CN104542144A - Resistance identification method for southern rice black-streaked dwarf virus variety - Google Patents
Resistance identification method for southern rice black-streaked dwarf virus variety Download PDFInfo
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- 241000756822 Southern rice black-streaked dwarf virus Species 0.000 title abstract description 13
- 241000176086 Sogatella furcifera Species 0.000 claims abstract description 133
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- 235000007164 Oryza sativa Nutrition 0.000 claims abstract description 89
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- 241000702634 Rice black streaked dwarf virus Species 0.000 claims description 53
- 201000010099 disease Diseases 0.000 claims description 53
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 53
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- A01G7/00—Botany in general
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The invention discloses a resistance identification method for southern rice black-streaked dwarf virus variety. The method comprises the steps of (a) acquiring sogatella furcifera, and separating and purifying a large amount of uniform and consistent 5-age sogatella furcifera nymphs; (b) breeding, separating and screening a large amount of uniform and consistent 2-age sogatella furcifera nymphs; (c) feeding virus to the obtained 2-age sogatella furcifera nymphs; (d) calculating the sogatella furcifera nymphs; (e) quantitatively co-breeding rice to be tested and the 2-age sogatella furcifera nymphs fed with virus, and spreading virus; (f) counting land symptomatic appearance, namely, transplanting the rice to be tested to a farm sogatella furcifera separating net chamber, investigating and counting the ratio of infected holes of southern rice black-streaked dwarf virus at the peak tillering phase, so as to accurately identify the resistance of the rice variety to be tested. With the adoption of the resistance identification method, the problems of the interference of antixenosis and the mass acquiring of sogatella furcifera can be overcome; the quantitative and reinforced inoculation identification of southern rice black-streaked dwarf virus can be achieved; the resistance level of the rice variety to southern rice black-streaked dwarf virus can be accurately evaluated.
Description
Technical field
The present invention relates to the sick varietal resistance authentication method of a kind of southern rice black-streaked dwarf, belong to plant disease authenticate technology.
Background technology
Southern rice black-streaked dwarf disease is found by Zhou Guohui etc., identify and named in Yangxi County of Guangdong Province of China in calendar year 2001 first." black-streaked dwarf " in disease title represents the classical symptom of disease, and the tubercle that namely diseased plant stunts and stem stalk surface has black bar to arrange is dashed forward." south " represents this disease Late Cambrian in South China, and mainly betides South Rice Region of China.Its field symptom is similar to black streaked dwarf virus of rice, and the latter mainly betides the lower wheat-paddy fields of temperature.The rice plant infecting this disease stunts, difficulty of not earing or ear, and seed is few and not full, causes heavy losses to output.Southern rice black-streaked dwarf virus is by southern rice black-streaked dwarf virus (Southern rice black-streaked dwarf virus, SRBSDV) cause, SRBSDV is the kind that Reoviridae Fijivirus belongs to, and belongs to together not of the same race with rice black-streaked dwarf virus.The black streaked dwarf virus of rice caused by rice black-streaked dwarf virus is propagated with persistent fashion primarily of small brown rice planthopper, natural host comprises the gramineous crops such as corn, paddy rice and wheat, and the southern rice black-streaked dwarf caused by southern rice black-streaked dwarf virus is sick primarily of the propagation of white-backed planthopper band poison, it is the main pathogen causing North Vietnam and south China provinces and regions rice dwarf virus disease in recent years.
In recent years, the ground population outbreak such as the sick Hainan at home of southern rice black-streaked dwarf, Guangdong and Guangxi Provinces, Hunan, Jiangxi, Jiangsu and Zhejiang Provinces and external Vietnam.According to incompletely statistics, within 2009, injured area is more than 300,000 hm, and about 6500hm paddy rice is lost and receives.2011, the Ministry of Agriculture set up the sick groupcontrol cooperative groups of southern rice black-streaked dwarf and expert instructs group, and strengthen the prevention and control dynamics to disease, onset area controls at about 24.67 ten thousand hm.Within 2012, disease is gone up to some extent, onset area about 400,000 hm.This disease economizes (autonomous region) semilate rice in China Guangdong, Guangxi, Hainan, Hunan, Hubei, Jiangxi, Yunnan and Guizhou etc. and late rice harm is the most serious, the common grave illness field that No kernels or seeds are gathered, as in a year of scarcity.Breaking out greatly of southern rice black-streaked dwarf disease bring serious harm to the production of south China paddy rice, and setback peasant's enthusiasm for production, brings great challenge to national food security.
As a kind of virus disease, lack the medicament effectively can preventing and treating this disease in the market, the prevention and control of southern rice black-streaked dwarf disease mainly adopt the measure of " controlling worm diseases prevention ", normally adopt insecticide to kill white-backed planthopper in early days and pass poison ring joint to block.But more than white-backed planthopper is moved into batch, insects sources is failed to understand, raises difficult questions for discussion to hold when moving into, and the biography poison characteristic of this worm is also not fully aware of.The disappearance of basic research makes to observe and predict difficulty to SRBSDV, prevention and control blindness is large, applicable measure few, long-term control effect is not good.
It is the most direct effective method that screening Resistant germplasm is applied to breeding for disease resistance production, and screening, excavation and innovation Resistant gerplasm are prerequisite and the basis of carrying out breeding for disease resistance.But black streaked dwarf virus of rice Resistance QTL detection so far and genetic effect analysis are also in the starting stage, the kind to SRBSDV immunity is not found, so carry out Large-scale Screening to southern rice black-streaked dwarf sick resistance kind matter and launch the sick genetics of resistance characteristic research of southern rice black-streaked dwarf seeming particularly urgent on producing yet.
The sick Resistance resource screening of current southern rice black-streaked dwarf only has employing land for growing field crops natural occurrence survey method, each identification of species or strain are planted in land for growing field crops with community form, about each community tens strain, land for growing field crops conventional cultivation management, pass virus mediator white-backed planthopper and naturally pass poison, treat that paddy rice manifests the cave incidence of disease of each identification of species of illness " Invest, Then Investigate " (family) community, differentiate the difference of rice varieties (family) anti-sense ability as standard.But the natural occurrence situation of each community of natural occurrence survey method, land for growing field crops is difficult to consistent, and the repeatability of same process different districts is poor, test data is difficult to analyze.Again because the resistance of plant to natural enemy is divided into tolerance to insects, antibiosis and antixenosis three kind, various resistance classification tool is by different resistance mechanisms.The difference of different rice varieties (family) Whitebacked Planthopper antixenosis makes the characteristics such as some physiology and morphology of different rice varieties (family), shows the difference taking food deflection of Whitebacked Planthopper.Therefore, identified the disease resisting rice obtained by natural appraisal method, pest-resistant (antixenosis is strong) paddy rice may be actually and be not disease resisting rice.But, if after carrying out the scale plantation of disease district with this pest-resistant not disease resisting rice improved variety, the disease-resistant performance of vacation that scale plantation causes biography virus mediator to be forced to select and paddy rice Whitebacked Planthopper is produced because arranging disappears, and may bring larger loss to Rice Production.
For getting rid of the impact of antixenosis enantiopathy qualification, artificial quantitatively inoculation band virus mediator being carried out to paddy rice and becomes a kind of inevitable.But, the Resistance Identification carrying out variety resources of rice needs a large amount of biography virus mediator white-backed planthoppers, and white-backed planthopper artificial feeding scale amount is little, and feeding cost is very high, be difficult to the demand of the scale amount meeting varietal resistance qualification, limit applying of artificial quantitatively inoculated identification.
White-backed planthopper quantity annual in the ecosystem of land for growing field crops, white-backed planthopper severely afflicated area is all very huge, artificial acquisition is also than being easier to, and laboratory rearing white-backed planthopper is used for variety source Resistance Identification and is difficult to the requirement meeting scale amount, if can design, a kind of method utilizes the white-backed planthopper of land for growing field crops ecosystem-scale amount quantitatively to strengthen paddy rice inoculated identification thus realize will be a kind of feasible selection to the accurate evaluation of variety resources of rice.
In land for growing field crops, insect and host crop were just living together in the very long period of history, dyadic interaction, to affect each other, mutual adjustment restriction, southern rice black-streaked dwarf disease waits insect to pass the large generation of virus mediator disease, just embodying long-term interaction between crop and insect, and the polytropy influenced each other by natural environment, ecotope between insect and crop restricts, for realizing the accurate evaluation to variety resources of rice, laboratory is needed to provide crop insect to make the stable and suitable ecotope studied mutually.In the simulation land for growing field crops ecosystem white-backed planthopper and paddy rice interaction, utilize the land for growing field crops ecosystem to solve an acquisition difficult problem, quantitative and strengthen southern rice black-streaked dwarf disease inoculated identification thus the accurate evaluation basic thinking of the present invention just realized variety resources of rice for scale white-backed planthopper.
This method is utilized to evaluate rice varieties to the resistance level of southern rice black-streaked dwarf disease, both the interference of antixenosis to qualification result can have been removed, a difficult problem for the required white-backed planthopper scale amount of varietal resistance qualification can be overcome again, and achieve the quantitatively sick inoculated identification of the southern rice black-streaked dwarf of strengthening.This authentication method, except can being applied to the excavation qualification of variety resources of rice, varietal resistance evaluation, disease-resistant variety seed selection, can also be applied to the various fields such as phenotypic evaluation and genetics of resistance law study of localization of disease resistance genes.
Summary of the invention
The white-backed planthopper scale amount that the object of the invention is for the antixenosis interference difficult problem existed in the sick Natural infection of southern rice black-streaked dwarf and indoors artificial inoculated identification obtains a difficult problem, there is provided a kind of southern rice black-streaked dwarf sick varietal resistance authentication method, this authentication method both can remove the interference of antixenosis to qualification result, solve again the difficult problem that the required white-backed planthopper scale amount of varietal resistance qualification obtains, and achieve the quantitatively sick inoculated identification of the southern rice black-streaked dwarf of strengthening, finally achieve the accurate evaluation to the sick varietal resistance of southern rice black-streaked dwarf.
The object of the invention is to solve by the following technical programs:
The sick varietal resistance authentication method of a kind of southern rice black-streaked dwarf, is characterized in that: described authentication method step is as follows:
(a), adopt enamel tray method to gather white-backed planthopper from field after by purifying screening plant obtain a large amount of 5 age white-backed planthopper nymph;
(b), screening is obtained 5 age white-backed planthopper nymph be inoculated into and the paddy rice cultivated in crop insect co-cultivation box educate altogether and breeds white-backed planthopper nymph of future generation, and utilize the purifying screening plant of adjustment to be separated further to obtain a large amount of neat and consistent 2 age white-backed planthopper nymph;
(c), by screening obtain 2 ages white-backed planthopper nymph move into plant have the seriously ill disaster area of southern rice black-streaked dwarf to draw materials tillering regularity paddy rice diseased plant on carry out raising poison;
(d), being weighed by precision electronic balance obtains the weight of single head 2 white-backed planthopper in age nymph, calculate required white-backed planthopper gross weight according to the worm determined/seedling ratio, and then weigh quantitatively;
(e), according to the worm determined/seedling ratio by raise after poison quantitative 2 age white-backed planthopper nymph be inoculated into the rice varieties rice shoot to be measured of cultivating in crop insect co-cultivation box educates altogether and pass poison;
(f), when white-backed planthopper half is dead by rice transplanting to be measured to land for growing field crops every worm solarium, conventional cultivation manages, the tillering regularity investigation statistics southern rice black-streaked dwarf sick cave incidence of disease, and the resistance against diseases of rice varieties to be measured is gone out according to cave incidence of disease precise Identification.
White-backed planthopper acquisition time in described step (a) paddy field at the beginning of annual 7 months turns the yellow phase by green grass or young crops.
When purifying screening plant in described step (a) uses, adopt aperture to be 18 orders and the combination of 22 object sub-sieves with effectively isolate 5 age white-backed planthopper nymph and part white-backed planthopper adult; Then by screening isolated white-backed planthopper pour in the dark bucket of inner wall smooth, bounce a barrel outer wall white-backed planthopper adult is flown away from and nymph be detained, can obtain a large amount of neat and consistent 5 age white-backed planthopper nymph.
In described step (b) 5 ages, white-backed planthopper nymph was inoculated on the paddy rice cultivated in crop insect co-cultivation box time, be 2 ~ 3 leaf phases with the paddy rice that 5 ages, white-backed planthopper nymph educated altogether.
The cultivation temperature of the crop insect co-cultivation box in described step (b) controls at 27 DEG C, the dark 12h of light 12h/.
After breeding white-backed planthopper nymph of future generation in crop insect co-cultivation box in described step (b), now screening plant overturns 90 ° and the vertical light direction that becomes is changed into horizontal direction, adopt aperture to be 30 orders with 32 object sub-sieves combinations be effectively separated obtain a large amount of neat and consistent 2 age white-backed planthopper nymph.
Tillering regularity paddy rice diseased plant in described step (c) is that upper one year is after drawing materials in the seriously ill disaster area of southern rice black-streaked dwarf in the refrigerator cold-storage of-20 DEG C, after diseased plant is thawed naturally, root immersion absorbent cotton parcel is wrapped up solid simulation field-transplanting mode with Polypropylence Sheet and is vertically fixed in crop insect co-cultivation box, then move into 2 age white-backed planthopper nymph raise poison 48 hours.
In described step (d) 2 ages white-backed planthopper nymph adopt light to be separated hairbrush brush from the impurity be mingled with during white-backed planthopper nymph and dead worm after raising poison, then transferase 12 0 ~ 50 white-backed planthopper nymph enters measuring cup, weighed by precision electronic balance, peeling calculates the quantity according to white-backed planthopper nymph in bottle after the gross weight of white-backed planthopper nymph in bottle, calculate the weight of single head white-backed planthopper nymph, determine to raise malicious white-backed planthopper nymph number according to paddy rice strain number to be measured and inoculation intensity, and then raise the gross weight of malicious white-backed planthopper nymph needed for calculating inoculation and weigh quantitatively.
Paddy growth to be measured in described step (e) was in for 1 ~ 2 leaf phase, and paddy rice to be measured is according to 20cm
2the ratio of/strain leaves and takes the healthy uniform rice seedling of growing way.
Paddy rice to be measured in described step (e) according to the ratio inoculation of 5 ~ 15 worms/seedling raise after poison 2 age white-backed planthopper nymph, the cultivation temperature of crop insect co-cultivation box control 27 DEG C, the dark 12h of light 12h/ educates that to pass poison dead to the white-backed planthopper naturally-aged of inoculation altogether.
The present invention has the following advantages compared to existing technology:
The present invention by purifying screening plant obtain source, a large amount of land for growing field crops 5 age white-backed planthopper nymph, and then by crop insect co-cultivation box bring out altogether a large amount of 2 age white-backed planthopper nymph, and again by purifying screening plant obtain a large amount of neat and consistent 2 age white-backed planthopper nymph, then by 2 age white-backed planthopper nymph educate altogether raise poison, rice quantitative to be measured educates the mode that inoculation passes poison altogether and makes rice pathogenesis to be measured, can go out the resistance against diseases of rice varieties to be measured by precise Identification finally by the investigation southern rice black-streaked dwarf sick cave incidence of disease.
Strengthening inoculation identification method of the present invention both can remove the interference of antixenosis to qualification result, the difficult problem that the required white-backed planthopper scale amount of varietal resistance qualification obtains can be overcome again, and achieve the quantitatively sick inoculated identification of the southern rice black-streaked dwarf of strengthening, the resistance level of rice varieties to southern rice black-streaked dwarf disease can be evaluated exactly; This authentication method, except can being applied to the excavation qualification of variety resources of rice, varietal resistance evaluation, disease-resistant variety seed selection, can also be applied to the various fields such as the phenotypic evaluation of localization of disease resistance genes and genetics of resistance law study, therefore suitablely promotes the use of.
Embodiment
Below in conjunction with embodiment, the present invention is further illustrated.
A kind of sick Resistance Identification method of southern rice black-streaked dwarf, this authentication method step is as follows: (a), the acquisition of land for growing field crops white-backed planthopper and separation and purification: at the beginning of annual 7 months, paddy field turns the yellow phase by green grass or young crops, when enamel tray method investigation white-backed planthopper 5 instar occupies the majority, enamel tray method is adopted to gather white-backed planthopper (wherein containing a large amount of impurity and assorted worm) from field, then purifying screening plant is utilized to carry out separation screening, for mainly obtain 5 age white-backed planthopper nymph, purifying screening plant adopts aperture to be 18 orders and the combination of 22 object sub-sieves, because sieve is got investigation and is found, 18 ~ 22 object sub-sieve combinations effectively can mask the dead worm of white-backed planthopper, impurity and nothing (weak) Phototactic insects, large build Phototactic insects, the interference of small body type Phototactic insects and white-backed planthopper children nymph in age, effectively isolate 5 age nymph and part white-backed planthopper adult, isolated white-backed planthopper is inserted in the dark bucket of inner wall smooth, bouncing a barrel outer wall makes white-backed planthopper adult fly away from and nymph delay, the a large amount of neat and consistent of final acquisition 5 age white-backed planthopper nymph, (b), 2 age white-backed planthopper nymph cultivation and separation screening: screening is obtained 5 age white-backed planthopper nymph be inoculated into and 2 ~ 3 leaf phase paddy rice cultivated in crop insect co-cultivation box educate altogether and breeds white-backed planthopper nymph of future generation, the cultivation temperature of crop insect co-cultivation box controls at 27 DEG C, the dark 12h of light 12h/, educate altogether and within about two weeks, can be observed the appearance of a large amount of white-backed planthopper nymph, treating that most nymph grew into for 2 length of times, being separated into plastic casing in by being gathered in Rice Leaf with a large amount of white-backed planthopper nymphs on stem stalk with the hairbrush of softness, for screen neat and consistent 2 age white-backed planthopper nymph, screening plant adjusts with reference to adult screening plant, the phototaxis of nymph is utilized to carry out sorting equally, sub-sieve combination selection 30 ~ 32 order, and due to nymph locomitivity more weak, now screening plant overturns 90 ° and arranges the vertical light direction that becomes is changed into horizontal direction, and nymph is placed in dark one end, the nymph light level that becomes is creeped, adopt 30 ~ 32 object sub-sieves combinations to be separated to obtain a large amount of neat and consistent 2 age white-backed planthopper nymph, c (), white-backed planthopper nymph raise poison: naturally thawed by the tillering regularity paddy rice diseased plant that upper one year draws materials in the seriously ill disaster area of southern rice black-streaked dwarf of the refrigerator and cooled Tibetan of-20 DEG C, diseased plant root immersion absorbent cotton is wrapped up and wraps up solid simulation field-transplanting mode with Polypropylence Sheet and is vertically fixed on (the illumination of only ventilating of crop insect co-cultivation box, do not sow, not water flowing) in, then move into 2 age white-backed planthopper nymph raise malicious 48h, make 2 age white-backed planthopper nymph band poison, (d) metering white-backed planthopper nymph: with the hairbrush of softness by raise after poison 2 age white-backed planthopper nymph be separated into plastic casing, do not establish sub-sieve, purifying screening plant is adopted to become light separation hairbrush brush from the impurity be mingled with during 2 white-backed planthopper nymph in age and dead worm, transferase 12 0 ~ 50 2 white-backed planthopper nymph in age enters measuring cup, precision electronic balance is weighed, peeling to calculate in bottle 2 age white-backed planthopper nymph gross weight, bottle in 2 age white-backed planthopper nymph naked eyes counting, calculate the weight of single head 2 white-backed planthopper in age nymph, to determine to raise malicious white-backed planthopper nymph number according to paddy rice strain number to be measured and inoculation intensity, and then raise the gross weight of malicious white-backed planthopper nymph needed for calculating inoculation and weigh quantitatively, (e), paddy rice to be measured and band malicious 2 age white-backed planthopper nymph quantitatively educate altogether and pass poison: when there are (many an age nymph) in a large number in white-backed planthopper nymph in the crop insect co-cultivation box in step (b), rice growing to be measured is cultivated in crop insect co-cultivation box, a rice varieties to be measured (family) sowed by every case, the growth of completing steps (c) rice seedling to be measured afterwards can be in for 1 ~ 2 leaf phase, according to 20cm
2the ratio of/strain eliminates weak seedling and uneven seedling to leave and take the healthy uniform rice seedling of growing way, according to 5 ~ 15 worms/seedling ratio by raise after poison quantitative 2 age white-backed planthopper nymph be inoculated into the rice varieties rice shoot to be measured of cultivating in crop insect co-cultivation box educates altogether and pass poison, the cultivation temperature of crop insect co-cultivation box control 27 DEG C, the dark 12h of light 12h/ educates that to pass poison dead to the white-backed planthopper half naturally-aged of inoculation altogether, (f), rice field to be measured show disease statistics: after the death of white-backed planthopper half by rice transplanting to be measured to land for growing field crops every worm solarium, conventional cultivation manages, the tillering regularity investigation statistics southern rice black-streaked dwarf sick cave incidence of disease, southern rice black-streaked dwarf disease symptoms main manifestations is plant dwarfing, blade is short wealthy, stiff, dark green leaf color, the vein of blade back and stem stalk show just wax, the billet warty protuberance of rear browning look, the statistical counting cave incidence of disease, and the resistance against diseases going out rice varieties to be measured according to southern rice black-streaked dwarf sick cave incidence of disease precise Identification.
Embodiment one
The freezing paddy rice diseased plant adopting the seriously ill disaster area of southern rice black-streaked dwarf to draw materials according to authentication method provided by the invention is as malicious source, the white-backed planthopper in source, inoculation land for growing field crops breeds next generation nymph, raise poison after 2 days by 2 ages white-backed planthopper nymph move into give birth to different rice varieties rice shoot (Wu-Yu-Geng 3, Kasalath, bright extensive 63,9311 and Mudgo) crop insect co-cultivation box in inoculate, inoculation intensity is 10 heads/plant, when white-backed planthopper lethality reaches half respectively by rice seedling planting in insect-proof net chamber; The same period is identified in southern rice black-streaked dwarf seriously ill disaster area natural occurrence these 5 rice varieties.To adopting the incidence of disease of the sick cave incidence of disease of the southern rice black-streaked dwarf of this method qualification and the qualification of severely afflicated area natural occurrence to add up, calculate the mutually antagonism sense ratio of the rice varieties cave incidence of disease compared with Wu-Yu-Geng 3 that two kinds of authentication methods obtain respectively simultaneously.
Can be found by table one, adopt this method qualification and the rice varieties cave incidence of disease of severely afflicated area natural occurrence and the sense of the antagonism mutually highly significant more most than difference respectively compared with Wu-Yu-Geng 3 thereof, the result of this technical appraisement of different cultivars paddy rice is more not identical than many with the resistance against diseases that severely afflicated area natural occurrence is identified, between kind, antagonism sense, than significant difference, may be that the antixenosis difference of different rice varieties causes mutually.
For studying the reason of the disease-resistant capacity variance of rice varieties of different authenticate technology qualification, we have done different cultivars antixenosis identification experiment.Rice paddy seed after vernalization is sowed in the plastic box of 65cm × 44cm × 14cm, each kind 1 row, often row 10 strain.Treat that seedling grows to 1.5 ~ 2.0 leaves, by the white-backed planthopper nymph in 3 ~ 4 ages of every strain 10 access, when every morning 8, afternoon 4 time investigate borer population on each individual plant, carry out expelling parasite after each investigation, make it be uniformly distributed as far as possible.Environmental temperature remains on 26 ± 2 DEG C, natural lighting.Within 5 days, calculate the average borer population on each individual plant of each kind afterwards, as driving property of row test value.
Can be found by table two, the antixenosis of five rice varieties Whitebacked Planthoppers is by being followed successively by the bright extensive 63> Wu-Yu-Geng 3 of Kasalath > Mudgo >9311> to weak by force.Can determine thus, the disease-resistant performance of the sick high disease-resistant variety Kasalath of the southern rice black-streaked dwarf identified at land for growing field crops natural appraisal is mainly from the forced-ventilated taxis of this kind, but plant matter self resistance against diseases well below Mudgo, in the sick resistive breeding work of later southern rice black-streaked dwarf, Kasalath can not be selected as anti-source.Also can find that the southern rice black-streaked dwarf disease morbidity performance of different rice varieties adopts the result of land for growing field crops natural occurrence qualification larger by the interference of white-backed planthopper antixenosis difference between kind by table one and table two.
Can be found by the present embodiment, the qualification result of this technology on the sick resistance against diseases of germ plasm resource southern rice black-streaked dwarf can get rid of the impact of antixenosis, and its result is more more accurate than land for growing field crops natural appraisal, is worth further genralrlization to utilize.
Above embodiment is only and technological thought of the present invention is described, can not limit protection scope of the present invention with this, every technological thought proposed according to the present invention, and any change that technical scheme basis is done, all falls within scope; The technology that the present invention does not relate to all is realized by prior art.
Claims (10)
1. the sick varietal resistance authentication method of southern rice black-streaked dwarf, is characterized in that: described authentication method step is as follows: (a), adopt enamel tray method to gather white-backed planthopper from field after by purifying screening plant obtain a large amount of 5 age white-backed planthopper nymph; (b), screening is obtained 5 age white-backed planthopper nymph be inoculated into and the paddy rice cultivated in crop insect co-cultivation box educate altogether and breeds white-backed planthopper nymph of future generation, and utilize the purifying screening plant of adjustment to be separated further to obtain a large amount of neat and consistent 2 age white-backed planthopper nymph; (c), by screening obtain 2 ages white-backed planthopper nymph move into plant have the seriously ill disaster area of southern rice black-streaked dwarf to draw materials tillering regularity paddy rice diseased plant on carry out raising poison; (d), being weighed by precision electronic balance obtains the weight of single head 2 white-backed planthopper in age nymph, calculate required white-backed planthopper gross weight according to the worm determined/seedling ratio, and then weigh quantitatively; (e), according to the worm determined/seedling ratio by raise after poison quantitative 2 age white-backed planthopper nymph be inoculated into the rice varieties rice shoot to be measured of cultivating in crop insect co-cultivation box educates altogether and pass poison; (f), when white-backed planthopper half is dead by rice transplanting to be measured to land for growing field crops every worm solarium, conventional cultivation manages, the tillering regularity investigation statistics southern rice black-streaked dwarf sick cave incidence of disease, and the resistance against diseases of rice varieties to be measured is gone out according to cave incidence of disease precise Identification.
2. the sick varietal resistance authentication method of southern rice black-streaked dwarf according to claim 1, is characterized in that: the paddy field at the beginning of annual 7 months of the white-backed planthopper acquisition time in described step (a) turns the yellow phase by green grass or young crops.
3. the sick varietal resistance authentication method of southern rice black-streaked dwarf according to claim 1, it is characterized in that: when purifying screening plant in described step (a) uses, adopt aperture to be 18 orders and the combination of 22 object sub-sieves with effectively isolate 5 age white-backed planthopper nymph and part white-backed planthopper adult; Then by screening isolated white-backed planthopper pour in the dark bucket of inner wall smooth, bounce a barrel outer wall white-backed planthopper adult is flown away from and nymph be detained, can obtain a large amount of neat and consistent 5 age white-backed planthopper nymph.
4. the sick varietal resistance authentication method of southern rice black-streaked dwarf according to claim 1, it is characterized in that: in described step (b) 5 ages, white-backed planthopper nymph was inoculated on the paddy rice cultivated in crop insect co-cultivation box time, be 2 ~ 3 leaf phases with the paddy rice that 5 ages, white-backed planthopper nymph educated altogether.
5. the sick varietal resistance authentication method of the southern rice black-streaked dwarf according to claim 1 or 4, is characterized in that: the cultivation temperature of the crop insect co-cultivation box in described step (b) controls at 27 DEG C, the dark 12h of light 12h/.
6. the sick varietal resistance authentication method of southern rice black-streaked dwarf according to claim 1, it is characterized in that: after breeding white-backed planthopper nymph of future generation in the crop insect co-cultivation box in described step (b), now screening plant overturns 90 ° and the vertical light direction that becomes is changed into horizontal direction, adopt aperture to be 30 orders with 32 object sub-sieves combinations be effectively separated obtain a large amount of neat and consistent 2 age white-backed planthopper nymph.
7. the sick varietal resistance authentication method of southern rice black-streaked dwarf according to claim 1, it is characterized in that: the tillering regularity paddy rice diseased plant in described step (c) is that upper one year is after drawing materials in the seriously ill disaster area of southern rice black-streaked dwarf in the refrigerator cold-storage of-20 DEG C, after diseased plant is thawed naturally, root immersion absorbent cotton parcel is wrapped up solid simulation field-transplanting mode with Polypropylence Sheet and is vertically fixed in crop insect co-cultivation box, then move into 2 age white-backed planthopper nymph raise poison 48 hours.
8. the sick varietal resistance authentication method of southern rice black-streaked dwarf according to claim 1, it is characterized in that: in described step (d) 2 ages white-backed planthopper nymph adopt light to be separated hairbrush brush from the impurity be mingled with during white-backed planthopper nymph and dead worm after raising poison, then transferase 12 0 ~ 50 white-backed planthopper nymph enters measuring cup, weighed by precision electronic balance, peeling calculates the quantity according to white-backed planthopper nymph in bottle after the gross weight of white-backed planthopper nymph in bottle, calculate the weight of single head white-backed planthopper nymph, determine to raise malicious white-backed planthopper nymph number according to paddy rice strain number to be measured and inoculation intensity, and then raise the gross weight of malicious white-backed planthopper nymph needed for calculating inoculation and weigh quantitatively.
9. the sick varietal resistance authentication method of the southern rice black-streaked dwarf according to claim 1 or 8, is characterized in that: the paddy growth to be measured in described step (e) was in for 1 ~ 2 leaf phase, and paddy rice to be measured is according to 20cm
2the ratio of/strain leaves and takes the healthy uniform rice seedling of growing way.
10. the sick varietal resistance authentication method of southern rice black-streaked dwarf according to claim 9, it is characterized in that: the paddy rice to be measured in described step (e) according to the ratio inoculation of 5 ~ 15 worms/seedling raise after poison 2 age white-backed planthopper nymph, the cultivation temperature of crop insect co-cultivation box control 27 DEG C, the dark 12h of light 12h/ educates that to pass poison dead to the white-backed planthopper naturally-aged of inoculation altogether.
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| CN111557215A (en) * | 2020-02-27 | 2020-08-21 | 四川省农业科学院植物保护研究所 | Identification method and application of rice variety resisting plant hopper and SRBSDV (sequence-specific locus-resistant virus) in adult stage |
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