CN101401562B - Acquiring method for special small brown rice planthopper for inoculating rice black-streaked dwarf virus - Google Patents
Acquiring method for special small brown rice planthopper for inoculating rice black-streaked dwarf virus Download PDFInfo
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- CN101401562B CN101401562B CN2008102350324A CN200810235032A CN101401562B CN 101401562 B CN101401562 B CN 101401562B CN 2008102350324 A CN2008102350324 A CN 2008102350324A CN 200810235032 A CN200810235032 A CN 200810235032A CN 101401562 B CN101401562 B CN 101401562B
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Abstract
The invention discloses a method for obtaining Laodelphax striatellus specially for inoculating black streaked dwarf virus of rice and application thereof in agriculture. The principle of the invention is to obtain Laodelphax striatellus without carrying stripe virus of rice by manual screening, feed the Laodelphax striatellus on rice carrying the black streaked dwarf virus without the stripe virus to obtain virus, transfer the Laodelphax striatellus to healthy rice sprouts without carrying viruses to feed for living through cycle period of the viruses, and further obtain the Laodelphax striatellus carrying the black streaked dwarf virus of the rice without the stripe virus of the rice. The established method for identifying the resistance of a rice variety with the black streaked dwarf virus can suppress interference due to that the Laodelphax striatellus simultaneously transmits the stripe virus of the rice, so that the identifying result is more objective and accurate, and the evaluation of resistance is more scientific and reasonable. The identifying method can be applied in discovery, identification, the evaluation of variety resistance, the breeding of disease-resistant variety, the research on resistance genetic development for rice variety resources and other fields.
Description
Technical field
The present invention relates to a kind of be exclusively used in preparation method of the small brown rice planthopper that inoculates rice black-streaked dwarf virus and the application on agricultural thereof, belong to the agricultural cience and farming techniques field.
Background technology
(Rice black-streaked dwarf virus is a kind of virus disease of being propagated by small brown rice planthopper RBSDV) to rice black-streaked dwarf virus, infects paddy rice and causes black streaked dwarf virus of rice, and paddy rice is in a single day susceptible just not to have results.This disease from 1963 since Yuyao county, China Zhejiang Province finds, mainly endanger in long-grained nonglutinous rice producing region, south, Zhejiang last century, and rarely have discovery in japonica rice main producing regions such as Jiangsu.Stripe disease is caused disaster in the outburst of provinces such as Jiangsu at the beginning of 2004, and the rapid large scale application of a collection of disease-resistant variety has been played crucial effect to the anti-stool control mode of the integral body of disease.Further studies show that, this class stripe disease disease-resistant variety is generally relatively poor to the resistance of RBSDV, since the MRDV that same cause of disease causes also be in recent years the popular phase for providing of disease sufficient malicious source, the increase year after year of small brown rice planthopper generating capacity in addition, plant protection scholarly forecast black streaked dwarf virus of rice have the potential risk that popular harm takes place.Really the RSV disease-resistant variety promotes that black streaked dwarf virus of rice just takes place popular after 2 years in japonica rice main producing regions such as Jiangsu,, the Huaibei coastal in this province by disease in 2007 according to Jiangsu Province's plant protection unit statistics and in the ground such as Yancheng, Lianyun Harbour, Taizhou, Huaian, Yangzhou and Xuzhou of going to river seriously take place, the area that harm takes place then reaches 30.8 ten thousand mu, the generation area of black streaked dwarf virus of rice in 2008 then surpasses 4,000,000 mu, and the situation of disease eruption and prevalence is triggered at any moment.
In the face of virus disease with sudden and explosive characteristics, it is the most direct effective method that the screening disease-resistant variety is applied to produce, and has increased many difficulties but for the resistance authentication method of black streaked dwarf virus of rice yet black streaked dwarf virus of rice and stripe disease are all the virus disease characteristics that rely on the small brown rice planthopper propagation.At first, the stripe disease owing to mostly occur greatly in current paddy rice main producing region, small brown rice planthopper can its cause of disease of transovarian transmission RSV, this makes the field small brown rice planthopper often carry RSV and RBSDV simultaneously, and rice varieties often is subjected to the interference of RSV when therefore carrying out the field evaluation to the resistance qualification result of black streaked dwarf virus of rice.Secondly, when adopting field collection small brown rice planthopper to carry out indoor inoculation, qualification result also can be subjected to same influence.In addition, adopt and indoorly raise malicious inoculation method identification of species resistance similar problem is also arranged, the small brown rice planthopper that gather in the field and raise the interference that malicious source that poison uses all is subjected to RSV easily.Simultaneously, small brown rice planthopper can not transovarian transmission RBSDV, and this makes that we can't be by obtaining to be exclusively used in the small brown rice planthopper of inoculation RBSDV with malicious adult offspring screening technique.Above-mentioned reason makes the resistant variety seed selection work of black streaked dwarf virus of rice be subjected to great restriction, press for and invent a kind of method that is exclusively used in the small brown rice planthopper used of inoculation rice black-streaked dwarf virus as early as possible, to assist to build up the resistance authentication method of black streaked dwarf virus of rice, excavate screening, breed breeding and genetic development research for the resource of the anti-black streak dwarf of paddy rice basic research means is provided.
Summary of the invention
The invention provides a kind of method that is exclusively used in the small brown rice planthopper that obtains the use of inoculation rice black-streaked dwarf virus.
Gathering small brown rice planthopper from the field feeds in the healthy paddy rice of not carrying rice stripe virus, the female worm of post-coitum single head lays eggs separately, adopt immunological method or molecular method to detect female worm and whether be with poison, keep and be not with malicious female worm offspring to raise separately to more than 2 ages, randomly draw in the colony small brown rice planthopper more than 10% and detect and confirm not to be with poison, and then screening obtains the small brown rice planthopper of not carrying rice stripe virus; The rice plant black streaked dwarf virus of rice symptom or that the artificial infection rice black-streaked dwarf virus obtains that having of gathering from the field again short strip shape blade fold that plant significantly stunt, was parallel to vein and white wax are dripped the shape projection, adopt immunological method or molecular method to confirm, and then obtain to carry rice black-streaked dwarf virus and the rice plant that do not carry rice stripe virus; The small brown rice planthopper of not carrying rice stripe virus that 1-3 age, above-mentioned screening obtained is not carried the carrying rice black-streaked dwarf virus of above-mentioned acquisition to be fed on the rice plant of rice stripe virus obtained poison in 2-3 days, the small brown rice planthopper that to obtain poison again be transferred to feed 10-13 days of tiding over virus on the healthy rice shoot that does not carry virus follow back the phase, can obtain only to carry rice black-streaked dwarf virus and the small brown rice planthopper of not carrying rice stripe virus.
The invention provides a kind of preparation method that is exclusively used in the small brown rice planthopper of inoculation rice black-streaked dwarf virus, utilize the varietal resistance authentication method of the black streaked dwarf virus of rice of its foundation can get rid of because small brown rice planthopper is propagated the interference that rice stripe virus causes simultaneously, make qualification result more objective and accurate, evaluation of resistance is more scientific and reasonable.This authentication method can be applied to the various fields such as excavation evaluation, varietal resistance evaluation, disease-resistant variety seed selection and the research of genetics of resistance rule of variety resources of rice.
Description of drawings
Fig. 1 is symptom figure (for infecting the rice black-streaked dwarf virus plant, the right is a normal plant in the bucket of the left side) after No. 6, the special small brown rice planthopper for inoculating rice black-streaked dwarf virus inoculation rice varieties China round-grained rice.
Specific implementation method
Embodiment 1, and inoculation RBSDV identifies the rice varieties resistance with small brown rice planthopper
1. do not carry the acquisition of the small brown rice planthopper of RSV.
The 4-5 month in 2005 was gathered the small brown rice planthopper nymph from grave illness district, Hai'an, Jiangsu, on educating No. 3, round-grained rice, susceptible variety force raises, the female worm of post-coitum single head lays eggs separately, randomly drawing in the colony 10% small brown rice planthopper adopts the spot immune combined techniques to detect female worm to be with malicious situation, keep not offspring with malicious female worm, later every generation is randomly drawed in the colony 10% small brown rice planthopper and is adopted the spot immune combined techniques to detect to guarantee not to be with poison, promptly obtain the small brown rice planthopper of not carrying RSV.
The acquisition of 2. carrying RBSDV and not carrying the rice plant of RSV.
Gather from experimental field, academy of agricultural sciences, Jiangsu in July, 2008 and have short strip shape blade fold and the white wax that plant significantly stunt, was parallel to vein and drip black streaked dwarf virus of rice symptom rice plants such as shape projection, the clip blade adopts the spot immune combined techniques to detect RSV and RBSDV respectively, filter out the plant of only carrying RBSDV and not carrying RSV, promptly obtain to carry RBSDV and the paddy rice of not carrying RSV.
3. small brown rice planthopper raises poison and follows back the phase
Choose 1-2 and 1. state small brown rice planthopper age for raising poison on the rice plant 2 days 2. stating, the phase of following back of raising behind the poison is 13 days, randomly drawing 10% small brown rice planthopper from colony adopts the spot immune combined techniques to detect RSV and RBSDV respectively, the result shows that detecting small brown rice planthopper does not all carry RSV, and have the part small brown rice planthopper to carry RBSDV, being with malicious rate is 55.2%.
4. the inoculation of rice varieties is identified
Rice varieties China round-grained rice is sowed at for No. 6 in the 1000ml beaker, and every glass of sowing 30 young plants repeat 1 time.Treat to inoculate evaluation when rice shoot grows to the 1.5-2.0 leaf, inoculate preceding 2 days and eliminate sick and weak seedling, select consistent seedling 25 strains of growth and be used for the inoculation evaluation.Calculate required worm amount by 3 of every young plants, insert and 3. state small brown rice planthopper.Catch up with every day twice of worm to guarantee that tested rice seedling evenly obtains poison.Remove whole small brown rice planthoppers after 48 hours, again rice seedling is transplanted to the land for growing field crops, note water and fertilizer management.Instituted an inquiry later in 7 days, investigated once continuous observation 5 times in per 3 days.The investigation standard is with reference to the classical symptom of black streaked dwarf virus of rice: have short strip shape blade fold and the white wax that plant significantly stunt, was parallel to vein and drip (Fig. 1) such as shape projections.The result shows that inoculation evaluation paddy rice has part to show the black streaked dwarf virus of rice symptom, and do not show the stripe disease symptom, this small brown rice planthopper that shows above-mentioned acquisition can be used to inoculate rice black-streaked dwarf virus, and can not be subjected to the interference of rice stripe virus, qualification result is effectively, can be used for the rice varieties evaluation of resistance.Result of the test such as table 1.
Table 1
The present invention mainly serves the resistance resource screening excavation of carrying out at black streaked dwarf virus of rice, the evaluation of disease-resistant variety and the seed selection of evaluation and resistant variety.This technology also can be used as biological detection method in the genetics of disease resistance of plant and the genetically modified research work of anti-rice black-streaked dwarf virus simultaneously, can accelerate the process of studying greatly.
Above-mentioned enforcement does not limit the present invention in any form.
Claims (1)
1. preparation method of small brown rice planthopper that is exclusively used in the inoculation rice black-streaked dwarf virus, it is characterized in that: gather small brown rice planthopper from the field and feed in the healthy paddy rice of not carrying rice stripe virus, the female worm of post-coitum single head lays eggs separately, adopt immunological method or molecular method to detect female worm and whether be with poison, keep and be not with malicious female worm offspring to raise separately to more than 2 ages, randomly draw in the colony small brown rice planthopper more than 10% and detect and confirm not to be with poison, and then screening obtains the small brown rice planthopper of not carrying rice stripe virus; The rice plant black streaked dwarf virus of rice symptom or that the artificial infection rice black-streaked dwarf virus obtains that having of gathering from the field again short strip shape blade fold that plant significantly stunt, was parallel to vein and white wax are dripped the shape projection, adopt immunological method or molecular method to confirm, and then obtain to carry rice black-streaked dwarf virus and the rice plant that do not carry rice stripe virus; The small brown rice planthopper of not carrying rice stripe virus that 1-3 age, above-mentioned screening obtained is not carried the carrying rice black-streaked dwarf virus of above-mentioned acquisition to be fed on the rice plant of rice stripe virus obtained poison in 2-3 days, the small brown rice planthopper that to obtain poison again be transferred to feed 10-13 days of tiding over virus on the healthy rice shoot that does not carry virus follow back the phase, can obtain only to carry rice black-streaked dwarf virus and the small brown rice planthopper of not carrying rice stripe virus.
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Families Citing this family (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101816273A (en) * | 2010-05-06 | 2010-09-01 | 江苏省农业科学院 | Identification method for indoor inoculation of anti-black-streaked dwarf disease of rice varieties |
| CN102578050A (en) * | 2012-03-02 | 2012-07-18 | 浙江大学 | Method for obtaining brown plant hoppers with rice ragged stunt virus and application of method |
| CN102630640A (en) * | 2012-03-02 | 2012-08-15 | 浙江大学 | Method for obtaining sogatella furcifera carrying SRBSDV (southern rice black-streaked dwarf virus) and application of sogatella furcifera |
| CN102870743B (en) * | 2012-10-12 | 2014-07-30 | 江苏省农业科学院 | Method for preserving rice black-streaked dwarf virus indoor living bodies |
| CN103749230A (en) * | 2014-01-27 | 2014-04-30 | 江苏徐淮地区连云港农业科学研究所 | Anti-rice black-streaked dwarf virus field inoculation identification method for rice varieties |
| CN104584890B (en) * | 2015-01-04 | 2017-10-31 | 甘肃恒基种业有限责任公司 | A kind of scale inoculation identification method of MRDV resistance |
| CN104830998B (en) * | 2015-05-20 | 2018-10-09 | 江苏焦点农业科技有限公司 | A kind of method for identifying molecules of black streaked dwarf virus of rice resistance |
| CN104871885B (en) * | 2015-06-18 | 2017-06-13 | 黄山丰乐谷生态农业综合开发有限公司 | A kind of prevention and controls of rice grub brown paddy plant hopper |
| CN105248168A (en) * | 2015-10-26 | 2016-01-20 | 江苏省农业科学院 | Maize rough dwarf virus variety resistance inoculation identification method established by taking wheat green dwarf virus plants as virus source |
| CN105746439B (en) * | 2016-03-09 | 2018-07-06 | 中国科学院微生物研究所 | Small brown rice planthopper is made to obtain the method and its man-made feeds of rice stripe virus |
| CN109456947A (en) * | 2018-11-29 | 2019-03-12 | 湖南农业大学 | A kind of method that efficient indoor winter saves propagation southern rice black-streaked dwarf virus |
| CN110192504B (en) * | 2019-04-19 | 2021-07-13 | 宁波大学 | Method for increasing incidence of rice black-streaked dwarf virus disease |
| CN110396503B (en) * | 2019-07-22 | 2021-07-30 | 中国农业大学 | Method for obtaining brassica yellows virus of potato leafroll virus by using green peach aphids |
| CN113693033B (en) * | 2021-09-14 | 2022-11-08 | 广西壮族自治区农业科学院 | Method for evaluating resistance of rice variety to southern rice black-streaked dwarf disease based on artificial inoculation |
| CN114946569A (en) * | 2021-12-30 | 2022-08-30 | 江苏省农业科学院 | Novel method for improving rice stripe virus infection efficiency through leaf inoculation |
| CN115005091A (en) * | 2022-04-28 | 2022-09-06 | 江苏丘陵地区镇江农业科学研究所 | Breeding method of rice black-streaked dwarf disease resistant breeding material |
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| CN101292640A (en) * | 2008-06-12 | 2008-10-29 | 江苏省农业科学院 | Process of fly louse with noxious ash by using freezing method |
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| 王朝辉等.从单头灰飞虱中检测水稻黑条矮缩病毒简单快速的方法.《上海交通大学学报（农业科学版）》.2002,(第4期),第340-343页. * |
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