CN110149950B

CN110149950B - Method for improving development quality of paphiopedilum albolanum seeds and test solution thereof

Info

Publication number: CN110149950B
Application number: CN201910492598.3A
Authority: CN
Inventors: 唐凤鸾; 赵健; 韦宇静; 郭丽君; 夏科; 吴巧芬
Original assignee: Guangxi Institute of Botany of CAS
Current assignee: Guangxi Institute of Botany of CAS
Priority date: 2019-06-06
Filing date: 2019-06-06
Publication date: 2021-12-03
Anticipated expiration: 2039-06-06
Also published as: CN110149950A

Abstract

The invention discloses a test solution for improving the development quality of paphiopedilum albolanum seeds, which comprises three special solutions, wherein the special solutions comprise the following components: ammonium nitrate, potassium nitrate, calcium chloride, magnesium sulfate heptahydrate, potassium dihydrogen phosphate, boric acid and gibberellin. The test solution is obtained through continuous experimental development on the basis of previous research. Accordingly, a method for improving the development quality of paphiopedilum albolanum seeds is also established, namely roots and leaves of paphiopedilum albolanum plants are watered with the special solutions 1 to 3. Researches show that the invention can enhance plant nutrition, promote capsule development and obtain paphiopedilum hirsutissimum seeds with high embryo rate and strong vitality. Therefore, the application of the invention can effectively improve the embryo development of paphiopedilum alboleraceum seeds, has great significance for solving the problems of low germination rate, slow seedling production and the like of paphiopedilum alboleraceum caused by no embryos or incomplete embryo development of the paphiopedilum alboleraceum seeds, has great significance for resource protection, development and utilization of wild paphiopedilum alboleraceum in a very small population, and has very wide application prospect.

Description

Method for improving development quality of paphiopedilum albolanum seeds and test solution thereof

Technical Field

The invention belongs to the technical field of paphiopedilum leucovorum reproduction and growth, and particularly relates to a method for improving the development quality of paphiopedilum leucovorum seeds and a test solution thereof.

Background

Paphiopedilum (Paphiopedilum emersonii) is a perennial semi-epiphytic or terrestrial plant of Paphiopedilum in the orchid family (Orchidaceae), a special species in China, and a national grade 1 protective plant. The paphiopedilum has very high ornamental value due to the characteristics of large flowers, peculiar flower types, pure and bright colors, flowery flavor and the like, and is regarded as a fine paphiopedilum product. The existing sources of paphiopedilum albopictus are only 7 colonies about 280 in the protected area of the cantonese wood theory and 6 colonies about 306 in the protected area of maoland Guizhou (spawn Wenyuan et al, 2012; Tantanning, 2009), and the colonies and the number of individuals are far lower than the minimum survivable number of a stable survival limit (the world recognizes that the herbaceous plant group is 10000 individuals) and are endangered at any time (Ren et al, 2012), which is the true "giant panda plant". Therefore, the establishment of paphiopedilum as one of the most preferred protective groups among paphiopedilum plants in our country by botanicologist ludwig et al (2003) has been included in "wild plant rescue and protection project planning for extremely small population in the whole country (2011-.

Paphiopedilum has a large number of seeds, and non-symbiotic germination of the seeds is a main means for obtaining paphiopedilum seedlings at present. Through researches on paphiopedilum micranthum for many years, the applicant finds that paphiopedilum micranthum capsules are easy to obtain no matter in field natural pollination or artificial pollination after introduction, but further electron microscope scanning and vitality detection find that most seeds have no embryo or are poor in development, and the TTC staining rate is only 1-3%. It can be seen that the development quality of paphiopedilum albolanum seeds is a main internal factor influencing the germination rate.

The strong fruiting plant is cultured, so that the flowering quantity and quality can be guaranteed, the resistance can be enhanced, the growth of capsules can be promoted, and the seed quality is improved. According to the inspection, no relevant report aiming at the nutrition and the capsule development of paphiopedilum albolanum plants is found.

Disclosure of Invention

The invention aims to solve the technical problem of providing a method for improving the development quality of paphiopedilum hirsutissimum seeds and a test solution thereof.

In order to solve the technical problems, the invention adopts the following technical scheme:

the test solution for improving the development quality of paphiopedilum albolanum seeds contains the following components: ammonium nitrate, potassium nitrate, calcium chloride, magnesium sulfate heptahydrate and potassium dihydrogen phosphate.

The tailored solution 1 contained the following ingredients: 2.0 to 3.5 g/L of ammonium nitrate, 0.3 to 0.5 g/L of potassium nitrate, 0.2 to 0.4 g/L of calcium chloride, 0.3 to 0.5 g/L of magnesium sulfate heptahydrate, and 0.3 to 0.5 g/L of monopotassium phosphate.

The tailored solution 1 contained the following ingredients: 2.8 g/L of ammonium nitrate, 0.4 g/L of potassium nitrate, 0.3 g/L of calcium chloride, 0.4 g/L of magnesium sulfate heptahydrate and 0.4 g/L of monopotassium phosphate.

The tailored solution 2 contained the following ingredients: 1.5 to 2.5 g/L of ammonium nitrate, 0.5 to 0.7 g/L of potassium nitrate, 0.2 to 0.4 g/L of calcium chloride, 0.3 to 0.5 g/L of magnesium sulfate heptahydrate, 0.5 to 0.7 g/L of monopotassium phosphate, and 0.005 to 0.012 g/L of boric acid.

The tailored solution 2 contained the following ingredients: 2.0 g/L of ammonium nitrate, 0.6 g/L of potassium nitrate, 0.3 g/L of calcium chloride, 0.4 g/L of magnesium sulfate heptahydrate, 0.6 g/L of monopotassium phosphate and 0.01 g/L of boric acid.

The tailored solution 3 contained the following ingredients: 1.5 to 2.5 g/l ammonium nitrate, 0.5 to 0.7 g/l potassium nitrate, 0.2 to 0.4 g/l calcium chloride, 0.3 to 0.5 g/l magnesium sulfate heptahydrate, 0.5 to 0.7 g/l potassium dihydrogen phosphate, 0.005 to 0.012 g/l boric acid, Gibberellin (GA)₃)50～100mg/L。

The tailored solution 3 had the following ingredients: 2.0 g/L of ammonium nitrate, 0.6 g/L of potassium nitrate, 0.3 g/L of calcium chloride, 0.4 g/L of magnesium sulfate heptahydrate, 0.6 g/L of monopotassium phosphate, 0.01 g/L of boric acid and Gibberellin (GA)₃)80mg/L。

A method for improving the development quality of paphiopedilum albolanum seeds comprises the steps of watering roots and spraying leaves on paphiopedilum albolanum plants by using special solutions 1-3; the tailored solution 1 contained the following ingredients: 2.0 to 3.5 g/L of ammonium nitrate, 0.3 to 0.5 g/L of potassium nitrate, 0.2 to 0.4 g/L of calcium chloride, 0.3 to 0.5 g/L of magnesium sulfate heptahydrate, and 0.3 to 0.5 g/L of monopotassium phosphate; the tailored solution 2 contained the following ingredients: 1.5 to 2.5 g/L of ammonium nitrate, 0.5 to 0.7 g/L of potassium nitrate, 0.2 to 0.4 g/L of calcium chloride, 0.3 to 0.5 g/L of magnesium sulfate heptahydrate, 0.5 to 0.7 g/L of monopotassium phosphate, and 0.005 to 0.012 g/L of boric acid; the tailored solution 3 contained the following ingredients: 1.5 to 2.5 g/l ammonium nitrate, 0.5 to 0.7 g/l potassium nitrate, 0.2 to 0.4 g/l calcium chloride, 0.3 to 0.5 g/l magnesium sulfate heptahydrate, 0.5 to 0.7 g/l potassium dihydrogen phosphate, 0.005 to 0.012 g/l boric acid, Gibberellin (GA)₃)50～100mg/L。

The method for improving the development quality of paphiopedilum albolanum seeds comprises the following steps:

(1) selecting healthy paphiopedilum plants planted in pot plants, watering roots and spraying leaves with a special solution 1 for 1 time each month;

(2) after the paphiopedilum albolense plant which is treated in the step (1) and successfully differentiates flower buds is treated, roots and leaves are watered with a special solution 1 and sprayed with leaves for 2 times each month;

(3) after the paphiopedilum albolense plant which is treated in the step (2) and has good flowers is opened, roots and leaves are watered with the special solution 2 and sprayed with the solution 2 for 2 times each month; meanwhile, carrying out artificial cross pollination on the flowers after the flowers bloom for 2-3 days;

(4) and (4) after the treatment of the step (3), successfully pollinating the paphiopedilum bungeanum plant with the expanded ovary, watering the root and spraying leaves with the special solution 3 for 2 times each month.

Aiming at the problems existing in the breeding of paphiopedilum albolanum at present, the inventor develops a test solution for improving the development quality of paphiopedilum albolanum seeds, which comprises three special solutions, wherein the three special solutions comprise the following components: ammonium nitrate, potassium nitrate, calcium chloride, magnesium sulfate heptahydrate, potassium dihydrogen phosphate, boric acid and gibberellin. The test solution is obtained through continuous experimental development on the basis of previous research. Accordingly, the inventor establishes a method for improving the growth quality of paphiopedilum albolanum seeds by taking paphiopedilum albolanum which is a very small population wild plant with good ornamental and scientific research values as a material, namely, performing root watering and leaf spraying treatment on paphiopedilum albolanum plants by using special solutions 1 to 3. Researches show that the paphiopedilum ariifolium seed can enhance plant nutrition, promote capsule development and obtain paphiopedilum hirsutissimum seeds with high embryo rate and strong vitality, the embryo rate of the paphiopedilum hirsutissimum seeds reaches 79.04%, the dyeing rate reaches more than 11.87%, and the seed quality and vitality are greatly improved. Therefore, the application of the invention can effectively improve the embryo development of paphiopedilum alboleraceum seeds, has great significance for solving the problems of low germination rate, slow seedling production and the like of paphiopedilum alboleraceum caused by no embryos or incomplete embryo development of the paphiopedilum alboleraceum seeds, has great significance for resource protection, development and utilization of wild paphiopedilum alboleraceum in a very small population, and has very wide application prospect.

Drawings

FIG. 1 is a graph of seed embryo development and TTC staining of example 1.

FIG. 2 is a graph of seed embryo development and TTC staining in example 2.

FIG. 3 is a graph of seed embryo development and TTC staining for comparative examples.

Detailed Description

Paphiopedilum plants used in the examples and the control examples were planted in orchid germplasm gardens of applicant units, and were in a natural state except for rain sheltering and sun shading, and were not subjected to special treatment.

Firstly, specially-made solution is tried

Through analysis of nutrient elements of primordial root soil and plant leaves of paphiopedilum micranthum, the nitrogen, phosphorus and potassium contents of paphiopedilum micranthum are high, particularly the total nitrogen content of the root soil averagely reaches 6.95g/kg, and thus the requirements of growth of paphiopedilum micranthum on major elements such as nitrogen, phosphorus and potassium are high. The nutrient element boron is proved to be capable of enhancing photosynthesis to promote carbohydrate formation, promoting pollen germination, stimulating pollen tube elongation and improving fertility rate and seed quality. In the tissue culture test, the improved MS culture medium is found to be beneficial to the culture of paphiopedilum, and the test-tube plantlet added with gibberellin has good flowering and fruiting performances after being transplanted.

To this end, the inventors designed a tailored solution for the treatment of paphiopedilum concordatum plants and performed a preliminary screening using different modes of use, as detailed in table 1.

TABLE 1 trial of the tailored solutions

In table 1, the methods for using the tailored solutions in 4 experiments are respectively:

tests 1 and 2 all use the same solution, and the application method, the amount and the time are the same as the corresponding steps of the method.

Run 3 used two solutions, where steps (1) and (2) used solution 1 and steps (3) and (4) used solution 2, in the same amounts and for the same time as the corresponding steps of the method of the invention described previously.

Test 4 three solutions were used, the application method, the amount, the time being the same as the corresponding steps of the method of the invention described previously.

As can be seen from Table 1, compared with the single use of the tailored solution 1 (test 1-2) or the simultaneous use of the tailored solutions 1 and 2 (test 3), the seed embryo rate and the staining rate of the paphiopedilum albolanum are optimal and the seed quality and the seed vitality can be improved more effectively when three different tailored solutions are used in different growth periods of the paphiopedilum albolanum (test 4).

Second, example

Example 1

(1) Selecting paphiopedilum albolense plants which are robust in growth, wide and thick in leaves, dark green in color, tall and straight in plant type and free of diseases and insect pests, and watering roots and spraying leaves with a special solution 1; the frequency of root watering and spraying is 1 time each month.

(2) Selecting paphiopedilum alboleracea plants which are treated in the step (1) and successfully differentiate flower buds, and watering roots and spraying leaves by using a special solution 1; the judgment method for successfully differentiating the flower buds comprises the following steps of observing at least 1 flower bud to grow by naked eyes; preparing a solution 1 in the same step (1); the frequency of root watering and spraying is 2 times per month.

(3) Selecting paphiopedilum aristata plants which are treated in the step (2) and have thick and straight stems, large flowers, regular flower shapes and pure white colors, watering roots and spraying leaves with a special solution 2, and performing artificial cross pollination on the flowers after the flowers bloom for 2-3 days; the frequency of root watering and spraying is 2 times per month.

(4) Selecting paphiopedilum albolens plants which are treated in the step (3) and successfully pollinated and whose ovaries begin to expand, watering roots and spraying leaves with a special solution 3; the frequency of root watering and spraying is 2 times per month.

The solution formula is as follows:

solution 1 was prepared: a mixed solution containing 2.8 g/L of ammonium nitrate, 0.4 g/L of potassium nitrate, 0.3 g/L of calcium chloride, 0.4 g/L of magnesium sulfate heptahydrate and 0.4 g/L of monopotassium phosphate;

solution 2 was made specifically: a mixed solution containing 2.0 g/l of ammonium nitrate, 0.6 g/l of potassium nitrate, 0.3 g/l of calcium chloride, 0.4 g/l of magnesium sulfate heptahydrate, 0.6 g/l of monopotassium phosphate and 0.01 g/l of boric acid;

solution 3 to make: contains ammonium nitrate 2.0 g/L, potassium nitrate 0.6 g/L, calcium chloride 0.3 g/L, magnesium sulfate heptahydrate 0.4 g/L, potassium dihydrogen phosphate 0.6 g/L, boric acid 0.01 g/L, and Gibberellin (GA)₃)80mg/L of mixed solution;

the experimental results are as follows: the embryo rate of the treated plant, capsule pollinated for 150 days, was 79.04% and the TTC staining rate was 11.87% (FIG. 1).

Example 2

The solution formula is as follows:

solution 1 was prepared: a mixed solution containing 2.0 g/l of ammonium nitrate, 0.3 g/l of potassium nitrate, 0.2 g/l of calcium chloride, 0.3 g/l of magnesium sulfate heptahydrate and 0.3 g/l of monopotassium phosphate;

solution 2 was made specifically: a mixed solution containing 1.5 g/l of ammonium nitrate, 0.5 g/l of potassium nitrate, 0.2 g/l of calcium chloride, 0.3 g/l of magnesium sulfate heptahydrate, 0.5 g/l of monopotassium phosphate and 0.005 g/l of boric acid;

solution 3 to make: contains ammonium nitrate 1.5 g/L, potassium nitrate 0.5 g/L, calcium chloride 0.2 g/L, magnesium sulfate heptahydrate 0.3 g/L, potassium dihydrogen phosphate 0.5 g/L, boric acid 0.005 g/L, and Gibberellin (GA)₃)50mg/L of mixed solution;

the experimental results are as follows: the embryo bearing rate of the treated plant, capsule pollinated for 150 days, was 62.63% and TTC staining rate was 7.9% (FIG. 2).

Example 3

The solution formula is as follows:

solution 1 was prepared: a mixed solution containing 3.5 g/l of ammonium nitrate, 0.5 g/l of potassium nitrate, 0.4 g/l of calcium chloride, 0.5 g/l of magnesium sulfate heptahydrate and 0.5 g/l of monopotassium phosphate;

solution 2 was made specifically: a mixed solution containing 2.5 g/l of ammonium nitrate, 0.7 g/l of potassium nitrate, 0.4 g/l of calcium chloride, 0.5 g/l of magnesium sulfate heptahydrate, 0.7 g/l of monopotassium phosphate and 0.012 g/l of boric acid;

solution 3 to make: contains 2.5 g/L ammonium nitrate, 0.7 g/L potassium nitrate, 0.4 g/L calcium chloride, 0.5 g/L magnesium sulfate heptahydrate, 0.7 g/L potassium dihydrogen phosphate, 0.012 g/L boric acid, and Gibberellin (GA)₃)100mg/L of mixed solution;

the experimental results are as follows: the embryo rate of the treated plant, capsule pollinated for 150 days, is 47.69 percent, and the TTC staining rate is 6.34 percent.

Third, comparative example

(1) Selecting paphiopedilum albolense plants which are strong in growth, wide and thick in leaves, dark green in color, tall and straight in plant type and free of plant diseases and insect pests, and applying 5g of slow release fertilizer for aolv 315S orchid produced by The Scotts Company in each pot.

(2) Selecting paphiopedilum alboleracea plants which are treated in the step (1) and successfully differentiate flower buds, namely, the plants with at least 1 flower bud can be seen by naked eyes; and (3) fertilizing for the second time 3 months after the fertilization in the step (1), and applying 5g of slow release fertilizer for ultragreen 315S orchid in each pot.

(3) Selecting paphiopedilum aristata plants which are treated in the step (2) and have thick and straight stems, large flowers, regular flower shapes and pure white colors, and carrying out artificial cross pollination on the flowers after the flowers bloom for 2-3 days; and (3) fertilizing for the third time 3 months after the fertilization in the step (2), and applying 5g of slow release fertilizer for ultragreen 315S orchid in each pot.

(4) Selecting paphiopedilum albolens plants which are processed in the step (3) and successfully pollinated, and whose ovaries begin to expand; and (4) fertilizing for the fourth time 3 months after the fertilization in the step (3), and applying 5g of slow release fertilizer for ultragreen 315S orchid in each pot.

The experimental results are as follows: the embryo bearing rate of the treated plant, capsule pollinated for 150 days, was 35.14% and TTC staining rate was 1.02% (FIG. 3).

Claims

1. a method for improving the quality of the development of the P. 3.5 g/L, potassium nitrate 0.3-0.5 g/L, calcium chloride 0.2-0.4 g/L, magnesium sulfate heptahydrate 0.3-0.5 g/L, potassium dihydrogen phosphate 0.3-0.5 g/L; the special solution 2 Contains the following ingredients: ammonium nitrate 1.5-2.5 g/L, potassium nitrate 0.5-0.7 g/L, calcium chloride 0.2-0.4 g/L, magnesium sulfate heptahydrate 0.3-0.5 g/L, potassium dihydrogen phosphate 0.5- 0.7 g/L, boric acid 0.005-0.012 g/L; the special solution 3 contains the following components: ammonium nitrate 1.5-2.5 g/L, potassium nitrate 0.5-0.7 g/L, calcium chloride 0.2-0.4 g/L, Magnesium sulfate heptahydrate 0.3-0.5 g/L, potassium dihydrogen phosphate 0.5-0.7 g/L, boric acid 0.005-0.012 g/L, gibberellin 50-100 mg/L; the method includes the following steps:

(1) Select robust P. chinensis plants, water the roots and spray the leaves with a special solution, once a month;

(2) through step (1) processing and successfully differentiated the Ph.

(3) the P. chinensis plants that are processed in step (2) and have well-opened flowers, water the roots and spray the leaves with a special solution 2, 2 times each month; at the same time, artificial cross-pollination is carried out 2 to 3 days after the flowers open;

(4) After the treatment in step (3) and the pollination was successful, the ovary of the P. chinensis plant was irrigated with the special solution 3 times and the leaves were sprayed twice a month.

2. the method for improving the quality of the growth of the P. g/L, magnesium sulfate heptahydrate 0.4 g/L, potassium dihydrogen phosphate 0.4 g/L.

3. the method for improving the quality of the development of the P. g/L, magnesium sulfate heptahydrate 0.4 g/L, potassium dihydrogen phosphate 0.6 g/L, boric acid 0.01 g/L.

4. the method for improving the quality of P. alba seed growth according to claim 1, is characterized in that: described special solution 3 contains following composition: ammonium nitrate 2.0 grams/liter, potassium nitrate 0.6 grams/liter, calcium chloride 0.3 g/L, magnesium sulfate heptahydrate 0.4 g/L, potassium dihydrogen phosphate 0.6 g/L, boric acid 0.01 g/L, gibberellin 80 mg/L.