CN107182945A - A kind of yellow sugarcane aphid indoors artificial captive breeding method - Google Patents
A kind of yellow sugarcane aphid indoors artificial captive breeding method Download PDFInfo
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- CN107182945A CN107182945A CN201710411546.XA CN201710411546A CN107182945A CN 107182945 A CN107182945 A CN 107182945A CN 201710411546 A CN201710411546 A CN 201710411546A CN 107182945 A CN107182945 A CN 107182945A
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- 241001179564 Melanaphis sacchari Species 0.000 title claims abstract description 77
- 238000009395 breeding Methods 0.000 title claims abstract description 34
- 231100000252 nontoxic Toxicity 0.000 claims abstract description 39
- 230000003000 nontoxic effect Effects 0.000 claims abstract description 39
- 240000000111 Saccharum officinarum Species 0.000 claims abstract description 36
- 235000007201 Saccharum officinarum Nutrition 0.000 claims abstract description 34
- 230000001488 breeding effect Effects 0.000 claims abstract description 20
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 5
- 238000000265 homogenisation Methods 0.000 claims abstract description 3
- 230000001954 sterilising effect Effects 0.000 claims abstract description 3
- 241000700605 Viruses Species 0.000 claims description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 5
- 238000003757 reverse transcription PCR Methods 0.000 claims description 5
- 239000000645 desinfectant Substances 0.000 claims description 4
- 238000001514 detection method Methods 0.000 claims description 4
- 238000005286 illumination Methods 0.000 claims description 4
- 238000011081 inoculation Methods 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- 238000012360 testing method Methods 0.000 claims description 3
- RCTYPNKXASFOBE-UHFFFAOYSA-M chloromercury Chemical compound [Hg]Cl RCTYPNKXASFOBE-UHFFFAOYSA-M 0.000 claims description 2
- 230000002045 lasting effect Effects 0.000 claims description 2
- 239000012528 membrane Substances 0.000 claims description 2
- 201000010099 disease Diseases 0.000 abstract description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 5
- 230000007613 environmental effect Effects 0.000 abstract description 2
- 244000005700 microbiome Species 0.000 abstract description 2
- 238000002474 experimental method Methods 0.000 description 5
- 241001124076 Aphididae Species 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 4
- 241000357694 Sugarcane yellow leaf virus Species 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- 241000238631 Hexapoda Species 0.000 description 3
- 240000008042 Zea mays Species 0.000 description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 3
- 235000005822 corn Nutrition 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241000607479 Yersinia pestis Species 0.000 description 2
- 239000002574 poison Substances 0.000 description 2
- 231100000614 poison Toxicity 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 235000015001 Cucumis melo var inodorus Nutrition 0.000 description 1
- 240000002495 Cucumis melo var. inodorus Species 0.000 description 1
- 241000258937 Hemiptera Species 0.000 description 1
- 241000819999 Nymphes Species 0.000 description 1
- 244000046109 Sorghum vulgare var. nervosum Species 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000003905 agrochemical Substances 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000002085 persistent effect Effects 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000001932 seasonal effect Effects 0.000 description 1
- 230000014599 transmission of virus Effects 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/90—Feeding-stuffs specially adapted for particular animals for insects, e.g. bees or silkworms
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Polymers & Plastics (AREA)
- Animal Husbandry (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Insects & Arthropods (AREA)
- Birds (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Food Science & Technology (AREA)
- Animal Behavior & Ethology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The present invention relates to a kind of yellow sugarcane aphid indoors artificial captive breeding method.Culture, the sterilization of yellow sugarcane aphid, virus-free yellow sugarcane aphid, sterileization, the homogenization that yellow sugarcane aphid indoors artificial captive breeding method includes sterile nontoxic sugarcane tissue-culture bottle seedling are cultivated, the virus-free sterileization Secondary Culture of yellow sugarcane aphid.The present invention is indoors raising medium using sterile nontoxic sugarcane tissue-culture bottle seedling, the sterile nontoxic yellow sugarcane aphid of hand-feeding propagation, yellow sugarcane aphid growth and breeding in closed culture vessel, germ-free condition is kept throughout the year, do not influenceed by environmental microorganism and natural enemy, also it will not migrate into external environment, it is to avoid propagation and the prevalence of corresponding disease to periphery crop cause of disease.It is low, easy to operate as medium, cost is raised with sterile nontoxic sugarcane tissue-culture bottle seedling, sufficient amount of nontoxic, homogeneous yellow sugarcane aphid worm sources can be provided with the anniversary.
Description
Technical field
The present invention relates to a kind of insect captive breeding method, and in particular to a kind of yellow sugarcane aphid indoors artificial captive breeding method,
Belong to insect captive breeding technical field.
Background technology
Sugarcane aphid（Melanaphis sacchari）There is yellow sugarcane aphidLongiunguis sacchari Zehntner（Also known as
Kaoliang aphid）, sugar cane cottony aphidCeratovacuna lanigeraZehntner（Also known as powder aphid）With thorn root aphidTetrancura bi rsuta Tri- kinds of Baker, is Homoptera, aphid section.Wherein yellow sugarcane aphid is sucked with adult or nymph cluster Sugarcane Leaves blade back
Juice, makes sugarcane leaf withered and yellow or even wilting, and a large amount of honeydews secreted out of can cause dark mildew, influences photosynthesis, causes yield to subtract
Production, sugar decline.Yellow sugarcane aphid is also to propagate the various plants such as corn mosaic virus, jowar mosaic virus and sugarcane yellow leaf virus
The communication media insect of virus, is propagated with non-persistent manner.
At present, yellow sugarcane aphid is as experiment materials such as viral transmission approach and resistant tests, and worm sources used are typically direct
Field host plant is picked up from, it is necessary to spend a large amount of manpower and materials, and under natural environment, the growth and breeding of yellow sugarcane aphid is by season
Section and environmental influence, it is impossible to which guarantee has enough experiment materials, and homogeneity is poor.Therefore, it is badly in need of solving yellow sugarcane aphid
Indoors artificial captive breeding technical problem, sets up simple and easy to apply, rapid, high volume reproduction technique system, be sugarcane resistance breeding to pests and
The research and development for preventing and treating the novel agrochemical of yellow sugarcane aphid provide the worm sources experiment material that reliable, homogeneity is good, be not subject to seasonal restrictions.
The content of the invention
, can be with convenient and swift week it is an object of the invention to provide a kind of method of indoors artificial amount reproduction yellow sugarcane aphid
Year persistently provides sufficient amount of virus-free, sterile, homogeneity good yellow sugarcane aphid.
A kind of yellow sugarcane aphid indoors artificial captive breeding method of the present invention, it is characterised in that indoors artificial captive breeding
Step is as follows：
（1）The culture of sterile nontoxic sugarcane tissue-culture bottle seedling：Cultivate, expand and numerous do not take viruliferous sterile nontoxic sugarcane tissue-culture after testing
Bottle seedling, and with sterile nontoxic sugarcane tissue-culture bottle seedling, it is used as the raising medium of yellow sugarcane aphid；
（2）The sterilization of yellow sugarcane aphid：The Sugarcane Leaves with yellow sugarcane aphid are gathered from field, in the yellow sugarcane aphid group of cluster together
Several body is extracted in body, determines, without carrying after virus, first to use sterilized water using molecular biology RT-PCR detection method
Rinse, aseptic filter paper suck dry moisture, then sterilize 3 ~ 5 min with disinfectant on superclean bench, then with aseptic water washing 3 ~ 5
It is secondary, aseptic filter paper suck dry moisture is used, sterile nontoxic yellow sugarcane aphid is obtained；
（3）Yellow sugarcane aphid is virus-free, sterileization, homogenization culture：In step（2）Sterile nontoxic yellow sugarcane aphid in, choose big
Small consistent several body, is inoculated in some sterile nontoxic sugarcane tissue-culture bottle seedlings, screws the bottle cap containing ventilated membrane, it is placed in 18 ~
30 DEG C, the hd of illumination 8 ~ 16-1, cultivate under the conditions of the Lx of light intensity 2000 ~ 10000；Using sterile nontoxic sugarcane tissue-culture bottle seedling as
The raising medium of yellow sugarcane aphid, during hand-feeding propagation, the sterile nontoxic sugarcane group more renewed at interval of a period of time
Train bottle seedling, you can uninterruptedly obtain virus-free, sterile, homogeneous yellow sugarcane aphid population；
（4）The virus-free sterileization Secondary Culture of yellow sugarcane aphid：Take step（2）Or（3）The yellow sugarcane aphid of acquisition is as worm sources, by step
Suddenly（3）Method for breeding, passage breeding, so as to keep the lasting passage of nontoxic sterile yellow sugarcane aphid to breed.
Wherein, step（2）Described disinfectant is the HgCl that concentration expressed in percentage by weight is 0.1 %2。
Step（3）The inoculation, refers to one sterile nontoxic yellow sugarcane aphid of every bottle of access.
Step（3）It is described at interval of a period of time, be at interval of 10 ~ 12 d.
Advantages of the present invention and usefulness：
1st, yellow sugarcane aphid indoors artificial captive breeding method of the invention, artificial using sterile nontoxic sugarcane tissue-culture bottle seedling indoors
The sterile nontoxic yellow sugarcane aphid of captive breeding, is not influenceed by natural conditions, can provide sufficient amount of nontoxic, homogeneous worm with the anniversary
Source.
2nd, sterile nontoxic yellow sugarcane aphid growth and breeding in closed culture vessel, will not migrate into external environment, keep away
The propagation to periphery crop cause of disease and the prevalence of corresponding disease are exempted from.
3rd, sterile nontoxic yellow sugarcane aphid growth and breeding in closed culture vessel, keeps germ-free condition, not by ring throughout the year
Border microorganism and natural enemy influence.
4th, it is low, easy to operate as medium, cost is raised with sterile nontoxic sugarcane tissue-culture bottle seedling.
Brief description of the drawings
Fig. 1：Yellow sugarcane aphid be inoculated with 2 days after sterile nontoxic sugarcane tissue-culture bottle seedling.
Fig. 2：Yellow sugarcane aphid raising up seed in sterile nontoxic sugarcane tissue-culture bottle seedling.
Fig. 3：Yellow sugarcane aphid growth and breeding curve in sterile nontoxic sugarcane tissue-culture bottle seedling.In figure, abscissa represents inoculation
Number of days afterwards, ordinate represents breeding rate.
Embodiment
For the yellow sugarcane aphid indoors artificial captive breeding method of the fully open present invention, with reference to embodiment to this hair
It is bright to be described in further detail.
Embodiment 1, a kind of yellow sugarcane aphid indoors artificial captive breeding method, comprise the following steps：
The sugarcane plant of good fortune agriculture 41 of robust growth, no disease and pests harm is taken from field, stem-segment with single bud is chopped to, the full bud of growth selection,
In 52 DEG C of constant temperature circulating hot waters bath 30 min of processing, 37 DEG C of vernalization are put, when bud height is not less than 5 cm, blade are taken, with reference to being published in
《Annals of Applied Biology》（Luo et al., 2016）With《Virus Genes》（Gao et al.,
2012）The RT-PCR detection method of corn mosaic virus, jowar mosaic virus and sugarcane yellow leaf virus described by document, it is determined that
Do not carry after virus, take bud, the bud s of 75 % ethanol disinfections 0 ~ 30, with 0.1 % HgCl215 min are sterilized, 3 ~ 5 are peelled off
Leaf sheath, cuts and is about stem apexs of 1 cm with growing point and is inoculated in culture medium A（MS + 6-BA 1.0mg/L + KT 0.5 mg/L
The g/L of+activated carbon 0.5+sugared 20 g/L）In, in 25 ± 2 DEG C, the hd of illumination 10 ~ 12-1, the Lx bars of light intensity 2000 ~ 3000
Cultivated in growth cabinet under part, every 20 d is transferred in the fresh culture of identical component, grows and culture is transferred to after clump bud
Base B（The mg/L of 1.0 mg/L+KT of the MS+6-BA 0.5+g/L of sugared 20 g/L+agar 5 ~ 7）In, every 20 d is transferred to phase
In congruent fresh culture, the sterile nontoxic sugarcane tissue-culture bottle seedling for growing to 3 ~ 5 cm height is situated between as the raising of yellow sugarcane aphid
Matter.
Field takes the Sugarcane Leaves with yellow sugarcane aphid, and 12 are randomly selected out from the yellow sugarcane aphid colony of cluster together
Individual, with reference to being published in《Annals of Applied Biology》（Luo et al., 2016）With《Virus Genes》
（Gao et al., 2012）The RT-PCR of corn mosaic virus, jowar mosaic virus and sugarcane yellow leaf virus described by document
Detection method, it is determined that without carrying after virus, the Sugarcane Leaves with remaining yellow sugarcane aphid first being used into aseptic water washing, then used
Aseptic filter paper suck dry moisture, carries out disinfection by sterile working mode on superclean bench, is inoculated with.Then concentration expressed in percentage by weight is used
For 0.1 % HgCl24 min are sterilized, then with after aseptic water washing 3 times, are placed on aseptic filter paper, band yellow sugarcane aphid
Blade be divided into the fritter blade for only carrying 1 yellow sugarcane aphid, access in sterile nontoxic sugarcane tissue-culture bottle seedling, as shown in figure 1,
One piece of blade of every bottle of inoculation, i.e., one sterile nontoxic yellow sugarcane aphid of every bottle access..
Inoculated sterile sugarcane tissue-culture bottle seedling is placed in 22 ± 1 DEG C, the hd of illumination 12-1, the Lx conditions of light intensity 6000
Yellow sugarcane aphid can be observed in lower culture, 48 h to move on aseptic plant.Yellow sugarcane aphid is raw on sterile nontoxic sugarcane plant
Long breeding, 3 ~ 5 d generation, generation overlap.As shown in Fig. 2 the process of sterile nontoxic yellow sugarcane aphid hand-feeding propagation indoors
In, with the time lengthening of hand-feeding propagation, yellow sugarcane aphid population quantity also gradually increases；As shown in figure 3, working as artificial feeding
Breed after 8 d, yellow sugarcane aphid population quantity is drastically expanded；During 10 d of hand-feeding propagation, average breeding rate is 29.8 times；
During 12 d of hand-feeding propagation, average breeding rate is 58.5 times.At this moment because yellow sugarcane aphid Population is excessive, some sugarcanes
There are the phenomena of mortality in yellow aphids individual.Therefore, clip carries the aseptic plant blade of yellow sugarcane aphid during 10 ~ 12 d of hand-feeding propagation
It is transferred in fresh sterile tissue-culture container seedling and continues to pass on breeding；Or choose the good yellow sugarcane aphid of homogeneity as pass poison experiment or
The worm sources of the experiments such as the resistance to the action of a drug.Above yellow sugarcane aphid detects by molecular biology RT-PCR, does not detect mosaic of sugarcane
Poison, jowar mosaic virus and sugarcane yellow leaf virus.
The foregoing is only presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, should all belong to the covering scope of the present invention.
Claims (4)
1. a kind of yellow sugarcane aphid indoors artificial captive breeding method, it is characterised in that as follows the step of indoors artificial captive breeding：
（1）The culture of sterile nontoxic sugarcane tissue-culture bottle seedling：Cultivate, expand and numerous do not take viruliferous sterile nontoxic sugarcane tissue-culture after testing
Bottle seedling, and with sterile nontoxic sugarcane tissue-culture bottle seedling, it is used as the raising medium of yellow sugarcane aphid；
（2）The sterilization of yellow sugarcane aphid：The Sugarcane Leaves with yellow sugarcane aphid are gathered from field, in the yellow sugarcane aphid group of cluster together
Several body is extracted in body, determines, without carrying after virus, first to use sterilized water using molecular biology RT-PCR detection method
Rinse, aseptic filter paper suck dry moisture, then sterilize 3 ~ 5 min with disinfectant on superclean bench, then with aseptic water washing 3 ~ 5
It is secondary, aseptic filter paper suck dry moisture is used, sterile nontoxic yellow sugarcane aphid is obtained；
（3）Yellow sugarcane aphid is virus-free, sterileization, homogenization culture：In step（2）Sterile nontoxic yellow sugarcane aphid in, choose big
Small consistent several body, is inoculated in some sterile nontoxic sugarcane tissue-culture bottle seedlings, screws the bottle cap containing ventilated membrane, it is placed in 18 ~
30 DEG C, the hd of illumination 8 ~ 16-1, cultivate under the conditions of the Lx of light intensity 2000 ~ 10000；Using sterile nontoxic sugarcane tissue-culture bottle seedling as
The raising medium of yellow sugarcane aphid, during hand-feeding propagation, the sterile nontoxic sugarcane group more renewed at interval of a period of time
Train bottle seedling, you can uninterruptedly obtain virus-free, sterile, homogeneous yellow sugarcane aphid population；
（4）The virus-free sterileization Secondary Culture of yellow sugarcane aphid：Take step（2）Or（3）The yellow sugarcane aphid of acquisition is as worm sources, by step
Suddenly（3）Method for breeding, passage breeding, so as to keep the lasting passage of nontoxic sterile yellow sugarcane aphid to breed.
2. a kind of yellow sugarcane aphid indoors artificial captive breeding method according to claim 1, it is characterised in that step（2）Institute
The disinfectant stated is the HgCl that concentration expressed in percentage by weight is 0.1 %2。
3. a kind of yellow sugarcane aphid indoors artificial captive breeding method according to claim 1, it is characterised in that step（3）Institute
Inoculation is stated, refers to one sterile nontoxic yellow sugarcane aphid of every bottle of access.
4. a kind of yellow sugarcane aphid indoors artificial captive breeding method according to claim 1, it is characterised in that step（3）Institute
State at interval of a period of time, be at interval of 10 ~ 12 d.
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| CN201710411546.XA CN107182945A (en) | 2017-06-05 | 2017-06-05 | A kind of yellow sugarcane aphid indoors artificial captive breeding method |
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| CN201710411546.XA CN107182945A (en) | 2017-06-05 | 2017-06-05 | A kind of yellow sugarcane aphid indoors artificial captive breeding method |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101401549A (en) * | 2008-11-04 | 2009-04-08 | 广州甘蔗糖业研究所 | Once-seedling forming quick propagating method for sugarcane tissue culture |
| CN102578047A (en) * | 2012-02-27 | 2012-07-18 | 中国农业科学院柑桔研究所 | Rearing method of non-toxic brown citrus aphids for researching virus transmission molecular mechanism |
| CN103081808A (en) * | 2013-02-18 | 2013-05-08 | 广西壮族自治区农业科学院甘蔗研究所 | Sugarcane tissue culture and rapid propagation production method adopting ex vitro rooting |
| CN105660546A (en) * | 2016-04-01 | 2016-06-15 | 云南省农业科学院花卉研究所 | Method for feeding frankliniella occidentalis indoors |
-
2017
- 2017-06-05 CN CN201710411546.XA patent/CN107182945A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101401549A (en) * | 2008-11-04 | 2009-04-08 | 广州甘蔗糖业研究所 | Once-seedling forming quick propagating method for sugarcane tissue culture |
| CN102578047A (en) * | 2012-02-27 | 2012-07-18 | 中国农业科学院柑桔研究所 | Rearing method of non-toxic brown citrus aphids for researching virus transmission molecular mechanism |
| CN103081808A (en) * | 2013-02-18 | 2013-05-08 | 广西壮族自治区农业科学院甘蔗研究所 | Sugarcane tissue culture and rapid propagation production method adopting ex vitro rooting |
| CN105660546A (en) * | 2016-04-01 | 2016-06-15 | 云南省农业科学院花卉研究所 | Method for feeding frankliniella occidentalis indoors |
Non-Patent Citations (1)
| Title |
|---|
| NUESSLY,GREGG S.L: "Susceptibility of stage IV canal point(CP) sugarcane clones to yellow sugarcane aphid(sipha flava(forbes)) feeding damage", 《JOURNAL OF THE AMERICAN SOCIETY OF SUGAR CANE TECHNOLOGISTS》 * |
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