CN110972844B - Greenhouse cultivation method for corn transgenic tissue culture seedlings - Google Patents
Greenhouse cultivation method for corn transgenic tissue culture seedlings Download PDFInfo
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- CN110972844B CN110972844B CN201911279711.6A CN201911279711A CN110972844B CN 110972844 B CN110972844 B CN 110972844B CN 201911279711 A CN201911279711 A CN 201911279711A CN 110972844 B CN110972844 B CN 110972844B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/20—Cereals
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C21/00—Methods of fertilising, sowing or planting
- A01C21/005—Following a specific plan, e.g. pattern
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
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Abstract
The invention discloses a greenhouse cultivation method of corn transgenic tissue culture seedlings, belonging to the technical field of transgenic plant cultivation. According to the greenhouse cultivation method of the corn transgenic tissue culture seedlings, through links such as specific control of transplanting, water and fertilizer management, pest control, growth environment control, pollination and the like, the transplanted corn tissue culture seedlings grow robustly and are coordinated in male and female, the maturing rate of the corn tissue culture seedlings is improved to over 95%, and the average maturing rate of each corn tissue culture seedling is over 160.
Description
Technical Field
The invention belongs to the technical field of transgenic plant cultivation, and particularly relates to a greenhouse cultivation method of corn transgenic tissue culture seedlings.
Background
Corn is not only an important grain, but also an important economic crop, and plays a very important role in national economy, so that the gene transformation research of the corn is one of the hot spots of the plant molecular biology research at home and abroad. The industrialization of corn transgenosis brings changes to the development of the global corn industry and the corn production mode, and provides important guarantee for the grain safety, the ecological safety and the income increase of farmers. The common selfing line for the corn transgenosis is an easily infected selfing line, so the common selfing line has poor resistance, is susceptible to diseases and is irregular in male and female, and the growth time in a genetic transformation tissue culture room is longer, so that the corn transgenic tissue culture seedlings are long in seedling age, poor in adaptability and low in fruiting rate. The development of corn functional genome and transgenic technology is severely limited by the corn tissue culture seedling obtained by the transgenic technology due to the lower seed setting rate and the number of harvested seeds. Therefore, it is important to improve the seed set rate and the seed set number of each plant of the tissue culture seedlings.
The existing efficient transplanting method of the corn transgenic tissue culture seedlings is managed according to the cultivation mode of the seedlings, although water and fertilizer are sufficient, the growth period of the transplanted tissue culture seedlings is shorter than that of the seedlings, the growth environment parameters of efficient photosynthetic product accumulation are absent, the requirements of vegetative growth and reproductive growth of the seedlings are difficult to meet, the materials are weak in growth vigor, poor in resistance, low in fruiting rate of about 60% -80%, and each plant can fruit 20-30 on average.
Disclosure of Invention
Therefore, the invention overcomes the defects of the prior art and provides a high-efficiency cultivation method of the corn transgenic tissue culture seedling.
Therefore, the technical scheme of the invention is as follows:
a greenhouse cultivation method of corn transgenic tissue culture seedlings comprises the following steps:
transplanting: hardening the corn tissue culture seedlings for 7-10 days, screening, transplanting into a matrix, and adding nutrient solution;
and (3) water and fertilizer management: watering once in 7-14 days of the seedling stage, watering once every 6-10 days in the silking and pollen scattering stage, and watering once in 5-8 days after pollination; the water amount is 5-7L/plant each time; and (3) topdressing three times of the tissue culture seedlings in the whole growth period: after 2 weeks of transplantation, 0.5-2g of additional fertilizer is applied for the first time; the second time is after 3-4 weeks of transplantation, 1-5g of additional fertilizer is applied; the third time is after 5-6 weeks of transplanting, before and after spinning, top dressing 3-7 g;
and (3) pest control: the prevention is taken as the main strategy, and the prevention and the treatment are combined;
in the prevention process, chemical agents are sprayed every 10 days; the chemical agents comprise an aphid preventing and controlling agent and a red spider mite preventing and controlling agent, and the agents with different functions are alternately used;
controlling the growth environment: controlling the temperature in the greenhouse at night to be 16-18 ℃, setting the temperature in the daytime along with the change of the intensity of the photosynthetic radiation, and controlling the relative humidity to be 30-60%; the daily average photosynthetically active radiation is not less than 70W/m2；
Pollination: artificial supplementary pollination is adopted, when the female ear is expanded to 3cm, the female ear is bagged, when the male ear reaches the pollen-scattering full period, the filaments are cut, the male ear bag is sleeved, and pollination is started at about 9 am on the next day.
In the method, the screening method comprises the following steps: the height of the selected plants is more than or equal to 15cm for transplanting, and the height of the selected plants is less than 15cm, so that the transplanting can be delayed until the next time.
In the above method, the method for transplanting to the substrate comprises: watering in the mixed matrix, digging a cylindrical transplanting hole with the depth of 7-10cm and the diameter of 7-10cm on the matrix, transplanting the screened seedling into the transplanting hole, backfilling with the matrix, and then pouring in nutrient solution.
In the above method, the substrate is: the turfy soil, the perlite and the vermiculite are uniformly mixed according to the volume ratio of 5:1:1, and 5L of chicken manure organic fertilizer, 10g of calcium hydroxide, 50g of potassium sulfate compound fertilizer and 50g of diammonium phosphate are added into each 100L of matrix.
In the above method, the method for setting the daytime temperature according to the intensity change of the photosynthetic radiation comprises: the temperature is set according to the following formula:
the temperature is 15+ photosynthetically active radiation/10, and the applicable photosynthetically active radiation range is 0-100W/m2；
The temperature is 25+ (photosynthetically active radiation-100)/100, and the applicable photosynthetically active radiation range is 100-2；
In the method, the aphid control medicament is more than one of aphid-mite cleaning missible oil, pirimicarb, imidacloprid or lufenuron; the agent for preventing and treating red spider mite is one or more of Icarin, dichlofenphos or abamectin.
In the method, preferably, 1g of the first top dressing is carried out in the three top dressing of the tissue culture seedlings in the whole growth period; topdressing for the second time by 3 g; and 5g of additional fertilizer is applied for the third time.
In the method, in the three additional fertilizers for the whole growth period of the tissue culture seedlings, the fertilizer is a macroelement water-soluble fertilizer which is a flower-free (20-20-20), wherein N-K2O-P2The O content was 20%, 20% and 20%, respectively.
In the method, the pollination method further comprises pollination of the same plant once more every other day and removal of tassels.
According to the greenhouse cultivation method of the corn transgenic tissue culture seedlings, the transplanted corn tissue culture seedlings are strong in plant growth and harmonious in male and female, the setting rate of the corn tissue culture seedlings is improved to more than 95%, and each plant sets more than 160 seeds on average.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to specific embodiments, and it should be understood that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, belong to the protection scope of the present invention. The materials and devices used in the present invention are commercially available unless otherwise specified.
The present invention is further illustrated by, but is not limited to, the following examples.
Example 1
(1) Preparing a matrix: the turfy soil, the perlite and the vermiculite are uniformly mixed according to the volume ratio of 5:1:1, 5L of chicken manure organic fertilizer (Beijing Yingtongxin agriculture development Co., Ltd.) is added into each 100L of substrate, and the chicken manure organic fertilizer contains pure nitrogen and phosphorus (P)2O5) Potassium (K)2O) about 1.63%, 1.54%, 0.85%), calcium hydroxide 10g (national drug group chemical reagent limited, active ingredient content is 99%), potassium sulfate compound fertilizer 50g (St. fertilizer industry GmbH, Anhui province, K)2O content of 45%) and diammonium phosphate 50g (Yunnan three-ring chemical fertilizer Co., Ltd., P)2O5And 64% of total N) and mixed evenly, 15L of matrix is filled in a 18L flowerpot, and 6L of matrix is watered for standby.
(2) Transplanting: and (6) screening 688 positive plants with the plant height of more than 15cm in 1 month and 1 day in 2019. Transplanting into the prepared flowerpot. Digging a cylindrical transplanting hole with the depth of 8cm and the diameter of 8cm on the matrix, transplanting the screened positive seedlings into the transplanting hole, then pouring 0.1 percent of flower-free nutrient solution, and pouring a circle of seedlings which are just transplanted, wherein about 1L of nutrient solution is needed, so that the matrix carried by the corn tissue culture seedlings is tightly combined with the matrix in the flowerpot.
(3) And (3) water and fertilizer management: watering for the first time after 15 days of transplanting, wherein each plant is watered with about 6L of water, applying fertilizer for the first time, and topdressing for flowers by 1 g; watering for the second time after transplanting for 25 days, wherein each plant is watered with about 6L of water, applying fertilizer for the second time, and topdressing flower (water-soluble fertilizer from Guangzhou Runjie agricultural science and technology Co., Ltd., N-K2O-P2The O content was 20%, 20% and 20%, respectively.
)3g of the total weight of the mixture; watering for the third time after 35 days of transplanting, wherein each plant is watered with about 6L of water, applying fertilizer for the third time, and topdressing 5 g; then, 6L of water was added every 7 days until harvest.
(4) And (3) pest control: the plant diseases and insect pests which are frequently generated by the corn tissue culture seedlings in the greenhouse are aphids and red spiders. Mainly adopts chemical reagent control. The medicament is sprayed once every 10 days and is divided into 6 combinations: 1. emamectin benzoate and aphid mite clear emulsifiable concentrate; 2. acarus, pirimicarb; 3. desmodium, abamectin and imidacloprid; 4. combretastatin, abamectin and lufenuron; 5. acarus, imidacloprid and lufenuron; 6. the pesticide comprises pyridaben, ivermectin, aphidicolin missible oil and pirimicarb; used sequentially, once every two months. And (3) finding out the red spiders to be serious plants, independently isolating the plants, cleaning the plants with clear water, and then spraying a medicament.
The aphid control medicament is one or more of aphid mite cleaning missible oil, pirimicarb, imidacloprid or lufenuron; the agent for preventing and treating red spider mite is one or more of Icarin, dichlofenphos or abamectin.
(5) Controlling the growth environment: the temperature in the greenhouse at night is 16-18 ℃, the temperature in the daytime changes along with the change of the intensity of photosynthetic radiation, the photosynthetic effective radiation is lower in Beijing 1 month, the highest temperature in the greenhouse in the white day does not exceed 27 ℃, and the average temperature in the day does not exceed 20 ℃. The relative humidity is 30-46%, and the average humidity is 35%.
Controlling the growth environment: controlling the temperature in the greenhouse at night to be 16-18 ℃, wherein the temperature in the daytime changes along with the change of the intensity of the photosynthetic radiation, and the specific formula is as follows:
the temperature is 15+ photosynthetically active radiation/10, and the applicable photosynthetically active radiation range is 0-100W/m2；
The temperature is 25+ (photosynthetically active radiation-100)/100, and the applicable photosynthetically active radiation range is 100-2；
The effective light radiation changes with time, the temperature value is obtained according to the measured value and formula, and then the required temperature is realized by manually controlling equipment such as a greenhouse skylight and the like.
For example, the effective light radiation is 80W/m at 8 am2Controlling the temperature of the greenhouse at 8 o' clock to be 23 ℃ at noonThe 12-point effective light radiation is 300W/m2And controlling the temperature in the greenhouse at 27 ℃. The temperature can be adjusted in real time by means of a device (Priva greenhouse control system) according to the active light radiation, or the time points can be selected, the detection and the adjustment of the temperature can be carried out at intervals of 1 or 2 hours, for example, 8, 10, 12, 14, 16 or 18 points can be selected.
(6) A pollination mode: the pollination mode of the tissue culture seedlings is artificial supplementary pollination. When the ear rises to 3cm, the ear is bagged to prevent silk from running. When the tassel is in the pollen-dispersing full-age period, cutting the filaments, covering the tassel bag, and pollinating at about 9 am the next day. Pollination is carried out again on the same plant every other day, and then tassels are removed.
According to the 688 corn transgenic tissue culture seedlings cultivated by the method, the transplanted corn plants grow robustly and are coordinated in male and female, the setting rate of the corn tissue culture seedlings is 97.8 percent, and each corn plant sets 181 seeds on average. In the prior art, the transplanting survival rate of the corn tissue culture seedlings is improved to more than 97 percent, the transplanted corn plants are vigorous in production, can normally blossom and fruit, and fruit 25-40 grains per plant on average.
The above examples are merely illustrative for clarity and are not intended to limit the embodiments. Other variations and modifications in the form described above will be apparent to those skilled in the art from the foregoing description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (8)
1. A greenhouse cultivation method of corn transgenic tissue culture seedlings is characterized by comprising the following steps:
transplanting: hardening the corn tissue culture seedlings for 7-10 days, screening, transplanting into a matrix, and adding a nutrient solution;
and (3) water and fertilizer management: watering once in 7-14 days of the seedling stage, watering once every 6-10 days in the silking and pollen scattering stage, and watering once in 5-8 days after pollination; the water amount is 5-7L/plant each time; topdressing three times in the whole growth period of the tissue culture seedlings: after 2 weeks of transplantation, 0.5-2g of additional fertilizer is applied for the first time; the second time is after 3-4 weeks of transplantation, 1-5g of additional fertilizer is applied; the third time is after 5-6 weeks of transplanting, before and after spinning, top dressing 3-7 g;
and (3) pest control: the prevention is taken as the main strategy, and the prevention and the treatment are combined;
in the prevention process, chemical agents are sprayed every 10 days; the chemical agents comprise an aphid preventing and controlling agent and a red spider mite preventing and controlling agent, and the agents with different functions are alternately used;
controlling the growth environment: controlling the temperature in the greenhouse at night to be 16-18 ℃, setting the temperature in the daytime along with the change of the intensity of the photosynthetic radiation, and controlling the relative humidity to be 30-60%; the daily average photosynthetically active radiation is not less than 70W/m2；
The method for setting the daytime temperature along with the intensity change of the photosynthetic radiation comprises the following steps: the temperature is set according to the following formula:
the temperature is 15+ photosynthetically active radiation/10, and the applicable photosynthetically active radiation range is 0-100W/m2；
The temperature is 25+ (photosynthetically active radiation-100)/100, and the applicable photosynthetically active radiation range is 100-2；
Pollination: artificial supplementary pollination is adopted, when the female ear is expanded to 3cm, the female ear is bagged, when the male ear reaches the pollen-scattering full period, the filaments are cut, the male ear bag is sleeved, and pollination is started at about 9 am on the next day.
2. The method of claim 1, wherein the screening is performed by: and screening the height of the plants to be more than or equal to 15cm for transplanting, and delaying the transplanting until the next time when the height of the plants is less than 15 cm.
3. The method of claim 1, wherein the method of transplanting to a substrate is: watering in the mixed matrix, digging a cylindrical transplanting hole with the depth of 7-10cm and the diameter of 7-10cm on the matrix, transplanting the screened seedling into the transplanting hole, backfilling with the matrix, and then pouring in nutrient solution.
4. The method of claim 1, wherein the substrate is: the turfy soil, the perlite and the vermiculite are uniformly mixed according to the volume ratio of 5:1:1, and 5L of chicken manure organic fertilizer, 10g of calcium hydroxide, 50g of potassium sulfate compound fertilizer and 50g of diammonium phosphate are added into each 100L of matrix.
5. The method according to claim 1, wherein the aphid control agent is one or more of aphidicolin, pirimicarb, imidacloprid or lufenuron; the agent for preventing and treating red spider mite is one or more of Icarin, dichlofenphos or abamectin.
6. The method as claimed in claim 1, wherein 1g of the first time of three topdressing of the tissue culture seedlings in the whole growth period; topdressing for the second time by 3 g; and 5g of additional fertilizer is applied for the third time.
7. The method as claimed in any one of claims 1 to 5, wherein the fertilizer is 20-20-20 flowers in three additional manuring operations throughout the whole growth period of the tissue culture seedling.
8. The method of claim 1 wherein pollinating further comprises pollinating the same plant once more every other day and then removing the tassel.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102119660A (en) * | 2010-12-16 | 2011-07-13 | 宜昌市科力生实业有限公司 | Method for rooting culture,seedling adaptation and transplantation of alpine rose tissue-cultured seedling in greenhouse |
| CN103858771A (en) * | 2014-04-02 | 2014-06-18 | 山东省农业科学院生物技术研究中心 | Maize transgenic tissue culture seedling transplanting method |
| CN109832188A (en) * | 2017-11-27 | 2019-06-04 | 中国农业大学 | A kind of preparation method of corn dihaploid |
-
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- 2019-12-13 CN CN201911279711.6A patent/CN110972844B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102119660A (en) * | 2010-12-16 | 2011-07-13 | 宜昌市科力生实业有限公司 | Method for rooting culture,seedling adaptation and transplantation of alpine rose tissue-cultured seedling in greenhouse |
| CN103858771A (en) * | 2014-04-02 | 2014-06-18 | 山东省农业科学院生物技术研究中心 | Maize transgenic tissue culture seedling transplanting method |
| CN109832188A (en) * | 2017-11-27 | 2019-06-04 | 中国农业大学 | A kind of preparation method of corn dihaploid |
Non-Patent Citations (5)
| Title |
|---|
| 光质对组培苗叶绿素含量和碳酸酐酶活性及生长的影响;赵玉国等;《现代农业科学》;20090420(第04期);第16-18页 * |
| 木本植物组培苗的温室炼苗技术;陈彩霞等;《林业实用技术》;20040515(第05期);第22-23页 * |
| 树莓组培苗温室驯化技术;杨玉春等;《新农业》;20091110(第11期);第27-28页 * |
| 玉米组织培养及植株再生研究进展;唐毅成等;《作物研究》;20071215;第629-633页 * |
| 玉米茎尖培养再生体系的建立;陈莉等;《淮阴师范学院学报(自然科学版)》;20060215(第01期);第64-68页 * |
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