CN105284388A - Blueberry softwood cutting seedling method - Google Patents

Abstract

The present invention discloses a blueberry softwood cutting seedling method. The method comprises the following steps: selecting and trimming cutting woods, soaking the trimmed woods with a rooting agent containing boron and IBA for 8-10 hours, and inserting the woods into a matrix, wherein the humidity of the matrix is kept at 80% or above, and the temperature is controlled at 20-28 DEG C. The method of the present invention is simple to study, high in operability, good in rooting effect and high in utility value, wherein the rooting rate reaches up to 91.18%, so that the callus formation rate and the rooting number can be significantly improved.

Description

A kind of blueberry green branch cutting seedling raising method

technical field

The invention belongs to the technical field of plant vegetative propagation, and in particular relates to a method for growing blueberry green branch cutting seedlings.

Background technique

At present, my country's blueberry planting industry is developing rapidly, the planting area continues to expand, the demand for seedlings is large, and the insufficient supply of seedlings has become a bottleneck hindering the development of my country's blueberry industry. However, because blueberries are shallow-rooted plants with underdeveloped root fibers and no root hairs, the requirements for soil pH, soil moisture, air permeability, drainage, climatic conditions, and management methods are relatively strict, so rooting is extremely difficult.

At present, tissue culture technology is mainly used for rapid propagation of blueberries in China, but some microorganisms are often carried on the surface and inside of explants. At the same time, the temperature, humidity, nutrition, pH value, etc. in the process of plant tissue culture are suitable for the growth of microorganisms. Once the microorganisms enter the culture container, they will multiply rapidly, and the plant materials will be produced through nutrient competition, erosion of plant materials, and secretion of toxic metabolites. Disease or death, resulting in failure of tissue culture. The occurrence of contamination in tissue culture is extremely common, and the induction of rooting is the bottleneck in the work of blueberry tissue culture, which largely limits the application of this technology in commercial production. Therefore, choosing the correct cutting method is an important means for rapid growth and reproduction of blueberries and ensuring the excellent properties of the variety.

In terms of production, the rapid breeding of blueberries mainly adopts the method of tissue culture propagation, and a small part adopts blueberry hard branch and green branch cutting technology, but mainly focuses on the separate use of various exogenous hormones (IAA, IBA, etc.) Promoting the rooting of blueberry twig cuttings, but the rooting rate is low, and the rooting effect is not ideal, and these technologies still cannot meet the market demand of blueberry large-scale propagation and carry out.

Contents of the invention

The object of the present invention is to provide a kind of blueberry green branch cuttage seedling raising method, by adding boron, sucrose, vitamin B of different concentration in IBA, apply self-made rooting agent to further improve blueberry cutting survival rate, for blueberry scale cutting breeding Provide new theoretical and practical basis.

The present invention is specifically realized through the following technical solutions:

A kind of blueberry green branch cutting seedling raising method specifically comprises the following steps:

1) Selection of cuttings: From March to April, select the middle and lower parts of vegetative branches that are 1-year-old with high hardness, good maturity and health as cuttings;

2) Treatment of cuttings: trim the branches to a length of 7-10cm, with 4-5 axillary buds and 1-2 leaves on the top of the cuttings.

3) Soaking of cuttings: Soak with a rooting agent of boron+IBA for 8-10 hours;

4) Cutting: Before cutting, pour the substrate with water to ensure humidity but no water accumulation. On the day of cutting, soak the base of the treated cuttings with 1000 times of fenflum for 10 seconds, and then insert the cuttings vertically into the substrate;

5) Moisture management: the humidity of the substrate is maintained above 80%, and the temperature is controlled at 20-28°C;

6) Management after cuttings: Gradually apply fertilizer after the cuttings take root. Fertilization is applied in the form of liquid fertilizer with a concentration of no more than 3%. Fertilize blueberry cuttings and green branches once a week.

further,

Rooting agent described in step (3) is made solvent with sterile water, and preparation rooting agent ensures that IBA concentration is 50mg/L, and the concentration of boron is 300mg/L;

The substrate is to use moss soil as the cutting substrate, adjust the pH value to 5.5, and spray the cutting bed substrate with 1000 times carbendazim one day before the cutting;

Specifically, the cuttings are inserted into the substrate at a depth of 2/3 of the entire cutting, and the distance is 5cm×10cm.

The beneficial effects of the present invention are as follows: 1) the method has undergone strict tests, and the result is accurate and reliable. The technology is simple and easy to learn, has strong operability, good rooting effect, high application value, and meets the production requirements of blueberry green branch cuttings; 2) The rooting rate of the present invention can reach up to 91.18%, and the average rooting number reaches 16.40, and the average The root length was 4.75, the rooting index was 1.83, and the cutting enzyme activity was significantly increased; 3) the callus formation rate and the number of roots could be significantly increased.

detailed description

The present invention will be further described below in conjunction with the embodiments. The following descriptions are only preferred embodiments of the present invention, and are not intended to limit the present invention to other forms. Changes to equivalent embodiments with equivalent changes. Any simple modifications or equivalent changes made to the following embodiments according to the technical essence of the present invention without departing from the solution content of the present invention fall within the protection scope of the present invention.

Example 1

The experiment was carried out in the greenhouse of the blueberry demonstration base in Gongcun, Baima Town, Meishan City, Sichuan Province, and the green branches of two varieties of Nangaobush blueberry, Lanfeng and Hebert, were selected from the blueberry planting base in Gongcun, Baima Town, Meishan City, Sichuan Province.

From March to April 2013, select southern highbush blueberry Herbert and Lanfeng's healthy green branches of the year and bring them back to the laboratory. The cuttings should be cut from trees that grow vigorously and are free of diseases and insect pests (it is better to choose hard, mature branches If there is a virus disease in the orchard, the cutting tree should be more than 15cm away from the diseased tree. Cutting branches are the best For one-year-old vegetative branches, the middle and lower parts of the branches should be selected for cutting as much as possible, and the branches should be trimmed into 7-10cm cuttings with 4-5 axillary buds and 1-2 leaves on the top. Use sterile water as a solvent to prepare a rooting agent to ensure that the concentration of IBA is about 50 mg/L and the concentration of boron is 300 mg/L. The cuttings of blueberry Herbert and Lanfeng are soaked in the rooting agent for 8 to 10 hours and then inserted into the matrix. It accounts for 2/3 of the entire cutting, and the spacing is 5cm×10cm. Before cutting, pour the substrate with water to ensure humidity but not water accumulation. On the day of cutting, soak the base of the cuttings with more than 1,000 chlorprofen for 10 seconds, and then insert the cuttings vertically. in the matrix. Use intermittent automatic spraying devices to water frequently, keep the humidity of the substrate on the cutting bed above 80%, and control the temperature at 20-28°C. If it is placed in the sun for too long, when the water temperature is high, you should wait for the water temperature to cool before watering, so as not to damage the seedlings. The most critical period for water management is from the beginning of May to the end of June. At this time, the leaves have unfolded, but the cuttings have not yet taken root. Insufficient water will easily cause the cuttings to die. When the top leaves start to turn green, the cuttings are rooted.

Fertilizer management: Do not apply any fertilizer to the matrix before cuttings and rooting, and gradually apply fertilizer after the cuttings take root to promote the growth of seedlings. Fertilization should be applied in the form of liquid fertilizer with a concentration of about 3%. Fertilize the green branches of blueberry cuttings once a week. In order to avoid damage to the leaves due to fertilization, spray water after each fertilization to rinse the fertilizer on the leaves.

Pest management: during rooting and seedling cultivation, ventilation treatment and removal of diseased plants are mainly used to control blueberry susceptible diseases. Therefore, greenhouse or greenhouse seedlings must be ventilated in time to reduce fungal diseases and reduce the temperature in the seedling room.

At the same time, a control group was set up, and the control group was administered with exogenous hormone IBA alone for rooting and raising seedlings.

According to the statistics of the applicant, compared with the conventional propagation and seedling raising method, the rooting rate and rooting efficiency in some periods are shown in Table 1 and Table 2, which shows that the self-made rooting agent can effectively improve the rooting rate and rooting efficiency of blueberries.

Table 1 The effect of boron compound IBA rooting agent on the rooting of Lanfeng at different rooting stages

Note: rooting efficiency Q=(P _n -P _n-1 )/rooting days (P _n is the rooting rate at the next rooting time point, and P _n-1 is the rooting rate at the previous rooting time point). (The same below).

Table 2 The effect of boron compound IBA rooting agent on rooting at different rooting stages of Herbert

As shown in Table 3 and 4, the rooting rate, average root number, average root length, and rooting power index of the two blueberry varieties were higher than those of the single IBA treatment after 70 days of cutting, and the rooting effect was obvious.

Table 3 Lanfeng final rooting situation

Note: rooting ability index = (average root length × number of roots)/total number of cuttings. (The same below).

Table 4 Herbert's final rooting situation

Through the application of this technology, the rooting rate of blueberry green branch cuttings has been greatly improved, which is beneficial to the rapid propagation of blueberry cuttings. The market prospect is broad, and it has great social and economic value.

In order to further illustrate the influence of the rooting agent on the propagation process of seedling rooting, the following uses conventional experiments to describe and analyze in detail.

1) Test material

The experiment was carried out in the greenhouse of the blueberry demonstration base in Gongcun, Baima Town, Meishan City, Sichuan Province, and the green branches of two varieties of Nangaobush blueberry, Lanfeng and Hebert, were selected from the blueberry planting base in Gongcun, Baima Town, Meishan City, Sichuan Province.

2) Overview of the test site

In October 2010, the Meishan Blueberry Ecological Base, with a total investment of 45 million yuan, entered the site smoothly and settled in Gong Village, the "No. 1 Village in Meishan". After more than half a year of careful cultivation, the first trial planting of blueberries in Meishan has been successful. The blueberry ecological base in Gong Village, Baima Town, the experimental site, currently has a scale of 100 mu, and three blueberry varieties have been successfully planted in the village. The experimental site has a large-scale greenhouse for blueberry cuttings and is equipped with a fully automatic spraying device, which can provide an environment that is conducive to the propagation of blueberry cuttings all year round.

3) Test equipment

Agilent1260 liquid chromatograph; rotary evaporator; pH acidity meter; centrifuge; shaker; constant temperature water bath; spectrophotometer; balance; mortar; test tube;

4) Test drugs

Hormone standard samples IAA and ABA are provided by SIGMA; chromatographic grade methanol and acetic acid; liquid nitrogen; polyvinylpyrrolidone (PVPP); analytically pure ethyl acetate and petroleum ether; di-tert-butyl-p-cresol (BHT); phosphoric acid Buffer solution (pH ₌ ^6.0 ); quartz sand: dichlorophenol; manganese chloride; ferric chloride: ethanol; concentrated sulfuric acid; .Mix before use and store in the dark. When using, add 4mL of reagent to 1mL sample).

5) Test method

On August 23, 2013, the green branches of the southern highbush blueberry Hebert and Lanfeng were selected and brought back to the laboratory, and the branches were trimmed to 7-10cm with 4-5 axillary buds and 1-2 leaves on the top. cuttings. IBA was formulated into 50mg·L ^-1 , 100mg·L ^-1 , and 200mg·L ^-1 rooting agents respectively, and water treatment was used as the control, a total of 4 treatments, repeated 3 times. For each variety, soak a bundle of 20 cuttings in different concentrations of IBA and water, take out a bundle every 2 hours (soaked for 10 hours in total), and use high-performance liquid chromatography to measure the thickness of the cortex within 1 to 2 cm from the base of the cuttings. The content of the source hormones IAA and ABA. The concentration and time of IBA treatment were determined by the highest IAA/ABA.

①Experimental design

The cutting time was from September 5, 2013 to November 29, 2013. The cutting substrate is moss soil with a pH value of 4.5. Trim the branches into 7-10cm cuttings with 4-5 axillary buds and 1-2 leaves on the top. The proportions of boron, sucrose, and vitamin B added to Lanfeng and Herbert rooting agents are shown in Table 1, and the IBA concentration is based on the highest endogenous IAA/ABA in the cuttings in Test 1. Each rooting agent formula is a treatment, and each treatment has 70 cuttings, which are repeated three times. Let the IBA treatment alone be CK ₁ , and the clean water treatment alone be CK ₂ . The treatment time of cuttings was the one corresponding to the highest ratio of endogenous hormones in Experiment 1. Cuttings were carried out on September 5, 2013, and 10 cuttings were randomly selected for 7 days per treatment (i.e. cutting 1d, 8th, 15d, 23d, 30d, 37d, 45d) to dynamically monitor the callus of cuttings And rooting situation, rooting efficiency Q=(Pn-Pn-1)/rooting days (Pn is the rooting rate of the next rooting time point, Pn-1 is the rooting rate of the previous rooting time). Because the root system of blueberry is weak and the root system develops slowly, the cuttings will not be taken out to observe and measure indicators after the cuttings are generally rooted (cutting 45d). The comprehensive rooting quality was evaluated by the rooting ability index of the root system, and the rooting ability index=(average root length×number of roots)/total number of cuttings.

Table 5 rooting agent formula

②Extraction method of endogenous hormone in cortex of cuttings

Liquid chromatography conditions: AgilentC18ZORBAX reversed-phase chromatographic column (150mm×4.6mm, 5μm); column temperature: 35°C; wavelength: 254nm; mobile phase is V (methanol): V (water): V (acetic acid) = 45: 54.4 : 0.6; flow rate: 1.0mL/min; injection volume: 20μl. Quantitative determination by external standard method. The endogenous hormone standard curve is as follows:

AreaIAA=21.254591X-9.7334905, R=0.99390;

AreaABA=21.0289715X-13.481436, R=0.99393.

The extraction method of endogenous hormone refers to Zhang Yongzhong. From September 5, 2013, every 15 days of treatment (i.e. the 1st day, 15th day, 30th day, and 45th day of cuttings) accurately weighed 1g of the cortex within the range of 0-2cm from the base of the cuttings. Grind and crush. Add 10 mL of 80% methanol pre-cooled at 4°C, extract overnight at 4°C, and centrifuge at 5000r·min ^-1 for 15min to absorb the supernatant. The residue was leached again overnight with 80% methanol, and the supernatants were combined. The supernatant was concentrated under reduced pressure at 35°C to 1/3 of the original volume, the concentrated solution was collected and adjusted to pH = 8.0 with 1mol L ^-1 Na ₂ HPO ₄ , and an equal volume of petroleum ether: ethyl acetate (1: 1, V/V) The mixture was extracted and decolorized 3 times, the ether phase was discarded, and 0.1 g of polyvinylpyrrolidone (PVPP) was added to the water phase to absorb phenolic substances. After shaking on a shaker at 4°C for 30 minutes, centrifuge at 10,000 r·min ⁻¹ at 4°C for 10 minutes and take the supernatant. The supernatant was adjusted to pH=3.0 with 2 mol·L ^-1 citric acid. The above solution was extracted three times with an equal volume of ethyl acetate, and the lipid phases were combined. Distill the lipid phase under reduced pressure at 35°C to less than 1 mL, draw the concentrated solution, wash the spinner flask several times with a small amount of chromatographically pure methanol, combine the concentrated solution and washing solution to 10 mL, pass through a 0.22 μm filter membrane for HPLC detection . The whole process is operated at low temperature and protected from light.

③Determination of indole acetic acid oxidase in the cortex of cuttings

From September 5, 2013, every 15 days of treatment (i.e. the 1st, 15th, 30th, and 45th day of cuttings) cut the fresh cortex within 0-2 cm of the base of the cuttings for the determination of IAAO enzyme activity, IAAO The activity assay was modified from Zhiliang Zhang's method. Enzyme activity is represented by the number of micrograms of IAA decomposed and destroyed per milligram of protein within 1 hour.

Weigh 1g of sample, add 5mL of 20mmol·L ^-1 pre-cooled phosphate buffer (pH=6.0), add a small amount of quartz sand, grind it in an ice bath to form a homogenate, and then add 1mL of extract to 100mg of fresh quality material , diluted with phosphate buffer, centrifuged (10000r·min ^-1 ) for 15min, and the supernatant enzyme solution was taken. Take 2 test tubes, add 1 mL of manganese chloride, 1 mL of dichlorophenol, 2 mL of IAA, 1 mL of enzyme solution, and 5 mL of phosphate buffered water into one of the test tubes, and mix well. In another test tube, except that the enzyme solution was replaced by concentrated phosphate buffer, the rest of the components were the same, and placed together in a constant temperature water bath at 30°C for 30 minutes. Pipette 2 mL of the reaction mixture, add 4 mL of indole acetic acid reagent A, shake well, and place in a dark place at 30°C for 30 min to develop color. The red reaction solution after color development was measured in a spectrophotometer with a wavelength of 530nm when measuring the absorbance. Find out the corresponding residual amount of indole acetic acid from the standard curve according to the reading. Subtract the amount of indole acetic acid remaining after the action of the enzyme from the amount of indole acetic acid added at the beginning to obtain the amount of indole acetic acid decomposed and destroyed by the enzyme. The enzyme activity is represented by the amount (μg) of indole acetic acid decomposed and destroyed within 1 hour per mL of enzyme solution.

Example 2 Effects of different self-made rooting agents on the rooting rate and rooting efficiency of southern highbush blueberry at different rooting stages

Table 6 and Table 7 show the change of rooting rate of southern highbush blueberries Herbert and Lanfeng at different rooting stages. It can be seen from Table 6 that the rooting rate of the cuttings treated with 200 mg·L ^-1 boron compound IBA of the Herbert variety was 11.67% when the cuttings were 23 days old, which was significantly higher than other treatments (P<0.01), and compared with the rooting rate of CK ₁ 10.00% increase, rooting did not appear in CK2 treatment; at 30d, the rooting rate of 200mg·L ^-1 boron compound IBA was still significantly higher than other treatments, 26.66% higher than CK ₁ ; when cutting 37d, 300mg·L The rooting rate of ^-1 boron compound IBA was the highest (the difference with 200mg·L ^-1 boron compound IBA treatment was not significant, respectively 46.67% and 43.33%), which was significantly increased by 31.67% compared with CK ₁ , and the rooting rate of CK ₁ was the lowest; When cutting for 45 days, the rooting rate of different concentrations of boron compound IBA treatment was significantly higher than that of other treatments (P<0.01), and the rooting rate of 300mg·L ^-1 boron compound IBA treatment was the highest, reaching 73.33%, CK ₁ The lowest rooting rate was 33.33%. From the perspective of different rooting stages, the rooting efficiency of 200mg·L ^-1 boron compound IBA treatment was significantly higher than other treatments in the early stage of adventitious root formation (23～30d) (P<0.01); during the adventitious root formation period (30～45d), 300mg ·L ^-1 boron and 100mg·L ^-1 boron combined with IBA had no significant difference in rooting efficiency, and were significantly higher than other treatments (P<0.01). It shows that 200 mg·L ^-1 boron promotes the rooting of cuttings at the early stage of adventitious root formation (23～30 days), while 300 mg·L ^-1 boron and 100 mg·L ^-1 boron promote Herbert rooting at the stage of adventitious root formation (30 days ~45d).

It can be seen from Table 7 that when Lanfeng was cutting for 23 days, the rooting rate of cuttings treated with 200mg·L ^-1 boron and 50mg·L ^-1 vitamin B compound IBA was 13.33%, which was significantly higher than other treatments (P<0.01 ), the rooting rate of CK ₁ was significantly lower than that of other treatments (P<0.01), and no rooting occurred in the treatment of CK ₂ ; when the cuttings were 30 days old, the cuttings treated with 200mg·L ^-1 boron and 50mg·L ^-1 vitamin B complex IBA The difference in rooting rate was not significant, they were 40% and 38.33%, which were significantly higher than other treatments (P<0.01), while the rooting rate of 300mg·L ^-1 boron compound IBA treatment was consistent with that of CK ₁ treatment, which was significantly lower than Other treatments (P<0.01); when cutting for 37 days, 200mg·L ^-1 boron compound IBA treatment had the highest rooting rate, which was 33.33% higher than CK ₁ , and CK ₁ had the lowest rooting rate, only 20%; when cutting for 45 days, 300mg·L -1 The rooting rate of cuttings treated with L ^-1 boron compound IBA was the highest, reaching 83.33%, followed by 200mg·L ^-1 boron (73.33%) and 100mg·L ^-1 boron (63.33%) compound IBA treatment, while the rest of the treatments were all very Significantly higher than CK ₁ (P<0.01). From the perspective of rooting efficiency at different stages, the rooting efficiency of cuttings treated with 200mg·L ^-1 boron and 50mg·L ^-1 vitamin B compounded with IBA was the highest at the initial stage of adventitious root formation (23-30 days), except for the treatment with 300mg·L ^-1 boron compounded Except for CK ₁ , all other treatments were significantly higher than CK ₁ (P<0.01); during the adventitious root formation period (30-45 days), the rooting efficiency of 300 mg·L ^-1 boron compound IBA treatment was significantly higher than that of other treatments ( P<0.01), CK ₁ had the lowest rooting efficiency. It shows that 300mg·L ^-1 boron compound IBA treatment promotes the rooting time of Lanfeng concentrated in the adventitious root formation period (30 ~ 45d), which is effective for the rapid rooting of cuttings.

Table 6 Effects of different rooting agents on rooting at different rooting stages of Herbert

Table 7 Effects of different rooting agents on the rooting of Lanfeng at different rooting stages

Example 3 Effects of different self-made rooting agents on the final rooting quality of southern highbush blueberry

Table 8 and Table 9 show the final rooting conditions of blueberry Hebert and Lanfeng cuttings treated with different rooting agents. It can be seen from Table 8 that the rooting statistics of the Herbert variety after 70 days of cutting (full cycle) show that the rooting rate from high to low is as follows: 5% sucrose>100mg L ^-1 boron>10% sucrose> ^{200mgL- 1} boron>2% sucrose>300mg·L ^-1 boron>CK ₁ >50mg·L ^-1 vitamin B>200mg·L ^-1 vitamin B>100mg·L ^-1 vitamin B. Among them, the rooting rate of 5% sucrose compound IBA is the highest, reaching 91.18%, and its average rooting number is also the highest, which is 16.40; the rooting rate of 100mg·L ^-1 boron compound IBA is the second, reaching 86.84%, and the third is 10%. With sucrose composite treatment, the rooting rate reached 86.49%. Cuttings treated with sucrose compounded with a certain concentration of IBA generally had the highest rooting rate, followed by boron compounded treatment; cuttings treated with boron compounded generally had the best rooting quality. Therefore, combined with the experimental results of dynamic rooting rate monitoring during the rooting period, the addition of boron can quickly and effectively increase the rooting rate, and the comprehensive rooting quality is higher; while sucrose can improve the rooting efficiency of cuttings in the late rooting period (45-70 days). The rooting effect of vitamin B complex IBA treatment was not ideal.

It can be seen from Table 9 that from the perspective of the rooting situation of the Lanfeng variety after 70 days of cuttings, the order of rooting rate from high to low is: 300mg·L ^-1 boron>100mg·L ^{- 1} boron>2% sucrose>200mg·L- ¹ boron>5% sucrose>200mg·L ^-1 vitamin B>50mg·L ^-1 vitamin B>CK ₁ >100mg·L ^-1 vitamin B>10% sucrose. Among them, the rooting rate of 300mg·L ^-1 boron complex IBA was the highest, reaching 91.18%, which was 33.12% higher than that of CK ₁ , and the average rooting rate was also the highest, 12.60; the rooting rate of 100 mg·L ^-1 boron complex IBA was second , reached 87.50%, and the third was the cuttings treated with 2% sucrose compound, and its rooting rate reached 85.71%. Cuttings treated with a certain concentration of boron and IBA generally have the highest rooting rate, the longest average root length, the largest average number of roots, and the highest rooting power index. But the effect of vitamin B complex IBA treatment was not obvious. Based on the experimental results of rooting dynamic observation, the addition of boron can rapidly improve the comprehensive rooting quality of Lanfeng cuttings.

Table 8 Herbert's final rooting investigation

Table 9 Lanfeng final rooting situation investigation

Difference analysis of endogenous hormones under the treatment of different rooting agents of the variety Herbert in embodiment 4

The difference analysis of the endogenous hormones IAA, ABA, and IAA/ABA of Herbert cuttings treated with different rooting agents is shown in Table 10, Table 11, and Table 12. It can be seen from Table 10 that on the 1st day of cutting, the mass fraction of IAA in the cortex of the cuttings treated with the IBA rooting agent added boron was significantly higher than that of the other treatments (P<0.01), while the IBA rooting agent added sucrose and vitamin B The cutting hormone content of treatment was significantly lower than that of CK (IBA treatment alone). On the 15th day of cutting, the hormone content of cuttings treated with IBA rooting agent added boron was still significantly higher than that of other treatments (P<0.01), while the IAA content of cuttings treated with IBA rooting agent added vitamin B was higher than that of CK; In the treatment with sucrose, except for 5% sucrose + 200mg·L ^-1 IBA, the hormone content of 2%, 10% sucrose + 200mg·L ^-1 IBA was still lower than that of CK. After cutting for 30 days, the IAA content of cuttings treated with 200mg·L ^-1 boron+200mg·L ^-1 IBA was significantly higher than that of other treatments (P<0.01), followed by 50mg·L ^-1 VB+200mg·L ^-1 IBA and Treatment with 200mg·L ^-1 VB+200mg·L ^-1 IBA. In the treatment of adding sucrose, except for 10% sucrose+200mg·L ^-1 IBA treatment, the IAA content of the cuttings treated with the other two concentrations of sucrose was lower than that of CK. When the cuttings were cut to 45 days, the hormone content of all cuttings treated with boron was significantly higher than that of other treatments (P<0.01), and the IAA content in all treatments from high to low was: 300mg·L ^-1 boron>100mg·L ^{- 1} Boron > 200mg·L ^-1 Boron > 100mg·L ^-1 VB > 50mg·L ^-1 VB > 200mg·L ^-1 VB > 5% sucrose > CK > 2% sucrose > 10% sucrose. The IAA content of cuttings treated with boron was significantly higher than that of other treatments (P<0.01). Through the dynamic detection of rooting rate of cuttings, it is found that 200mg·L ^-1 boron promotes the rooting of cuttings at the early stage of adventitious root formation (23~30d), while 300mg·L ^-1 boron and 100mg·L ^-1 boron promote the time of Herbert's rooting Concentrate on the adventitious root formation period (30 ~ 45d). This is related to the increase of endogenous IAA content in the adventitious root formation period of southern highbush blueberry cuttings with the addition of boron.

The influence of table 10 different rooting agents on Herbert's endogenous IAA content

It can be seen from Table 11 that on the 1st day of cutting, the ABA content of cuttings treated with boron-added IBA rooting agent was higher than that of other treatments, and the order of ABA content from high to low was: 200 mg·L ^-1 boron> 100mg·L ^-1 boron > 300mg·L ^-1 boron, the ABA content was the lowest under CK treatment. On the 15th day of cutting, the ABA content of cuttings treated with boron-added IBA rooting agent was still higher than that of other treatments, except for 200mg·L ^-1 VB+200mg·L ^-1 IBA treatment, the hormone content of all treatments was significantly higher than that of CK (P<0.01). On the 30th day of cutting, the ABA content of cuttings under the treatment of 100mg·L ^-1 boron + 200mg·L ^-1 IBA was significantly higher than that of all treatments (P<0.01), followed by 2% sucrose + 200mg·L ^-1 IBA, 5% sucrose +200mg·L ^-1 IBA and CK treatment, 300mg·L ^-1 boron+200mg·L ^-1 IBA and 200mg·L ^-1 boron+200mg·L ^-1 IBA were significantly lower than all treatments ( P<0.01). On the 45th day of cutting, the ABA content of cuttings treated with 50mg·L ^-1 VB+200mg·L ^-1 IBA, 100mg·L ^-1 VB+200mg·L ^-1 IBA and CK was significantly higher than that of other treatments (P<0.01) , the ABA content of cuttings treated with IBA rooting agent added boron and sucrose was generally lower than that treated with vitamin B and IBA alone at 45 days after cutting. It shows that the addition of boron can promote the mass rooting of southern highbush blueberry in the period of adventitious root formation (30-45 days), which is related to the reduction of ABA content in the cortex of cuttings.

Table 11 Effects of different rooting agents on Herbert's endogenous ABA content

It can be seen from Table 12 that on the 1st day of cutting, the ratio of cutting hormone IAA/ABA under CK treatment was significantly higher than that of other treatments (P<0.01). On the 15th day of cutting, the hormone ratios of all cuttings treated with vitamin B-added IBA rooting agent, 200mg·L ^-1 boron+200mg·L ^-1 IBA, 300mg·L ^-1 boron+200mg·L ^-1 IBA higher than CK. On the 30th day of cutting, the hormone ratio of 200mg·L ^-1 boron+200mg·L ^-1 IBA was significantly higher than that of other treatments (P<0.01), followed by 50mg·L ^-1 VB+200mg·L ^-1 IBA and 300mg ·L ^-1 boron+200mg·L ^-1 IBA treatment. After 45 days of cutting, the IAA/ABA ratios of cuttings under the treatment of all boron-added IBA rooting agents were significantly higher than those of other treatments (P<0.01), and the order of hormone ratios in all treatments from high to low was: 300mg·L ^-1 boron> 100mg·L ^-1 boron>200mg·L ^-1 boron>200mg·L ^-1 VB>100mg·L ^-1 VB>2% sucrose>50mg·L ^-1 VB>5% sucrose>CK>10% sucrose. The results showed that the addition of boron IBA rooting agent treatment significantly increased the IAA content of the cutting cortex during the Herbert adventitious root formation period (30-45 days), while reducing the ABA content, greatly increasing the ratio of IAA/ABA to facilitate rooting.

Table 12 Effects of different rooting agents on Herbert's endogenous IAA/ABA

Example 5 Differential analysis of endogenous hormones under different rooting agents for variety Lanfeng

The difference analysis of the endogenous hormones IAA, ABA, and IAA/ABA of Lanfeng cuttings treated with different rooting agents is shown in Table 13, Table 14, and Table 15. It can be seen from Table 13 that the IAA content of all cuttings treated with boron-added IBA rooting agent on the first day of cutting was generally higher than that of other treatments, and the hormone content was the highest under the treatment of 300mg·L ^-1 boron + 50mg·L ^-1 IBA ; On the 15th day of cutting, the IAA content of cuttings treated with boron and vitamin B IBA rooting agent was significantly higher than that of sucrose and IBA alone (P<0.01), and the IAA content of cuttings under CK treatment was the lowest among all treatments; On the 30th day of cutting, the IAA content of cuttings treated with 200mg·L ^-1 boron+50mg·L ^-1 IBA was significantly higher than that of other treatments (P<0.01), followed by 100mg·L ^-1 boron+50mg·L ^-1 IBA and 10% sucrose + 50mg·L ^-1 IBA treatment, all IAA content of cuttings treated with sucrose, boron, and vitamin B rooting agents were higher than those of CK; The order of treatment to the lowest level is: 300mg·L ^-1 boron>200mg·L ^-1 boron>2% sucrose>200mg·L ^-1 VB>100mg·L ^-1 boron>100mg·L ^-1 VB>50mg·L ^{- 1} VB > 5% sucrose > 10% sucrose > CK. Combined with the results of dynamic observation and analysis on the rooting rate of cuttings of variety Lanfeng, the IBA rooting agent added with 300 mg·L ^-1 boron can rapidly increase the rate of adventitious root formation in cuttings of Lanfeng during the adventitious root formation period (30-45 days), rather than increase the The content of IAA in the cortex of cuttings was closely related.

Table 13 Effects of different rooting agents on the content of endogenous IAA in Lanfeng

It can be seen from Table 14 that on the 1st day of cutting, the ABA content of all cuttings treated with boron-added IBA rooting agent was significantly higher than that of CK, while the ABA content of cuttings treated with sucrose-added IBA rooting agent was significantly lower than that of CK (P <0.01); on the 15th day of cutting, the ABA content of the cuttings treated with boron was still significantly higher than that of the other treatments (P<0.01); on the 30th day of cutting, 5% sucrose+50mg·L ^-1 IBA and 100mg·L The ABA content of cuttings treated with L ^-1 VB+50mg·L ^-1 IBA was significantly higher than that of other treatments (P<0.01), while the ABA content of cuttings treated with boron was significantly lower than that of CK (P<0.01); On the 45th day of cutting, the treatment order of ABA content from high to low was: 50mg·L ^-1 VB＞200mg·L ^-1 VB＞CK＞100mg·L ^-1 boron＞100mg·L ^-1 VB＞5% sucrose＞ 300mg·L ^-1 boron > 2% sucrose > 200mg·L ^-1 boron > 10% sucrose. Through the investigation of the final rooting situation of Lanfeng cuttings, it was found that the rooting effect of cuttings treated with IBA rooting agent added with vitamin B was not obvious, which is consistent with the increase of ABA content in the cortex of Lanfeng cuttings during the adventitious root formation period (30-45d) by vitamin B related.

Table 14 Effects of different rooting agents on the content of endogenous ABA in Lanfeng

It can be seen from Table 15 that on the first day of cutting, the IAA/ABA of cuttings treated with 200mg·L ^-1 VB+50mg·L ^-1 IBA was significantly higher than that of other treatments (P<0.01), followed by 10% sucrose+50mg·L ^{- 1} IBA and 5% sucrose + 50mg·L ^-1 IBA; on the 15th day of cutting, the IAA/ABA of cuttings treated with vitamin B-added IBA rooting agent was generally significantly higher than that of other treatments (P<0.01), except Except for 300mg·L ^-1 boron+50mg·L ^-1 IBA treatment, the IAA/ABA of cuttings treated with CK was the lowest; on the 30th day of cutting, the IAA/ABA of cuttings treated with 200mg·L ^-1 boron+50mg·L ^-1 IBA Significantly higher than other treatments (P<0.01), followed by 50mg·L ^-1 VB+50mg·L ^-1 IBA and 100mg·L ^-1 boron+50mg·L ^-1 IBA, and all added sucrose, The IAA/ABA of cuttings treated with boron and vitamin B was significantly higher than that of CK (P<0.01); when the cuttings reached 45 days, the order of IAA/ABA from high to low was: 10% sucrose>300mg·L ^-1 boron >200 mg·L ^-1 boron>2% sucrose>100 mg·L ^-1 boron>5% sucrose>200 mg·L ^-1 VB>100 mg·L ^-1 VB>50 mg·L ^-1 VB>CK. Because the ABA content of cuttings treated with 300mg·L ^-1 boron-combined IBA was significantly higher than that of 10% sucrose-combined IBA, the IAA/ABA ratio of cuttings was lower than that of 10% sucrose. By monitoring the rooting dynamics of Lanfeng cuttings, it was found that on the 45th day of cutting, the cuttings treated with 300 mg·L ^-1 boron compound IBA had the highest rooting rate, and all the cuttings treated with boron had higher rooting rates than those treated with sucrose . It shows that the IBA rooting agent added with boron significantly increases the IAA content of the cortex of Lanfeng cuttings, which is the main reason for promoting its rooting.

Table 15 Effects of different rooting agents on Lanfeng endogenous IAA/ABA

Claims (4)

1. a blueberry green branch cutting seedling raising method is characterized in that comprising the following steps: 1) Selection of cuttings: From March to April, select the middle and lower parts of vegetative branches that are 1-year-old with high hardness, good maturity and health as cuttings; 2) Treatment of cuttings: trim the branches to a length of 7-10cm, with 4-5 axillary buds and 1-2 leaves on the top of the cuttings. 3) Soaking of cuttings: Soak with a rooting agent of boron+IBA for 8-10 hours; 4) Cutting: Before cutting, pour the substrate with water to ensure humidity but no water accumulation. On the day of cutting, soak the base of the treated cuttings with 1000 times of fenflum for 10 seconds, and then insert the cuttings vertically into the substrate; 5) Moisture management: the humidity of the substrate is maintained above 80%, and the temperature is controlled at 20-28°C; 6) Management after cuttings: Gradually apply fertilizer after the cuttings take root. Fertilization is applied in the form of liquid fertilizer with a concentration of no more than 3%. Fertilize blueberry cuttings and green branches once a week. 2. a kind of blueberry green branch cutting seedling raising method according to claim 1 is characterized in that: the rooting agent described in the step (3) is made solvent with sterile water, and preparation rooting agent, guarantees that IBA concentration is 50mg/L , the concentration of boron is 300mg/L. 3. a kind of blueberry green branch cutting seedling raising method according to claim 1, it is characterized in that: described substrate is to utilize moss soil as cutting substrate, adjust pH value to be 5.5, the cutting bed substrate is used more than 1000 times the day before cutting. Bacillus spray treatment. 4. A method for raising seedlings by cuttings of blueberries and green branches according to claim 1, characterized in that: the depth of the cuttings inserted into the matrix accounts for 2/3 of the entire cuttings, and the spacing is 5cm×10cm.