CN112314232B - Method for regulating and controlling blue flower danshen period - Google Patents
Method for regulating and controlling blue flower danshen period Download PDFInfo
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Abstract
The invention provides a method for regulating and controlling a blue flower danshen period, belonging to the technical field of plant tissue culture or cultivation. The method does not need to add any plant hormone, polyamine, plant growth regulator and fertilizer, and comprises the following specific steps: (1) selecting a blue flower red lead tissue culture seedling or a seedling; (2) carrying out the following temperature change treatment on the plants in the step (1): the blue flower and the red flower are alternately treated for 12h/d at the temperature of 25-35 ℃ and the temperature of 30-35 ℃ respectively, and the treatment time is 27-38 d, so that the regulation and control of the blue flower and the red flower period are realized. The regulating and controlling method can effectively regulate and control the flowering period of the blue flower red lead, so that the blue flower red lead can bloom according to the needs, the blue flower red lead can be regulated and controlled to bloom at any time in the unnatural flowering period, the crown width and the color of the flowering finished seedling flower are good, and great market benefits can be obtained.
Description
Technical Field
The invention belongs to the technical field of plant tissue culture or cultivation, relates to a method for artificially regulating and controlling a plant flowering phase, and particularly relates to a method for regulating and controlling a blue flower danshen flowering phase.
Background
The blue flower red lead (Plumbago auriculata lam.) belongs to the genus Plumbaceae (Plumbaginaceae), is a evergreen and popular ornamental shrub, and is originally produced in south Africa with blue-purple flowers. The plant has extremely high ornamental value due to unique flower type and gorgeous flower color, becomes one of the most popular tropical crops, and is increasingly introduced worldwide as an ornamental plant for cultivation and application in landscape architecture and indoor ornamental. The blue flower red lead is a plant which flowers for more than one year in the original place. The plant flowering all year round in Sichuan area is changed to only concentrate in summer flowering due to extremely high ornamental value, and the flowering phase is concentrated in 6 to 10 months for 5 months, so that the ornamental value of the blue flower red lead is greatly reduced, and the ornamental value and the landscaping popularization and application are influenced.
In recent years, related researches on the field of blue flower red lead are carried out in China. In 2016, zhao Yunfang et al studied the flower bud differentiation characteristics of Plumbum Preparatium, and studied the biological characteristics of Plumbum Preparatium by observing the biological characteristics of 6 years of seedlings of Plumbum Preparatium, dividing the processes of flowers and carrying out correlation analysis of morphological dissection and physiological substance change on flower bud differentiation of different processes, and found that the flower bud differentiation of Plumbum Preparatium is about 60d in 5 ten days of 3 months and preliminary flowers are at the end of 5 months to the beginning of 6 months; at the same time, the phenomenon of flowering in the bottle of the occasional group-cultivated plant is also found.
Zhou Jie et al studied the flowering phase regulation of micro China rose (Rosa chinensis minima). Experiments show that under the conditions that the illumination intensity is 20000lx and the temperature change treatment is carried out for 16h (25 ℃) to 8h (16 ℃), the rose flowering phase is remarkably advanced and flowers are intensively opened, but the crown width and the plant height of the rose become smaller, and the chlorophyll content and the leaf area of the leaves are reduced. According to the method, the flowering phase of the China rose is regulated by researching illumination, temperature and the like through the LED light source, so that a control-promoting cultivation technology is formed, and the method has important theoretical and practical guiding significance for large-scale cultivation of potted China rose, but has great influence on normal growth of the China rose.
Chen Fengling exogenous hormone for regulating and controlling flowering phase of butterfly orchid is obtained by extracting inflorescence to form flower budHormone spray treatment of the plants of (a) showed that: when gibberellin (GA 3) is sprayed, the concentration is 100-200 mg.L -1 Can bloom 10-17 days in advance.
By looking up a great deal of prior art, the research on the regulation means of the blue-flower danshen period is still blank, and the reference is only limited to the regulation means of the flowering period of other plants, but the characters shown by the same regulation means on different plants can be greatly different due to the diversity of plant varieties. The existing researches relate to means for regulating and controlling the flowering period of other plants, but most of the existing researches adopt regulating and controlling means such as plant hormones, polyamines, plant growth regulators or fertilizers, and the like, under the regulating and controlling means, the flowering period of corresponding plants can be advanced, but experiments show that the regulating and controlling means are unsuitable for regulating and controlling the flowering period of the blue flower red lead, have poor regulating and controlling effects, can not effectively change the flowering time of the blue flower red lead, and are easy to cause adverse effects on the growth of the blue flower red lead.
Therefore, for cultivation and marketing of the blue flower red lead, how to perform effective flowering phase regulation and control to ensure that the blue flower red lead can bloom according to requirements, and can perform regulation and control treatment at any period of the non-natural flowering phase of the blue flower red lead, so that the blue flower red lead bloom in the non-natural flowering phase, and the blue flower red lead becomes a technical problem to be solved urgently in expanding the application market of the blue flower red lead.
Disclosure of Invention
The invention aims to solve the technical problems, and provides a method for regulating and controlling the blue flower danshen period. The method does not need to add plant hormone, polyamine, plant growth regulator or fertilizer, abandons the regulation and control method commonly adopted in the prior art, and does not have adverse effect on the growth of the blue flower red lead. Meanwhile, the regulating and controlling method can ensure that the flowering of the blue flower red lead in Sichuan areas is not limited by time and season, and can obtain great market benefit.
In order to achieve the above object, the present invention provides a method for regulating and controlling the danshen-hua period, which does not need any plant hormone, polyamine, plant growth regulator and fertilizer, and comprises the following specific steps:
(1) Selecting blue flower dan tissue culture seedlings (namely sterile seedlings growing for 1 month in a tissue culture bottle) or seedlings (three-month-old cultivation seedlings);
(2) The plants in the step (1) are subjected to the following temperature changing treatment: the blue flower red flower is alternately treated for 12h/d at the temperature of 25-35 ℃ and the temperature of 30-35 ℃ for 27-38 d, so that the blue flower red flower blooms in the unnatural flowering period.
According to the flowering phase regulating method provided by the invention, substances such as plant hormone, polyamine, plant growth regulator or fertilizer and the like are not required to be added, and only the culture temperature is required to be changed, so that the nutrition growth period can be greatly shortened by regulating the temperature according to the method provided by the invention, the flowering of the blue flower red lead in Sichuan areas is not limited by seasons, and the flowering can be realized in the unnatural flowering phase. The flowering seedling of the blue flower red lead obtained by the regulating and controlling method has short production period, is not affected by the nodes, has good crown width and color of the finished flower, is simple and easy to operate, and does not need to add any hormone, fertilizer or other chemical reagents. The method has a certain practical significance for the production and sales of commercial seedlings, and the flowering in the bottle has important scientific value for the research and breeding of the mechanism of the heterogenic self-incompatibility of the commercial seedlings.
Further, the temperature change treatment in the step (2) is performed under the following conditions: the treatment is carried out alternately for 12h/d and 29 to 36d at the temperature of 25 ℃ and 30 ℃.
As a more preferable scheme, the temperature change treatment condition in the step (2) is as follows: and respectively and alternately treating for 12h/d and 27-34 d at the temperature of 30 ℃ and the temperature of 35 ℃.
As a more preferable mode, the temperature change treatment condition in the step (2) is as follows: and respectively and alternately treating for 12h/d and 30-38 d at the temperature of 25 ℃ and the temperature of 35 ℃.
In another preferred treatment mode, in the step (2), the tissue culture seedling or the seedling is further subjected to the following illumination treatment: the illumination intensity is 200 mu mol.m -2 ·s -1 The illumination time is 12h/d.
Further, the environmental humidity is maintained to be 70-90% in the treatment process of the step (2).
Further, the method for obtaining the tissue culture seedling comprises the following steps:
(1) Collecting mature, full and healthy blue flower dan seeds, soaking the seeds with the seed coats removed in distilled water at 30 ℃ and under dark conditions for 2d, and then washing with running water for 2h;
(2) Immersing the seeds in 75% w/v ethanol solution for 45s, taking out the seeds, and washing the seeds with sterile distilled water for 5-6 times;
(3) Immersing the seeds in 0.1% w/v mercuric chloride solution for 4min, washing with sterile distilled water for 5-6 times, and repeating the steps;
(4) Seed was inoculated at 30 g.L -1 Sucrose and 6.5 g.L -1 And (3) culturing agar on an MS culture medium with the pH value of 5.8-6.0 to obtain the tissue culture seedlings.
Further, the method for obtaining the seedlings comprises the following steps: collecting mature, full and healthy blue flower dan seeds, sowing the seeds on a peat soil matrix for germination culture for three months, and obtaining seedlings with consistent growth conditions.
Further, 800 times of carbendazim is sprayed on the Lan Huadan plants 800 liquid and irrigated pot soil every two weeks during germination culture.
Compared with the prior art, the invention has the beneficial effects that:
(1) The method provided by the invention combines the tissue culture seedling and the seedling of the blue flower, screens a set of preferable scheme which can be suitable for regulating and controlling the flowering of the tissue culture seedling and the seedling of the blue flower at the same time, has short period of flowering seedling production and small influence by seasons, has good crown width and color of the finished product seedling flower, and can realize the flowering of the blue flower in Sichuan areas according to requirements;
(2) The regulation and control method provided by the invention is simple and easy to operate, does not need to add any hormone or fertilizer, greatly reduces the cost, can realize the sales of flowering seedlings all the year round, and has higher economic benefits for production and sellers; flowering in the bottle has a certain scientific value for researching the plant reproduction and system evolution mechanism;
(3) The method provides technical reference for flowering phase regulation of the blue flower red lead and the same genus plants, and is beneficial to developing markets and applications of the same genus plants.
Detailed Description
In order that the objects, technical solutions and advantages of the present invention will become more apparent, the following detailed description of the present invention will be made with reference to the examples, which are given by way of illustration and explanation only, and are not intended to limit the present invention. Some non-essential modifications and adaptations of the invention according to the foregoing summary will still fall within the scope of the invention.
Example 1
(one) obtaining tissue culture seedlings
Tissue culture seedlings obtained by aseptic seeding of seeds. Seeds were collected from the artificial population of the basic blue flower red lead developed in modern agriculture in Chongzhou, sichuan agricultural university in China. The specific method for obtaining the tissue culture seedling comprises the following steps: mature, full and healthy seeds were soaked in distilled water at 30℃and in the dark for 2d, and then washed under running water for 2h. They were then immersed in an ethanol solution (75% w/v) for 45s and washed with sterile distilled water (5 or 6 times) before they were immersed in mercuric chloride (HgCl) 2 0.1% w/v) for 4 minutes and washed with sterile distilled water (5 or 6 times) and this step is repeated. Finally, seed is inoculated at 30 g.L -1 Sucrose and 6.5 g.L -1 Agar, on Murashige-Skoog (MS) medium (one or two seeds/flasks) at pH 5.8 to 6.0. Plantlets derived from the sterile germination of seeds were then used to explore the flowering phase regulation of tissue culture seedlings.
Method for controlling tissue culture Miao Huaqi of blue flower red lead
The method comprises the following steps: taking healthy blue flower red lead tissue culture seedlings with the same growth vigor, respectively placing the healthy blue flower red lead tissue culture seedlings in a plurality of same illumination incubators (RTOP, topinstrovent, zhejiang, china), setting corresponding temperatures according to the temperature regulation means in the following table 1, and setting all other conditions to be the same, wherein the conditions comprise 12h/d illumination and 200 mu mol photon m -2 ·s -1 The influence of the temperature regulation means on the tissue culture Miao Huaqi of the blue flower red lead was studied at 70-90% of the artificial light intensity and the air humidity, and the obtained results are shown in Table 1.
TABLE 1
Note that: ++ indicates growth the situation is good and the quality of the product is good, ++ indicates that the growth situation is general, + indicates poor growth, and blank indicates poor growth. Lower case letters following the values in the tables represent significant differences at the 5% level; l, a, b take the value range [ -100,100], L is close to 100 representing the higher the color brightness, is close to-100 representing the lower the brightness, a is close to 100 representing the thicker the red, is close to-100 representing the thicker the green; b near 100 represents more intense yellow and near-100 represents more intense blue; the following is the same.
Example 2
Obtaining seedlings
Mature, full and healthy seeds collected from the artificial population of the basic blue flower red lead are developed from modern agriculture in Chongzhou of Sichuan university of China, then the seeds are sown on a peat soil substrate for germination culture, and three-month-old seedling of the real blue flower red lead with consistent growth condition is selected for experiment. And (5) spraying 800 liquid and irrigation basin soil on Lan Huadan plants by 800 times of carbendazim every two weeks during cultivation.
Method for regulating flowering phase of blue flower red lead seedling
The method comprises the following steps: taking healthy three-month blue flower pill seedlings with the same growth vigor, respectively placing the seedlings in a plurality of same illumination incubators (RTOP, topinstrument, zhejiang, china), setting corresponding temperatures according to the temperature regulation means in the following table 2, and setting all other conditions to be the same, wherein the conditions comprise 12h/d illumination and 200 mu mol photon m -2 ·s -1 The influence of the temperature regulation means on the tissue culture Miao Huaqi of the blue flower red lead was studied at 70-90% of the artificial light intensity and the air humidity, and the obtained results are shown in Table 2.
TABLE 2
Note that: ++ indicates growth the situation is good and the quality of the product is good, ++ indicates that the growth situation is general, + indicates poor growth, and blank indicates poor growth.
As can be seen from the above tables 1 and 2, the tissue culture seedlings and the seedlings of Plumbum Preparatium were cultured in a fixed light environment (12 h/d, light intensity 200. Mu. Mol. M -2 ·s -1 ) The flowering time of the carthamus tinctorius can be advanced by adopting a variable temperature treatment mode (the treatment time reaches the onset of flowering of the carthamus tinctorius) with the temperature of 25 ℃/30 ℃ for 12h/d, the temperature of 25 ℃/35 ℃ for 12h/d and the temperature of 30 ℃/35 ℃ for 12h/d, wherein the flowering time of the carthamus tinctorius obtained by adopting the variable temperature treatment mode with the temperature of 25 ℃/35 ℃ for 12h/d is fastest, and the tissue culture seedlings and the seedlings are 31.85 +/-0.31 d and 30.27+/-0.71 d respectively; the flowering rate is highest and is 91.11+/-0.72% and 94.11+/-0.52% respectively; the flowering period cannot be effectively shortened by other temperature regulation and control modes, particularly, the flowering time of the tissue culture seedlings and the seedlings of the blue flower obtained by the variable temperature treatment mode of each treatment for 12h/d at the temperature of 15 ℃/20 ℃ is 87.18 +/-1.87 c and 84.54 +/-2.65 c respectively, so that the flowering period cannot be effectively shortened, and the flowering time is delayed; the flowering time can be shortened to a certain extent by the temperature change treatment mode of each treatment for 12h/d at the temperature of 20 ℃/25 ℃, but the difference is obvious compared with the three temperature change treatment modes, and the flowering-period regulation effect of the blue flower red is poor.
Further, according to the research results in the tables 1 and 2, when the temperature is 15 ℃ and the treatment condition is 24 hours/d, the blue flower red lead can not bloom, and the delay of the blue flower red lead period can be regulated; and the flower has no distortion compared with the flower which is naturally opened, and the crown width and the petal color of the flower are normal. Compared with the blue-flower danshen period in the natural state without treatment, the method for regulating and controlling the blue-flower danshen period is longer, can realize the node-meeting flowering of the tropical introduced flowers, can realize the flowering only by carrying out the variable temperature treatment of 25 ℃/35 ℃ for 12h/d for the shortest 30 days when regulating and controlling the flowering period in advance, forms the cultivated seedlings with market value, and can realize the rapid seedling formation of the blue-flower danshen tissue cultivated seedlings and the seedlings; the method has the advantages that the cultivation temperature is kept at 15 ℃ under the condition of 24 hours/d after the flowering period is regulated, the blue flower red lead can not bloom, the effect is that the blue flower red lead seedlings can keep a nutrition growth state in order, the flower bud differentiation is not generated, and the method is favorable for regulating and controlling the flowering period in a large quantity and obtaining plants with a long-term flowering period in a large quantity. The whole method for regulating and controlling the blue flower danshen period does not need to add any plant hormone, polyamine, plant growth regulator and fertilizer, only needs to change the culture temperature, has short production period and small influence of seasons, is simple and easy to operate, and greatly reduces the cost. Meanwhile, the method has important significance for protecting germplasm resources, accelerating popularization and application.
Comparative example 1
Reference 1 (Nitsch C, nitsch J P.the induction of flowering in vitro in stem segments of Plumbago indica L. -I.the production of vegetative buds [ J)]Planta,1967,72 (4): 355-370.): in 1967, nitsch and Nitsch successfully regulated the flowering phase of Plumbum Preparatium (Plumbago indica) of Plumbum by inoculating sterilized Plumbum Preparatium stem into MS medium containing Kinetin (KT) 1mg/L, adenine (Adenine) 300mg/L and 2.5% sucrose, and treating at 25deg.C, light intensity and cycle of 200μmol.m -2 ·s -1 The 9h/d results found that treatment for a minimum of about 4 weeks induced flowers of Plumbum Preparatium (method one).
The method one in the reference is used for regulating the flowering phase of the blue flower red lead tissue culture seedling. The tissue culture seedlings of the blue flower dan are obtained in the same way as in the example 1, and tender and healthy stem segments are selected for tissue culture. The stem segments were soaked in distilled water for 5min and washed 5 times. It was then immersed in an ethanol solution (75% w/v) for 45s and washed with sterile distilled water (5 or 6 times). Subsequently, they are immersed in mercury chloride (HgCl 2 0.1% w/v) for 4 minutes and washed with sterile distilled water (5 or 6 times) and this step is repeated. Inoculating the stem of Plumbum Preparatium on MS+2.5% sucrose+6.5 g/agar+kinetin (KT) and Adenine (Adenine) culture medium, standing at 25deg.C, and setting light intensity and period to 200μmol.m -2 ·s -1 In a 9h/d light incubator, the growth environment was maintained consistent with that in scheme one. The flowering phase test method for regulating and controlling the blue flower red lead tissue culture seedlings comprises the following steps: after the blue flower dan seeds germinate and root in the culture medium, dividing the same number of blue flower dan tissue culture seedlings into 10 experimental groups, and observing the methodThe regulation and control effect of the flowering phase of the tissue culture seedlings of the blue flower red flower is shown in table 3.
TABLE 3 Table 3
Note that: n represents non-flowering. The following is the same.
And (II) the method one in the reference is used for regulating the flowering phase of the blue flower dansheng seedlings. The seedlings were obtained in the same manner as in example 2, mature, full and healthy seeds were collected, and then the seeds were sown on a peat soil substrate for germination culture, and three-month blue flower pellet seedlings with consistent growth conditions were selected. The plants Lan Huadan and the irrigated pot soil were sprayed with 800 times carbendazim every two weeks during cultivation. Placing the treated blue flower red lead seedling at the ambient temperature of 25 ℃ and setting the light intensity and period of 200 mu mol.m -2 ·s -1 In a 9h/d illumination incubator, the growth environment is kept consistent with the first scheme. The flowering phase test method for regulating and controlling the blue flower red lead seedlings comprises the following steps: dividing the equal quantity of the blue flower dan seedlings into 10 experimental groups, and spraying cytokinin with different concentrations to plants of different experimental groups once a week under the condition of original culture soil: kinetin (KT) and Adenine (Adenine). The regulation and control effects of the method on the flowering phase of the blue flower red lead seedlings are observed, and the test results are shown in Table 4.
TABLE 4 Table 4
Using the method pair of the above references for flowering phase regulation, it was found that either the tissue-cultured seedlings or the seedlings of the blue flower red lead could not reach the four-week flowering described in method one in the unnatural flowering phase and had flowering. The regulation method has a large difference from the effect of regulating the red lead, and in conclusion, the regulation of the blue lead period is ineffective.
Comparative example 2
The method in reference 2 (Li Ru, li Zhilin, bai Jiankun, et al.2 effects of polyamines on flowering in dendrobium candidum bottles [ J ]. Tropical crop journal, 2020, 041 (004): 709-714.): li Ru and the like take dendrobium candidum as test materials, the regulation and control of exogenous putrescine (Put) and spermine (Spm) with different concentrations on the flowering phase of the dendrobium candidum are studied, and the flowering rate can be improved by adding proper amounts of Put and Spm into the culture medium. Culturing dendrobium nobile flowers by using 1/2MS+6-BA 0.2mg/L+NAA0.2 mg/L+banana 150 g/L+agar 7 g/L+sucrose 30 g/L+activated carbon 0.5 g/L+Put and Spm with different concentrations, wherein the culture conditions are that the pH of the culture medium is 5.8, RH (75+/-5)%, the temperature is (22+/-0.2) DEG C, the illumination intensity is 1800-2500 Lx, and the illumination period is 14h/d. When the Put concentration in the culture medium is 0.2mg/L, the initial flowering period of the dendrobium candidum is the shortest, 83.33d, and the ornamental period is the longest, 43.33d, so that the flowering period can be advanced, and the ornamental period can be prolonged. When the Put concentration is 0.4mg/L, the highest flowering rate in the dendrobium candidum bottle is 30.47%; when the concentration of Spm is 0.2mg/L, the highest flowering rate in the dendrobium candidum bottle is 22.26%. (method II)
The second method in the above document was used for controlling the flowering period of Plumbum Preparatium, and the method was referred to in comparative example 1, except that the treated matters were different, and it was found that neither the tissue culture seedlings nor the seedlings of Plumbum Preparatium were able to flower for 83 days as described in the second method in the unnatural flowering period. In conclusion, the second method in the prior art is ineffective in regulating and controlling the blue flower danshen period.
Comparative example 3
The method in reference 3 (Jiang Lin. Penta flower tissue culture and tube flowering study [ D ]) is described: jiang Lin plant growth regulators were used: paclobutrazol (PP 333) regulates the flowering phase of penta flowers (Pentas lanceolata (forsk.) k.schum.) which are perennial plants of the genus penta of the family rubiaceae. The research takes tender stem segments without plant diseases and insect pests of the five-star flowers, and establishes a five-star flower flowering regulation and control technical system with higher flowering efficiency as a test material after sterilization treatment. (1) The culture environment is as follows: the temperature is 23+/-2 ℃, the illumination intensity is 2500-3000lux, and the illumination duration is 14h/d; (2) the culture medium is as follows: PP3330.5 mg/L or PP3330.2 mg/L is added to MS+KT0.5 mg/L+IBA0.8 mg/L+4% sucrose. The five-star flower can be made to bloom at about 51d by the above conditional culture. (method three)
The third method in the above document was used for controlling the flowering period of Plumbum Preparatium, and was found to be different from comparative example 1 except that paclobutrazol (PP 333) was not capable of promoting flowering of Plumbum Preparatium and inhibiting overground growth of Plumbum Preparatium.
Comparative example 4
Reference 4 (MOHAMED et al, the role ofplant growth regulators in the development of in vitro flowering, histology and ultrastructural studies in Impatiens balsamina cv.Dwarfbush [ J)]Planta daniha.2018.), the method described in: mohamed used gibberellin (GA 3) and Kinetin (KT) to regulate the flowering phase of impatiens balsamina, and found that impatiens balsamina was flowering at the sixth week when impatiens balsamina was treated with gibberellin 1.0mg/L and kinetin 0.5mg/L. Healthy and full impatiens balsamina seeds were selected, then carefully washed under running tap water for 30 minutes, then soaked with Dettol and two drops of Tween sterilant for 5 minutes, and then washed three times with distilled water. Seeds were treated with various concentrations of surface sterilized sodium hypochlorite solutions (70%, 50% and 30%) for 5 minutes. Each seed was thoroughly washed with distilled water. Finally, the seeds are treated with 70% ethanol for 1 minute, rinsed with sterile distilled water, and finally inoculated on MS+gibberellin (GA 3) +kinetin (KT) medium with pH adjusted to 5.8, the temperature is set to 23-26 ℃, the illumination is carried out for 16 hours, and the photosynthetic photon flux density is 60 mu mol.m -2 ·s -1 . (method four)
The fourth method in the above-mentioned document was used for controlling the flowering phase of Plumbum Preparatium, and was found to be ineffective for controlling the flowering phase of Plumbum Preparatium by gibberellin (GA 3) and Kinetin (KT) with reference to comparative example 1, except that the treated substances were different, and it was found that the use of gibberellin 1.0mg/L and kinetin 0.5mg/L for the treatment of Plumbum Preparatium was not flowering either in the tissue culture or the seedlings of Plumbum Preparatium in the unnatural flowering phase, which was significantly different from the effect of controlling the impatiens.
Comparative example 5
The method in reference 5 (Jiang Lin. Penta flower tissue culture and tube flowering study [ D ]) is described: jiang Lin auxin was used: naphthalene Acetic Acid (NAA) and indolebutyric acid (IBA) regulate the flowering phase of five-star flowers. The research takes tender stem segments without plant diseases and insect pests of the five-star flowers, and establishes a five-star flower flowering regulation and control technical system with higher flowering efficiency as a test material after sterilization treatment. (1) The culture temperature is 23+/-2 ℃, the illumination intensity is 2500-3000lux, and the illumination duration is 14h/d; (2) the culture medium is MS basic culture medium, adding sucrose 3%, agar 0.6%, adjusting pH to 6.0, and adding NAA0.1mg/L or IBA 0.5mg/L. The five-star flower can bloom at about 47d by the above condition culture. (method five)
The fifth method in the above document was used for controlling the florescence of blue flower dan, and the method thereof was found to be ineffective for controlling the florescence of blue flower dan by referring to comparative example 1, naphthalene Acetic Acid (NAA) and indolebutyric acid (IBA).
Comparative example 6
The method in reference 6 (Zhu Lingli. Effect of photoperiod on the flowering effects of 'good luck-bee' malachite and 'thai mountain-orange' marigold [ D ]. University of agriculture in Sichuan, 2016.): two cultivars of marigold are used as research materials, namely, two varieties of marigold (Tagetes patula 'BonanzaBees') and Tagetes Orange marigold (Tagetes eresta 'Mount Tai Orange') are used as research materials, experimental seeds are soaked in warm water at 25 ℃ for 8 hours, naturally dried, sowed in a plug tray, cultivated by garden soil, the pH value of soil is about 6.5, the plug tray is cultivated for 20d, plants are placed on a pot, and experiments are started after seedlings are slowly grown for 10d, wherein the experimental temperature is kept at 25+/-2 ℃ and the illumination intensity is 150 mu molm-2s-1 in the whole experimental process. The effect of different photoperiod (8/16 h, 10/14h, 12/12 h) conditions on the flowering of malachite and marigold was mainly studied. Under the treatment of 8/16h, 10/14h and 12/12h, the primary flowering period of the 'good luck-bee' malachite is 54, 53 and 50d respectively; the initial flowering period of the 'Taishan-orange yellow' marigold under the treatment of 8/16h, 10/14h and 12/12h is respectively 45, 45 and 44d, wherein the 'Hongyo-bee' malachite and the 'Taishan-orange yellow' marigold under the treatment of 12/12h are earliest in flowering. (method six)
The method six in the above document is used for regulating and controlling the flowering period of the blue flower red lead, and the method refers to the comparative example 1, and the blue flower red lead is treated by using different photoperiod of the method six, and as a result, the effect in the method six is not achieved no matter whether the tissue culture seedling or the seedling flowering time of the blue flower red lead is, and the difference between the tissue culture seedling or the seedling flowering time of the blue flower red lead is larger. 25+ -2deg.C, 12/12h, 150. Mu. Mol.m -2 ·s -1 Under the condition of (1), the blue flower dan tissue culture seedling cannot bloom, the time from the seedling treatment to the initial flowering is 65 days, the blue flower dan tissue culture seedling cannot bloom after about 48 days of the treatment described in the method six, and the method six has poor regulation effect on the blue flower dan period.
Comparative example 7
The method of reference 7 (Chen Ying. Magdi paphiopedilum cultivation and flowering phase control research [ D ]. 2017.): the paphiopedilum (Cypripedium tibeticum) is a terrestrial, semi-epiphytic or epiphytic herbaceous plant, chen Yingzhen can bloom about 40d after being treated by using a flower fertilizer No. 2 and 1.0g/L, firstly, a group of seedlings of the paphiopedilum test tube seedlings obtained by aseptic seeding with regular growth vigor are moved to a greenhouse for tissue culture seedling hardening in 8 months, and the culture medium is bark: planting gold stones (1:1), hardening seedlings for 10d, and performing experiments; fertilize 1 time per week. The leaf surface is sprayed with clear water for 1 time every day, and the leaf surface is slightly formed into water drops. (method seven)
The method seven in the above document is used for regulating and controlling the flowering phase of the blue-flower dan, is ineffective for regulating and controlling the flowering phase of the blue-flower dan, and is beneficial to increasing the flowering quantity.
Claims (6)
1. A method for regulating and controlling the flowering phase of blue flowers, which is characterized by comprising the following specific steps of:
(1) Selecting blue flower red lead tissue culture seedlings or seedlings;
(2) The plants in the step (1) are subjected to the following temperature changing treatment:
alternately treating for 12h/d at 25 ℃ and 30 ℃ for 29-35 d;
or alternatively treating for 12h/d at 30 ℃ and 35 ℃ respectively, wherein the treatment time is 27-34 d;
or alternatively treating at 25 ℃ and 35 ℃ for 12h/d respectively, wherein the treatment time is 30-38 d;
and (3) regulating and controlling the flowering phase of the blue flower according to the temperature-changing treatment mode, so as to realize flowering in the unnatural flowering phase.
2. The method of modulating a blue-flower danshen period according to claim 1, characterized in that in step (2) the plants are also subjected to the following light treatments: the illumination intensity is 200 mu mol.m -2 ·s -1 The illumination time is 12h/d.
3. The method of claim 1, wherein the humidity of the environment is maintained at 70-90% during the step (2).
4. The method for regulating and controlling the flowering phase of blue flowers according to claim 1, wherein the method for obtaining the tissue culture seedlings is as follows:
(1) Collecting mature, full and healthy blue flower dan seeds, soaking the seeds with the seed coats removed in distilled water at 30 ℃ and under dark conditions for 2d, and then washing with running water for 2h;
(2) Immersing the seeds in 75% w/v ethanol solution for 45s, taking out the seeds, and washing the seeds with sterile distilled water for 5-6 times;
(3) Immersing the seeds in 0.1% w/v mercuric chloride solution for 4min, washing with sterile distilled water for 5-6 times, and repeating the steps;
(4) Seed was inoculated at 30 g.L -1 Sucrose and 6.5 g.L -1 And (3) culturing agar on an MS culture medium with the pH value of 5.8-6.0 to obtain the tissue culture seedlings.
5. The method for regulating and controlling the flowering phase of blue flowers according to claim 1, wherein the method for obtaining seedlings is as follows: collecting mature, full and healthy blue flower dan seeds, sowing the seeds on a peat soil matrix for germination culture for three months, and obtaining seedlings with consistent growth conditions.
6. The method of modulating a flowering phase of blue flowers according to claim 5, wherein the Lan Huadan plants and the watering pot are sprayed with 800 times the carbendazim every two weeks during the germination culture.
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