CN108254510B - Method for rapidly identifying young bud-grafting clonal seedlings of rubber trees - Google Patents
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Abstract
The invention relates to a method for rapidly identifying young bud-grafting clonal seedlings of a rubber tree, which comprises the steps of seedling cultivation, index measurement, index difference analysis, frequency distribution graph drawing, identification index determination and result judgment; the method can be used for quickly identifying the old and young bud-grafting clone seedlings of the same genotype of the rubber tree, is quick and simple, does not need additional equipment and instruments, is low in cost and strong in operability, can quickly and accurately identify whether the bud-grafting clone of the same genotype of the rubber tree is a young bud-grafting clone only from the morphological characteristics of leaves and the extraction angle between scion sprouting and stock, has objective and effective identification result and high accuracy, and is beneficial to improving the purity and the quality of the bud-grafting clone seedlings of the rubber tree; in addition, the method has low requirements on background knowledge and actual operation of operators, and is beneficial to implementation of the identification method.
Description
Technical Field
The invention relates to a method for identifying young bud-grafted clonal seedlings, in particular to a method for quickly identifying young bud-grafted clonal seedlings of a rubber tree.
Background
The Hevea brasiliensis (Hevea brasiliensis) is the species currently known to produce the most gum, native to the Heira river basin, Brazilian in south America. The natural rubber produced by the milk duct is a restricted resource and an important strategic material, and has irreplaceable effects on national defense and economic construction.
When the planting variety is determined in the production of rubber trees, the survival, growth, stress resistance, gum yield and the like of the nursery stocks can be influenced to different degrees by selecting different types of planting materials. Therefore, each planting area is particularly important for selecting high-quality planting materials. In the history of natural rubber development in decades in China (China), the following planting materials have been used in sequence to different degrees: seedlings with or without soil of different sizes; seedling stumps or bud grafting stumps; high-cut stem, medium-cut stem seedling or bud-grafting seedling; the bud grafting belt is dry and passes through the winter bud grafting pile; field bud grafting seedlings; and seedling bud grafting seedling and the like. When the seedlings are used as planting materials, the other used bud-grafting seedlings are old bud-grafting seedlings. The related research shows that the growth speed, the stress resistance, the gum yield and the like of the young plants of the rubber trees are all stronger than those of the old plants. However, the research and utilization of the clone of the young rubber tree still stay in the experimental stage, and the large-scale popularization and planting are not seen. At present, the research and utilization of old young clone of rubber tree still have many problems, such as how to distinguish and link between old young states, the problem that the identification standard and index of old young bud clone are single, chaotic, etc. has not been solved, so the reliability of the identification result is not strong, and the identification process is tedious, and it also needs to rely on instrument and equipment. Therefore, it is highly desirable to find a method for rapidly identifying young bud-grafted clonal seedlings of rubber trees.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a method for rapidly identifying young rubber tree bud clone seedlings, so as to solve the problems of single and disordered identification standard and index, complicated identification process and the like of old and young rubber tree bud clone seedlings, obviously improve the identification speed and enable the identification result to be objective and reliable.
The invention realizes the purpose through the following technical scheme:
(1) and (3) seedling cultivation: under certain standing conditions suitable for rubber tree growth, selecting a rubber tree clone variety, collecting anthers, performing tissue culture to obtain a young root clone, selecting 5 plants with consistent growth vigor after nursery stocks grow for 1 year, and performing shoot budding to obtain a young bud clone; collecting the scions of the old-state bud-grafting clone from the corresponding variety 15-year-old clone big tree, selecting 3 plants for picking, and culturing to obtain the old-state bud-grafting clone;
(2) measurement of the index: measuring by taking plants as units, and respectively measuring and recording the emergence angles of the bud seedlings in the old and young states when the first bud leaves of the bud seedlings are stable; after the seedlings sprout and survive for 4-5 months, respectively adopting middle and lower leaves of the same variety of old young bud seedlings and the same stable leaf fleabane, and measuring and recording the leaf indexes of the seedlings;
(3) and (3) index difference analysis: performing difference analysis on the indexes measured in the step (2), and analyzing whether the differences of the measured indexes among the old and young bud clone are obvious or not;
(4) and (3) drawing a frequency distribution diagram: regarding the indexes with significant difference in the step (3), respectively drawing a frequency distribution graph of old and young bud-grafting clone seedlings of the same variety on each index by taking the index measurement value and the plant frequency distribution as a vertical axis and a horizontal axis;
(5) determination of the authentication index: deeply analyzing the frequency distribution map drawn in the step (4), and selecting indexes which are discontinuously distributed or slightly crossed in the frequency distribution map as indexes for identifying the seedling of the young bud-grafting clone of the rubber tree;
(6) and (4) judging a result: and (5) using the analysis result in the step (5) as a text description as a method for identifying the seedling of the same genotype seedling of the rubber tree in the young bud grafting clone, namely, the seedling of the young bud grafting clone has doubled leaf length and leaf area compared with the seedling of the old bud grafting clone, the leaf vein distribution is thinned, and the extraction angle is less than 25 degrees.
Further, the old and young bud seedling in the step (2) is a plant which grows for more than 4 months and has more than 3 or more than 3 leafy froglets within 1 year after the scion sprouts.
Further, the stable erigeron breviscapus in the step (2) is all erigeron breviscapus except for the erigeron breviscapus leaf and the 1 st erigeron breviscapus leaf from bottom to top.
Further, the extraction angle in the step (2) is an included angle between the scion bud and the stock, and the measuring method is measuring by using a protractor; leaf indices include leaf length, leaf width, leaf area, width base distance/leaf length, vein width/leaf width, vein density and water content.
The invention also aims to provide application of the identification method in quickly identifying the seedling of the young bud-grafting clone of the rubber tree.
Specifically, the application method comprises the following steps: comparing the leaf length, leaf area, leaf vein density and extraction angle of the same genotype old and young bud-grafting clone plant, the young bud-grafting clone plant has longer leaf length, larger leaf area, more sparse leaf vein distribution or extraction angle less than 25 degrees, otherwise, is an old bud-grafting clone seedling.
The invention has the beneficial effects that:
the method can quickly and simply identify the old and young bud-grafting clone seedlings of the same genotype of the rubber tree, does not need additional equipment and instruments, is low in cost and strong in operability, can quickly and accurately identify whether the bud-grafting clone of the same genotype of the rubber tree is a young bud-grafting clone or not only from the morphological characteristics of leaves and the extraction angle between scion sprouting and stock, has objective and effective identification result and high accuracy, and is favorable for improving the purity and the quality of the bud-grafting clone seedlings of the rubber tree.
Drawings
FIG. 1 is a frequency distribution diagram of a rubber tree clone variety hot-reclaimed 525 young bud graft clone (A is a leaf length frequency distribution diagram, B is a leaf area frequency distribution diagram, C is a leaf vein density frequency distribution diagram, and D is a extraction angle frequency distribution diagram);
FIG. 2 is a frequency distribution diagram of the rubber tree clone variety RRII105 old young bud clone plant (A is a leaf length frequency distribution diagram, B is a leaf area frequency distribution diagram, C is a leaf vein density frequency distribution diagram, and D is a drawing angle frequency distribution diagram);
FIG. 3 is a frequency distribution diagram of old and young bud clone plant of the clone variety Yunchang 73-477 of rubber tree (A is a leaf length frequency distribution diagram, B is a leaf area frequency distribution diagram, C is a leaf density frequency distribution diagram, and D is a drawing angle frequency distribution diagram);
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described with reference to the accompanying drawings.
All rubber trees in the following examples were planted in the scenic flood rubber tree germplasm resource nursery of Ministry of agriculture.
Example 1
(1) Selecting a rubber tree clone variety for hot reclamation 525, marking as No. 182, collecting anthers of integrated aged trees for tissue culture to obtain a clone obtained from a root juvenile state, selecting 5 plants with consistent growth vigor after the clone from the root juvenile state grows to 1 year, and collecting strips for bud grafting to obtain a juvenile state bud grafting clone, marking as 182-5; the control is an old bud clone which is bred by cutting and bud grafting from 3 15-year-old big trees and is marked as 182-2. 300 seedlings are budded in each bud of the old and young clone;
(2) when the first canopy leaves of the bud-grafted seedlings are stable, measuring the sprouting angle between the scion and the stock by taking the plants as a unit and using a protractor, and recording in detail, wherein 200 healthy seedlings are selected for measuring for the old and young bud-grafted clone;
(3) after the bud-grafted seedling sprouts and survives for 4 months, measuring the leaf form index by taking plants as units, namely selecting more than 15 healthy plants with more than 3 or more than 3 leafy froglets from each clone, taking (from bottom to top) 10 leaves at the middle-lower part of other leafy froglets except for the roof froglets and the 1 st leafy froglets from each plant, taking the plants back to a laboratory to measure the leaf length, the leaf width, the leaf area, the wide base distance/the leaf length, the vein left width/the leaf width, the density and the water content of the vein one by one, and recording the measured values;
(4) and (3) carrying out difference analysis on the data measured in the steps (2) and (3), and analyzing whether the difference of the measured indexes among the bud clones in the old and young states of the hot reclamation 525 is obvious or not. The results show that 4 indexes of the young bud clone of the hot reclamation 525, such as leaf length, leaf area, leaf vein density, extraction angle and the like, are all remarkably greater than the old bud clone, and the difference among other indexes is not remarkable;
(5) regarding the indexes (leaf length, leaf area, leaf vein density and extraction angle) with significant difference in the step (4), respectively drawing a frequency distribution graph of the thermally cultivated 525 old and young bud clone plants in each index by taking the index measurement numerical value and the plant frequency distribution as a vertical axis and a horizontal axis;
(6) and (5) deeply analyzing the frequency distribution map drawn in the step (5) to find out indexes of discontinuous distribution or a few crossed indexes in the frequency distribution map as indexes for identifying the seedling of the thermally reclaimed 525 seedling bud graft. The results show that the old and young bud clone has discontinuous distribution among 3 indexes such as leaf length, leaf area, leaf vein density and the like, has no cross, and can be used as the identification index of the seedling of the hot-reclaimed 525 young bud clone; the extraction angle has only a very small amount (6.25%) of crossing, and can also be used as an index for identifying the seedling of the young bud grafting clone of the hot reclamation 525. As shown in fig. 1.
As can be seen from FIG. 1, there is no crossing in leaf length, leaf area and leaf density between the young bud clone and the old bud clone of hot-cultivated 525, and the leaf length and leaf area of the young bud clone are much larger than those of the old bud clone, and a discontinuous threshold can be found, respectively 19.5cm and 130cm2(ii) a The density of the veins of the young bud clone is far less than that of the old bud clone, and a discontinuous threshold value can be found out and is 95/m; from the perspective of extraction, although the extraction angle of the young bud clone is slightly crossed with that of the old bud clone, the extraction angle of the young bud clone is more than 80% of the plants with the extraction angle of less than 25 °, and the extraction angle of the old bud clone is more than 90% of the plants with the extraction angle of more than 30 °. Therefore, an extraction angle of less than 25 ° can be used as a criterion for the young bud clone.
(7) The method for identifying the thermally reclaimed 525 young bud clone seedlings can be concluded from the analysis result of the step (6), namely the young bud clone seedlings are longer in leaves and are multiplied in leaf area compared with the old bud clone seedlings, the leaf vein distribution is thinned, and the extraction angle is less than 25 degrees.
Example 2
(1) Selecting rubber tree clone variety RRII105, marking as No. 223, adopting anther of integrated age tree to make tissue culture to obtain clone obtained from root juvenile state, after the clone obtained from root juvenile state is grown to 1 year, selecting 5 plants with identical growth vigor, adopting stripe bud grafting to obtain juvenile state bud grafting clone, marking as 223-5; the control is an old bud clone cultured by cutting and bud grafting from 3 15-year-old big trees and is marked as 223-2. 300 seedlings are budded in each bud of the old and young clone;
(2) when the first canopy leaves of the bud-grafted seedlings are stable, measuring the sprouting angle between the scion and the stock by taking the plants as a unit and using a protractor, and recording in detail, wherein 200 healthy seedlings are selected for measuring for the old and young bud-grafted clone;
(3) after the bud-grafted seedling sprouts and survives for 4 months, measuring the leaf form index by taking plants as units, namely selecting more than 15 healthy plants with more than 3 or more than 3 leafy froglets from each clone, taking (from bottom to top) 10 leaves at the middle-lower part of other leafy froglets except for the roof froglets and the 1 st leafy froglets from each plant, taking the plants back to a laboratory to measure the leaf length, the leaf width, the leaf area, the wide base distance/the leaf length, the vein left width/the leaf width, the density and the water content of the vein one by one, and recording the measured values;
(4) and (3) carrying out difference analysis on the data measured in the steps (2) and (3), and analyzing whether the difference of the measured indexes among the RRII105 old and young bud clone is obvious or not. The result shows that 4 indexes of the young bud clone of the RRII105, such as leaf length, leaf area, leaf vein density, extraction angle and the like, are all obviously greater than the old bud clone, and the difference among other indexes is not obvious;
(5) and (3) drawing a frequency distribution diagram: regarding the indexes (leaf length, leaf area, leaf vein density and extraction angle) with significant difference in the step (4), respectively drawing a frequency distribution graph of the RRII105 old and young bud-grafting clone plants in each index by taking the index measurement value and the plant frequency distribution as a vertical axis and a horizontal axis;
(6) and (5) deeply analyzing the frequency distribution map drawn in the step (5) to find out indexes of discontinuous distribution or a few crossed indexes in the frequency distribution map as indexes for identifying the seedlings of the RRII105 young bud grafting clone. The result shows that 4 indexes of the young bud clone and the old bud clone of the RRII105 are discontinuously distributed on the indexes of leaf length, leaf area, leaf vein density and extraction angle, have no cross and can be used as the identification index of seedlings of the young bud clone of the RRII 105. As shown in fig. 2.
It can be seen from FIG. 2 that there is no crossover in leaf length, leaf area, leaf density, and angle of emergence between the young and old budded clones of RRII 105. The young bud clone has a length and area far greater than those of the old bud clone, and can find out a discontinuous limit value of 16.5cm and 90cm2(ii) a The density of the veins of the young bud clone is far less than that of the old bud clone, and a discontinuous threshold value can be found out and is 110/m; from the perspective of sprouting, the sproutsThe extraction angle of the grafted clone is less than 25 degrees, so that the extraction angle of less than 25 degrees can also be used as the discrimination standard of the seedling of the young bud grafted clone.
(7) The identification method of the seedling of the RRII105 young bud grafting clone can be concluded from the analysis result of the step (6), namely the seedling of the young bud grafting clone has doubled leaf length and leaf area compared with the seedling of the old bud grafting clone, the leaf vein distribution is thinned, and the extraction angle is less than 25 degrees.
Example 3
(1) Selecting a rubber tree clone variety Yunyan 73-477, marking as No. 113, collecting anthers of the tree of the integrated age for tissue culture to obtain a clone obtained from a root juvenile, selecting 5 plants with consistent growth vigor after the root juvenile clone grows to 1 year, collecting strips and carrying out bud grafting to obtain a juvenile bud graft clone, marking as 113-5; the control is an old bud clone which is bred by cutting and bud grafting from 3 15-year-old big trees and is marked as 113-2. 300 seedlings are budded in each bud of the old and young clone;
(2) when the first canopy leaves of the bud-grafted seedlings are stable, measuring the sprouting angle between the scion and the stock by taking the plants as a unit and using a protractor, and recording in detail, wherein 200 healthy seedlings are selected for measuring for the old and young bud-grafted clone;
(3) after the bud-grafted seedling sprouts and survives for 4 months, measuring the leaf form index by taking plants as units, namely selecting more than 15 healthy plants with more than 3 or more than 3 leafy froglets from each clone, taking (from bottom to top) 10 leaves at the middle-lower part of other leafy froglets except for the roof froglets and the 1 st leafy froglets from each plant, taking the plants back to a laboratory to measure the leaf length, the leaf width, the leaf area, the wide base distance/the leaf length, the vein left width/the leaf width, the density and the water content of the vein one by one, and recording the measured values;
(4) and (3) carrying out difference analysis on the data measured in the steps (2) and (3), and analyzing whether the difference of the measured indexes among the old and young bud clone of the Yunyao 73-477 is obvious or not. The result shows that 4 indexes of the young bud clone of the Yunyan 73-477, such as leaf length, leaf area, leaf vein density, extraction angle and the like, are all obviously greater than the old bud clone, and the difference among other indexes is not obvious;
(5) regarding the indexes (leaf length, leaf area, leaf vein density and extraction angle) with significant differences in the step (4), respectively drawing a frequency distribution diagram of the old and young bud-grafting clone plants of the cloud research 73-477 by taking the index measurement value and the plant frequency distribution as a vertical axis and a horizontal axis;
(6) and (5) deeply analyzing the frequency distribution map drawn in the step (5) to find out indexes of discontinuous distribution or a few crossed indexes in the frequency distribution map as indexes for identifying the seedlings of the cloud research 73-477 young bud grafting clone. The result shows that 3 indexes of the young bud clone and the old bud clone of the Yunchao 73-477 are all discontinuously distributed and have no cross and can be used as the identification indexes of seedlings of the young bud clone of the Yunchao 73-477; the leaf vein density has only a little crossover, and can also be used as the identification standard of the seedlings of the bud clone of the cloud research 73-477 young state. As shown in fig. 3.
As can be seen from FIG. 3, there was no crossing in the leaf length, leaf area and emergence angle between the young and old bud clones of Yun Jiang 73-477, and the leaf length and leaf area of the young bud clone were larger than those of the old bud clone, and a discontinuous threshold was found, 15.5cm and 95cm, respectively2And 25 °; the density of the veins of the young bud clone is far less than that of the old bud clone, only a small amount (6.0%) of intersections exist on a frequency distribution diagram, and a discontinuous threshold value can be found, wherein the threshold value is 125/m;
(7) the analysis result in the step (6) can conclude the identification method of the young bud-grafted clone seedlings of the Yunyan 73-477, namely the young bud-grafted clone seedlings have doubled leaf length and leaf area compared with the old bud-grafted clone seedlings, the leaf vein distribution is diluted, and the extraction angle is less than 25.
In summary, the young bud-grafting clonal seedlings can be quickly and accurately identified by comparing the leaf length, leaf area, leaf vein density and extraction angle of old and young bud-grafting clonal plants of the same genotype of the rubber tree. Namely, if the leaves of the plant are longer, the leaf vein distribution is more sparse, the leaf area is larger or the extraction angle is less than 25 degrees, the seedling is the seedling of the seedling bud clone, otherwise, the seedling of the seedling bud clone is the seedling of the old seedling bud clone.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (6)
1. A method for rapidly identifying young bud-grafted clonal seedlings of a rubber tree is characterized by mainly comprising the following steps:
(1) and (3) seedling cultivation: selecting a rubber tree clone variety, collecting anthers, carrying out tissue culture to obtain a young clone from roots, selecting 5 plants with consistent growth vigor after the seedlings grow for 1 year, and carrying out stripe bud grafting to obtain a young bud grafting clone; collecting the scions of the old-state bud-grafting clone from the corresponding variety 15-year-old clone big tree, selecting 3 plants for picking, and culturing to obtain the old-state bud-grafting clone;
(2) measurement of the index: measuring by taking plants as units, and respectively measuring and recording the emergence angles of the bud seedlings in the old and young states when the first bud leaves of the bud seedlings are stable; after the seedlings sprout and survive for 4-5 months, respectively adopting middle and lower leaves of the same variety of old young bud seedlings and the same stable leaf fleabane, and measuring and recording the leaf indexes of the seedlings;
(3) and (3) index difference analysis: performing difference analysis on the indexes measured in the step (2), and analyzing whether the differences of the measured indexes among the old and young bud clone are obvious or not;
(4) and (3) drawing a frequency distribution diagram: regarding the indexes with significant difference in the step (3), respectively drawing a frequency distribution graph of old and young bud-grafting clone seedlings of the same variety on each index by taking the index measurement value and the plant frequency distribution as a vertical axis and a horizontal axis;
(5) determination of the authentication index: deeply analyzing the frequency distribution map drawn in the step (4), and selecting indexes which are discontinuously distributed or slightly crossed in the frequency distribution map as indexes for identifying the seedling of the young bud-grafting clone of the rubber tree;
(6) and (4) judging a result: and (5) using the analysis result in the step (5) as a text description as a method for identifying the seedling of the same genotype seedling of the rubber tree in the young bud grafting clone, namely, the seedling of the young bud grafting clone has doubled leaf length and leaf area compared with the seedling of the old bud grafting clone, the leaf vein distribution is thinned, and the extraction angle is less than 25 degrees.
2. The method for rapidly identifying seedlings of young rubber tree bud clones as claimed in claim 1, wherein the old young bud seedlings in step (2) are plants within 1 year which grow for more than 4 months after scion sprouting and have more than 3 leafy frogs.
3. The method for rapidly identifying seedlings of young bud-grafted clones of rubber trees as claimed in claim 1, wherein the stable erigeron breviscapus in step (2) is all erigeron breviscapus except for erigeron breviscapus leaf and erigeron breviscapus leaf from bottom to top.
4. The method for rapidly identifying seedlings of rubber tree scion-grafted clones as claimed in claim 1, wherein the extraction angle in step (2) is the angle between scion shoot and rootstock, and the measurement method is by protractor; the leaf indexes include leaf length, leaf width, leaf area, ratio of width base distance to leaf length, ratio of vein left width to leaf width, vein density and water content.
5. The use of the method of any one of claims 1 to 4 for the rapid identification of seedlings of rubber tree juvenile bud clones.
6. The method for rapidly identifying seedlings of a rubber tree juvenile bud-grafting clone according to claim 5, wherein the young bud-grafting clone has longer leaf length, larger leaf area, more sparse leaf vein distribution and less than 25 degrees of extraction angle compared with the leaf length, leaf area, leaf vein density and extraction angle of the same genotype old juvenile bud-grafting clone, and is an old bud-grafting clone seedling.
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| CN103503699A (en) * | 2013-07-08 | 2014-01-15 | 云南省热带作物科学研究所 | Reproduction method for rejuvenizing excellent mature individual plant of rubber tree |
| CN103688768A (en) * | 2013-12-20 | 2014-04-02 | 云南省热带作物科学研究所 | Rubber tree tender bud cultivating and budding method |
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| CN103503699A (en) * | 2013-07-08 | 2014-01-15 | 云南省热带作物科学研究所 | Reproduction method for rejuvenizing excellent mature individual plant of rubber tree |
| CN103688768A (en) * | 2013-12-20 | 2014-04-02 | 云南省热带作物科学研究所 | Rubber tree tender bud cultivating and budding method |
| CN103743915A (en) * | 2014-01-20 | 2014-04-23 | 云南省热带作物科学研究所 | Identification method for high-quality seedling of rubber tree |
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