CN115211321A - Early identification method for sex of male and female poplar hybrid progeny - Google Patents

Early identification method for sex of male and female poplar hybrid progeny Download PDF

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CN115211321A
CN115211321A CN202211005961.2A CN202211005961A CN115211321A CN 115211321 A CN115211321 A CN 115211321A CN 202211005961 A CN202211005961 A CN 202211005961A CN 115211321 A CN115211321 A CN 115211321A
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seedlings
mass concentration
male
poplar
female
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CN115211321B (en
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丁昌俊
董玉峰
张伟溪
张晓艳
苏晓华
李善文
黄秦军
王延平
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Research Institute of Forestry of Chinese Academy of Forestry
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Research Institute of Forestry of Chinese Academy of Forestry
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Abstract

The invention provides an early identification method of male and female sex of poplar hybrid filial generation, belonging to the technical field of early identification of female and male sex of forest trees. The method promotes the rapid flower bud differentiation of the poplar hybrid progeny single plant through comprehensive measures such as drought stress, plant management, hormone spraying, low-temperature treatment and the like, determines the sex of the hybrid progeny through flower bud observation by utilizing the characteristics of the poplar male and female heteroplants, has simple and effective operation and wider application value, can be used for evaluating the excellent germplasm of poplar hybrid breeding and screening excellent varieties, can greatly shorten the breeding period of the improved variety of the poplars without flying cotton and improve the breeding efficiency. Meanwhile, the method has important reference significance for identifying the sex of the male and female trees of other tree species.

Description

Early identification method for sex of male and female of poplar hybrid progeny
Technical Field
The invention belongs to the technical field of early sex identification of female and male woods, and particularly relates to an early sex identification method for poplar hybrid progeny.
Background
Poplar is a perennial woody plant, generally a female and male heteroplant, forms flower buds in the first growing season under the action of a series of internal and external factors after vegetative growth for 6-10 years, flowers in the second growing season after hibernation, and flowers in the following growing period in spring every year. At present, most varieties planted in poplar production are female plants, and female plant seeds generally float after maturing from middle and last ten days of 4 months to 5 months of each year, namely the float wadding of poplar commonly known by the public. Currently, with the improvement of the requirements of people on environmental conditions and life quality, people have higher and higher call for governing the flying cotton of poplar trees.
There are several methods for treating poplar catkins, such as cutting off and injection of hormone, among which the most economical and effective way is to cultivate and plant poplar species without catkins (male plants). At present, the breeding of new poplar varieties is mainly characterized in that cross pollination is manually controlled, and then evaluation screening and region determination are carried out on filial generations hybridized by different parent combinations, so that new varieties without flying floc are bred. However, poplar is a perennial plant, and generally can bloom in 6-10 years, and how to identify male and female filial generation of flying-floc-free hybrids in the early stage is a technical problem which needs to be solved urgently.
At present, many scholars at home and abroad explore early sex identification of plants from different aspects, mainly including morphological identification, physiological and biochemical identification, isozyme identification, chromosome identification, nucleic acid identification, molecular marker identification and the like. However, in practical application, methods such as morphological and physiological and biochemical identification are mostly focused on the research on sex differences between male and female plants of adult trees, the work on early sex identification at the seedling stage is not enough, the methods such as molecular marking have the problems of complex operation, high technical difficulty, high cost and the like, the large-area popularization and application in the current production are difficult, and the early sex identification technology of plants needs to be further improved.
Disclosure of Invention
In view of this, the invention aims to provide an early identification method for sex of male and female poplar filial generation, and the method provided by the invention is simple and effective to operate, greatly shortens the breeding period of a flying-cotton-free poplar variety and improves the breeding efficiency.
In order to achieve the above purpose, the invention provides the following technical scheme:
the invention provides an early identification method of sex of male and female filial generations of a poplar, which comprises the following steps:
1) Transplanting the filial generation nursery stock of the current-year poplar at the end of 10 months in the current year or at the beginning of 3 months in the second year;
2) Carrying out water control treatment on the poplar hybrid progeny seedlings at the beginning of 6 months in the next year until the growth season is finished, and keeping the soil water content at 25-30%;
3) Removing 1/3-2/3 of newly germinated lateral buds at the lower part of the seedling stem at the beginning of 6 months in the second year, and pinching the new tips at the upper part of the seedling stem; spraying the liquid medicine on the nursery stock for 1 time every 7-10 days after ten days in the middle of 6 months, and simultaneously performing winching at 1/5 of the upper part and the lower part of the nursery stock to reach the xylem;
the liquid medicine contains paclobutrazol with the mass concentration of 0.05-0.1%, boron with the mass concentration of 0.02-0.05%, cytokinin with the mass concentration of 0.02-0.05% and calcium superphosphate with the mass concentration of 0.1-0.2%;
4) Removing all leaves of the seedlings in the last 7 months of the next year, refrigerating at the temperature of minus 3-5 ℃ and the ambient humidity of 60-70% for 20-30 days, and then refrigerating at the temperature of 3-5 ℃ and the ambient humidity of 60-70% for 5-10 days;
5) In 8 th late month of the next year, the seedlings are watered thoroughly and naturally lose water until the soil water content is kept at 30-40%, the seedlings are sprayed with the solution for 1 time, then the seedlings are sprayed for 1 time every 7-10 days, and the male and female sexes of the filial generations of the poplar hybrid are identified according to the flower bud differentiation condition of the seedlings;
the solution contains brassinolide with the mass concentration of 0.002-0.003 percent, compound sodium nitrophenolate with the mass concentration of 0.03-0.05 percent, gibberellin with the mass concentration of 0.05-0.1 percent, monopotassium phosphate with the mass concentration of 0.2-0.5 percent and boron with the mass concentration of 0.02-0.05 percent.
Preferably, the rice diameter of the filial generation seedlings of the poplar hybrid in the current year in the step 1) is 1.5-2.5 cm.
Preferably, the rooting agent is used for irrigating the roots for 1 time after the transplanting in the step 1), the rooting agent is obtained by mixing an indoleacetic acid solution with the concentration of 100-150 mg/L and a naphthylacetic acid solution with the concentration of 100-150 mg/L according to a volume ratio of 1, and 800-1000 ml of the rooting agent is used for irrigating the roots for 1 time.
Preferably, the nursery stock in the step 4) is refrigerated after being sealed by a plastic bag.
Preferably, the illumination time during the refrigeration treatment in the step 4) is 10-12 h/d, and the illumination intensity is 3000-3500 lx.
Preferably, the step 3) is performed by using iron wires.
Preferably, when no flower bud is differentiated in the step 5), repeating the steps 2) and 3) until the seedlings overwinter after natural leaf falling, and identifying the sex of the filial generation of the poplar according to the flower bud differentiation condition of the seedlings in 3 months in the third year.
The beneficial effects of the invention are as follows:
the method promotes the rapid flower bud differentiation of the poplar hybrid progeny single plant through comprehensive measures such as drought stress, plant management, hormone spraying, low-temperature treatment and the like, determines the sex of the hybrid progeny through flower bud observation by utilizing the characteristics of the poplar male and female heteroplants, has simple and effective operation and wider application value, can be used for evaluating the excellent germplasm of poplar hybrid breeding and screening excellent varieties, can greatly shorten the breeding period of the improved variety of the poplars without flying cotton and improve the breeding efficiency. Meanwhile, the method has important reference significance for identifying the sex of the male and female trees of other tree species.
Drawings
FIG. 1 shows pot growth 6 months prior;
FIG. 2 shows nursery stocks in a continuous drought treatment;
FIG. 3 shows the comparison of the nursery stock treated by continuous drought and the normal nursery stock;
FIG. 4 shows that the flower buds of the treated seedlings have differentiated.
Detailed Description
The invention provides an early identification method of sex of male and female filial generations of a poplar, which comprises the following steps:
1) Transplanting the filial generation seedlings of the current-year poplar at the end of 10 months or 3 months in the second year;
2) Carrying out water control treatment on the poplar hybrid progeny seedlings at the beginning of 6 months in the next year until the growth season is finished, and keeping the soil water content at 25-30%;
3) Erasing newly-germinated lateral buds 1/3-2/3 of the lower part of the seedling stem in the first 6 months of the second year, and pinching the new tips of the upper part of the seedling stem; spraying the liquid medicine on the nursery stock for 1 time every 7-10 days after ten days in the middle of 6 months, and simultaneously performing winching at 1/5 of the upper part and the lower part of the nursery stock to reach the xylem;
the liquid medicine contains paclobutrazol with the mass concentration of 0.05-0.1%, boron with the mass concentration of 0.02-0.05%, cytokinin with the mass concentration of 0.02-0.05% and calcium superphosphate with the mass concentration of 0.1-0.2%;
4) Removing all leaves of the seedlings in the last 7 months of the next year, refrigerating at the temperature of minus 3-5 ℃ and the ambient humidity of 60-70% for 20-30 days, and then refrigerating at the temperature of 3-5 ℃ and the ambient humidity of 60-70% for 5-10 days;
5) In 8 th late month of the next year, the seedlings are watered thoroughly and naturally lose water until the soil water content is kept at 30-40%, the seedlings are sprayed with the solution for 1 time, then the seedlings are sprayed for 1 time every 7-10 days, and the male and female sexes of the filial generations of the poplar hybrid are identified according to the flower bud differentiation condition of the seedlings;
the solution contains brassinolide with the mass concentration of 0.002-0.003 percent, compound sodium nitrophenolate with the mass concentration of 0.03-0.05 percent, gibberellin with the mass concentration of 0.05-0.1 percent, monopotassium phosphate with the mass concentration of 0.2-0.5 percent and boron with the mass concentration of 0.02-0.05 percent.
The method transplants the filial generation seedlings of the current-year poplar hybrid at the end of 10 months or 3 months in the second year.
In the invention, the rice diameter of the filial generation seedlings of the current-year poplar hybrid is preferably 1.5-2.5 cm, the specification is convenient for pot culture operation and transportation in the later test process, and meanwhile, the seedlings have good lignification degree, certain resistance and convenience for development of resistance tests, and have advantages. In the invention, the root of the seedling of the current-year poplar hybrid progeny is preferably provided with a soil ball, the diameter of the soil ball is slightly smaller than the inner diameter of a container into which the seedling is moved, and the height of the soil ball is preferably 6-10 cm. The container of the present invention is not particularly limited, and a conventional container such as a lightweight plastic container may be used. In the invention, the bottom and the periphery of the container are preferably filled with grass carbon and soil in a volume ratio of 2. After the transplanting is finished, preferably cutting off the top of the part with weak lignification degree of the selected part of the seedling, and keeping the height of the seedling at 2.0-2.5 m so as to improve the transplanting survival rate.
The method carries out water control treatment on the poplar hybrid progeny seedlings at the beginning of 6 months in the next year until the growth season is finished, and keeps the soil moisture content at 25-30%. The method preferably places the seedlings in the container in a ventilated shed which can shade sun and rain at the beginning of 6 months, controls water for the potted seedlings, keeps the water content of soil at 25% -30%, ensures that young tips and leaves of the hybrid progeny transplanted seedlings droop at a rate of 14-00, and recovers to a normal state after sunset, wherein the water control is to simulate soil drought stress and induce the plants to transform from vegetative growth to reproductive development.
In the invention, newly-germinated lateral buds at 1/3-2/3 of the lower part of a seedling stem are removed at the beginning of 6 months in the second year, and the new tips at the upper part of the seedling stem are picked; spraying the liquid medicine on the nursery stock for 1 time at intervals of 7-10 days after the middle ten days of 6 months until the whole growing season is finished, and simultaneously twisting the nursery stock at 1/5 of the lower part of the height of the nursery stock to the xylem; the liquid medicine contains paclobutrazol with the mass concentration of 0.05-0.1%, boron with the mass concentration of 0.02-0.05%, cytokinin with the mass concentration of 0.02-0.05% and calcium superphosphate with the mass concentration of 0.1-0.2%. The invention preferably uses iron wire for the twisting. The treatments of drug application, low temperature and the like are all to promote reproductive development and flower bud differentiation.
In 7-month last ten days of the next year, all leaves of nursery stocks are removed, the nursery stocks are refrigerated for 20-30 days at the temperature of minus 3-5 ℃ and the ambient humidity of 60-70%, and then refrigerated for 5-10 days at the temperature of 3-5 ℃ and the ambient humidity of 60-70%, low-temperature treatment is a necessary condition for flower bud differentiation, low temperature in winter is artificially simulated, and flower bud differentiation is induced to be advanced. In the invention, the nursery stock is preferably refrigerated after being sealed by a plastic bag. In the present invention, the cold storage treatment is pre-selected in a modified atmosphere. In the present invention, the light irradiation time during the cold storage treatment is preferably 10 to 12h/d, and the light irradiation intensity is preferably 3000 to 3500lx.
In 8 th late month of the next year, the seedlings are watered thoroughly and naturally lose water until the water content of the soil is kept at 30-40%, the seedlings are sprayed with the solution for 1 time, then the seedlings are sprayed for 1 time every 7-10 days, and the male and female sexes of the filial generations of the poplar hybrid are identified according to the flower bud differentiation condition of the seedlings; the solution contains brassinolide with the mass concentration of 0.002-0.003 percent, compound sodium nitrophenolate with the mass concentration of 0.03-0.05 percent, gibberellin with the mass concentration of 0.05-0.1 percent, monopotassium phosphate with the mass concentration of 0.2-0.5 percent and boron with the mass concentration of 0.02-0.05 percent.
In the invention, when no flower bud differentiation exists, the steps 2) and 3) are repeated until the nursery stock overwinter after natural leaf falling, and in 3 months in the third year, the sex of the filial generation of the poplar is identified according to the flower bud differentiation condition of the nursery stock. The present invention preferably repeats steps 2) and 3) in ten days of 9 months.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
In 2018, a cuttage and grafting clone line of hybrid combined filial generation super seedlings with red poplar as a female parent and European black poplar No. 6 as a male parent is used as a material, an early identification test of male and female sex is carried out, the hybrid filial generation reference seedling is 12 seedlings, and the specific steps are as follows:
(1) Preparing nursery stocks: taking the current-year seedlings of the poplar hybrid offspring seedlings to be subjected to sex identification for field cuttage, selecting cuttage seedlings with the rice diameter of 1.8cm, transplanting the seedlings at the beginning of 10 th month, carrying soil balls, wherein the diameter of the soil balls is slightly smaller than the inner diameter and the height of a container to be transplanted, and 6-10 cm, filling grass peat and soil which are mixed according to the volume ratio of 2. After the transplanting is finished, the part with weak lignification degree of the selected part of the seedling is cut off, and the height of the seedling is kept at 2.5m, so that the transplanting survival rate is improved. And (5) normal seedling culture measure management.
(2) Controlling water for nursery stocks: and (3) placing the container seedlings in a ventilated shed capable of shading sun and rain at the beginning of 6 months, controlling water of the potted seedlings, keeping the water content of soil at 25%, allowing young tips and leaves of the hybrid offspring transplanted seedlings to droop at a rate of 14%, and recovering to a normal state after sunset.
(3) And (3) seedling management: pruning the cuttage seedlings from the beginning of 6 months, timely erasing 1/3-2/3 of newly germinated lateral buds at the lower parts of the seedlings, and simultaneously picking new shoots at the upper parts of the seedlings. Spraying liquid medicine to the seedlings every 7 th in the middle of 6 months, wherein the liquid medicine comprises paclobutrazol (with the mass concentration of 0.05%), boron (with the mass concentration of 0.02%), cytokinin (with the mass concentration of 0.02%) and calcium superphosphate (with the mass concentration of 0.1%), and twisting the seedlings by iron wires at 1/5 of the height and the bottom, deep and xylem of the seedlings in the middle of 6 months.
(4) Low-temperature treatment: in the last 7 th month, removing all leaves on the potted seedlings, sealing the potted seedlings by using plastic bags, and transferring the potted seedlings into a modified atmosphere storage, wherein the temperature of the modified atmosphere storage is kept at minus 3 ℃, the humidity is kept at 60%, the temperature of the modified atmosphere storage is increased to 3 ℃, the temperature of the modified atmosphere storage is kept for 5 days, the illumination time of the modified atmosphere storage is kept at 10h/d, and the illumination intensity is 3500lx; and in the middle and last ten days of 8 months, the potted nursery stock is moved out of the refrigeration house and then is put into a ventilated shed for shading sun and rain.
(5) Field management and male and female observation: after the nursery stocks in late 8 months are put into a ventilated shed for shading sun and rain, the potted nursery stocks are watered thoroughly, then the water content of the soil is naturally maintained at 30% -40%, and liquid medicine is sprayed on the nursery stocks, wherein the liquid medicine is brassinolide (with the mass concentration of 0.002%%), compound sodium nitrophenolate (with the mass concentration of 0.03%), gibberellin (with the mass concentration of 0.05%), potassium dihydrogen phosphate (with the mass concentration of 0.2%) and boron (with the mass concentration of 0.02%), and then the liquid medicine is sprayed every 7 days. Meanwhile, the flower bud differentiation conditions of the seedlings of the hybrid filial generation of different poplars are observed, and the sex of the male and female of the hybrid filial generation can be identified according to the female inflorescence and the male inflorescence.
(6) And (3) low-temperature management: and (3) for the seedlings of the poplar hybrid progeny without flower bud differentiation, spraying the leaves, controlling water, twisting with iron wires from the last 9 th month according to the operation in the step (3), watering thoroughly immediately after the seedlings grow to normal fallen leaves, burying a pot culture container in soil, enabling the seedlings to normally overwinter outdoors, and identifying the sex of the seedlings according to differentiated male and female flower buds when the seedlings germinate in the last 3 rd month of the seedlings in the spring of the next year.
The results were: 9 male and female sexes are identified, wherein the identification percentage of the female plant is 75 percent, the male plant is 4, and the female plant is 5.
Example 2
In 2018, a cuttage and grafting clone line of hybrid combined filial generation super seedlings with L324 poplar as a female parent and Lulin No. 16 poplar as a male parent is used as a material, an early identification test of sex of male and female is carried out, the filial generation reference seedling is 14 plants, and the specific steps are as follows:
(1) Preparing nursery stocks: the method comprises the following steps of taking the current-year seedlings of poplar hybrid offspring to be subjected to sex identification to be subjected to field cuttage, selecting cuttage seedlings with the diameter of 1.8cm, transplanting the seedlings at the beginning of 10 th, carrying soil balls, wherein the diameter of the soil balls is slightly smaller than the inner diameter and the height of a container to be transplanted to be 6-10 cm, filling grass peat and soil which are mixed according to the volume ratio of 2. After the transplanting is finished, the part with weak lignification degree of the selected part of the seedling is cut off, and the height of the seedling is kept to be 2.0-2.5 m, so that the transplanting survival rate is improved. And (5) normal seedling culture measure management.
(2) Controlling water for nursery stocks: placing the seedlings in the container in a ventilated shed capable of shading sun and rain at the beginning of 6 months, controlling water for the potted seedlings, keeping the water content of soil at 25% -30%, leading young tips and leaves of the hybrid offspring transplanted seedlings to droop at a ratio of 14-00, and restoring to a normal state after sunset.
(3) And (3) seedling management: pruning the cuttage seedlings from the beginning of 6 months, timely erasing 1/3-2/3 of newly germinated lateral buds at the lower parts of the seedlings, and simultaneously picking new shoots at the upper parts of the seedlings. Spraying liquid medicine to the nursery stock every 7-10 days from the middle of 6 months, wherein the liquid medicine is paclobutrazol (mass concentration of 0.05%), boron (mass concentration of 0.02%), cytokinin (mass concentration of 0.02%), calcium superphosphate (mass concentration of 0.1%), and twisting with iron wires from the middle of 6 months to the 1/5 part of the height and depth of the nursery stock to the xylem.
(4) Low-temperature treatment: in the last 7 th month, removing all leaves on the potted seedlings, sealing the potted seedlings by using plastic bags, and transferring the potted seedlings into a modified atmosphere storage, wherein the temperature of the modified atmosphere storage is kept at minus 3 ℃, the humidity is kept at 60%, the temperature of the modified atmosphere storage is increased to 5 ℃, the temperature of the modified atmosphere storage is kept for 7 days, the illumination time of the modified atmosphere storage is kept at 10h/d, and the illumination intensity is 3500lx; and in the middle and last ten days of 8 months, the potted nursery stock is moved out of the refrigeration house and then is put into a ventilated shed for shading sun and rain.
(5) Field management and male and female observation: after the nursery stocks in late 8 months are put into a ventilated shed for shading sun and rain, the potted nursery stocks are watered thoroughly, then the water content of the soil is naturally maintained at 30%, and the nursery stocks are sprayed with liquid medicine, wherein the liquid medicine is brassinolide (with the mass concentration of 0.002%%), compound sodium nitrophenolate (with the mass concentration of 0.03%), gibberellin (with the mass concentration of 0.05%), potassium dihydrogen phosphate (with the mass concentration of 0.2%) and boron (with the mass concentration of 0.02%), and then the liquid medicine is sprayed every 7 days. Meanwhile, the flower bud differentiation conditions of the seedlings of the hybrid filial generation of different poplar trees are observed, and the male and female sexes of the hybrid filial generation can be identified according to the female inflorescence and the male inflorescence.
(6) And (3) low-temperature management: and (3) for the seedlings of the filial generation of the poplar without flower bud differentiation, spraying on leaves, controlling water and twisting with iron wires from the last ten days of 9 months according to the operation in the step (3), immediately watering until the seedlings grow to normal leaves, burying a pot culture container in the soil, enabling the seedlings to normally overwinter outdoors, and identifying the sex of the seedlings according to the differentiated male and female flower buds when the seedlings germinate in the last 3 months of the seedlings in the next spring.
The results were: the sex of 11 male and female plants is identified, wherein the identification percentage of the female plant is 78.6 percent, the identification percentage of the male plant is 5 percent and the female plant is 6 percent.
Example 3
In 2018, a cuttage and grafting clone material of hybrid combined filial generation super seedlings with silver gland poplar No. 1 as a female parent and Xinjiang poplar as a male parent is used, an early identification test of male and female sex is carried out, the hybrid filial generation reference seedlings are 14 plants, and the method specifically comprises the following steps:
(1) Preparing nursery stocks: taking the current-year seedlings of the poplar hybrid offspring seedlings to be subjected to sex identification for field cuttage, selecting cuttage seedlings with the rice diameter of 1.8cm, transplanting the seedlings at the beginning of 10 th, carrying soil balls, wherein the diameter of the soil balls is slightly smaller than the inner diameter and the height of a container to be transplanted to the container to be 6cm, filling grass peat and soil mixed according to the volume ratio of 2. After the transplanting is finished, the part with weak lignification degree of the selected part of the seedling is cut off, and the height of the seedling is kept to be 2.0-2.5 m, so that the transplanting survival rate is improved. And (5) normal seedling culture measure management.
(2) Controlling water for nursery stocks: placing the container seedlings in a ventilated shed capable of shading sun and rain at the beginning of 6 months, controlling water of the potted seedlings, keeping the water content of soil at 25% -30%, enabling young tips and leaves of the hybrid progeny seedlings to droop at 14% and enabling the seedlings to recover to a normal state after sunset.
(3) And (3) seedling management: pruning the cuttage seedlings from the beginning of 6 months, timely erasing 1/3-2/3 of newly-germinated lateral buds at the lower parts of the seedlings, and simultaneously picking up new tips at the upper parts of the seedlings. Spraying liquid medicine to the nursery stock every 7-10 days from the middle of 6 months, wherein the liquid medicine is paclobutrazol (mass concentration of 0.05%), boron (mass concentration of 0.02%), cytokinin (mass concentration of 0.02%), calcium superphosphate (mass concentration of 0.1%), and twisting with iron wires from the middle of 6 months to the 1/5 part of the height and depth of the nursery stock to the xylem.
(4) Low-temperature treatment: in the last 7 th month, removing all leaves on the potted seedlings, sealing the potted seedlings by using plastic bags, and then transferring the potted seedlings into a modified atmosphere storage, wherein the temperature of the modified atmosphere storage is kept at minus 3 ℃, the humidity is kept at 60%, the temperature of the modified atmosphere storage is increased to 3 ℃, the temperature of the modified atmosphere storage is kept for 5 days, the illumination time of the modified atmosphere storage is kept at 10h/d, and the illumination intensity is 3200lx; and in the middle and last ten days of 8 months, the potted nursery stock is moved out of the refrigeration house and then is put into a ventilated shed for shading sun and rain.
(5) Field management and male-female observation: after the seedlings are put into a ventilated shed for shading sun and sheltering rain again in late 8 months, the potted seedlings are watered thoroughly, then the seedlings naturally lose water until the water content of the soil is kept between 30% and 40%, and liquid medicine is sprayed on the seedlings, wherein the liquid medicine is brassinolide (with the mass concentration of 0.002%), compound sodium nitrophenolate (with the mass concentration of 0.03%), gibberellin (with the mass concentration of 0.05%), potassium dihydrogen phosphate (with the mass concentration of 0.2%) and boron (with the mass concentration of 0.02%), and then the liquid medicine is sprayed once every 7 days. Meanwhile, the flower bud differentiation conditions of the seedlings of the hybrid filial generation of different poplar trees are observed, and the male and female sexes of the hybrid filial generation can be identified according to the female inflorescence and the male inflorescence.
(6) And (3) low-temperature management: and (3) for the seedlings of the poplar hybrid progeny without flower bud differentiation, spraying the leaves, controlling water, twisting with iron wires from the last 9 th month according to the operation of the step (4), watering thoroughly immediately after the seedlings grow to normal fallen leaves, burying a pot culture container in soil, enabling the seedlings to normally overwinter outdoors, and identifying the sex of the seedlings according to differentiated male and female flower buds when the seedlings germinate in the last 3 rd month of the seedlings in the spring of the next year.
The results were: and 12 male and female sexes are identified, wherein the identification percentage of the female plant is 85.7 percent, and the identification percentage of the male plant is 5.
Example 4
In 2018, a cuttage and grafting clone system of hybrid combined filial generation super seedlings with the silver gland poplar No. 2 as a female parent and Xinjiang poplar as a male parent is used as a material, an early identification test of male and female sex is carried out, the hybrid filial generation tested seedlings are 16 plants, and the method specifically comprises the following steps:
(1) Preparing nursery stocks: taking a current-year seedling of a poplar hybrid offspring seedling to be subjected to sex identification for field cuttage, selecting a cuttage seedling with the diameter of 1.5-2.5 cm, planting at the bottom of 10, carrying a soil ball, wherein the diameter of the soil ball is slightly smaller than the inner diameter and the height of a container to be transplanted to 6-10 cm, filling the bottom and the periphery of the container with grass carbon and soil which are mixed according to the volume ratio of 2. And after the transplanting is finished, cutting off the top of the part with weak lignification degree of the selected part on the seedling, and keeping the height of the seedling to be 2.0-2.5 m so as to improve the transplanting survival rate. And (5) normal seedling culture measure management.
(2) Controlling water for nursery stocks: placing the container seedlings in a ventilated shed capable of shading sun and rain at the beginning of 6 months, controlling water of the potted seedlings, keeping the water content of soil at 25% -30%, enabling young tips and leaves of the hybrid progeny seedlings to droop at 14% and enabling the seedlings to recover to a normal state after sunset.
(3) And (3) seedling management: pruning the cuttage seedlings from the beginning of 6 months, timely erasing 1/3-2/3 of newly-germinated lateral buds at the lower parts of the seedlings, and simultaneously picking up new tips at the upper parts of the seedlings. Spraying liquid medicine to the nursery stocks every 7-10 days from the middle of 6 months, wherein the liquid medicine is paclobutrazol (mass concentration is 0.05%), boron (mass concentration is 0.02%), cytokinin (mass concentration is 0.02%), calcium superphosphate (mass concentration is 0.1%), and the liquid medicine is stranded on 1/5 of the lower part of the seedling height and deep to xylem of the nursery stocks by using iron wires at the same time from the middle of 6 months.
(4) Low-temperature treatment: in the last 7 th month, removing all leaves on the potted seedlings, sealing the potted seedlings by using plastic bags, and then transferring the potted seedlings into a modified atmosphere storage, wherein the temperature of the modified atmosphere storage is kept at minus 3 ℃, the humidity is kept at 60%, the temperature of the modified atmosphere storage is increased to 3 ℃, the temperature of the modified atmosphere storage is kept for 5 days, the illumination time of the modified atmosphere storage is kept at 10h/d, and the illumination intensity is 3200lx; and in the middle and last ten days of 8 months, the potted nursery stock is moved out of the refrigeration house and then is put into a ventilated shed for shading sun and rain.
(5) Field management and male and female observation: after the nursery stocks in late 8 months are put into a ventilated shed for shading sun and rain, the potted nursery stocks are watered thoroughly, then the water content is naturally reduced to 30% -40%, and the nursery stocks are sprayed with liquid medicine, wherein the liquid medicine is brassinolide (with the mass concentration of 0.002%), sodium nitrophenolate (with the mass concentration of 0.03%), gibberellin (with the mass concentration of 0.05%), potassium dihydrogen phosphate (with the mass concentration of 0.2%) and boron (with the mass concentration of 0.02%), and then the liquid medicine is sprayed every 7 days. Meanwhile, the flower bud differentiation conditions of the seedlings of the hybrid filial generation of different poplars are observed, and the sex of the male and female of the hybrid filial generation can be identified according to the female inflorescence and the male inflorescence.
(6) And (3) low-temperature management: and (3) for the seedlings of the poplar hybrid progeny without flower bud differentiation, spraying the leaves, controlling water, twisting with iron wires from the last 9 th month according to the operation of the step (4), watering thoroughly immediately after the seedlings grow to normal fallen leaves, burying a pot culture container in soil, enabling the seedlings to normally overwinter outdoors, and identifying the sex of the seedlings according to differentiated male and female flower buds when the seedlings germinate in the last 3 rd month of the seedlings in the spring of the next year.
The results were: 13 male and female sexes are identified in total, wherein 8 female plants, 5 male plants and the identification percentage of the male and female plants is 81.3 percent.
The embodiment can show that the identification method provided by the invention obviously shortens the identification period and improves the breeding efficiency.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims (7)

1. The method for early identifying the sex of the male and female parents of the poplar hybrid offspring is characterized by comprising the following steps of:
1) Transplanting the filial generation seedlings of the current-year poplar at the end of 10 months or 3 months in the second year;
2) Carrying out water control treatment on the poplar hybrid progeny seedlings at the beginning of 6 months in the next year until the growth season is finished, and keeping the soil water content at 25-30%;
3) Erasing newly-germinated lateral buds 1/3-2/3 of the lower part of the seedling stem in the first 6 months of the second year, and pinching the new tips of the upper part of the seedling stem; spraying the liquid medicine on the nursery stock for 1 time every 7-10 days after ten days in the middle of 6 months, and simultaneously performing winching at 1/5 of the upper part and the lower part of the nursery stock to reach the xylem;
the liquid medicine contains paclobutrazol with the mass concentration of 0.05-0.1%, boron with the mass concentration of 0.02-0.05%, cytokinin with the mass concentration of 0.02-0.05% and calcium superphosphate with the mass concentration of 0.1-0.2%;
4) Removing all leaves of the seedlings in the last 7 months of the next year, refrigerating at the temperature of minus 3-5 ℃ and the ambient humidity of 60-70% for 20-30 days, and then refrigerating at the temperature of 3-5 ℃ and the ambient humidity of 60-70% for 5-10 days;
5) In 8 th late month of the next year, the seedlings are watered thoroughly and naturally lose water until the soil water content is kept at 30-40%, the seedlings are sprayed with the solution for 1 time, then the seedlings are sprayed for 1 time every 7-10 days, and the male and female sexes of the filial generations of the poplar hybrid are identified according to the flower bud differentiation condition of the seedlings;
the solution contains brassinolide with the mass concentration of 0.002-0.003%, compound sodium nitrophenolate with the mass concentration of 0.03-0.05%, gibberellin with the mass concentration of 0.05-0.1%, monopotassium phosphate with the mass concentration of 0.2-0.5% and boron with the mass concentration of 0.02-0.05%.
2. The early stage identification method according to claim 1, wherein the rice diameter of the offspring seedling of the current-year poplar hybrid in step 1) is 1.5-2.5 cm.
3. The early identification method according to claim 1, wherein the rooting agent is used for root irrigation for 1 time after the transplanting in the step 1), the rooting agent is obtained by mixing an indoleacetic acid solution with the concentration of 100-150 mg/L and a naphthylacetic acid solution with the concentration of 100-150 mg/L according to the volume ratio of 1.
4. The early stage identification method according to claim 1, wherein the nursery stock in step 4) is sealed with a plastic bag and then subjected to a cold storage treatment.
5. The early stage identification method according to claim 1, wherein the light irradiation time during the cold storage treatment in step 4) is 10 to 12h/d, and the light irradiation intensity is 3000 to 3500lx.
6. The early identification method according to claim 1, wherein the step 3) comprises performing the twist-off using a wire.
7. The early stage identification method according to claim 1, wherein when no flower bud is differentiated in the step 5), the steps 2) and 3) are repeated until the seedling overwinter after natural leaf falling, and in the third year for 3 months, the male and female sex of the filial generation of the poplar is identified according to the flower bud differentiation condition of the seedling.
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