CN101113977A - Method for rapidly identifying resistance of transgenic Bt sugarcane to borer - Google Patents
Method for rapidly identifying resistance of transgenic Bt sugarcane to borer Download PDFInfo
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Abstract
The invention relates to a rapid identification method of transgenic Bt sugarcane resistance to Stem Borers by artificial inoculating. The method includes planting sugarcane in buckets in greenhouse, choosing insect source, artificial inoculating and evaluation of resistance. The experimental insect source of the same batch transgenic Bt sugarcane strain is the newly hatched larvae of the same striped sugarcane borer, or equally divides the newly hatched larvae of 2-3 female moth born to each sugarcane to be analyzed. The result can be investigated after 5 days of artificial inoculating to the sugarcanes planted in buckets in greenhouse and the pest resistance of the sugarcane strain is evaluated comprehensively according to the larval survival rate and sugarcane damage level. The most notable characteristics of the invention are: 1. the consistency and unity of inoculating time are guaranteed to the utmost extent to ensure the accuracy of identification; 2. The periodicity of pest resistance identification is so short that 7-10 days is enough to get the result of the experiment; 3. Without the influence of the natural enemy in the field or special weather (e.g. rainstorm), the experiment result is reliable; 4.the method has better maneuverability.
Description
Technical field
The present invention relates to crop molecular breeding and resistance breeding technical field, especially transgenic Bt sugarcane insect resistace authentication method.
Technical background
Sugarcane moth borer is that China sugarcane district extensively takes place and the most serious class sugarcane insect of causing harm.Cause harm because of sugarcane moth borer and to cause sugarcane loss generally to reach 10%～25%, heavy person can reach 40%～60%, also causes the sugarcane sugar loss simultaneously and brings out some stem's disease.
Sugarcane moth borer is a lepidopterous class borer pest.The common sugarcane moth borer in China sugarcane district has bar snout moth's larva, chilo infuscatellus, yellow snout moth's larva, pink rice borer and white snout moth's larva etc., and wherein, bar snout moth's larva (Proceras venosatum Walker) is the most serious a kind of sugarcane moth borer of causing harm.Article sugarcane borer all has generation in each sugarcane district, the whole nation.The bar snout moth's larva is caused harm and is caused the withered heart seedling stage sugarcane, causes borer pest joint, spike top (dead tail), long lateral bud in a large number behind the sugarcane jointing, and formation windbreak stem causes sugarcane yield and sugar heavy losses when meeting strong wind.
Because sugarcane itself lacks anti-source, the anti-snout moth's larva breeding of sugarcane does not obtain big progress, does not have real anti-snout moth's larva kind in the production.Though relatively lighter kind of borer pest is arranged, be mostly that fiber part is higher and be unfavorable for that snout moth's larva gets the kind of food that its process character is generally also relatively poor.The Sugarcane Superior Variety of the high sugar of most high yields also is the heavier kind of borer pest.For a long time, the sugarcane moth borer control is mainly based on pesticide control.Long-term a large amount of use of insecticide had both increased production cost, also caused environmental pollution, destroyed the ecologic equilibrium.It should be noted that the generation of snout moth's larva and cause harm and do not obtain ideal control because of use a large amount of pesticides year after year.By the molecular breeding means, import the external source anti insect gene, the pest-resistant sugar cane breed of seed selection is also applied with suiting measures to local conditions, is expected to control effectively the population growth of sugarcane moth borer, reduces applications of pesticide amount and application times significantly, alleviates the loss that insect pest causes.This undoubtedly to increasing both production and income, reducing production costs and reducing environmental pollution all has significant meaning.
Bacillus thuringiensis (Bacillus thuringiensis, be called for short Bt) can produce the parasporal crystal of property of protein, protozoans such as various insects, nematode, mite class are had special insecticidal activity to the person poultry harmless, free from environmental pollution, thereby be widely used in the biological control of insect.Wherein the delta-endotoxin of CryI series is effective especially to Lepidoptera (Lepidoptera) insect, thereby the many genes among the CryI have been widely used in the anti insect gene engineering of plant.The Bt gene is imported the cultivation sugarcane, can solve the problem that sugarcane self anti insect gene source lacks, breed pest-resistant cultivar.
People such as Arencibia A (Mol Breeding 1997,3:247-255) that cryIAb is gene constructed in CaMV35S promoter downstream, transform sugarcane, the expression of CryIAb albumen in transgenic sugarcane is not high, can't resist the sugarcane borer efficiently.This may be since the CaMV 35S promoter that is applicable to dicotyledon in monocotyledon, express not good (Adang MJ, Plant Mol Biol 1993,21:1131-1145).People such as Dawen (PlantScience 2003,165:743-750) report, and PUbi promoter in transgenic sugarcane (corn ubiquitin promoter) can drive gus gene and express, but does not detect the expression of gus gene under the CaMV 35S promoter.We also find in the transgenosis moment expression test of sugarcane, the PUbi promoter is than CaMV 35S promoter strong 5～6 times (Weng, Li-Xing etc., Pest Management Science, 2005 In Press), obtain the transgenic Bt sugarcane strain of a collection of high expressed.
The insect resistace evaluation is one important procedure of transgenic pest-resistant breeding.The transgenic Bt sugarcane insect resistace can detect by marker gene, genes of interest detects, toxalbumin detects and method such as biologicall test is estimated, but finally all to promptly to estimate with the pest-resistant foundation that shows as of growing sugarcane under the natural conditions through the test of field condition or simulation field condition.The test period that the field insect resistace is identified is long, be subjected to effect of natural conditions (ring as NATURAL DISTRIBUTION, natural enemy interference and the wind in worm source, rain shadow etc.) big, much the factor that can not artificially control causes testing the inconsistency of condition between processing and the comparability of test findings, often need the result of many tests for many years to go to do comprehensive evaluation, therefore be difficult to obtain at short notice reliable test findings.Manually connect worm and identify the influence to avoid the field uncontrollable factor effectively, but and obtain the result within a short period of time.But manually connect worm in the past and identified it is to gather worm's ovum, and indoorly hatched larva and be inoculation worm source with the field.Snout moth's larva ovum natural enemy nature parasitic rate height (reach about 90%, even higher), being difficult to obtain enough larvas and supplying needing of test, strain particularly to be identified is more for a long time.Simultaneously because source, test worm source is inconsistent and connect the worm time and be difficult to unification and influence measurement result.
Article sugarcane borer is that the most general, the most serious snout moth's larva of causing harm is taken place, and larva is got food at lobus cardiacus, and the 2nd day just visible its in invasion back got the food trace (being commonly called as " floral leaf ") that food stays, and heavy, poisoning manifestations is also fast.The present invention is representative snout moth's larva kind with the article sugarcane borer, the major technique feature: the snout moth's larva kind that (1) is used to test has representative preferably, and the damage symptom appearance is fast, obvious as seen, and specificity is arranged, and observes easily and investigates; (2) source and the quantity tool comparability of the larva on every strain sugarcane to be identified received with to connect the worm time consistent in source, test worm source, and with the larva in same source at one time the section mean allocation to each strain sugarcane plant to be measured; (3) Shi Yi test sugarcane breeding method and anti-snout moth's larva evaluation method can obtain test findings in 7～10 days after connecing worm.
Summary of the invention
Advantages such as the objective of the invention is to propose the rapid identification method of a kind of transgenic Bt sugarcane to the snout moth's larva resistance, it can carry out under indoor environment, has the test period weak point, can get rid of the influence of field natural enemy and special weather (as storm), and test findings is reliable.
Purpose of the present invention can realize by following approach:
A kind of transgenic Bt sugarcane is characterized in that comprising the steps: to the rapid identification method of snout moth's larva resistance
A. greenhouse barrel plant sugarcane;
B. the worm source is selected: the bar snout moth's larva larva with first hatching is test worm source;
C. manually connect worm: the worm source of step B is manually connect worm to the barrel plant sugarcane of steps A;
D. evaluation of resistance: through step C manually meet 7 days " Invest, Then Investigate " results of worm, according to the participate in the experiment insect resistace of sugarcane strain of the extent of injury comprehensive evaluation of the survival rate of larva, sugarcane.
Wherein among the step B, to test worm source with a collection of transgenic sugarcane strain to be evaluated, the newly hatched larvae that can be laid eggs for same the female moth of article sugarcane borer, also can be newly hatched larvae that 2～3 female moths are produced respectively mean allocation to every strain sugarcane strain to be measured.
The step that obtains test worm source among the step B is as follows:
B1. gather the pupa of article sugarcane borer in the sugarcane field, place in the box, under indoor conditions, sprout wings, allow female, male moth natural mating;
B2. select the female moth of successful post-coitum, put into transparent plastic bottle, every bottle of 1 female moth, treat that it laid eggs back 4 days in, with scissors or pocket knife pieces of an egg are taken off one by one together with the plastic bottle wall, under stereoscope, calculate every ovum grain number, and pieces of an egg are placed in the double dish that is placed with thieving paper preserve moisture;
The total ovum grain number that produces according to same female moth and waiting connects the strain number of the test sugarcane of worm, plans the quantity that every strain sugarcane connects worm;
As optimization, connect determining of worm time among the step C: when finding in the ovum pore to be arranged, promptly carry out the preparation that connects worm, begin to connect worm when the larva hatching is arranged immediately.
As optimization,
In the steps A, barrel plant is cultivated the transgenic Bt sugarcane strain and the acceptor kind thereof of pending anti-snout moth's larva evaluation in the greenhouse, every barrel of 4 the sugarcane buds of sowing, and Cheng Miaohou stays seedling 2～4 strains for every barrel; The sugarcane of selecting 1～1.5 meter of plant height is as the suitable strain age of testing;
Among the step C, the quantity that the larva that just incubates is connect worm according to plan is connected to the lobus cardiacus portion of every strain test sugarcane, and the lobus cardiacus of every strain sugarcane connects 10 first and incubates bar snout moth's larva larva, and connecing the worm duration is 1～2 day;
Among the step D, connect worm can be observed lobus cardiacus after the 2nd day the shape of being injured, existence situation of 5 days " Invest, Then Investigate " larvas and sugarcane strain extent of injury are pressed the resistance rank that " pest-resistant sugarcane strain bar snout moth's larva infringement grading standard " evaluates the strain of participating in the experiment.
As further optimization,
Among the step C, require every strain sugarcane of same test to insert the newly hatched larvae from same female moth of equal number on the same day.
Compared with prior art, the present invention has following outstanding substantive distinguishing features and marked improvement:
(1) be the representative insects with article sugarcane borer (Proceras venosatum Walker), its take place in China sugarcane district the most general, cause harm the most seriously, invasion is got food back symptom and is occurred fast and obviously;
(2) to adopt bar snout moth's larva pupa in indoor emergence, obtaining worm's ovum under controlled conditions, serves as test worm source with the bar snout moth's larva larva (first-instar young) of first hatching, has solved the problem of ovum insect parasite influence, worm source acquisition methods workable.
(3) each test is the worm source with the newly hatched larvae of the ovum of same female moth product only, or the newly hatched larvae that 2～3 female moths produce is distinguished mean allocation arrive every strain sugarcane strain to be measured, reduce the difference between the individuality of test worm source to greatest extent, guarantee the accuracy of insect resistace qualification result;
(4) the test period weak point can obtain test findings in 7～10 days after connecing worm.
(5) got rid of the influence of field natural enemy and special weather (as the wind and rain that exposes to the sun), test findings is reliable.
Description of drawings
Fig. 1 transgenic sugarcane and contrast (non-transgenic) connect behind the worm the 7th Heaven's commandments snout moth's larva larval growth situation and sugarcane extent of injury relatively
Among the figure, A: medium resistance, the obvious diapause of larva; B: it is promptly dead that strong resistance, larva are got the food back on a small quantity; C: contrast (non-transgenic), larval growth is normal; The larva in D:7 flying worm age, a left side is the larva of contrast strain, the right side is the larva of transgenosis sugarcane strain; E: the sugarcane stem situation of being injured, a left side is the transgenosis strain, rightly is the contrast strain.
Fig. 2 connects manually behind the worm that sugarcane is injured shape relatively after 15 days
Among the figure, a left side: contrast (non-transgenic), all the sugarcane strain is killed, the withered heart, long lateral bud.Right: change the Bt strain, snout moth's larva fails to invade, and the sugarcane strain does not have insect pest.
Embodiment
Below the present invention is further illustrated with a specific embodiment:
A kind of transgenic Bt sugarcane is to the rapid identification method of snout moth's larva resistance, and it comprises the steps:
Steps A. plantation transgenic Bt sugarcane plant
To contain transformed plant Guangdong sugar 79-177 series 5 strains (numbering is respectively Y1, Y2, Y3, Y4 and Y5) of Bt gene and sugared 8 strains of No. 16 series (numbering is respectively T1, T2, T3, T4, T5, T6, T7 and the T8) barrel plant of new platform in the greenhouse, every barrel of 4 the sugarcane buds of sowing, Cheng Miaohou stays seedling 2～4 strains for every barrel.And the while is planted not genetically modified sugar cane breed Guangdong sugar 79-177 (Yck) in the greenhouse and new platform sugar No. 16 (Tck) contrasts, and does pest-resistant evaluation and uses.The sugarcane strain of selected plant up to 1～1.5 meter is as the test sugarcane.
Step B. obtains test worm source
The snout moth's larva that the present invention uses is the newly hatched larvae of article sugarcane borer.
(1) gathers the pupa of article sugarcane borer in the sugarcane field mid-July, put in the carton of suitable size, under indoor conditions, sprout wings, allow female, male moth natural mating.
(2) the female moth of the successful post-coitum of selection is put into transparent plastic bottle, 1 female moth of each bottle.Treat that it laid eggs back 4 days in, pieces of an egg are taken off one by one together with the plastic bottle wall, under stereoscope, calculate every ovum grain number, and pieces of an egg are placed in the double dish that is placed with thieving paper preserve moisture with scissors or pocket knife.The total ovum grain number that produces according to same female moth and waiting connects the strain number of the test sugarcane of worm, plans the quantity that every strain sugarcane connects worm, treats that doing the pest-resistant test of transgenic Bt sugarcane after its hatching uses the worm source.
Step C. connects worm
Every morning 8～9 o'clock observation worm's ovum, finding has pore in the ovum, shows to be about to hatching, has the larva hatching promptly to begin to connect worm.Newly hatched larvae is inoculated on the lobus cardiacus of sugarcane.With acceptor sugar cane breed (not transgenosis) is contrast.The lobus cardiacus of every strain sugarcane connects 10 first and incubates the article sugarcane borer larva.
If the larva that 1 female moth produces is not enough to a batch of material and connects worm, the larva that available 2～3 female moths produce does to mix the worm source, this moment should with from the newly hatched larvae of same female moth separately mean allocation to each strain sugarcane strain to be measured.
Step D. investigation and evaluation of resistance
Connect worm and can observe the shape of being injured of lobus cardiacus on the 2nd day, existence situation of 5 days " Invest, Then Investigate " larvas and sugarcane strain extent of injury are formulated grade scale by pest-resistant sugarcane strain bar snout moth's larva infringement degree, evaluate the resistance rank of the strain of participating in the experiment.
Insect resistace is estimated: according to the insect resistace of larval mortality and sugarcane borer pest degree comprehensive evaluation commentaries on classics Bt sugarcane.According to test findings, the extent of injury of sugarcane strain wants big with the relation of survival larva survival state than larval mortality.Larvae alive is few in some sugarcane strain, but growth is normal, and being injured of sugarcane strain is still more serious; And some sugarcane strain larvae alive is more, but shows dullly, and food ingestion is little, and invasive is not strong, and sugarcane strain extent of injury is lighter.In view of the above, the extent of injury of sugarcane strain can be with the growth conditions of larva, get the seating in a restaurant or dining hall and put and eat the normative reference (table 1) that indexs such as the trace order of severity are made grading evaluation.In table 1 grade scale, 1 grade and 2 grades is only got the food back on a small quantity for larva is dead, and the sugarcane strain shows very strong insect resistace; 3 grades have the obvious growth hysteresis phenomenon for larva: appetite is little, growth is slow, getting the seating in a restaurant or dining hall, to put transfer velocity slow; 4 grades for larva has lighter diapause phenomenon, the plant insect resistace a little less than; 5 grades then at when investigation and the basic indifference of check variety.
The pest-resistant sugarcane strain of table 1 bar snout moth's larva infringement grading standard (connect worm after investigation in the 7th day)
| The borer pest rank | Describe |
| 1 2 3 4 5 | Lobus cardiacus has only a small amount of pin mark shape to get the food vestige, there is not perforation, no excrement bits lobus cardiacus is eaten the hole on a small quantity, see the argol bits, but nothing is newly eaten mark raw and excrement bits lobus cardiacus is eaten hole food mark (floral leaf) in a large number, less or do not have and newly to eat mark and worm excrement raw, larva is still got food on the lobus cardiacus blade, the little floral leaf of the individual obviously comparison of survival larva photograph is serious, 1/3 above larva has changed leaf sheath portion over to and has got food, the close floral leaf of survival larva polypide or worm age and contrast is serious, and most larvas are got food at leaf sheath and sugarcane stem tissue, and the borer pest joint is arranged |
Qualification result and analysis
Test findings shows: 2 check varieties, i.e. and Yck and Tck, the larva of access all survives; In the trans Bt gene strain, except that Y1, Y5, T3 and T4, other 9 strains all show stronger insect resistace, 7 strains such as Y2, Y3, Y4, T1, T2, T5 and T6 wherein, the 7th day larval mortality reaches 100% after connecing worm, and T7 and T8 reach 83.3% and 75% respectively, see Table 2.The larva of invading food Bt sugarcane mainly shows as dead and the progress of growing is slowed down, and the sugarcane strain shows as that the food trace is few, " floral leaf " or " floral leaf " symptom not occur lighter, does not have or the few borer pest joint and the withered heart, as depicted in figs. 1 and 2.
The 7th Heaven's commandments snout moth's larva larval mortality and borer pest rank behind the table 2 greenhouse test sense worm
| Strain | Mortality ratio (%) | The borer pest rank | Strain | Mortality ratio (%) | The borer pest rank | |
| Y1 | 0 | 5 | T1 | 100 | 2 | |
| Y2 | 100 | 1 | T2 | 100 | 2 | |
| Y3 | 100 | 2 | T3 | 0 | 4 | |
| Y4 | 100 | 2 | T4 | 0 | 4 | |
| Y5 | 0 | 4 | T5 | 100 | 3 | |
| Yck | 0 | 5 | T6 | 100 | 1 | |
| T7 | 83.3 | 3 | ||||
| T8 | 75 | 3 | ||||
| Tck | 0 | 5 |
Claims (6)
1. a transgenic Bt sugarcane is characterized in that comprising the steps: to the rapid identification method of snout moth's larva resistance
A. greenhouse barrel plant sugarcane;
B. the worm source is selected, and serves as test worm source with the bar snout moth's larva larva of first hatching;
C. manually connect worm, the worm source of step B is manually connect worm to the barrel plant sugarcane of steps A;
D. evaluation of resistance, through step C manually meet 5 days " Invest, Then Investigate " results of worm, according to the participate in the experiment insect resistace of sugarcane strain of the extent of injury comprehensive evaluation of the survival rate of larva, sugarcane;
Be the newly hatched larvae that the female moth of same top snout moth's larva is laid eggs to test worm source among the step B wherein with a collection of transgenic Bt sugarcane strain to be evaluated.
2. a transgenic Bt sugarcane is characterized in that comprising the steps: to the rapid identification method of snout moth's larva resistance
A. greenhouse barrel plant sugarcane;
B. the worm source is selected, and serves as test worm source with the bar snout moth's larva larva of first hatching;
C. manually connect worm, the worm source of step B is manually connect worm to the barrel plant sugarcane of steps A;
D. evaluation of resistance, through step C manually meet 5 days " Invest, Then Investigate " results of worm, according to the participate in the experiment insect resistace of sugarcane strain of the extent of injury comprehensive evaluation of the survival rate of larva, sugarcane;
Wherein among the step B to the test worm source with a collection of transgenic Bt sugarcane strain to be evaluated be newly hatched larvae that the female moth of 2～3 top snout moth's larvas is produced respectively mean allocation to every strain sugarcane strain to be measured.
3. transgenic Bt sugarcane as claimed in claim 1 or 2 is characterized in that to the rapid identification method of snout moth's larva resistance the step in acquisition test worm source among the step B is as follows:
B1. gather the pupa of article sugarcane borer in the sugarcane field, place in the box, under indoor conditions, sprout wings, allow female, male moth natural mating;
B2. select the female moth of successful post-coitum, put into transparent plastic bottle, every bottle of 1 female moth, treat that it laid eggs back 4 days in, with scissors or pocket knife pieces of an egg are taken off one by one together with the plastic bottle wall, under stereoscope, calculate every ovum grain number, and pieces of an egg are placed in the double dish that is placed with thieving paper preserve moisture.
4. transgenic Bt sugarcane as claimed in claim 3 is characterized in that manually connecing determining of worm time among the step C to the rapid identification method of snout moth's larva resistance: when finding in the ovum pore to be arranged, promptly carry out the preparation that connects worm, begin to connect worm when the larva hatching is arranged immediately.
5. transgenic Bt sugarcane as claimed in claim 1 or 2 is characterized in that the rapid identification method of snout moth's larva resistance:
In the steps A, barrel plant is cultivated the transgenic Bt sugarcane strain and the acceptor kind thereof of pending anti-snout moth's larva evaluation in the greenhouse, every barrel of 4 the sugarcane buds of sowing, and Cheng Miaohou stays seedling 2～4 strains for every barrel; The sugarcane of selecting 1～1.5 meter of plant height is as the suitable strain age of testing;
Among the step C, the quantity that the larva that just incubates is connect according to plan worm is connected to the lobus cardiacus portion that sugarcane is tested in every strain, incubates bar snout moth's larva larva at the beginning of the lobus cardiacus of every strain sugarcane connects 10, and connecing the worm duration is 1～2 day;
Among the step D, connect worm is observed lobus cardiacus after the 2nd day the shape of being injured, existence situation of 5 days " Invest, Then Investigate " larvas and sugarcane strain extent of injury are pressed the resistance rank that " pest-resistant sugarcane strain bar snout moth's larva infringement grading standard " evaluates the strain of participating in the experiment.
6. transgenic Bt sugarcane as claimed in claim 5 is characterized in that the rapid identification method of snout moth's larva resistance:
Among the step C, require every strain sugarcane of same test to insert the newly hatched larvae from same female moth of equal number on the same day.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101785443A (en) * | 2010-02-11 | 2010-07-28 | 江苏省农业科学院 | Inoculation method for improving accuracy of rice stem borer test |
| CN102577897A (en) * | 2012-02-16 | 2012-07-18 | 扬州大学 | Method for ecological safety evaluation on nilaparvata lugens by Bt genetically modified rice based on intervention of pesticides |
| CN104365314A (en) * | 2014-01-26 | 2015-02-25 | 江西省农业科学院植物保护研究所 | Method for identifying thrip resistance of asparagus |
| CN112034097A (en) * | 2019-07-31 | 2020-12-04 | 广西壮族自治区植保站 | Method for determining trapping and killing effect of prodenia litura adults |
| CN115413551A (en) * | 2022-07-26 | 2022-12-02 | 广西壮族自治区农业科学院 | Sugarcane germplasm material field resistance to borer evaluation method |
Family Cites Families (3)
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| CN1183558A (en) * | 1997-10-16 | 1998-06-03 | 山东农业大学 | Transgenic cotton insect-resistant test method |
| CN1318286A (en) * | 2000-04-14 | 2001-10-24 | 浙江大学 | Breeding process of hydrid rice borer resisting Bt sterile line and Bt restoring line |
| CN100501398C (en) * | 2006-03-16 | 2009-06-17 | 崔金杰 | Fast inspection of transgenic cotton against pests |
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- 2007-03-27 CN CN2007100273028A patent/CN101113977B/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101785443A (en) * | 2010-02-11 | 2010-07-28 | 江苏省农业科学院 | Inoculation method for improving accuracy of rice stem borer test |
| CN101785443B (en) * | 2010-02-11 | 2011-12-21 | 江苏省农业科学院 | Inoculation method for improving accuracy of rice stem borer test |
| CN102577897A (en) * | 2012-02-16 | 2012-07-18 | 扬州大学 | Method for ecological safety evaluation on nilaparvata lugens by Bt genetically modified rice based on intervention of pesticides |
| CN104365314A (en) * | 2014-01-26 | 2015-02-25 | 江西省农业科学院植物保护研究所 | Method for identifying thrip resistance of asparagus |
| CN112034097A (en) * | 2019-07-31 | 2020-12-04 | 广西壮族自治区植保站 | Method for determining trapping and killing effect of prodenia litura adults |
| CN112034097B (en) * | 2019-07-31 | 2022-06-07 | 广西壮族自治区植保站 | Method for determining trapping and killing effect of prodenia litura adults |
| CN115413551A (en) * | 2022-07-26 | 2022-12-02 | 广西壮族自治区农业科学院 | Sugarcane germplasm material field resistance to borer evaluation method |
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