CN113917082A - Method for identifying influence of local commercial crop pollination on exotic commercial crops - Google Patents
Method for identifying influence of local commercial crop pollination on exotic commercial crops Download PDFInfo
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- 230000010152 pollination Effects 0.000 title claims abstract description 27
- 238000000034 method Methods 0.000 title claims abstract description 23
- 241000238631 Hexapoda Species 0.000 claims abstract description 32
- 235000013399 edible fruits Nutrition 0.000 claims abstract description 16
- 235000013500 Melia azadirachta Nutrition 0.000 claims description 53
- 235000004298 Tamarindus indica Nutrition 0.000 claims description 27
- 241000196324 Embryophyta Species 0.000 claims description 11
- 238000011218 seed culture Methods 0.000 claims description 8
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 claims description 8
- XOSXWYQMOYSSKB-LDKJGXKFSA-L water blue Chemical compound CC1=CC(/C(\C(C=C2)=CC=C2NC(C=C2)=CC=C2S([O-])(=O)=O)=C(\C=C2)/C=C/C\2=N\C(C=C2)=CC=C2S([O-])(=O)=O)=CC(S(O)(=O)=O)=C1N.[Na+].[Na+] XOSXWYQMOYSSKB-LDKJGXKFSA-L 0.000 claims description 7
- 240000004584 Tamarindus indica Species 0.000 claims description 6
- 239000013598 vector Substances 0.000 claims description 6
- 238000004820 blood count Methods 0.000 claims description 4
- 238000009395 breeding Methods 0.000 claims description 3
- 230000001488 breeding effect Effects 0.000 claims description 3
- 240000005343 Azadirachta indica Species 0.000 claims 1
- 239000000834 fixative Substances 0.000 claims 1
- 238000010186 staining Methods 0.000 claims 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims 1
- 241000894007 species Species 0.000 abstract description 15
- 238000002955 isolation Methods 0.000 abstract description 2
- 230000001850 reproductive effect Effects 0.000 abstract description 2
- 244000237986 Melia azadirachta Species 0.000 description 52
- 241000596504 Tamarindus Species 0.000 description 22
- 238000004043 dyeing Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000008901 benefit Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000005094 fruit set Effects 0.000 description 3
- 238000005034 decoration Methods 0.000 description 2
- 230000035784 germination Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 210000001672 ovary Anatomy 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 229920000018 Callose Polymers 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000000749 insecticidal effect Effects 0.000 description 1
- 238000007427 paired t-test Methods 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 238000012353 t test Methods 0.000 description 1
- 229940046536 tree pollen allergenic extract Drugs 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/0098—Plants or trees
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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
- A01G7/00—Botany in general
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N2015/1022—Measurement of deformation of individual particles by non-optical means
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N2015/1024—Counting particles by non-optical means
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Abstract
The invention provides an identification method for influence of local economic crop pollination on an exotic economic crop, and relates to the technical field of forestry planting. The invention utilizes the reproductive isolation among different species, if the flower stigmas receive heterogeneous pollen, the species can not be fertilized and fruited normally, and the seed yield is reduced finally. The comprehensive judgment is carried out on the basis of 3 indexes which are gradually increased layer by layer, namely the pollen proportion carried by the entomophile, the plant stigma pollen proportion and the fruit setting rate measured after the seed setting period. For a certain foreign economic crop, if mixedCIs significantly lower than pureC(mixed: mixed forest; pure: pure forest; C is fruit setting rate), and mixed KBook (I)(K: insects; this: native pollen ratio) and mix ZBook (I)(Z: stigma) are all more than 50%, then the local channel is judgedEconomic crops have a significant negative impact on pollination and seed set of foreign commercial crops and are an essential factor in reducing seed yield of foreign commercial crops.
Description
Technical Field
The invention belongs to the technical field of forestry planting, and particularly relates to an identification method for influence of local economic crop pollination on an exotic economic crop.
Background
Neem (Azadirachta indica a. juss.) is a multi-purpose tree species of native south asian next continent, the seeds of which contain a variety of biological insecticidal active substances, and is internationally recognized as a resource plant most suitable for the development of biological pesticides. Neem is introduced into the dry-hot valley areas in the southwest of China for large-scale planting in the last 90 th century, and the area of the neem currently reaches 35 million hectares. However, in the southwest dry-hot valley area, the economic forest of neem introduced and cultivated in a large area is subjected to large-scale mixed planting with native fruit Tamarindus indica. When local enterprises collect neem seeds, the local enterprises often find that even under the same cultivation technical conditions, the average seed yield of each plant of pure neem forest is greatly higher than the average seed yield of each plant of mixed neem and tamarind, but scientific explanation cannot be given, so that a new method needs to be designed to judge the influence relationship between foreign tree seeds and local tree seeds.
Disclosure of Invention
In view of the above, the present invention provides a method for identifying influence of local economic crop pollination on an external economic crop, which determines positive or negative influence between the local economic crop and the external economic crop, so as to provide specific cultivation measures for the external economic forest, increase seed yield and improve economic benefit.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides an identification method for foreign economic crops affected by local economic crop pollination, which comprises the following steps:
(1) respectively designing a pure variety sample plot of the foreign economic crops and a mixed variety sample plot of the foreign economic crops and the local economic crops;
(2) capturing a plurality of insect vectors in a mixed sample plot in the full-bloom period, and counting the proportion of local economic crop pollen carried by the insect vectors to the total amount of the carried pollen, namely mixed KBook (I)；
(3) Randomly collecting stigmas of the pollinated foreign economic crops in the mixed breeding plot, and counting the proportion of local economic crop pollen on the stigmas of the foreign economic crops, namely mixed ZBook (I)；
(4) Randomly selecting equal amount of foreign channels from pure and mixed sample plotEconomic crops, respectively counting the fruit setting rate C of the exotic economic cropsPureAnd CMixing of；
When C is presentMixing ofIs significantly lower than CPureAnd mixed with KBook (I)And mixed ZBook (I)Both of which are greater than 50%, it is determined that the local commercial crop has a significant negative impact on pollination and fruiting of the foreign commercial crop.
Preferably, the pure seed culture plots and the mixed seed culture plots in the step (1) are each provided in 3 or more blocks, and the intervals between the pure seed culture plots and the mixed seed culture plots exceed 2 km.
Preferably, 30 flowering insects are captured in step (2), and pollen from each flowering insect is eluted on a blood count plate according to the number of the same and the insect type for observation and statistics.
Preferably, the observation comprises microscopic observation.
Preferably, the counting in the step (3) comprises randomly collecting 60 stigmas on the foreign economic crops after pollination, fixing the stigmas by FAA fixing solution, dyeing by using water-soluble aniline blue, and observing under a microscope.
Preferably, the foreign economic crop and the local economic crop are insect-pollinated plants.
Preferably, the foreign commercial crop comprises neem and the local commercial crop comprises tamarind.
Has the advantages that: the invention provides a method for identifying the influence of local commercial crop pollination on exotic commercial crops, which utilizes reproductive isolation among different species, for example, if heterogenous pollen is accommodated on flower stigmas, the species can not be fertilized and fruited normally, and finally, the seed yield is reduced. The comprehensive judgment is carried out on the basis of 3 indexes which are gradually increased layer by layer, namely the pollen proportion carried by the entomophile, the plant stigma pollen proportion and the fruit setting rate measured after the seed setting period. For a certain exotic commercial crop, if CMixing ofIs significantly lower than CPure(mixed: mixed forest; pure: pure forest; C is fruit setting rate), and mixed KBook (I)(K: insects; this: native pollen ratio) and mix ZBook (I)(Z: stigma) are all more than 50 percent, the local commercial crop is judged to generate obvious negative for pollination and seed setting of the foreign commercial cropThe method has the following advantages that the method has the following effects and is an essential factor causing the reduction of the seed yield of the exotic economic crops. Otherwise, it indicates that no obvious negative effect is caused.
Detailed Description
The invention provides an identification method for foreign economic crops affected by local economic crop pollination, which comprises the following steps:
(1) respectively designing a pure variety sample plot of the foreign economic crops and a mixed variety sample plot of the foreign economic crops and the local economic crops;
(2) capturing a plurality of insect vectors in a mixed sample plot in the full-bloom period, and counting the proportion of local crop pollen carried by the insect vectors, namely' mixed KBook (I)"(K: insects; this: local pollen ratio);
(3) randomly collecting postpollination stigmas of the foreign economic crops in the mixed breeding plot, and counting the local pollen proportion of the foreign economic crops, namely' mixed ZBook (I)"(Z: stigma);
(4) randomly selecting an equal amount of the foreign economic crops from the pure variety sample plot and the mixed sample plot, and respectively counting the fruit setting rate C of the foreign economic cropsPureAnd CMixing of；
When C is presentMixing ofIs significantly lower than CPureAnd "mix KBook (I)"and" mix ZBook (I)"both are greater than 50%, it is judged that the local commercial crop has a significant negative impact on pollination and fruiting of the foreign commercial crop.
The exotic economic crops and the local economic crops are preferably all insect-borne flower plants, and insects are mainly used for pollination. In the invention, when setting sample plots, 3 sample plots or more are preferably arranged for each design, and the interval of each sample plot is ensured to exceed 2km so as to reduce the mutual interference of pollinating insects among the sample plots. In the embodiment of the invention, preferably, neem is used as a foreign economic crop, tamarind is used as a local economic crop to perform an experiment, 3 blocks of neem pure forest sample plots and 3 blocks of neem + tamarind mixed forest sample plots are respectively arranged (namely, each block plot is repeated for 3 times to eliminate mathematical statistical errors), the area of each block sample is 5 mu, and the block samples are numbered, and when the block plots are arranged, the space between every two blocks is ensured to be more than 2km to reduce the mutual interference of pollinating insects between the block plots.
When the method is used for measuring the proportion of pollen carried by the entomophilous insect, the pollen can be distinguished according to the difference of the surface form and the number of germination groove holes, for example, the shape of the neem pollen is a sphere-like 3 groove hole, and the tamarind pollen is a wedge-shaped 5 groove hole. In the present embodiment, it is preferable to capture 30 test flowering insects in the mixed sample at the full-bloom stage, and the pollen from each test flowering insect is eluted on a blood count plate according to the number of the sample and the type of the insect, and observed under a microscope in a laboratory.
When counting the proportion of the alien pollen of the exotic economic crops, the method preferably comprises randomly collecting 60 stigma of the exotic economic crops after pollination, fixing the stigma by FAA fixing liquid, dyeing by water-soluble aniline blue, and observing under a microscope. The water-soluble aniline blue provided by the invention can anchor the pollen grains on the stigma, cannot be rinsed off, is convenient for rinsing the stigma, and can compare the number of the pollen grains on the stigma before and after rinsing. In addition, pollen and callose of pollen tube wall are combined with aniline blue and induced to produce fluorescence, and the germination of pollen grains on stigma and the elongation growth of pollen tube can be observed under a fluorescence microscope. In the embodiment of the invention, preferably, 60 stigma on each flower of neem after pollination (the stigma after pollination is provided with anther, so that the stigma is easy to distinguish) are randomly collected, fixed by FAA stationary liquid and then taken back to the laboratory for dyeing by water-soluble aniline blue, observed under a microscope, pollen is distinguished according to the shape and the ditch hole of each tree pollen, and the types and the proportion of the same-species pollen and the heterogeneous pollen on the neem stigma of the neem pure forest and the neem of the mixed forest are respectively counted. Wherein the pollen of the same species is neem pollen on the neem stigma; alien pollen, i.e., native pollen, that is tamarind pollen on the stigma of neem;
the proportion of heterogeneous pollen is equal to the number of tamarind pollen deposited on the neem stigma/the total number of pollen, and is shown in a formula I.
When the fruit setting rate of the exotic economic crops is counted, preferably 20 exotic economic crop sample plants are randomly marked in each sample area, the number of flowers blooming is counted, and when the ovary is enlarged, the fruit setting rate of the exotic economic crops of pure forest and mixed forest is counted for the marked plants according to sample area numbers.
The fruit setting rate (fruit setting number/total flowering number) is multiplied by 100 percent, and the formula II is shown.
When the method is used for evaluation, the difference significance of the fruit setting rate is preferably p <0.05 level, and a pairing t test method in the traditional mathematical statistics is adopted.
The method for identifying foreign economic crop affected by local economic crop pollination provided by the invention is explained in detail with the following examples, but they should not be construed as limiting the scope of the invention.
Example 1
1. Sample plot setting
3 blocks of the neem pure forest plot and the neem + tamarind mixed forest plot are respectively arranged (namely, each block of the plot is repeated for 3 times to eliminate mathematical statistical errors), the area of each block of the plot is 5 mu and is numbered, and when the plots are arranged, the spacing between the plots is ensured to be larger than 2km, so that the mutual interference of pollinating insects among the plots is reduced.
2. Statistics of pollen carried by pollinating insects in mixed forest
In the full-bloom stage, 30 different flowering insects were captured in the mixed forest of neem and tamarind, and the pollen was eluted on a blood count plate according to the number of the insects and the type of the insects, and observed under a microscope in a laboratory.
TABLE 1 comprehensive statistics of pollens carried by pollinating insects in Azadirachta indica + tamarind mixed forest plot
3. Statistics of tamarind pollen quantity on neem flower stigma in mixed forest
In the neem and tamarind mixed forest plot, randomly collecting 60 stigmas on the neem flowers after pollination (the stigmas after pollination are provided with anthers, so that the stigmas are easy to distinguish), fixing the neem flowers by FAA fixing solution, then returning the neem flowers to a laboratory for water-soluble aniline blue dyeing, observing the neem flowers under a microscope, and counting the types and the proportions of the same-species pollen and the different-species pollen on the neem stigmas of the mixed forest.
TABLE 2 pollen composition and ratio of neem stigma in mixed neem and tamarind forest
| Neem pollen | Tamarind pollen | Other pollen |
| 18.91％ | 79.68％ | 1.41％ |
4. Statistics of fruit setting rate of pure forest plot and mixed forest plot after fructification period
In the pure neem forest, the mixed forest of neem and tamarind, randomly marking 20 neem-like plants in each sample, counting the number of flowers blooming, and when the ovary of the plant is enlarged, respectively carrying out the neem fruit setting rate statistics of the pure forest and the mixed forest according to the sample number.
TABLE 3 statistics of difference between fruit set percentage of Neem in pure Neem forest and mixed Neem and Tamarindus indica forest
According to Table 3, the average of the fruit set rates of 3 blocks of pure Neem forest is about 2.91%, the average of the fruit set rates of the mixed Neem of Neem and tamarind is about 0.82%, and the P value of the two trees is 0.013(P <0.05) by using the paired t test algorithm in IBM SPSS Statistics 24 statistical software (or other similar mathematical statistical software), which indicates that there is a significant level difference between the two trees. In combination with the fact that the tamarind pollen carried by pollinating insects in the mixed forest in table 1 is more than 50%, and the tamarind pollen carried by the neem stigma in the mixed forest in table 2 is also more than 50%, it is further demonstrated that tamarind tree species in the mixed forest have a significant negative effect on neem. When the nim is mixed with the local tamarind seeds, the fruit setting rate of the nim is greatly reduced compared with the pure nim forest, and the seed yield is obviously reduced, which is the reason that the nim is influenced by the pollination of the tamarind trees. Mainly, when the neem and tamarind are mixed, pollination insects tend to be more selective to pollinate tamarind of a local tree species, and pollination insects of neem of an external tree species are correspondingly reduced. Meanwhile, when part of insects pollinate the neem, a large amount of tamarind pollen carried by the body falls on the stigma of the neem, so that heterogeneous pollen interference is caused, and the non-pollination of the neem is aggravated.
Accordingly, for the case that the foreign tree species are affected by the local tree species pollination, the proposal is that when the foreign tree species and the local tree species are configured in a mixed mode, the mutual influence of the foreign tree species and the local tree species can be greatly reduced by adopting a block mixed mode. For example, during afforestation, neem may be first configured in 10 × 10 rows and columns to form a large pure forest, tamarind trees may be configured in 5 × 5 rows and columns in the same way to form a small pure forest, and the two blocks may be arranged in one afforestation community, so as to realize the block mixed mode of mixed tree species.
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. A method for identifying foreign economic crops affected by local economic crop pollination, comprising the steps of:
(1) respectively designing a pure variety sample plot of the foreign economic crops and a mixed variety sample plot of the foreign economic crops and the local economic crops;
(2) capturing a plurality of insect vectors in a mixed sample plot in the full-bloom period, and counting the proportion of local economic crop pollen carried by the insect vectors to the total amount of the carried pollen, namely mixed KBook (I)；
(3) Randomly collecting stigmas of the pollinated foreign economic crops in the mixed breeding plot, and counting the proportion of local economic crop pollen on the stigmas of the foreign economic crops, namely mixed ZBook (I)；
(4) Randomly selecting an equal amount of the foreign economic crops from the pure variety sample plot and the mixed sample plot, and respectively counting the fruit setting rate C of the foreign economic cropsPureAnd CMixing of；
When C is presentMixing ofIs significantly lower than CPureAnd mixed with KBook (I)And mixed ZBook (I)Both of which are greater than 50%, it is determined that the local commercial crop has a significant negative impact on pollination and fruiting of the foreign commercial crop.
2. The method according to claim 1, wherein the pure seed culture plots and the mixed seed culture plots in step (1) are each provided in 3 or more blocks, and the spacing between the pure seed culture plots and the mixed seed culture plots exceeds 2 km.
3. The method of claim 1, wherein 30 individual flowering insects are captured in step (2), and pollen from each flowering insect is eluted on a blood count plate according to the number of the same plot and the insect type, and observed and counted.
4. The method of claim 3, wherein said observing comprises observing under a microscope.
5. The method of claim 1, wherein said counting in step (3) comprises randomly collecting 60 stigmas from the pollinated foreign commercial crop, fixing with FAA fixative, staining with water soluble aniline blue, and observing under a microscope.
6. The method of claim 1, wherein the foreign and local commercial crops are insect-pollinated plants.
7. The method of claim 1 or 6, wherein the foreign commercial crop comprises neem and the local commercial crop comprises tamarind.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114774378A (en) * | 2022-04-26 | 2022-07-22 | 中国林业科学研究院 | Thioredoxin for regulating and controlling identification of plant pollen tube and stigma and preparation method thereof |
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| US20090000188A1 (en) * | 2007-06-27 | 2009-01-01 | Pioneer Hi-Bred International, Inc. | Method and apparatus of high-throughput pollen extraction, counting, and use of counted pollen for characterizing a plant |
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| US20090000188A1 (en) * | 2007-06-27 | 2009-01-01 | Pioneer Hi-Bred International, Inc. | Method and apparatus of high-throughput pollen extraction, counting, and use of counted pollen for characterizing a plant |
| CN103868842A (en) * | 2014-02-11 | 2014-06-18 | 中国农业科学院蜜蜂研究所 | Method for detecting quantity of stigma pollens after pollination of flowering plants |
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| Title |
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| CN114774378A (en) * | 2022-04-26 | 2022-07-22 | 中国林业科学研究院 | Thioredoxin for regulating and controlling identification of plant pollen tube and stigma and preparation method thereof |
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