NL2027805B1 - Breeding method of adansonia digitata - Google Patents

Abstract

The present disclosure provides a breeding method of Adansonia digitata, and belongs to the technical field of cuttage seedling. The method includes the following steps: step 1, disinfecting a culture substrate with benzalkonium.bromide solution to obtain a disinfected substrate; and step 2, disinfecting a cutting of A. digitata with ampicillin.soluticw1to obtain51disinfected.cutting, soaking the bottom end of the disinfected cutting in fulvic acid solution for 20—40 s, and.inserting the cutting in.the disinfected.substrate obtained.in step 1 for cultivation. The survival rate of cuttings of A. digitata is improved. by using the breeding' method. provided. by the present disclosure.

Description

BREEDING METHOD OF ADANSONIA DIGITATA

TECHNICAL FIELD The present disclosure relates to the technical field of cuttage seedling, and in particular to a breeding method of Adansonia digitata.

BACKGROUND Adansonia digitata is a large perennial deciduous tree that belongs to the family Malvaceae and the genus Adansonia. The tree is native to tropical arid areas in Africa. A. digitata is a tree species with high economic value and extensive use. Dried leaves of A. digitata are rich in carotene and vitamin A, leaflets contain 4% protein, and leaves further serve as a seasoning. Juicy fruit pulp can serve as a fruit to eat directly or be used to brew fruit wine and beverages. Seed can be eaten raw as snack, or as a thickener or flavoring agent or after baking; high-quality edible oil can be extracted from seeds, which is faint yellow, with an oil content of ~15%. Barks, rich in fiber, can be used for papermaking, mat knitting with coarse cloth, ropemaking, and red dye making. The shell can be carved into various ladles and containers. In medicine, roots and leaves are indicated for antiphlogosis, pyretolysis, antimalarial, heat clearing and detoxification, tranquilizing and allaying excitement. In addition, because adult tree has a robust trunk and an exquisite and unique tree shape, A.

digitata has superb ornamental value in gardening and landscape architecture. In recent years, seedling demand is rising. A. digitata is not naturally distributed in China, but only introduced on a small scale in a few areas in Taiwan, Fujian, Hainan, Guangdong, Guangxi, and Xishuangbanna, Yunnan.

According to a report from the place of origin in Africa (SCUC. Baobab Manual, Field Manual for Extension Workers and Farmers [M], Southampton: University of Southampton, 2006.), the survival rate of cuttings was 20% when soaking shoots of A. digitata in IBA solution, but the survival rate of cuttage seedlings without soaking in hormone was only 2%.

SUMMARY In view of this, an objective of the present disclosure is to provide a breeding method of A. digitata. The breeding method provided by the present disclosure may be used to improve the survival rate of A. digitata.

To achieve the above objective, the present disclosure provides the following technical solution: The present disclosure provides a breeding method of A. digitata, including the following steps: step 1, disinfecting a culture substrate with benzalkonium bromide solution to obtain a disinfected substrate; and step 2, disinfecting a cutting of A. digitata with ampicillin solution to obtain a disinfected cutting, soaking the bottom end of the disinfected cutting in fulvic acid solution for 20-40 s, and inserting the cutting in the disinfected substrate obtained in step 1 for cultivation.

Preferably, the mass percent of the benzalkonium bromide solution in step 1 may be 0.05-0.1%.

Preferably, the ampicillin solution in step 2 may have a concentration of 10-50 mg/L.

Preferably, the fulvic acid solution in step 2 may have a concentration of 0.1-0.2 g/L.

Preferably, the bottom end of the disinfected cutting in step 2 may be soaked in the fulvic acid solution for 30 s.

Preferably, the cultivation in step 2 may be conducted at 22-32°C with an ambient humidity of 60-753.

Preferably, the moisture content of the disinfected substrate may be held at 55-65% during the cultivation in step

2.

Preferably, a shading net may be used for shading during the cultivation in step 2, and the shading net may have a light transmittance of 50-70%.

Preferably, the culture substrate in step 1 may be prepared as follows: mixing eucalyptus bark, Pinus kesiva bark, and oak bark in a mass ratio of 6:3:1 for decomposition to obtain the culture substrate. Preferably, the cutting of A. digitata in step 1 may be 7-10 cm in length and 0.3-0.8 cm in diameter. The present disclosure provides a breeding method of A. digitata. The method includes the following steps: step 1, disinfecting a culture substrate with benzalkonium bromide solution to obtain a disinfected substrate; and step 2, disinfecting a cutting of A. digitata with ampicillin solution to obtain a disinfected cutting, soaking the bottom end of the disinfected cutting in fulvic acid solution for 20-40 s, and inserting the cutting in the disinfected substrate obtained in step 1 for cultivation. The mechanism underlying the improvement of the survival rate of A, digitata in the present disclosure is that: the fulvic acid is used to promote the rooting and survival of the cutting of A. digitata, and the survival of the cutting is improved by disinfection with benzalkonium bromide plus ampicillin.

DETAILED DESCRIPTION The present disclosure provides a breeding method of A. digitata, including the following steps: step 1, disinfecting a culture substrate with benzalkonium bromide solution to obtain a disinfected substrate; and step 2, disinfecting a cutting of A. digitata with ampicillin solution to obtain a disinfected cutting, soaking the bottom end of the disinfected cutting in fulvic acid solution for 20-40 s, and inserting the cutting in the disinfected substrate obtained in step 1 for cultivation. In the present disclosure, the culture substrate may preferably be prepared as follows: mixing eucalyptus bark, Pinus kesiva bark, and oak bark in a mass ratio of 6:3:1 for decomposition to obtain the culture substrate. In the present disclosure, the culture substrate may preferably be prepared in accordance with a method for preparing an organic seedling light substrate using Pinus kesiya bark as a raw material disclosed by CN Patent Application No. CN201210498290.8, and the Pinus kesiya bark may be replaced with the eucalyptus bark,

Pinus kesiya bark, and oak bark. In the present disclosure, the culture substrate is air-permeable without waterlogging, prone to root growth of A. digitata, and further improve the survival rate of A. digitata.

In the present disclosure, the mass percent of the benzalkonium bromide solution may preferably be 0.05-0.1%. In the present disclosure, the culture substrate may preferably be disinfected by conventional spraying method. In the present disclosure, the ampicillin solution may preferably have a concentration of 10-50 mg/L. In the present disclosure, the cutting of A. digitata may preferably be disinfected by spraying method.

In the present disclosure, the cutting of A. digitata may preferably be 7-10 cm in length and 0.3-0.8 cm in diameter.

In the present disclosure, standards for the cutting of A. digitata may further preferably include: growing robustly; having apical buds; the cut at the bottom end of the cutting being wedged and tapered; reserving two or three young leaves at the apex of the cutting; and cutting the remaining blade off the cutting along the petiole, instead of cutting 1/2 to 3/4 of the blade along the outer edge or base of the blade, in order to reduce the degree of mechanical damage to the cutting.

The bottom end of the disinfected cutting may preferably be soaked in the fulvic acid solution for 30 s.

In the present disclosure, the cultivation may preferably be conducted at 22-32°C with an ambient humidity of 60-75%. In the present disclosure, the cultivation may preferably be conducted at 32°C in the daytime and at 22°C in the evening.

In the present disclosure, the moisture content of the disinfected substrate may preferably be held at 55-65% during the cultivation. In the present disclosure, a shading net may preferably be used for shading during the cultivation, and the shading net may preferably have a light transmittance of 50-70%.

The technical solution provided by the present disclosure will be described in detail below with reference to examples,

but they should not be construed as limiting the claimed scope of the present disclosure.

The experimental site is located in Nali Advanced Science and Technology Demonstration Park, Yuanyang County, Yunnan 5 Province, with an elevation of 500 m. The site belongs to subtropical monsoon climate. Annual average temperature is 21-23°C, mean temperature of the warmest quarter is 28.3-29.7°C, and mean temperature of the coldest quarter is 15.9-17.0°C; annual precipitation is 900 mm, and rainy season is from May to October. The sample size of each example or comparative example is 100, with three duplicates.

Example 1 (1) Selection and treatment of bedded land: The bedded land was weeded and leveled; a layer of sand or gravel was spread on the ground to make the seedbed bottom 3 cm above the ground; a seedbed was arranged around by red bricks.

(2) Preparation and disinfection of nutrition bag: Bark-based seedling substrates (raw materials of the bark-based seedling substrates included eucalyptus bark, Pinus kesiya bark, and oak bark in a mass ratio of 6:3:1, prepared in accordance with a method for preparing an organic seedling light substrate using Pinus kesiva bark as a raw material disclosed by CN Patent Application No. CN201210498290.8) were bagged in nutrition bags with a diameter of 6 cm and a height of 12 cm. Well-bagged nutrition bags were placed in the seedbed in sequence, and the substrates were disinfected by spraying

0.05 wt% benzalkonium bromide solution.

(3) Shoot collection and cutting: Robust shoots with apical buds, 0.4 cm in diameter, were selected as cuttings; the shoots were sprayed with water immediately after cutting, placed in a foam box, and covered over with wet towels for moisture retention. The cutting length was 7 cm, and the cut at the bottom end of the cutting was wedged and tapered. Two or three young leaves were reserved at the apex of the cutting, and the remaining blade was cut off the cutting along the petiole, instead of cutting 1/2 to 3/4 of the blade along the outer edge or base of the blade, in order to reduce the degree of mechanical damage to the cutting.

(4) Cuttage: The cuttings obtained in step (3) were subjected to spray disinfection with 10 mg/L ampicillin solution; subsequently, the bottom end of each cutting was soaked in 0.1 g/L fulvic acid solution for 30 s and inserted into the seedling substrates in the nutrition bag; after that, the nutrition bags were sprayed with water until drenched.

{5) Post-cuttage management: Keels were arranged every 50 cm on the seedbed, a 40-60 cm high plastic film arch shed was built, and a layer of shading net with 50-70% light transmittance was covered over to reduce the loss of water from the cuttings due to transpiration. The temperature and humidity in shed were checked every 24 h; the temperature in shed was held at 22°C (in the evening) and 32°C (in the daytime), and the air humidity in shed was 60-75%. Regular watering was based on the condition of substrates in the nutrition bag, and substrate moisture content was held at 55-65%.

(6) Transplantation: After the cuttings took root, the plastic film and the shading net were uncovered; regular watering and management proceeded until seedlings grew to 20-25 cm, followed by outplanting and transplantation. Results are shown in Table 1.

Example 2 {1) Selection and treatment of bedded land: The bedded land was weeded and leveled; a layer of sand or gravel was spread on the ground to make the seedbed bottom 2 cm above the ground; a seedbed was arranged around by red bricks.

(2) Preparation and disinfection of nutrition bag: Bark-based seedling substrates (raw materials of the bark-based seedling substrates included eucalyptus bark, Pinus kesiya bark, and oak bark in a mass ratio of 6:3:1, prepared in accordance with a method for preparing an organic seedling light substrate using Pinus kesiya bark as a raw material disclosed by CN Patent Application No. CN201210498290.8) were bagged in nutrition bags with a diameter of 5 cm and a height of 10 cm. Well-bagged nutrition bags were placed in the seedbed in sequence, and the substrates were disinfected by spraying

0.075 wt% benzalkonium bromide solution.

(3) Shoot collection and cutting: Robust shoots with apical buds, 0.5cm in diameter, were selected as cuttings; the shoots were sprayed with water immediately after cutting, placed in a foam box, and covered over with wet towels for moisture retention. The cutting length was 8 cm, and the cut at the bottom end of the cutting was wedged and tapered. Two or three young leaves were reserved at the apex of the cutting, and the remaining blade was cut off the cutting along the petiole, instead of cutting 1/2 to 3/4 of the blade along the outer edge or base of the blade, in order to reduce the degree of mechanical damage to the cutting.

(4) Cuttage: The cuttings obtained in step (3) were subjected to spray disinfection with 30 mg/L ampicillin solution; subsequently, the bottom end of each cutting was soaked in 0.15 g/L fulvic acid solution for 30 s and inserted into the seedling substrates in the nutrition bag; after that, the nutrition bags were sprayed with water until drenched.

(5) Post-cuttage management: Keels were arranged every 50 cm on the seedbed, a 40-60 cm high plastic film arch shed was built, and a layer of shading net with 50-70% light transmittance was covered over to reduce the loss of water from the cuttings due to transpiration. The temperature and humidity in shed were checked every 24 h; the temperature in shed was held at 22°C (in the evening) and 32°C (in the daytime), and the air humidity in shed was 60-75%. Regular watering was based on the condition of substrates in the nutrition bag, and substrate moisture content was held at 55-65%.

(6) Transplantation: After the cuttings took root, the plastic film and the shading net were uncovered; regular watering and management proceeded until seedlings grew to 20-25 cm, followed by outplanting and transplantation. Results are shown in Table 1.

Example 3 {1) Selection and treatment of bedded land: The bedded land was weeded and leveled; a layer of sand or gravel was spread on the ground to make the seedbed bottom 3 cm above the ground;

a seedbed was arranged around by red bricks.

(2) Preparation and disinfection of nutrition bag: Bark-based seedling substrates (raw materials of the bark-based seedling substrates included eucalyptus bark, Pinus kesiya bark, and oak bark in a mass ratio of 6:3:1, prepared in accordance with a method for preparing an organic seedling light substrate using Pinus kesiya bark as a raw material disclosed by CN Patent Application No. CN201210498290.8) were bagged in nutrition bags with a diameter of 6 cm and a height of 12 cm. Well-bagged nutrition bags were placed in the seedbed in sequence, and the substrates were disinfected by spraying

0.1 wt% benzalkonium bromide solution.

(3) Shoot collection and cutting: Robust shoots with apical buds, 0.8 cm in diameter, were selected as cuttings; the shoots were sprayed with water immediately after cutting, placed in a foam box, and covered over with wet towels for moisture retention. The cutting length was 10 cm, and the cut at the bottom end of the cutting was wedged and tapered. Two or three young leaves were reserved at the apex of the cutting, and the remaining blade was cut off the cutting along the petiole, instead of cutting 1/2 to 3/4 of the blade along the outer edge or base of the blade, in order to reduce the degree of mechanical damage to the cutting.

{4) Cuttage: The cuttings obtained in step (3) were subjected to spray disinfection with 50 mg/L ampicillin solution; subsequently, the bottom end of each cutting was soaked in 0.2 g/L fulvic acid solution for 30 s and inserted into the seedling substrates in the nutrition bag; after that, the nutrition bags were sprayed with water until drenched.

(5) Post-cuttage management: Keels were arranged every 50 cm on the seedbed, a 40-60 cm high plastic film arch shed was built, and a layer of shading net with 50-70% light transmittance was covered over to reduce the loss of water from the cuttings due to transpiration. The temperature and humidity in shed were checked every 24 h; the temperature in shed was held at 22°C (in the evening) and 32°C (in the daytime), and the air humidity in shed was 60-75%. Regular watering was based on the condition of substrates in the nutrition bag, and substrate moisture content was held at 55-65%. {6) Transplantation: After the cuttings took root, the plastic film and the shading net were uncovered; regular watering and management proceeded until seedlings grew to 20-25 cm, followed by outplanting and transplantation. Results are shown in Table 1. Comparative Example 1 As done in Example 3, fulvic acid was merely replaced with ABT rooting agent with the same concentration. Results are shown in Table 1. Comparative Example 2 As done in Example 3, fulvic acid was merely replaced with indolebutyric acid (IBA) with the same concentration. Results are shown in Table 1. Comparative Example 3 As done in Example 3, neither plant growth regulator nor root promoter was used in step 4. Results are shown in Table

1. Comparative Example 4 As done in Example 3, the seedling substrate in the nutrition bag was replaced with river sand, which was disinfected by spraying 75% (v/v) alcohol. The cuttings were further disinfected by spraying 75% (v/v) alcohol. Fulvic acid was replaced with ABT rooting agent with the same concentration. Results are shown in Table 1. Comparative Example 5 As done in Example 3, the seedling substrate in the nutrition bag was replaced with red clay, which was disinfected by spraying 0.2 wt% commercially available carbendazim solution. The cuttings were disinfected with 0.1 wt% potassium permanganate solution. Fulvic acid was replaced with IBA with the same concentration. Results are shown in Table 1. Comparative Example 6 As done in Example 3, the bark-based seedling substrate in the nutrition bag was disinfected by spraying 0.5 wt% potassium permanganate solution. The cuttings were disinfected by spraying 0.1 wt? carbendazim solution. Neither plant growth regulator nor root promoter was used in step 4. Results are shown in Table 1. Comparative Example 7 The seedling substrate in the nutrition bag was replaced with commercially available humus soil. Neither plant growth regulator nor root promoter was used in step 4. The disinfection method and other steps were the same as those in Example 3. Results are shown in Table 1.

Table 1 Cuttage results Survival Average rate of number of Average Days of | cuttings lateral basal callus | | 30 days roots with diameter formation after a length growth cuttage of >0.5 cm Example 1 | 14 | 69.38 | 6 | 0.12 em Example 2 | 14 | 72.7 | 6 0.19 en Example 3 a 0.24 cm Comparative

45.4% 5 0.21 cm Example 1 Comparative 17 31.2% 2 0.18 cm Example 2 Comparative 7

8.54 1 0.07 cm Example 3 Comparative 18 36.1% 4 0.19 cm Example 4 Comparative 19 29.0% 2 0.17 cm Example 5 Comparative 7 22 7.7% 1 0.06 cm Example 6 Comparative 22 1 0.06 cm Example 7

From the above examples and comparative examples, Example 3 is superior to Comparative Examples 1, 2, and 3, indicating that the fulvic acid as root promoter in the present disclosure has a better effect than conventional hormone; Comparative Example 1 is superior to Comparative Example 4, indicating that the combination of the substrate and disinfectants in the present disclosure is superior to that of conventional river sand and alcohol; Comparative Example 2 is superior to Comparative Example 5, indicating that the combination of the substrate and disinfectants in the present disclosure is superior to that of conventional red clay, carbendazim, and potassium permanganate; Comparative Example 3 is superior to Comparative Example 6, indicating that the combination of the disinfectants in the present disclosure is superior to that of conventional carbendazim and potassium permanganate; Comparative Example 3 is superior to Comparative Example 7, indicating that the selection of the substrate in the present disclosure is superior to that of conventional humus soil. In summary, the survival rate of cuttings of A. digitata is improved by using the breeding method provided by the present disclosure.

The above descriptions are merely preferred implementations of the present disclosure. It should be noted that several improvements and modifications may also be made by a person of ordinary skill in the art without departing from the principle of the present disclosure, and such improvements and modifications should be deemed as falling within the protection scope of the present disclosure.

Claims (10)

CONCLUSIONS A method for culturing Adansonia digitata, comprising the following steps: step 1: disinfecting a cultivation substrate with a benzalkonium bromide solution to obtain a disinfected substrate; and step 2: disinfecting a cutting of Adansonia digitata with an ampicillin solution to obtain a disinfected cutting, soaking the underside of the disinfected cutting in a fulvic acid solution for 20-405, and placing it in the disinfected cutting substrate cuttings of the cutting obtained in step 1 for cultivation. The culture method according to claim 1, wherein the mass percentage of the benzalkonium bromide solution in step 1 is 0.05-0.13. The culture method of claim 1, wherein the ampicillin solution in step 2 has a concentration of 10-50 mg/L. The culture method of claim 1, wherein the fulvic acid solution in step 2 has a concentration of 0.1-0.2 g/L. The culture method of claim 1, wherein the underside of the disinfected cutting is soaked in the fulvic acid solution in step 2 for 30 seconds. The cultivation method according to claim 1, wherein the cultivation in step 2 is carried out at 22-32°C with a humidity of 60-75%. The cultivation method according to claim 1, wherein the humidity level of the disinfected substrate is maintained at 55-65% during cultivation in step 2. A breeding method according to claim 1, wherein a shade net for shade is used during the cultivation in step 2, and the shade net has a light transmittance of 50-70%. The cultivation method of claim 1, wherein the cultivation substrate is prepared in step 1 as follows: mixing eucalyptus bark, Pinus kesiya bark, and oak bark in a mass ratio of 6:3:1 for decomposition to obtain the cultivation substrate. The cultivation method of claim 1, wherein the cutting of Adansonia digitata in step 1 is 7-10 cm in length and 0.3-0.8 cm in diameter, -0-0-0-0-