CN115530073A - Ulmus parvifolia grafting propagation method through callus adventitious bud regeneration - Google Patents
Ulmus parvifolia grafting propagation method through callus adventitious bud regeneration Download PDFInfo
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- CN115530073A CN115530073A CN202211164664.2A CN202211164664A CN115530073A CN 115530073 A CN115530073 A CN 115530073A CN 202211164664 A CN202211164664 A CN 202211164664A CN 115530073 A CN115530073 A CN 115530073A
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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
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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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
- A01G2/00—Vegetative propagation
- A01G2/30—Grafting
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/22—Improving land use; Improving water use or availability; Controlling erosion
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/40—Afforestation or reforestation
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Abstract
The invention discloses a ulmus parvifolia grafting propagation method through callus adventitious bud regeneration, which comprises the following steps of: step one, collecting dormant branches of ulmus parvifolia as scions, wherein the scions do not need buds; step two, before grafting, soaking the upper surface of the scion in callus induction liquid; taking the ulmus parvifolia seedlings as stocks, and grafting the scions in the step two; in the binding process, the scion is completely wrapped by the grafting film; after grafting is finished, carrying out heat preservation and light-shading treatment on the scion by using a heat preservation and light-shading material; step five, removing the heat-preservation shading material to induce callus to differentiate adventitious buds 15-30 days after grafting; and step six, after the adventitious bud differentiation is completed, timely cutting a grafting film on the top of the scion to promote the bud growth. The invention innovates the regeneration mode of the scion in the grafting process and gets rid of the dependence of the existing asexual propagation technologies such as grafting, cuttage and the like on the original bud body of the scion.
Description
Technical Field
The invention belongs to the field of forest seedling cultivation, relates to a grafting propagation method, and particularly relates to a ulmus parvifolia grafting propagation method through callus adventitious bud regeneration.
Background
Ulmus parvifolia Jacq, ulmaceae Ulmus L, is a big tree of fallen leaves, mainly distributed in China, selands in Korea and Japan, is an important greening tree species in the south of China, and meanwhile, the wood is one of the hard materials in China. The domestic Ulmus parvifolia resource is mainly used for bonsai manufacture and greening decoration, is directly obtained by field excavation, and is added with urban development and agricultural production activities, so that the field survival quantity is sharply reduced, and the protection, development and utilization are urgently needed. Therefore, the development of the elm resource protection, fine breed cultivation and other works is particularly urgent. The improved variety breeding of Ulmus parvifolia has been carried out abroad, for example, in the 40 th generation of North America, ulmus parvifolia and Ulmus parvifolia are introduced, which are mainly used for carrying out disease-resistant breeding of Ulmus parvifolia, and 11 fast-growing, dry and straight-through Ulmus parvifolia varieties with beautiful leaf shapes or strong disease resistance are successfully cultivated at present. The Luzhou et al (2020) determine the seedling height and the ground diameter of 20 Ulmus parvifolia grafting clone in Jiangsu region through the seedling stage, the differences reach extremely remarkable levels, and the clone JN-02 and JD-08 have abundant genetic variation, and can provide materials for afforestation tests. The asexual propagation of garden burnet mainly adopts technologies such as grafting, cuttage and the like, for example, the study on hard branch cuttage of garden burnet is carried out by Chenpisten et al (2015), and the study on tender branch cuttage of garden burnet is carried out by Zhoujie et al (2021), which shows that the cuttage technology can be used as a technical means for the asexual propagation of garden burnet.
However, compared with grafting propagation, cutting propagation has higher requirements on tissue materials and facility conditions, and the growth amount of the regenerated plants is small, so that the cutting propagation is not used in seedling growth at present. The traditional asexual propagation techniques such as grafting and cutting have high requirements on scions (cutting slips), and full buds with complete development are required to germinate regenerated plants. In the actual production process, particularly in the field excellent germplasm resource collection process, due to the fact that the small branches of the Ulmus parvifolia droop, the materials suitable for grafting or cuttage are difficult to collect, and the asexual propagation and preservation of excellent germplasm are hindered. Therefore, the grafting propagation method which does not need full axillary buds but is differentiated by adventitious buds is developed, the problem of scion collection is solved, the rapid growth of asexual propagation regeneration plants is guaranteed, the dependence of asexual propagation on scion quality is solved, and the grafting propagation method has important significance on asexual propagation of ulmus parvifolia and other tree species.
Disclosure of Invention
The invention provides a ulmus parvifolia grafting propagation method through callus adventitious bud regeneration, which aims to solve the problems that scions are required to have full buds and the like in the prior art.
In order to achieve the purpose, the invention provides a ulmus parvifolia grafting propagation method through callus adventitious bud regeneration, which is characterized in that: the method comprises the following steps:
step one, collecting dormant branches of ulmus parvifolia as scions, wherein the scions do not need buds;
step two, before grafting, soaking the upper surface of the scion in callus induction liquid;
taking the ulmus parvifolia seedlings as stocks, and taking the scions in the second step for grafting; in the binding process, the scion is completely wrapped by the grafting film;
after grafting is finished, carrying out heat preservation and light-shading treatment on the scion by using a heat preservation and light-shading material;
step five, removing the heat-preservation shading material to induce callus to differentiate adventitious buds 15-30 days after grafting;
and step six, after the adventitious bud differentiation is completed, timely cutting off a grafting film at the top of the scion to promote the bud growth.
Further, the invention provides a ulmus parvifolia grafting propagation method through callus adventitious bud regeneration, and the method can also have the following characteristics: wherein, the time for collecting the branches in the first step is 1 to 2 months per year.
Further, the invention provides a ulmus parvifolia grafting propagation method through callus adventitious bud regeneration, and the method can also have the following characteristics: wherein, the grafting time in the third step is 3-4 months in the current year.
Further, the invention provides a ulmus parvifolia grafting propagation method through callus adventitious bud regeneration, and the method can also have the following characteristics: wherein, in the first step, the length of the scion is 12-15 cm, and the thickness is 0.5-2.0 cm.
Further, the invention provides a ulmus parvifolia grafting propagation method through callus adventitious bud regeneration, and the method can also have the following characteristics: wherein, in the step one, the collected branches are sealed by wax and then stored at 4 ℃.
Further, the invention provides a ulmus parvifolia grafting propagation method through callus adventitious bud regeneration, and the method can also have the following characteristics: wherein in the second step, the callus induction liquid is a mixed liquid of 0.5ppm to 5.0ppm of naphthylacetic acid and 0.5ppm to 5.0ppm of 6-benzylaminopurine.
Further, the invention provides a ulmus parvifolia grafting propagation method through callus adventitious bud regeneration, which can also have the following characteristics: wherein, in the second step, the soaking time is 30 min-2 h.
Further, the invention provides a ulmus parvifolia grafting propagation method through callus adventitious bud regeneration, and the method can also have the following characteristics: wherein, in the third step, the Ulmus parvifolia seedlings are 1-2 years old seedlings, and the ground diameter is 0.6-3.0 cm.
Further, the invention provides a ulmus parvifolia grafting propagation method through callus adventitious bud regeneration, which can also have the following characteristics: wherein, in the third step, the scion is cut into 5.0 cm-10.0 cm, preferably 6cm.
Further, the invention provides a ulmus parvifolia grafting propagation method through callus adventitious bud regeneration, which can also have the following characteristics: in the fourth step, the scion is bound by a plastic bag, and the outside is wrapped by shading materials (such as newspaper, black plastic paper and the like) in a shading way.
The invention has the beneficial effects that: the invention provides a ulmus parvifolia grafting propagation method through callus adventitious bud regeneration, which realizes grafting survival through the soaking of callus induction liquid and heat-preservation and light-resistant treatment. After the callus is generated, adventitious buds can be differentiated under natural illumination, the prior grafting technology needs complete buds to germinate to form plants, the buds grow badly, the grafting process is damaged or the grafting is inactivated, so that the grafting fails. Compared with the prior art, the ulmus parvifolia scion selected by the invention does not need buds in the market, and adventitious bud regeneration plants are differentiated from the upper section of the scion through callus, so that the scion regeneration mode in the grafting process is innovated, and the dependence of the conventional asexual propagation technologies such as grafting, cuttage and the like on the original buds of the scion is eliminated.
Drawings
FIG. 1 is a photograph of a living Ulmus parvifolia grafted with a bud-free scion by the method of the present invention;
FIG. 2 is a photograph of a living Ulmus parvifolia with bud scions grafted by conventional grafting methods.
Detailed Description
The invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the description of the embodiments is only for illustrating the present invention and should not be taken as limiting the invention as detailed in the claims.
Experimental materials: the stock is a container seedling of annual Ulmus parvifolia (container specification of 30cm multiplied by 20 cm) in the Experimental forest farm of scientific research institute of forestry in Jiangsu province (Nanjing Jiangning); the scion wood is collected branches of a big ulmus parvifolia tree grown for more than 30 years.
The test method comprises the following steps: in 2022, 1-2 months, surveying and collecting robust branches of excellent wild germplasm resources of Ulmus parvifolia as scions in Yangzhou Jiangdu, wuxi Yangzhou, nanjing Jiangning and the like, sealing with wax, and storing in a cold store at 4 ℃; in 3 middle of the month in 2022, grafting is started when winter buds of Ulmus parvifolia seedlings sprout.
Comparative example
The comparative example adopts the existing grafting propagation method to carry out grafting propagation on ulmus parvifolia.
The specific method comprises the following steps: and 3, in 2022, 16 days after 3 months, respectively selecting scions with full buds and no buds to perform a grafting test, adopting a grafting method without other treatments, treating 30 plants each, and observing the survival condition of the scions in different states for grafting the ulmus parvifolia. And (5) unwinding the bound grafting membrane for 15 days after 4 months, finding that the scions are healed with the rootstocks, and the scions have activity, wherein the germination regeneration plant conditions are shown in table 1.
TABLE 1 grafting situation table with or without bud
As can be seen from table 1, the scions without buds can not be grafted and survived by the conventional grafting propagation method, and only the scions with plump buds can be grafted, i.e. the conventional grafting propagation method has high requirements on the scions.
Examples
The embodiment provides a ulmus parvifolia grafting propagation method through callus adventitious bud regeneration, which comprises the following steps: step one, collecting dormant branches of ulmus parvifolia as scions, wherein the scions do not need buds; step two, before grafting, soaking the upper surface of the scion in callus induction liquid; the callus inducing liquid is a mixed liquid of 0.5ppm to 5.0ppm of naphthylacetic acid (NAA) and 0.5ppm to 5.0ppm of 6-benzylaminopurine (6-BA), and the soaking time is 30min to 2h; taking the ulmus parvifolia seedlings as stocks, and grafting the scions in the step two; in the binding process, the scion is completely wrapped by the grafting film; after grafting is finished, carrying out heat preservation and light-shading treatment on the scion by using a heat preservation and light-shading material; step five, removing the heat-preservation shading material to induce callus to differentiate adventitious buds 13-17 days after grafting; and step six, after the adventitious bud differentiation is completed, timely cutting off a grafting film at the top of the scion to promote the bud growth.
Specifically, the present example observes the effect of different exogenous hormone concentration treatments on the survival of the bud-free scion grafting through four groups of experiments:
setting process 1: NAA 0.5ppm +6-BA 2.0ppm, treatment 2: NAA 0.5ppm +6-BA 0.5ppm, treatment 3: NAA 2.0ppm +6-BA 0.5ppm, and using clear water as Control (CK), soaking the upper end of scion for 30min, grafting, and treating 30 plants each, and making total 120 plants. The test is carried out in 2022, 3 months and 16 days, the scion after grafting is bound by a plastic bag for heat preservation, and the outside is wrapped by newspaper for shading. After 20 days, the newspaper and the plastic bag are untied, the callus generation condition is observed, after 45 days, the callus differentiation bud condition is observed, an opening is cut on the grafting binding grafting film, and the grafting survival rate is counted in 15 days after 6 months, and the results are shown in table 2.
TABLE 2 Effect of different exogenous hormone concentration treatments on the survival of non-budding scions by grafting
According to the results shown in table 2, the heat preservation and light shielding measures can promote the formation of callus on the upper surface of the buddless scion of ulmus parvifolia, and the plants are regenerated through adventitious bud differentiation to realize grafting survival. In addition, the hormone ratio has a large influence on the generation of callus and the regeneration of plants, wherein the callus is formed on the upper surface of the scion with the length of 28 percent in the treatment 2, the generation rate of the callus is up to 93.3 percent, the rate of the callus adventitious bud differentiation regeneration plant is 82.1 percent, and the rates are respectively improved by 7.1 times and 3.3 times compared with a control.
Fig. 1 shows that the ulmus parvifolia survived grafted by the bud-free scion according to the method of the embodiment, fig. 2 shows that the ulmus parvifolia survived grafted by the bud-containing scion according to the conventional grafting method, and as can be seen from fig. 1 and 2, the effect of the method is consistent with that of grafting the bud-containing scion.
Claims (10)
1. A ulmus parvifolia grafting propagation method through callus adventitious bud regeneration is characterized by comprising the following steps:
the method comprises the following steps:
step one, collecting dormant branches of ulmus parvifolia as scions, wherein the scions do not need buds;
step two, before grafting, soaking the upper surface of the scion in callus induction liquid;
taking the ulmus parvifolia seedlings as stocks, and grafting the scions in the step two; in the binding process, the scion is completely wrapped by the grafting film;
after grafting is finished, carrying out heat preservation and light shading treatment on the scion by using a heat preservation and light shading material;
step five, removing the heat-preservation shading material to induce callus to differentiate adventitious buds 15-30 days after grafting;
and step six, after the adventitious bud differentiation is completed, timely cutting off a grafting film at the top of the scion to promote the bud growth.
2. The Ulmus parvifolia grafting propagation method by callus adventitious bud regeneration according to claim 1, wherein:
wherein the time for collecting the branches in the first step is 1-2 months per year.
3. The Ulmus parvifolia grafting propagation method by callus adventitious bud regeneration according to claim 2, wherein:
wherein, the grafting time in the third step is 3-4 months in the current year.
4. The Ulmus parvifolia grafting propagation method by callus adventitious bud regeneration according to claim 1, wherein:
wherein, in the first step, the length of the scion is 12-15 cm, and the thickness is 0.5-2.0 cm.
5. The grafting propagation method of ulmus parvifolia by callus adventitious bud regeneration according to claim 1, characterized in that:
in the first step, the collected branches are sealed by wax and then stored at 4 ℃.
6. The Ulmus parvifolia grafting propagation method by callus adventitious bud regeneration according to claim 1, wherein:
wherein in the second step, the callus induction liquid is a mixed liquid of 0.5ppm to 5.0ppm of naphthylacetic acid and 0.5ppm to 5.0ppm of 6-benzylaminopurine.
7. The Ulmus parvifolia grafting propagation method by callus adventitious bud regeneration according to claim 1, wherein:
wherein, in the second step, the soaking time is 30 min-2 h.
8. The Ulmus parvifolia grafting propagation method by callus adventitious bud regeneration according to claim 1, wherein:
wherein, in the third step, the Ulmus parvifolia seedlings are 1-2 years old seedlings, and the ground diameter is 0.6-3.0 cm.
9. The Ulmus parvifolia grafting propagation method by callus adventitious bud regeneration according to claim 1, wherein:
wherein, in the third step, the scion is cut into 5.0 cm-10.0 cm, preferably 6cm.
10. The grafting propagation method of ulmus parvifolia by callus adventitious bud regeneration according to claim 1, characterized in that:
and in the fourth step, the scion is bound by a plastic bag, and the outside is wrapped by a shading material in a shading way.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1799318A (en) * | 2005-12-16 | 2006-07-12 | 江苏阳光生态农林开发股份有限公司 | Multi-sprout long-tube sleeve joint method for ulmis pumilal |
| CN102640665A (en) * | 2012-04-21 | 2012-08-22 | 桐城市景艺苗木有限公司 | Method improving nursery stock grafting survival rate |
| CN105494014A (en) * | 2015-12-25 | 2016-04-20 | 成都苗夫现代苗木科技有限公司 | Grafting and shaping method of big ulmus parvifolia tree |
| CN106305157A (en) * | 2016-08-29 | 2017-01-11 | 盛林蓝莓集团股份有限公司 | Blueberry grafting method |
| US20190069459P1 (en) * | 2017-08-28 | 2019-02-28 | Southern Selections, Llc | Uamtf |
| CN113207695A (en) * | 2021-06-17 | 2021-08-06 | 江苏省林业科学研究院 | Formula for tissue culture of ulmus parvifolia |
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- 2022-09-23 CN CN202211164664.2A patent/CN115530073B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1799318A (en) * | 2005-12-16 | 2006-07-12 | 江苏阳光生态农林开发股份有限公司 | Multi-sprout long-tube sleeve joint method for ulmis pumilal |
| CN102640665A (en) * | 2012-04-21 | 2012-08-22 | 桐城市景艺苗木有限公司 | Method improving nursery stock grafting survival rate |
| CN105494014A (en) * | 2015-12-25 | 2016-04-20 | 成都苗夫现代苗木科技有限公司 | Grafting and shaping method of big ulmus parvifolia tree |
| CN106305157A (en) * | 2016-08-29 | 2017-01-11 | 盛林蓝莓集团股份有限公司 | Blueberry grafting method |
| US20190069459P1 (en) * | 2017-08-28 | 2019-02-28 | Southern Selections, Llc | Uamtf |
| CN113207695A (en) * | 2021-06-17 | 2021-08-06 | 江苏省林业科学研究院 | Formula for tissue culture of ulmus parvifolia |
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