CN108811869A - A kind of rice Adult plant confrontation accurate identification method of brown plant-hopper resistance scale - Google Patents
A kind of rice Adult plant confrontation accurate identification method of brown plant-hopper resistance scale Download PDFInfo
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- 235000007164 Oryza sativa Nutrition 0.000 title claims abstract description 152
- 235000009566 rice Nutrition 0.000 title claims abstract description 152
- 241000196324 Embryophyta Species 0.000 title claims abstract description 97
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- 241001556089 Nilaparvata lugens Species 0.000 title claims abstract description 45
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- 241000238631 Hexapoda Species 0.000 claims description 15
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 230000002093 peripheral effect Effects 0.000 claims description 12
- 238000005070 sampling Methods 0.000 claims description 11
- 230000012010 growth Effects 0.000 claims description 10
- 238000012216 screening Methods 0.000 claims description 9
- 241000607479 Yersinia pestis Species 0.000 claims description 7
- 241000746966 Zizania Species 0.000 claims description 6
- 235000002636 Zizania aquatica Nutrition 0.000 claims description 6
- 238000002955 isolation Methods 0.000 claims description 6
- 238000007789 sealing Methods 0.000 claims description 6
- 239000002689 soil Substances 0.000 claims description 6
- 238000013401 experimental design Methods 0.000 claims description 5
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- 201000010099 disease Diseases 0.000 claims description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 4
- 238000002054 transplantation Methods 0.000 claims description 2
- 238000005516 engineering process Methods 0.000 abstract description 8
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- 239000000523 sample Substances 0.000 description 18
- 108090000623 proteins and genes Proteins 0.000 description 17
- 241001498622 Cixius wagneri Species 0.000 description 12
- 230000008447 perception Effects 0.000 description 11
- 230000000306 recurrent effect Effects 0.000 description 6
- 239000003550 marker Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 238000001514 detection method Methods 0.000 description 4
- 238000012252 genetic analysis Methods 0.000 description 4
- 244000118056 Oryza rufipogon Species 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 238000013507 mapping Methods 0.000 description 3
- 239000002917 insecticide Substances 0.000 description 2
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- 239000002028 Biomass Substances 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 235000019082 Osmanthus Nutrition 0.000 description 1
- 241000333181 Osmanthus Species 0.000 description 1
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- 235000013399 edible fruits Nutrition 0.000 description 1
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- 238000004904 shortening Methods 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
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Abstract
The invention discloses a kind of rice Adult plants to fight the accurate identification method of brown plant-hopper resistance scale, belongs to Genetic and breeding in rice field.Rice Adult plant of the invention fights the accurate identification method of brown plant-hopper resistance scale, includes the following steps：(1) prepare rice stub sample；(2) rice stub sample is planted；(3) excessive to connect worm identification；(4) management after connecing worm；(5) evaluation of qualification result.The present invention establishes " rice stub identification method ", provides efficient Resistance Identification technology for the Rice Resistance brown plant-hopper breeding of various scales.The resistance breeding material of general large, medium and small various scales, both can disposably identify, identification can also be repeated several times, can also result to seedling stage assay in Adult plant carry out repetitive identified, qualification result is accurate, to effectively facilitate the progress of genetics of resistance breeding project.
Description
Technical field
The invention belongs to Genetic and breeding in rice fields, and in particular to a kind of rice Adult plant confrontation brown plant-hopper resistance rule
The accurate identification method of modelling.
Background technique
Rice is one of the staple food crop in China, and seriously threatening for brown plant-hopper is faced during Rice Production.Rice
Brown paddy plant hopper is a kind of rice grub explosive, harm is big, and the annual planthopper of China's rice region (mainly brown paddy plant hopper) occurs at present
Area is 20,000,000 hectares or more, and directly paddy loss is up to 2,800,000 tons or more caused by endangering because of planthopper.For a long time, brown
The prevention and treatment of plant hopper is mainly by application chemical insecticide, a large amount of applications of insecticide, often leads to the destruction, a large amount of of ecological environment
Brown paddy plant hopper natural enemy is killed to be improved with the drug resistance of pest, and then causes " rampant again " of brown paddy plant hopper, is brought to prevention and treatment bigger
It is difficult.It facts have proved that cultivating resistant rice varieties is method the most economic, effective, safe, ecological in brown paddy plant hopper integrated control.
Resistance breeding mainly import resistant gene into excellent rice breed, wherein Resistance Identification be resistant gene excavate and
Committed step in resistance breeding, by obtaining reliable and stable resistance offspring, therefrom selecting to breeding progeny Resistance Identification
Resistant rice varieties；By carrying out Resistance Identification to hereditary segregating population, genetic analysis and the molecule mark of resistant gene can be carried out
Note positioning.Brown plant-hopper is caused harm in production, is mostly occurred in rice Adult plant, but rice Adult plant reflects to the resistance of brown plant-hopper
Surely the problems such as always existing low determination rates, qualification result poor reliability, in resistance breeding and in the building of genetic analysis group
Deng application it is few, therefore, solves the problems, such as efficient, quick, the accurate identification technology of rice Adult plant with important practical significance.
Summary of the invention
The purpose of the present invention is establish rice Adult plant to efficient, the accurate identification technology system of brown plant-hopper resistance and be somebody's turn to do
Application of the technology in anti-brown plant-hopper rice breeding and genetic analysis informative population.
The purpose of the present invention is what is be achieved through the following technical solutions：
A kind of rice Adult plant confrontation accurate identification method of brown plant-hopper resistance scale, includes the following steps：
(1) prepare rice stub sample；
By the Rice Population identified plantation in crop field, field pipe is carried out to rice using general planting method
Reason；Next band root tiller is shelled from each rice plant for the group for needing to identify in Adult plant, each tiller is intercepted with scissors
Lower part rice stub is as identification sample；
(2) rice stub sample is planted；
Rice stub succeeding planting is compareed in the support basin for filling shallow soil, and with sense worm kind, and every basin plants 1 row reference substance
Kind；
(3) excessive to connect worm identification；
It carries out connecing worm identification in 3 days after rice stub plantation, 20 or more 2-3 age rice brown planthopper nymphs is connect to each rice stub；
(4) management after connecing worm；
I, keeps insect preventing net cover sealing after connecing worm, prevents worm from escaping；
The water layer of 1cm depth is remained no greater than in II, basin；There is water body isolation outside basin, prevent ant from causing harm；
III, support basin is placed in the authentication room of sunshade net sunshade or the place culture that sunlight is faint, prevents 32 DEG C or more
High temperature；
When IV, meets high temperature and strong sunlight direct projection, shady and cool place is moved on in time；
The place of V, authentication room or outdoor placement identification basin will reduce air-flow flowing, when meeting obvious air-flow, shut identification
Wind deflector will be arranged in room windward side windshield window, outdoor identification；
(5) evaluation of qualification result；
1~2 day after feeling the whole death of worm check variety, Resistance Identification is recorded as a result, carrying out evaluation note by 9 grade standards
It carries, first defines the level to each seedling, then calculate the average mined grade of each part material：
1 grade (1.0~1.9) is highly resistance (HR), aggrieved unobvious；
3 grades (2.0~3.9) are anti-(R), and peripheral first leaf sheath turns yellow；
5 grades (4.0~5.9) be in resist (MR), peripheral second leaf sheath turns yellow, and plant strain growth gesture obviously dies down；
7 grades (6.0~7.9) are middle sense (MS), and plant starts wilting；
9 grades (8.0~9.0) are high sense (HS), and plant is dead；
(6) repetitive identified；
Repetitive identified can be carried out according to actual needs；
(1) instant repetitive identified takes repetition rice stub according to number of repetition when sampling, carried out 2 times by experimental design or more
Repetitive identified；
(2) repetitive identifieds, if necessary, can spirit before heading stage with reference to qualification result after primary identification
Sampling living carries out repetitive identified.
As a preferred option of the technical scheme, each tiller uses scissors to intercept lower part rice stub as identification sample in the step (1)
This, intercepted length 10-20cm.
As a preferred option of the technical scheme, require the tiller peeled from different plants in the same size in the step (1), growth
Stalwartness, no disease and pests harm.
As a preferred option of the technical scheme, sense worm kind described in the step (2) is TN1,9311 or OryzasativaLcv.Nipponbare.
As a preferred option of the technical scheme, plant spacing is 4-8cm in the step (2).
As a preferred option of the technical scheme, in the step (3) after connecing worm in 3 days, for may the insufficient support basin of worm sources,
Adjunction worm sources are wanted to reach enough insect density.
As a preferred option of the technical scheme, the rice stub on basin is being ask to cover insect preventing net cover after worm is connect in the step (3).
As a preferred option of the technical scheme, the identification method can be identified at any time in Adult plant, and the Adult plant is
Start to heading stage within 20 days after rice transplantation.
Rice Adult plant of the invention fights the accurate identification method of brown plant-hopper resistance scale, in brown planthopper resistant rice
Genetic breeding, genetic research or the application of wild rice Resistance resource screening aspect.
The invention has the advantages that：
1, the present invention establishes " rice stub identification method ", provides efficiently for the Rice Resistance brown plant-hopper breeding of various scales
Resistance Identification technology.The resistance breeding material of general large, medium and small various scales, both can disposably identify, can also be repeated several times
Identification, can also result to seedling stage assay in Adult plant carry out repetitive identified, qualification result is accurate, to effectively facilitate resistance
The progress of breeding project.
2, " rice stub identification method " of the invention is suitable for the genetic analysis research and genetic marker positioning of Bph genes
The mapping population of research constructs, especially when conventional Adult plant identification method be not suitable for large-scale mapping population building and
Precisely when analysis, this technology more effectively can provide accurate identification technology for the building and pedigree analysis of extensive mapping population.
3, in rice brown planthopper resistant breeding process, when resistant gene source is unclear, genetic background is complicated, resistance trait
When the molecular labeling not being applicable in, or when marker assisted selection is difficult to precise Identification resistant plant, technical method of the invention
Provide a kind of effective resistant plant screening alternative.By being detected to breeding material, that is, can determine whether by sample material
Whether there is Brown Planthopper Resistance, and then quickly screen pest-resistant cultivar or strain for rice breeding, this is easy to detect quickly, helps
In the breeding cycle for shortening rice varieties, breeding efficiency is effectively improved.
4, rice Adult plant growth period is long, and the present invention can carry out Resistance Identification in rice Adult plant at any time, i.e., from rice
Start to heading stage within 20 days after transplanting, optimal time or optimal conditions can be selected according to the actual situation to be identified.
5, rice stub reduces significantly relative to plant, biomass, may be implemented excessively to connect worm, keeps ability dead rapidly,
Make qualification cycle and pest development cycle synchronisation, improves the accuracy of qualification result, and can be identified within a short period of time
As a result.
6, the present invention can take more rice stubs to carry out repetitive identified each plant, and 1 rice stub represents 1 repetition, thus
Improve the accuracy of qualification result.
If 7, carrying out seedling stage assay, resistant plant obtained, this season can be carried out Adult plant repetitive identified and test
Card, improves the accuracy and timeliness of qualification result.
Detailed description of the invention
Fig. 1 is the upgrowth situation figure that the identification of rice stub method strain-forming period resistance connects 4 days rice stubs after worm, and perceptual plant viability is obvious
Weaken and start turn yellow [every part of 1 row of material kind, the 2nd row of the left side rise be respectively：The anti-source Ptb33, candidate Bph3 near isogenic lines 1,
TN1 (perception control), candidate Bph3 near isogenic lines 2,9311 (recurrent parent), candidate Bph3 near isogenic lines 3].
Fig. 2 is rice stub upgrowth situation figure after the identification identification of rice stub method strain-forming period resistance, perception control and perception identification
Material is all dead, and [every part of 1 row of material kind is respectively from left to right：The anti-source Ptb33, candidate Bph3 near isogenic lines 1, TN1 (perception
Control), candidate Bph3 near isogenic lines 2,9311 (recurrent parent), candidate Bph3 near isogenic lines 3].
Fig. 3 is the upgrowth situation figure that conventional method plant Resistance Identification (control) connects rice plant before worm is identified.
Fig. 4 is the upgrowth situation figure of rice plant after conventional method plant Resistance Identification (control) is identified, [every part of material
Expect kind of two rows, is respectively from left to right：The anti-source Ptb33, candidate Bph3 near isogenic lines 1, TN1 (perception control), candidate Bph3 are closely etc.
Gene line 2,9311 (recurrent parent), candidate Bph3 near isogenic lines 3].
Rice stub method Resistance Identification (Fig. 2) is identical with conventional plant Resistance Identification (control) (Fig. 4) result, and Ptb33 is anti-
Anti-, remaining material shows as high sense during source shows as highly resistance, candidate Bph3 near isogenic lines 3 is shown as, and assessment resistance difference is bright
It is aobvious, show rice stub method Resistance Identification the result is that reliable, easy and efficient.
Specific embodiment
The present invention is further illustrated below by specific embodiment, so that advantages and features of the invention are easier to be managed
Solution, it should be understood that following embodiment is only preferred version of the invention, rather than limiting the invention.
Embodiment 1
A kind of rice Adult plant confrontation accurate identification method of brown plant-hopper resistance scale, includes the following steps：
(1) prepare rice stub sample；
By the Rice Population identified plantation in crop field, field pipe is carried out to rice using general planting method
Reason；Next band root tiller is shelled from each rice plant for the group for needing to identify in Adult plant, it is desirable that is peeled from different plants
Tiller it is in the same size, robust growth, no disease and pests harm, each tiller use scissors interception lower part rice stub as identify sample, section
Taking length is 20cm；
(2) rice stub sample is planted；
Rice stub succeeding planting is compareed in the support basin for filling shallow soil, and with sense worm kind TN1, and every basin plants 1 row pair
According to kind；When plantation, plant spacing is 4-6cm；
(3) excessive to connect worm identification；
It carries out connecing worm identification in 3 days after rice stub plantation, 20 or more 2-3 age rice brown planthopper nymphs is connect to each rice stub,
The rice stub on basin is being ask to cover insect preventing net cover after connecing worm；After connecing worm in 3 days, for the insufficient support basin of possible worm sources, adjunction
Worm sources reach enough insect density；
(4) management after connecing worm；
I, keeps insect preventing net cover sealing after connecing worm, prevents worm from escaping；
The water layer of 1cm depth is remained no greater than in II, basin；There is water body isolation outside basin, prevent ant from causing harm；
III, support basin is placed in the authentication room of sunshade net sunshade or the place culture that sunlight is faint, prevents 32 DEG C or more
High temperature；
When IV, meets high temperature and strong sunlight direct projection, shady and cool place is moved on in time；
The place of V, authentication room or outdoor placement identification basin will reduce air-flow flowing, when meeting obvious air-flow, shut identification
Wind deflector will be arranged in room windward side windshield window, outdoor identification；
(5) evaluation of qualification result；
1~2 day after feeling the whole death of worm check variety, Resistance Identification is recorded as a result, carrying out evaluation note by 9 grade standards
It carries, first defines the level to each seedling, then calculate the average mined grade of each part material：
1 grade (1.0~1.9) is highly resistance (HR), aggrieved unobvious；
3 grades (2.0~3.9) are anti-(R), and peripheral first leaf sheath turns yellow；
5 grades (4.0~5.9) be in resist (MR), peripheral second leaf sheath turns yellow, and plant strain growth gesture obviously dies down；
7 grades (6.0~7.9) are middle sense (MS), and plant starts wilting；
9 grades (8.0~9.0) are high sense (HS), and plant is dead；
(6) repetitive identified；
Repetitive identified can be carried out according to actual needs；
(1) instant repetitive identified takes repetition rice stub according to number of repetition when sampling, carried out 2 times by experimental design or more
Repetitive identified；
(2) repetitive identifieds, if necessary, can spirit before heading stage with reference to qualification result after primary identification
Sampling living carries out repetitive identified.
Technical application verifying one：The evaluation and screening of Bph3 near isogenic lines
Test material includes the anti-source Ptb33 of planthopper, and perceptual check variety TN1, perceptual parent 9311 and 3 parts are to be identified
Bph3 near isogenic lines, these Bph3 near isogenic lines to be identified be Ptb33 hybridize with 9311, offspring it is continuous with 9311 again
Mostly generation backcrossing is bred as.It is transplanted to after material seeding and seedling raising in the planting bed of brown plant-hopper authentication room, every part of 2 row of material kind, often
8 section of Hang Zhong, normal cultivation management after plantation.25 days after transplanting, the material of every part of Adult plant selects 10 sections at random, and every section takes 1 rice
Stake, plantation is in holding in the palm in basin, every part of 1 row rice stub of material kind.
Then selected materials are identified using identification method described in the present embodiment.Plant expert evidence is after connecing worm
Sense worm control TN1 plant is all dead within 10th day, and rice stub expert evidence is all dead in the 6th day sense worm control TN1 plant.Two
Processing carries out the investigation of resistance result after TN1 plant is all dead respectively, while the sampling of each part material carries out Bph3 gene mark
Note detection, test result are shown in Fig. 1 and table 1.
Experiments have shown that (1) plant Resistance Identification result and rice stub Resistance Identification result are completely the same；(2) anti-source Ptb33 because
It is most strong to the resistance of brown plant-hopper containing multiple resistant genes, reach 1 grade of highly resistance；(3) perception control TN1, recurrent parent
9311 and candidate Bph3 near isogenic lines 1 and 2 is because all without carrying resistant gene, resistance is 9 grades high sense；(4) it detects
Candidate 3 resistance of Bph3 near isogenic lines for carrying resistant gene Bph3 reaches water resistant in 5 grades and puts down.The result shows that rice stub identification method
The result is that reliable, qualification time is short, few, high-efficient with worm amount.
1 rice material Adult plant plant of table and rice stub are to the Resistance Identification result of brown plant-hopper and resistant gene Bph3
Markers for Detection
Note："+" mark contains Bph3 genetic marker, and "-" mark does not contain Bph3 genetic marker.
Embodiment 2
A kind of rice Adult plant confrontation accurate identification method of brown plant-hopper resistance scale, includes the following steps：
(1) prepare rice stub sample；
By the Rice Population identified plantation in crop field, field pipe is carried out to rice using general planting method
Reason；Next band root tiller is shelled from each rice plant for the group for needing to identify in Adult plant, it is desirable that is peeled from different plants
Tiller it is in the same size, robust growth, no disease and pests harm, each tiller use scissors interception lower part rice stub as identify sample, section
Taking length is 10cm；
(2) rice stub sample is planted；
Rice stub succeeding planting is compareed in the support basin for filling shallow soil, and with sense worm kind 9311, and every basin plants 1 row pair
According to kind；When plantation, plant spacing is 6-8cm；
(3) excessive to connect worm identification；
It carries out connecing worm identification in 3 days after rice stub plantation, 25 or more 2-3 age rice brown planthopper nymphs is connect to each rice stub,
The rice stub on basin is being ask to cover insect preventing net cover after connecing worm；After connecing worm in 3 days, for the insufficient support basin of possible worm sources, adjunction
Worm sources reach enough insect density；
(4) management after connecing worm；
I, keeps insect preventing net cover sealing after connecing worm, prevents worm from escaping；
The water layer of 1cm depth is remained no greater than in II, basin；There is water body isolation outside basin, prevent ant from causing harm；
III, support basin is placed in the authentication room of sunshade net sunshade or the place culture that sunlight is faint, prevents 32 DEG C or more
High temperature；
When IV, meets high temperature and strong sunlight direct projection, shady and cool place is moved on in time；
The place of V, authentication room or outdoor placement identification basin will reduce air-flow flowing, when meeting obvious air-flow, shut identification
Wind deflector will be arranged in room windward side windshield window, outdoor identification；
(5) evaluation of qualification result；
1~2 day after feeling the whole death of worm check variety, Resistance Identification is recorded as a result, carrying out evaluation note by 9 grade standards
It carries, first defines the level to each seedling, then calculate the average mined grade of each part material：
1 grade (1.0~1.9) is highly resistance (HR), aggrieved unobvious；
3 grades (2.0~3.9) are anti-(R), and peripheral first leaf sheath turns yellow；
5 grades (4.0~5.9) be in resist (MR), peripheral second leaf sheath turns yellow, and plant strain growth gesture obviously dies down；
7 grades (6.0~7.9) are middle sense (MS), and plant starts wilting；
9 grades (8.0~9.0) are high sense (HS), and plant is dead；
(6) repetitive identified；
Repetitive identified can be carried out according to actual needs；
(1) instant repetitive identified takes repetition rice stub according to number of repetition when sampling, carried out 2 times by experimental design or more
Repetitive identified；
(2) repetitive identifieds, if necessary, can spirit before heading stage with reference to qualification result after primary identification
Sampling living carries out repetitive identified.
Technical application verifying two：The scale application of " rice stub identification method "
Respectively by Rice Resistance source 05BPH16 (qBph30 containing resistant gene (t)), 08BPH327 (qBph29 containing resistant gene
(t)) respectively with rice hair restorer 9311, wide extensive 998, osmanthus 99, survey 253 etc. and hybridized, selected altogether in Offspring F2 segregating generation
The plant excellent to 500 characters is selected, F3 carries out plant plantation, in Adult plant by method described in the present embodiment to selected rice
Carry out Resistance Identification, every plant takes 1 section, and every section takes 3 rice stubs to carry out 3 repetitive identifieds, while to two cross combinations after
In generation, takes rice leaf sample extraction DNA to carry out genetic marker analysis verifying, detection qBph30 (t) specific molecular mark QY14 (primer
Sequence 5 '-AGCCCAAATGGAACAAACAA-3 ', 5 '-GCTCACGGTTAAGCAATGGT-3 '), detection qBph29 (t) is with tightly
Close chain molecular labeling YM54 (primer sequence：5 '-CTCTCCCCTCCAATCCTTTT-3 ', 5 '-
AGCACCCTGGAAAGCAGTAG-3').It identifies and that detects the results are shown in Table 2, it is equal through detecting to identify the partial resistance plant come
Contain homozygous or heterozygosis resistant gene, the results showed that, " rice stub identification method " is highly suitable for scale Resistance Identification, identification knot
Fruit is entirely accurate.
The application of table 2 " rice stub identification method " identifies brown plant-hopper resistance to Rice Progenies
Embodiment 3
Technical identification three：The screening of wild rice Resistance resource
Wild rice is the valuable source in China, containing many resistant genes, including anti-with important utility value
Brown plant-hopper gene.Resistance resource is screened from wild rice, can provide research material for the anti-brown plant-hopper new gene of discovery.But
It is that wild rice sowing is difficult, carries out anti-brown plant-hopper with seed and identify extremely inconvenient, inefficiency.This test " rice stub mirror
Determine method " brown plant-hopper identification is carried out to Exploration of Wild Rice Germplasm Resources, achieve beneficial effect.
A kind of rice Adult plant confrontation accurate identification method of brown plant-hopper resistance scale, includes the following steps：
(1) prepare rice stub sample；
Garden sampling identification is saved from Guangxi University's common wild-rice, every part of Exploration of Wild Rice Germplasm Resources takes 3 tender tillers of children with root
Make 3 repetitive identified materials'uses, each tiller uses the lower part rice stub of scissors interception 15cm or so length as identification sample, originally
It criticizes identification and takes 800 parts of materials altogether；
(2) rice stub sample is planted；
Rice stub succeeding planting is compareed in the support basin for filling shallow soil, and with sense worm kind TN1, and every basin plants 1 row pair
According to kind；When plantation, plant spacing is 5-8cm；80 parts of 8 rows of kind of material to be identified.800 parts of materials, 3 repetitions are total to kind of 30 basins；
(3) excessive to connect worm identification；
It carries out connecing worm identification in 3 days after rice stub plantation, 30 or more 2-3 age rice brown planthopper nymphs is connect to each rice stub,
The rice stub on basin is being ask to cover insect preventing net cover after connecing worm；After connecing worm in 3 days, for the insufficient support basin of possible worm sources, adjunction
Worm sources reach enough insect density；
(4) management after connecing worm；
I, keeps insect preventing net cover sealing after connecing worm, prevents worm from escaping；
The water layer of 1cm depth is remained no greater than in II, basin；There is water body isolation outside basin, prevent ant from causing harm；
III, support basin is placed in the authentication room of sunshade net sunshade or the place culture that sunlight is faint, prevents 32 DEG C or more
High temperature；
When IV, meets high temperature and strong sunlight direct projection, shady and cool place is moved on in time；
The place of V, authentication room or outdoor placement identification basin will reduce air-flow flowing, when meeting obvious air-flow, shut identification
Wind deflector will be arranged in room windward side windshield window, outdoor identification；
(5) evaluation of qualification result；
1~2 day after feeling the whole death of worm check variety, Resistance Identification is recorded as a result, carrying out evaluation note by 9 grade standards
It carries, first defines the level to each seedling, then calculate the average mined grade of each part material：
1 grade (1.0~1.9) is highly resistance (HR), aggrieved unobvious；
3 grades (2.0~3.9) are anti-(R), and peripheral first leaf sheath turns yellow；
5 grades (4.0~5.9) be in resist (MR), peripheral second leaf sheath turns yellow, and plant strain growth gesture obviously dies down；
7 grades (6.0~7.9) are middle sense (MS), and plant starts wilting；
9 grades (8.0~9.0) are high sense (HS), and plant is dead；
(6) repetitive identified；
The resource for being 1 grade and 3 grades to brown plant-hopper resistance for identifying above-mentioned primary dcreening operation middle acquisition, by above-mentioned (1)~(5)
Method carries out repeating screening.
Test result：
In 800 parts of O.rufipogon resources, 12 parts, 3 grades 16 parts of resource of 1 grade of Resistance resource are obtained, Resistance resource accounting
It is low (as shown in table 3).Secondary screening is carried out to 1 grade and 3 grades of resources, 12 parts of resource secondary screening whole resistances that primary dcreening operation is 1 grade are 1 grade, just
Having 1 part when 16 parts of resource secondary screenings that sieve is 3 grades is 1 grade, remaining is 3 grades.The result shows that the Resistance Identification result precision of primary dcreening operation
Very high, 1 grade of Resistance resource obtained can make anti-source and utilize.
Resistance primary dcreening operation qualification result of the 3 common wild-rice seedling stem rice stub of table to brown plant-hopper
Embodiment 4
A kind of rice Adult plant confrontation accurate identification method of brown plant-hopper resistance scale, includes the following steps：
(1) prepare rice stub sample；
By the Rice Population identified plantation in crop field, field pipe is carried out to rice using general planting method
Reason；Next band root tiller is shelled from each rice plant for the group for needing to identify in Adult plant, each tiller is intercepted with scissors
Lower part rice stub is as identification sample, intercepted length 18cm；
(2) rice stub sample is planted；
Rice stub succeeding planting is compareed in the support basin for filling shallow soil, and with sense worm kind OryzasativaLcv.Nipponbare, and every basin plants 1 row
Check variety；
(3) excessive to connect worm identification；
It carries out connecing worm identification in 3 days after rice stub plantation, 30 or more 2-3 age rice brown planthopper nymphs is connect to each rice stub；
(4) management after connecing worm；
I, keeps insect preventing net cover sealing after connecing worm, prevents worm from escaping；
The water layer of 1cm depth is remained no greater than in II, basin；There is water body isolation outside basin, prevent ant from causing harm；
III, support basin is placed in the authentication room of sunshade net sunshade or the place culture that sunlight is faint, prevents 32 DEG C or more
High temperature；
When IV, meets high temperature and strong sunlight direct projection, shady and cool place is moved on in time；
The place of V, authentication room or outdoor placement identification basin will reduce air-flow flowing, when meeting obvious air-flow, shut identification
Wind deflector will be arranged in room windward side windshield window, outdoor identification；
(5) evaluation of qualification result；
1~2 day after feeling the whole death of worm check variety, Resistance Identification is recorded as a result, carrying out evaluation note by 9 grade standards
It carries, first defines the level to each seedling, then calculate the average mined grade of each part material：
1 grade (1.0~1.9) is highly resistance (HR), aggrieved unobvious；
3 grades (2.0~3.9) are anti-(R), and peripheral first leaf sheath turns yellow；
5 grades (4.0~5.9) be in resist (MR), peripheral second leaf sheath turns yellow, and plant strain growth gesture obviously dies down；
7 grades (6.0~7.9) are middle sense (MS), and plant starts wilting；
9 grades (8.0~9.0) are high sense (HS), and plant is dead；
(6) repetitive identified；
Repetitive identified can be carried out according to actual needs；
(1) instant repetitive identified takes repetition rice stub according to number of repetition when sampling, carried out 2 times by experimental design or more
Repetitive identified；
(2) repetitive identifieds, if necessary, can spirit before heading stage with reference to qualification result after primary identification
Sampling living carries out repetitive identified.
The present invention establishes " rice stub identification method ", provides efficient resist for the Rice Resistance brown plant-hopper breeding of various scales
Property identification technology.The resistance breeding material of general large, medium and small various scales, both can disposably identify, mirror can also be repeated several times
It is fixed, can also the result to seedling stage assay carry out repetitive identified in Adult plant, qualification result is accurate, educates to effectively facilitate resistance
The progress of kind project.
It is identified using method of the invention, obtains identifying FIG. 1 to FIG. 4 as follows.Fig. 1 is rice stub method strain-forming period resistance mirror
Surely connect the upgrowth situation figure of 4 days rice stubs after worm, perceptual plant viability obviously weaken and start to turn yellow [every part of 1 row of material kind, it is left
The 2nd row of side, which rises, is respectively：The anti-source Ptb33, candidate Bph3 near isogenic lines 1, TN1 (perception control), candidate's Bph3 near isogene
It is 2,9311 (recurrent parents), candidate Bph3 near isogenic lines 3].
Fig. 2 is rice stub upgrowth situation figure after the identification identification of rice stub method strain-forming period resistance, perception control and perception identification
Material is all dead, and [every part of 1 row of material kind is respectively from left to right：The anti-source Ptb33, candidate Bph3 near isogenic lines 1, TN1 (perception
Control), candidate Bph3 near isogenic lines 2,9311 (recurrent parent), candidate Bph3 near isogenic lines 3].
Fig. 3 is the upgrowth situation figure that conventional method plant Resistance Identification (control) connects rice plant before worm is identified.Fig. 4 is normal
The upgrowth situation figure of rice plant after rule method plant Resistance Identification (control) is identified, [every part of two row of material kind, divides from left to right
It is not：The anti-source Ptb33, candidate Bph3 near isogenic lines 1, TN1 (perception control), (wheel of candidate Bph3 near isogenic lines 2,9311
Return parent), candidate Bph3 near isogenic lines 3].
Rice stub method Resistance Identification (Fig. 2) is identical with conventional plant Resistance Identification (control) (Fig. 4) result, and Ptb33 is anti-
Anti-, remaining material shows as high sense during source shows as highly resistance, candidate Bph3 near isogenic lines 3 is shown as, and assessment resistance difference is bright
It is aobvious, show rice stub method Resistance Identification the result is that reliable, easy and efficient.
Claims (9)
1. a kind of rice Adult plant fights the accurate identification method of brown plant-hopper resistance scale, it is characterised in that：Including walking as follows
Suddenly：
(1) prepare rice stub sample；
By the Rice Population identified plantation in crop field, field management is carried out to rice using general planting method；?
Adult plant shells next band root tiller from each rice plant for the group for needing to identify, each tiller intercepts lower part rice with scissors
Stake is as identification sample；
(2) rice stub sample is planted；
Rice stub succeeding planting is compareed in the support basin for filling shallow soil, and with sense worm kind, and every basin plants 1 row check variety；
(3) excessive to connect worm identification；
It carries out connecing worm identification in 3 days after rice stub plantation, 20 or more 2-3 age rice brown planthopper nymphs is connect to each rice stub；
(4) management after connecing worm；
I, keeps insect preventing net cover sealing after connecing worm, prevents worm from escaping；
The water layer of 1cm depth is remained no greater than in II, basin；There is water body isolation outside basin, prevent ant from causing harm；
III, support basin is placed in the authentication room of sunshade net sunshade or the place culture that sunlight is faint, prevents 32 DEG C or more of high temperature；
When IV, meets high temperature and strong sunlight direct projection, shady and cool place is moved on in time；
The place of V, authentication room or outdoor placement identification basin will reduce air-flow flowing, when meeting obvious air-flow, to shut authentication room and meet
Wind deflector will be arranged in wind face windshield window, outdoor identification；
(5) evaluation of qualification result；
1~2 day after feeling the whole death of worm check variety, Resistance Identification is recorded as a result, carrying out evaluation record by 9 grade standards, first
It defines the level to each seedling, then calculates the average mined grade of each part material：
1 grade (1.0~1.9) is highly resistance (HR), aggrieved unobvious；
3 grades (2.0~3.9) are anti-(R), and peripheral first leaf sheath turns yellow；
5 grades (4.0~5.9) be in resist (MR), peripheral second leaf sheath turns yellow, and plant strain growth gesture obviously dies down；
7 grades (6.0~7.9) are middle sense (MS), and plant starts wilting；
9 grades (8.0~9.0) are high sense (HS), and plant is dead.
2. rice Adult plant according to claim 1 fights the accurate identification method of brown plant-hopper resistance scale, feature
It is：Each tiller uses scissors to intercept lower part rice stub as identification sample, intercepted length 10-20cm in the step (1).
3. rice Adult plant according to claim 1 fights the accurate identification method of brown plant-hopper resistance scale, feature
It is：Require the tiller peeled from different plants in the same size in the step (1), robust growth, no disease and pests harm.
4. rice Adult plant according to claim 1 fights the accurate identification method of brown plant-hopper resistance scale, feature
It is：The plant spacing of rice stub succeeding planting is 4-8cm in the step (2)；The sense worm kind is TN1,9311 or Japan
It is fine.
5. rice Adult plant according to claim 1 fights the accurate identification method of brown plant-hopper resistance scale, feature
It is：In the step (3) after connecing worm in 3 days, for the insufficient support basin of possible worm sources, adjunction worm sources to reach enough worms
Mouth density.
6. rice Adult plant according to claim 1 fights the accurate identification method of brown plant-hopper resistance scale, feature
It is：The rice stub on basin is being ask to cover insect preventing net cover after connecing worm in the step (3).
7. rice Adult plant according to claim 1 fights the accurate identification method of brown plant-hopper resistance scale, feature
It is：The identification method is identified that the Adult plant started for 20 days after rice transplantation to heading stage in Adult plant at any time.
8. rice Adult plant according to claim 1 fights the accurate identification method of brown plant-hopper resistance scale, feature
It is：It further include repetitive identified, specially：
(1) instant repetitive identified takes repetition rice stub according to number of repetition when sampling, by experimental design progress 2 times or more weights
Multiple identification；
(2) repetitive identifieds, with reference to qualification result, if necessary, can flexibly take after primary identification before heading stage
Sample carries out repetitive identified.
9. the application of the rice Adult plant confrontation accurate identification method of brown plant-hopper resistance scale as described in claim 1,
It is characterized in that：Its application in terms of brown planthopper resistant Genetic and breeding in rice and genetic research or wild rice Resistance resource screening.
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| CN111264320A (en) * | 2020-03-06 | 2020-06-12 | 湖南杂交水稻研究中心 | Method for accurately identifying resistance of brown planthopper in adult stage of rice |
| CN114271158A (en) * | 2021-12-30 | 2022-04-05 | 华智生物技术有限公司 | Method for identifying resistance of brown planthopper of rice in field adult stage |
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| CN114271158A (en) * | 2021-12-30 | 2022-04-05 | 华智生物技术有限公司 | Method for identifying resistance of brown planthopper of rice in field adult stage |
| CN114271158B (en) * | 2021-12-30 | 2022-12-23 | 华智生物技术有限公司 | Method for identifying resistance of brown planthopper of rice in field adult stage |
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