from Table 2, the transplant survival rate of ginger seedlings grown using the tissue culture method described in this invention is relatively the highest.

[0096] In summary, the tissue culture method of this invention, which includes disinfection, optimized preparation of various culture media, and rational application of foliar fertilizer and fungicide, can significantly improve the germination rate, proliferation coefficient, and rooting rate of ginger rhizomes. It can also further enhance the growth vigor and disease resistance of ginger seedlings, ensure transplant survival rate, and provide reliable technical support for the rapid propagation and large-scale cultivation of ginger.

[0097] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A tissue culture method for promoting the germination of ginger truncatum, characterized in that, Includes the following steps: (1) The truncated ginger rhizomes were used as explants, and were cleaned and disinfected to obtain sterile explants; (2) Sterile explants were sequentially inoculated into induction medium and proliferation medium to obtain adventitious bud sterile seedlings; (3) After inoculating the aseptic seedlings of adventitious buds into the rooting medium and culturing for 15-20 days, the seedlings are acclimatized and hardened to obtain truncated ginger seedlings; (4) When the truncated ginger seedlings have grown 3-5 strong roots, transplant them into a mixed substrate with a volume ratio of vermiculite: peat moss: raw red soil = (1-1.5):(1-1.8):2 to obtain the finished truncated ginger seedlings.

2. The tissue culture method according to claim 1, characterized in that, Step (1) Use dish soap to clean and soak for 5-15 minutes.

3. The tissue culture method according to claim 1, characterized in that, In step (1), alcohol and mercuric chloride are used for disinfection in sequence; The volume concentration of the alcohol is 70-80%, and the alcohol disinfection time is 10-20 seconds. The mercuric chloride concentration is 0.08-0.12%, and the mercuric chloride disinfection time is 10-15 minutes.

4. The tissue culture method according to claim 1, characterized in that, The induction medium in step (2) is: MS + 0.8-1.5 mg / L zeatin + 0.2-0.4 mg / L NAA + 40-60 mg / L activated carbon + 0.8-1.2 g / L PPA + 20-30 g / L sucrose + 4-6 g / L agar; pH value is 5.7-5.9; Induce and culture in induction medium until the shoot length reaches 2-4 cm.

5. The tissue culture method according to claim 1, characterized in that, The proliferation medium described in step (2) is: MS + 0.8-1.8 mg / L zeatin + 0.1-0.3 mg / L NAA + 20-30 g / L sucrose + 4-6 g / L agar; pH value is 5.7-5.9; Cultured in proliferation medium until the adventitious shoots reach a length of 4-6 cm.

6. The tissue culture method according to claim 1, characterized in that, The rooting medium in step (3) is: MS + 0.1-0.2 mg / L IBA + 0.1-0.3 mg / L IAA + 20-30 g / L sucrose + 4-6 g / L agar; pH value is 5.7-5.

9.

7. The tissue culture method according to claim 1, characterized in that, The cultivation temperatures in steps (2) and (3) are independently 23-28℃, the light intensity is independently 1200-1500 Lux, and the light duration is independently 8-12 h / d.

8. The tissue culture method according to claim 1, characterized in that, The acclimatization and hardening process described in step (3) is as follows: after acclimatizing the seedlings for 4-5 days with the covers off, move them to natural light and harden them at 24-26℃ for 5-7 days.

9. The tissue culture method according to claim 1, characterized in that, It also includes applying foliar fertilizer and / or pesticide after culturing for 20-30 days in step (4), wherein the foliar fertilizer is a 0.2-0.3% potassium dihydrogen phosphate solution and the pesticide is a 60-70% methyl thiophanate wettable powder.

10. The tissue culture method according to claim 9, characterized in that, The application frequency is 5-7 days / time, and the number of applications is 2-3 times.

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