Method for promoting rooting of vitis davidii cuttings

Abstract

The invention relates to the technical field of vitis davidii cuttage, in particular to a method for promoting rooting of vitis davidii cuttings. The invention provides a method for promoting vitis davidii cuttings to root, which comprises the following steps: (1) cutting cuttings from grape branches, and storing in sand; (2) soaking the cuttings obtained in the step (1) in water, a growth regulator and rooting powder in sequence; (3) cutting the cuttings obtained in the step (2) into a substrate for culture, and applying a nutrient solution to obtain cutting seedlings; and (4) transplanting the cutting seedlings obtained in the step (3) to a field. According to the method for promoting rooting of the vitis davidii cuttings, germination and rooting of the vitis davidii cuttings can be effectively promoted, the cuttings have the very high rooting amount, the survival rate of grape cutting seedling raising is greatly increased, and healthy growth of grape plants in the later period is promoted.

Description

Technical Field

[0001] The invention relates to the technical field of thorny grape cuttings, in particular to a method for promoting rooting of thorny grape cuttings. Background Art

[0002] Grape (Vitis vinifera L.) has a long history of cultivation and has developed very rapidly. It is one of the fruits with the best economic benefits. Its wild germplasm is rich and has strong resistance. It can be used as resistance breeding materials or rootstocks. Some wild grapes can be directly promoted and utilized in production. Among them, thorny grapes are one of the most typical and characteristic wild grape resources. Thorny grapes are mainly distributed in the southern region. They have strong resistance to anthracnose, powdery mildew, white rot, gray mold and black pox, and are an important resource of moisture-resistant and disease-resistant germplasm. Thorny grapes have large berries, high content of anthocyanins and resveratrol, rich nutrients, strong antioxidant capacity, good quality, growth and yield, and are used in fresh eating, juicing and winemaking and other related industries, with good development prospects. Thorny grapes are also a promising source of high-quality anthocyanins, with great application potential in the food, pharmaceutical and cosmetics industries.

[0003] Vitis thornii is a woody plant. The propagation of seedlings mainly depends on cutting propagation. However, due to the properties of Vitis thornii itself, it is difficult for it to take root, which seriously restricts the promotion of the Vitis thornii industry. Cutting seedlings are currently the most widely used method for grape seedlings. Adventitious roots are formed by inducing wounds. Its survival rate is related to the variety, cutting quality, treatment method, cutting environment, etc., and the factors that affect the rooting of cuttings include variety characteristics, cutting maturity and substrate ratio. The substrate has a significant effect on the occurrence of adventitious roots, and the propagation medium has the greatest impact on rooting.

[0004] Therefore, how to propagate Vitis thornii on a large scale by means of cuttings and screen suitable cutting media for Vitis thornii are problems that those skilled in the art need to solve urgently. Summary of the invention

[0005] The purpose of the present invention is to provide a method for promoting rooting of thorny grape cuttings. The present invention uses thorny grapes as materials, studies the effects of different matrices on the rooting of thorny grape cuttings, combines transcriptome and hormone-targeted metabolome, analyzes the rooting mechanism of thorny grape cuttings, clarifies the role of hormones in it, and determines the key genes that play a role in the process.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The present invention provides a method for promoting rooting of thorny grape cuttings, comprising the following steps:

[0008] (1) Cut cuttings from grape branches and store them in sand;

[0009] (2) soaking the cuttings obtained in step (1) in water, a growth regulator and rooting powder in sequence;

[0010] (3) inserting the cuttings obtained in step (2) into a substrate for cultivation, and applying a nutrient solution to obtain cutting seedlings;

[0011] (4) Transplanting the cuttings obtained in step (3) into a field.

[0012] Preferably, in step (1), each cutting retains 1 to 3 buds, the upper incision is 2 to 4 cm away from the nearest bud eye, and the lower incision is 4 to 6 cm away from the nearest bud eye.

[0013] Preferably, the sand storage time in step (2) is 60 to 70 days.

[0014] Preferably, in step (2), the cuttings are soaked in water for 14 to 18 hours; the cuttings are soaked in a growth regulator for 16 to 20 hours; and the cuttings are soaked in rooting powder for 3 to 5 hours.

[0015] Preferably, the growth regulator in step (2) is: IBA with a final concentration of 700-900 mg / L + NAA solution with a final concentration of 140-180 mg / L.

[0016] Preferably, the rooting powder in step (2) is ABT1 rooting powder with a mass concentration of 30-40%.

[0017] Preferably, the matrix in step (3) is one or more of perlite, rice husk carbon or river sand.

[0018] Preferably, the nutrient solution in step (3) is Hoagland nutrient solution with a volume concentration of 1.0 to 1.5 g / L; the nutrient solution is irrigated once every 4 to 6 days, and the amount used each time is 1 to 1.5 L / m 2 .

[0019] Preferably, the culturing time in step (3) is 35 to 45 days.

[0020] Preferably, in step (4), after transplanting into the field, the cuttings are topped when they have grown 7 to 8 leaves and tied to the poles.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] In this study, the comprehensive score of the cuttings under the condition of perlite (T12) was the highest, with a germination rate of 80% and a rooting rate of up to 90%. It can be used as a good matrix for inducing rooting of thorny grape cuttings that are difficult to root. Compared with other matrices, the adventitious roots produced by cuttings on the perlite matrix are the longest, and the total root length and the number of lateral roots are the highest. However, perlite does not have nutrients and is not suitable for long-term cultivation of plants as a single component. Therefore, other matrices such as rice husk biochar (RHB) need to be added for use. In the present invention, the growth of the root system and the growth of the cuttings are good when rice husk carbon (T1 and T2) is added to coarse river sand and perlite respectively, which may be related to the fact that rice husk carbon is breathable, sterilizing, and rich in nutrients.

[0023] The method for promoting rooting of thorny grape cuttings provided by the invention can effectively promote germination and rooting of thorny grape cuttings, not only making the cuttings have a high rooting amount, but also greatly improving the survival rate of grape cutting seedlings, and promoting the robust growth of grape plants in the later stage. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying creative work.

[0025] Figure 1 This shows the germination and rooting conditions of the thorny grape cuttings in groups T1 to T4.

[0026] Figure 2 This shows the germination and rooting conditions of T5 to T8 group thorny grape cuttings.

[0027] Figure 3 This shows the germination and rooting conditions of the T9 to T13 group of thorny grape cuttings.

[0028] Figure 4 It is the growth index of cuttings under different substrate conditions, Tn (germination rate, rooting rate, callus formation rate, average root length and number of roots). DETAILED DESCRIPTION

[0029] The present invention provides a method for promoting rooting of thorny grape cuttings, comprising the following steps:

[0030] (1) Cut cuttings from grape branches and store them in sand;

[0031] (2) soaking the cuttings obtained in step (1) in water, a growth regulator and rooting powder in sequence;

[0032] (3) inserting the cuttings obtained in step (2) into a substrate for cultivation, and applying a nutrient solution to obtain cutting seedlings;

[0033] (4) Transplanting the cuttings obtained in step (3) into a field.

[0034] In the present invention, in step (1), each cutting retains 1 to 3 buds, the upper incision is 2 to 4 cm away from the nearest bud eye, and the lower incision is 4 to 6 cm away from the nearest bud eye; preferably, each cutting retains 2 buds, the upper incision is 3 cm away from the nearest bud eye, and the lower incision is 5 cm away from the nearest bud eye.

[0035] In the present invention, the sand storage time in step (2) is 60 to 70 days; preferably 62 to 68 days; more preferably 64 to 66 days; and more preferably 65 days.

[0036] In the present invention, the cuttings in step (2) are soaked in water for 14 to 18 hours, preferably 15 to 17 hours, and more preferably 16 hours.

[0037] In the present invention, the cuttings are immersed in the growth regulator for 16 to 20 hours, preferably 17 to 19 hours, and more preferably 18 hours.

[0038] In the present invention, the cuttings are soaked in the rooting powder for 3 to 5 hours, preferably 4 hours.

[0039] In the present invention, the growth regulator in step (2) is: a final concentration of 700-900 mg / L IBA + a final concentration of 140-180 mg / L NAA solution; preferably a final concentration of 740-860 mg / L IBA + a final concentration of 150-170 mg / L NAA solution; further preferably a final concentration of 780-820 mg / L IBA + a final concentration of 160 mg / L NAA solution; more preferably a final concentration of 800 mg / L IBA + a final concentration of 160 mg / L NAA solution.

[0040] In the present invention, the rooting powder in step (2) is a rooting powder with a mass concentration of 30-40% ABT1; preferably 32-38% ABT1 rooting powder; further preferably 34-36% ABT1 rooting powder; more preferably 35% ABT1 rooting powder.

[0041] In the present invention, the matrix in step (3) is one or more of perlite, rice husk carbon or river sand; preferably perlite, rice husk carbon and river sand, with a mass ratio of 1:1:1.

[0042] In the present invention, the nutrient solution in step (3) is Hoagland's nutrient solution (Qingdao Haibo Biological Hoagland's nutrient solution) with a volume concentration of 1.0 to 1.5 g / L; preferably 1.1 to 1.5 g / L; further preferably 1.2 to 1.5 g / L; more preferably 1.5 g / L.

[0043] In the present invention, the nutrient solution in step (3) is irrigated once every 4 to 6 days, and the amount used each time is 1 to 1.5 L; preferably 1.1 to 1.5 / m 2 . ; More preferably, it is 1.2 to 1.5 L / m 2 More preferably, 1.5 L / m 2 .

[0044] In the present invention, the culturing time in step (3) is 35 to 45 days; preferably 37 to 43 days; more preferably 39 to 41 days; and more preferably 40 days.

[0045] In the present invention, in step (4), after transplanting into the field, the cutting seedlings are topped when they have grown 7 to 8 leaves and tied to the vertical poles; preferably, the topping is performed when they have grown 8 leaves.

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

[0047] Example 1

[0048] A method for promoting rooting of thorny grape cuttings, the steps are as follows:

[0049] 1. During winter pruning, cut the annual grape branches from the tree, cutting every 2 buds. The upper incision should be 2 cm away from the nearest bud, and the lower incision should be cut obliquely, with the lower incision 4 cm away from the nearest bud.

[0050] 2. After 60 days of storage in the sand, take the cuttings out of the sand and treat them with treatment agents. First, soak the cuttings in treatment agent 1 for 14 hours, then soak the cuttings in treatment agent 2 for 16 hours, and finally soak the roots in treatment agent 3 for 3 hours.

[0051] Among them, treatment agent 1 is water, treatment agent 2 is a growth regulator with a final concentration of 700 mg / L IBA + a final concentration of 140 mg / L NAA solution, and treatment agent 3 is 30% ABT1 rooting powder;

[0052] 3. After treatment with the treatment agent, choose to plant the cuttings in a matrix composed of rice husk carbon + perlite (1:1) according to the soil conditions of the nursery base;

[0053] 4. Every 4 days, use 1.0g / L Hoagland nutrient solution to irrigate once, with 1L used each time.

[0054] 5. 35 days after cutting, transplant the cuttings with the substrate to the field and irrigate after transplanting;

[0055] 6. When the cuttings grow 7 leaves, pinch off the top and tie them to the pole.

[0056] Example 2

[0057] A method for promoting rooting of thorny grape cuttings, the steps are as follows:

[0058] 1. During winter pruning, cut the annual grape branches from the tree, cutting every 2 buds. The upper incision should be 4 cm away from the nearest bud, and the lower incision should be cut obliquely, with the lower incision 6 cm away from the nearest bud.

[0059] 2. After 70 days of sand storage, the cuttings were taken out of the sand and treated with treatment agents. First, they were soaked in treatment agent 1 for 18 hours, then in treatment agent 2 for 20 hours, and finally in treatment agent 3 for 5 hours.

[0060] Among them, treatment agent 1 is water, treatment agent 2 is a growth regulator with a final concentration of 900 mg / L IBA + a final concentration of 180 mg / L NAA solution, and treatment agent 3 is 40% ABT1 rooting powder;

[0061] 3. After treatment with the treatment agent, choose to plant the cuttings in a matrix composed of rice husk carbon + river sand (1:1) according to the soil conditions of the nursery base;

[0062] 4. Every 6 days, use 1.3g / L Hoagland nutrient solution to irrigate once, with 1.3L used each time.

[0063] 5. 40 days after cutting, transplant the cuttings with the substrate to the field and irrigate after transplanting;

[0064] 6. When the cuttings grow 8 leaves, pinch off the top and tie them to the pole.

[0065] Example 3

[0066] A method for promoting rooting of thorny grape cuttings, the steps are as follows:

[0067] 1. During winter pruning, cut the annual grape branches from the tree, cutting every 2 buds. The upper incision should be 3 cm away from the nearest bud, and the lower incision should be cut obliquely, with the lower incision 5 cm away from the nearest bud.

[0068] 2. After 65 days of storage in the sand, take the cuttings out of the sand and treat them with treatment agents. First, soak the cuttings in treatment agent 1 for 16 hours, then soak the cuttings in treatment agent 2 for 18 hours, and finally soak the roots in treatment agent 3 for 4 hours.

[0069] Among them, treatment agent 1 is water, treatment agent 2 is a growth regulator with a final concentration of 800 mg / L IBA + a final concentration of 160 mg / L NAA solution, and treatment agent 3 is 35% ABT1 rooting powder;

[0070] 3. After treatment with the treatment agent, choose to plant the cuttings in a matrix composed of perlite, rice husk carbon and river sand, with a mass ratio of 1:1:1, according to the soil conditions of the nursery base;

[0071] 4. Every 5 days, use 1.5g / L Hoagland nutrient solution to irrigate once, with 1.5L used each time.

[0072] 5. 40 days after cutting, transplant the cuttings with the substrate to the field and irrigate after transplanting;

[0073] 6. When the cuttings grow 8 leaves, pinch off the top and tie them to the pole.

[0074] Experimental Example 1

[0075] The single factor experiment was based on the method of Example 3 to investigate the effects of different cutting media on the growth of cuttings.

[0076] 1. Substrate formula: T1: rice husk ash + coarse river sand (1:1); T2: rice husk ash + perlite (1:1); T3: rice husk ash + perlite + vermiculite (1:1:1); T4: rice husk ash + perlite + sandy soil (1:1:1); T5: perlite + vermiculite (1:1); T6: perlite + vermiculite + sandy soil (1:1:1); T7: perlite + sandy soil (1:1); T8: vermiculite + sandy soil (1:1); T9: rice husk ash; T10: vermiculite; T11: sandy soil; T12: perlite; T13: coarse river sand. Three groups for each treatment, 10 plants in each group.

[0077] 2. Growth indicator survey

[0078] The germination rate, leaf expansion rate, survival rate, rooting rate and number of rooted branches were counted 14d, 28d and 42d after branch cuttings. The growth of callus tissue of all cuttings and the growth status of tender shoots were observed, and the root length and number of roots were measured.

[0079] Germination rate (%) = number of germinations / total number of buds × 100; leaf expansion rate (%) = number of expanded leaves / total number of buds × 100; survival rate (%) = number of surviving seedlings / total number of cuttings × 100; rooting rate (%) = number of rooted seedlings / total number of cuttings × 100; callus formation rate (%) = number of seedlings generating callus / total number of cuttings × 100.

[0080] result:

[0081] 1. The effects of different substrates on the growth of cuttings Figure 1 to Figure 4The growth indicators of thorny grape cuttings under 13 substrate conditions are shown in Table 1.

[0082] Table 1 Questionnaire on growth status of different groups

[0083]

[0084]

[0085] From the records in the table, we can see that after the cuttings of the thorny grapes were cut, the germination rate was the fastest in T1 and T5 substrates, reaching 53.33% and 43.33% in 14 days (Table 1). The germination rate was the highest in T5 and T10, which were both vermiculite components, at 90%. The next highest were T1, T8, and T12, all above 80%. The leaf expansion and germination conditions under each substrate condition were basically the same. The flowering rate of thorny grapes was the highest under T4 conditions, at 30%, followed by T13 (20%).

[0086] Overall, perlite (T12) is the most conducive to rooting, with a rooting rate of 90% and a callus formation rate of 100%, followed by T2, T3, and T13, with rooting rates of 60% and callus formation rates of 83.33%, 76.67%, and 90%, respectively. However, the average root length of T12 is only 4.8 cm, and the average number of roots is 3.22. Adding rice husk ash (T2) to T12 increased the root length by 2.33 cm and the number of roots by 2.58, but the rooting rate decreased by 30%; adding vermiculite (T5) to T12 increased the root length by 1.33 cm and the number of roots by 1.21, but the rooting rate decreased by 66.67%. The rooting rate of T11 is 0, but the callus formation rate is 80%. T1 has the longest average root length, which is 9.46cm. The root lengths of T6, T7 and T13 are all above 8.0cm. T13 and T1 both contain river sand. The medium contains river sand, which is easy to grow long roots. Compared with T13, T1 added rice husk ash, which increased the germination rate by 16.67%, the average root length by 1.39cm, and the number of roots by 2.6, but the rooting rate decreased by 20%. T1 has the largest average number of roots, which is 5.83, followed by T2 (5.80) and T10 (5.25).

[0087] When rice husk ash (T9), vermiculite (T10), sand (T11), perlite (T12), and coarse river sand (T13) were used alone, the germination rates were 75%, 90%, 65%, 80%, and 70%, respectively, and the corresponding rooting rates were 20%, 40%, 0, 90%, and 60%. Therefore, vermiculite is helpful for germination, and perlite and coarse river sand are helpful for rooting. When rice husk ash was used alone (T9), the rooting rate was only 20%, and the callus formation rate was 55%; after adding river sand (T1), it reached 40%, and the callus formation rate was 76.67%; after adding perlite (T2) and perlite + vermiculite (T3), the rooting rate reached 60%, and the callus formation rates also reached 83.33% and 76.67%. When sand was used alone (T11), the rooting rate was 0, and the germination rate was 65%. When added to other formulas, it would reduce the germination rate and rooting rate of the original formula. Perlite (T12) and perlite + sand (T7), vermiculite (T10) and vermiculite + sand (T8), perlite + vermiculite (T5) and perlite + vermiculite + sand (T6), rice husk ash + perlite (T2) and rice husk ash + perlite + sand (T4), the germination rate was reduced by 13.33%, 6.67%, 16.67%, 16.67% respectively compared with the case without sand (the latter was lower than the former), and the rooting rate was also reduced by 83.33%, 36.67%, 3.33%, and 50% respectively. In addition, from Figure 1 to Figure 3 It can be seen that the formula containing sand after the substrate (T4 T6T7 T8) will cause the substrate to be airtight and the cuttings to appear black.

[0088] 2. PCA analysis of important growth indicators under different substrate conditions

[0089] PCA analysis was performed on the six indicators under 13 substrate conditions, and the cumulative contribution rates of the three principal components were 43.253%, 27.678%, and 24.004%, respectively, with a cumulative contribution rate of 94.934%. Through comprehensive score analysis, the effects of the 13 substrates on the cuttings of Vitis vinifera were ranked as follows: T12, T1, T2, T13, T10, T5, T3, T6, T9, T8, T11, T4, and T7.

[0090] Table 2 PCA analysis table

[0091]

[0092]

[0093] Table 3 Scoring matrix

[0094] Ingredients 1 Ingredient 2 Ingredient 3 Germination rate 0.715 -0.624 0.304 Leaf expansion rate 0.738 -0.539 0.397 Rooting rate 0.65 0.668 0.245 Callus formation rate 0.227 0.687 0.653 Average root length 0.632 0.19 -0.705 Average number of roots 0.816 0.163 -0.455 Cumulative Contribution Rate 43.25％ 70.93％ 94.93％

[0095] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for promoting rooting of thorny grape cuttings, characterized in that: The steps include: (1) Cut cuttings from grape branches and store them in sand; (2) soaking the cuttings obtained in step (1) in water, a growth regulator and rooting powder in sequence; (3) inserting the cuttings obtained in step (2) into a substrate for cultivation, and applying a nutrient solution to obtain cutting seedlings; (4) Transplanting the cuttings obtained in step (3) into a field.

2. The method according to claim 1, characterized in that Step (1) Each cutting retains 1 to 3 buds, the upper incision is 2 to 4 cm away from the nearest bud eye, and the lower incision is 4 to 6 cm away from the nearest bud eye.

3. The method according to claim 1, characterized in that The sand storage time in step (2) is 60 to 70 days.

4. The method according to claim 1, characterized in that: In step (2), the cuttings are soaked in water for 14 to 18 hours; the cuttings are soaked in a growth regulator for 16 to 20 hours; and the cuttings are soaked in rooting powder for 3 to 5 hours.

5. The method according to claim 1, characterized in that The growth regulator in step (2) is: a final concentration of 700-900 mg / LIBA + a final concentration of 140-180 mg / LNAA solution.

6. The method according to claim 1, characterized in that The rooting powder in step (2) is ABT1 rooting powder with a mass concentration of 30-40%.

7. The method according to claim 1, characterized in that The matrix in step (3) is one or more of perlite, rice husk carbon or river sand.

8. The method according to claim 1, characterized in that The nutrient solution in step (3) is a Hoagland nutrient solution with a volume concentration of 1.0 to 1.5 g / L; the nutrient solution is irrigated once every 4 to 6 days, and the amount used each time is 1 to 1.5 L / m 2 .

9. The method according to claim 1, characterized in that: The culturing time in step (3) is 35 to 45 days.

10. The method according to claim 1, characterized in that Step (4) After transplanting to the field, the cuttings are topped when they grow 7 to 8 leaves and tied to the vertical poles.

Images