CN1326671A - Breeding method for anti-rice blast rice breed - Google Patents
Breeding method for anti-rice blast rice breed Download PDFInfo
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- CN1326671A CN1326671A CN 00114616 CN00114616A CN1326671A CN 1326671 A CN1326671 A CN 1326671A CN 00114616 CN00114616 CN 00114616 CN 00114616 A CN00114616 A CN 00114616A CN 1326671 A CN1326671 A CN 1326671A
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Abstract
The present invention relates to a breeding method of new crop variety. The present invention features that through the young hybrid embryo salvage, nursing and culture of cultivated rice and wild rice and 4-6 generation backcross of the rotonning plant with cultivated rice, rice blast rice breed is obtained. The culture medium used has complexed germanium in 0.5-5 g/L and cane sugar in 20-50 g/L. Its identification relates to the identification of resistance gene fitting fingerprint and rice blast strain inoculation.
Description
The invention belongs to the creative method category of new crop varieties new material, its international Patent classificating number is A01H1/02.
In the rice disease that China's rice workspace takes place, rice blast is one of main disease that influences rice high yield, stable yields.The rice disease integrated control facts have proved that the genetic improvement that carries out the rice varieties blast resisting is the most economical valid approach of control rice blast.Rice blast is the fungal disease that is caused by pathogen Pyricularia grisea, and it can produce pathogenic variation by multiple modes such as sudden changes.So the new disease-resistant variety of promoting was generally just lost its specialization resistance in the production in 3 or five years, the speed that brings out new resistant variety often is unable to catch up with the pathogenic pace of change of pathogen.Application with anti-source of rice blast of durable resistance will bring opportunity for the forfeiture that overcomes the rice varieties blast resisting.Qualification result to lasting anti-rice blast rice kind shows both at home and abroad: the durable resistance kind is being suitable for keeping its disease resistance to reach decades under the popular environmental condition of disease generation.Domestic and international research proves, desires to breed high anti-and lasting disease-resistant variety, at first needs excellent anti-source.The many important agronomic shape of wild rice as lasting blast resisting, can be applied in the breeding of paddy rice.But because the isolation of reproduction and heredity, it is very difficult that wild rice exchanges with heredity between the cultivated rice, and this has caused obstacle to the application of wild rice in breeding.Along with the development of modern biotechnology, break the isolation between wild rice and cultivated rice, realize between them that the heredity interchange has become possibility thereby make.China science retainer of a big family grain husk is used hybrid method in nineteen twenty-nine and has just been obtained hybrid between common wild-rice (O.rufipogon) and the cultivated rice, and therefrom seed selection obtains in the kind mountain No. 1, obtain the large-area (Lin Shicheng that applies aborning, the Min Shaokai chief editor, 1971, rice in China kind and pedigree thereof, Shanghai science tech publishing house, P254-256).The scientist of International Rice Research Institute (IRRI) once obtained the hybrid between Ni Wala wild rice (O.nivana) and cultivated rice, and cultivate series of products (KushGS, Disease and insect resistance in rice, 1977 of anti-careless short disease, Adv Agron, 29:265-341).The nineteen ninety people have transferred to (JenaKK et al in the cultivated rice with the characteristic of against-plant hopper in the oryza officinalis (O.officinalis) again, Introgression of genes from Oryza officinalis wall ex watt to cultivated riceOryza sativa L., Theor Appl Genet 80:737-745).Afterwards, Chinese scholar has been transferred to the characteristic of against-plant hopper in the tight fringe wild rice (O.eichingeri) (face splendidness etc., 1997, the research of the brown planthopper resistance importing cultivated rice of tight fringe wild rice, Acta Genetica Sinica, 24 (5): 424-431) in the cultivated rice again.
In the past, be the kind of the lasting blast resisting of seed selection, people adopt breeding methods such as pyramiding breeding and anti-pest gene add up more.But there is not satisfied lasting anti-pest material to be created out so far as yet.Tracing it to its cause, is because the lasting anti-pest proterties that is complexity, and it relates to main the effect and little effect two big resistance systems, only is difficult to achieve the goal from unilaterally setting about.The chromosome set of granule wild rice (O.minuta) is AACC.The granule wild rice shows durable resistance to rice blast, simultaneously bacterial leaf-blight is also had resistance.Because reproduction is isolated, the existing conventional breeding method is to be difficult to the hereditary basis of granule wild rice and cultivated rice is attached to one.The present invention adopts the granule wild rice as anti-source, and the method that adopts cell engineering to combine with Protocols in Molecular Biology, and the durable resistance genetic mechanism (the main effect and little effect) of granule wild rice is transferred in the cultivated rice.
The objective of the invention is to overcome the deficiencies in the prior art, propose a kind of method of seed selection anti-rice blast rice breed, to improve the efficient of rice blast resistance breeding.
The present invention implements by the following technical solutions: with the granule wild rice is female parent, cultivated rice is the male parent sexual hybridization, get 16-18 days the rataria in hybridization back and place the MS medium that is added with 0.3-6g/l complexing germanium and 20-50g/l sucrose, doing nurse with endosperm cultivates, regeneration plant was backcrossed for 4-6 generations with cultivated rice in the field afterwards, and the hybrid plant that is obtained is identified with the rice blast fungi isolates inoculation, and carry out the dna fingerprint analysis, obtain the breeding material of anti-rice blast rice at last.
Detail of the present invention is: with the granule wild rice is female parent, and cultivated rice is a male parent, sexual hybridization.Get 16-18 days the rataria in hybridization back and on medium (collective media+0.5mg/L2,4-D+1mg/L methyl+1mg/L kinetin+10ml/L caseinhydrolysate .+0.5-5g/L complexing germanium+20-50g/L sucrose .), cultivate, and carry out plant regeneration.The seedling that differentiates carries out culture of rootage on the collective media of no hormone.Seedling is long is transplanted in the soil when high to 10 centimetres, adopts resistant gene match (RGA) primer CLLR to carry out the amplification of DNA, identifies true and false hybrid.Backcross with cultivated rice and true hybrid.In backcross process, adopt resistant gene match (RGA) primer CLLR to carry out the dna fingerprint analysis, and adopt wide spectrum rice blast experimental strain that elected plant is inoculated evaluation.Backcross and carry out 4-6 time.
The lasting anti-rice blast rice material OMI-1 that obtains by cultivated rice and granule wild rice described in the present invention is brand-new lasting blast resisting germplasm for obtaining first.Since cell engineering and Protocols in Molecular Biology combine and conventional method has improved breeding efficiency than greatly, for the durable resistance breeding provides novel method.What is particularly worth mentioning is that, in backcross process, to carrying lasting blast resisting plant really regularly, the method that we have adopted inoculation rice blast experimental strain and molecular labeling to combine, this is to lack in the former breeding method.Simultaneously, the creation of lasting anti-rice blast rice material OMI-l will play important impetus to the seed selection of lasting anti-rice blast rice kind.
Embodiment 1
With the granule wild rice is female parent, and cultivated rice is a male parent, hybridizes.Hybridization back 10,12,14,16,18,20 days, get rataria respectively in medium (collective media+0.5mg/L2,4-D+1mg/L methyl+1mg/L kinetin+10ml/L caseinhydrolysate .+0.5-5g/L complexing germanium+20-50g/L sucrose .) on cultivate, carry out plant regeneration.The result shows: there is material impact the hybridization back to the rataria culture success ratio sample time, to hybridize the culture success ratio height of the rataria of back after 16-18 days.Sampling too early because embryo is too little, is difficult for cultivating successfully; Sampling is too then because dying young of embryo can influence success rate equally.
Embodiment 2
With the granule wild rice is female parent, and cultivated rice is a male parent, hybridizes.Hybridized back 16 days, and got rataria respectively and on the medium described in the embodiment 1, cultivate, carry out plant regeneration.Sucrose concentration in the medium is set to 20,25,30,35,40,50,55,60g/L.Concentration be set to 0.1,0.5,1,2,5,7,10g/L result shows: complexing germanium in the medium and sucrose concentration are very big to the influence of culture efficiency.The concentration of complexing germanium is being combined as with 30-50g/L with 0.5-5g/L and sucrose concentration.On the medium of optimizing, the culture efficiency of rataria improves 12 times approximately than routine.
Embodiment 3
With the granule wild rice is female parent, and cultivated rice is a male parent, hybridizes.Hybridized back 16 days, and got rataria respectively and on the medium described in the embodiment 1, cultivate, carry out plant regeneration.The endosperm that we are provided with land-reclaimable 58 S of cultivated rice in medium is done nurse and is cultivated, and has made blank (CK).The result shows: the endosperm nurse is cultivated has significant effects to culture efficiency.The culture efficiency of blank only is 3%, only is 20.4% and do the culture efficiency of nursing cultivation with endosperm
Embodiment 4
The OMI-1 plant is adopted spray-on process inoculation wide spectrum rice blast fungi isolates 1366,0258,0386,1672, with the 1-5 level as standard of perfection.From 1997-2000, identify their blast resisting continuously, qualification result is as shown in table 1.The blast resisting qualification result of table 1 1997-2000 OMI-1 (average sick level) time wide spectrum rice blast fungi isolates
1366 0258 0386 1672?1997 1.2 1 1 1.1?1998 1 1.3 1.2 1.5?1999 1.3 1.8 1 1.2?2000 2 1.6 2.2 1.4
The land-reclaimable 58S of check variety CO39 and cultivated rice parent their average sick level of all high sense in four annual bearings is 5 and 4.2.The result of table 1 shows: the average sick level of OMI-1 is 1.36, belong to resistance level, and performance is stable between year, that is, OMI-1 has obtained lasting blast resisting to a certain extent.
Compare with traditional breeding method, advantage main manifestations of the present invention in the following areas:
1) method of immature embryo culture described in the invention can improve the efficient of embryo rescue greatly.
2) in breeding process, can effectively identify and control the plant of carrying the durable resistance hereditary basis.
3) can effectively the resistance to rice blsat in the granule wild rice be transferred in the cultivated rice.
Claims (3)
1. the method for a seed selection anti-rice blast rice breed, it is characterized in that: with the granule wild rice is female parent, cultivated rice is a male parent, the hybrid rataria that obtains is cultivated in sexual hybridization, adopts endosperm to do nurse in the described incubation and cultivates, regeneration plant, backcross 4-6 generation with cultivated rice in the field afterwards, the hybrid plant of acquisition is identified with the rice blast fungi isolates inoculation, and is carried out the dna fingerprint analysis.
2. according to the seed selection anti-rice blast rice breed method described in claims 1, it is characterized in that, adopt the rataria of granule wild rice and cultivated rice hybridization back after 16-18 days to place on the MS medium that is added with 0.3-6g/L complexing germanium and 20-50g/L sucrose and cultivate, comprise in the described incubation that doing nurse with endosperm cultivates.
3. according to the seed selection anti-rice blast rice breed method described in claims 1, it is characterized in that, in backcross process, adopt resistant gene match (RGA) to carry out the fingerprint analysis of material, and adopt the wide spectrum rice blast fungi isolates that elected plant is inoculated evaluation.
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| CN 00114616 CN1326671A (en) | 2000-06-05 | 2000-06-05 | Breeding method for anti-rice blast rice breed |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101824418B (en) * | 2009-12-10 | 2011-12-21 | 中国水稻研究所 | Coding region of rice blast resistant gene Pi 25 and application thereof |
| CN102668973A (en) * | 2012-06-04 | 2012-09-19 | 广西壮族自治区农业科学院水稻研究所 | Novel method for improving achievement rate of first backcross hybrid generation of oryza sativa and oryza minuta |
| CN103229674A (en) * | 2013-05-21 | 2013-08-07 | 江苏胜田农业科技发展有限公司 | Method for simply and rapidly identifying resistance of pyricularia grisea |
| CN104381125A (en) * | 2014-11-19 | 2015-03-04 | 长江大学 | Authentication method of rice blast disease resistant breeding material |
| CN105104048A (en) * | 2015-09-28 | 2015-12-02 | 江苏丘陵地区镇江农业科学研究所 | Method for inducing, identifying and screening rice blast resistance breeding materials |
-
2000
- 2000-06-05 CN CN 00114616 patent/CN1326671A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101824418B (en) * | 2009-12-10 | 2011-12-21 | 中国水稻研究所 | Coding region of rice blast resistant gene Pi 25 and application thereof |
| CN102668973A (en) * | 2012-06-04 | 2012-09-19 | 广西壮族自治区农业科学院水稻研究所 | Novel method for improving achievement rate of first backcross hybrid generation of oryza sativa and oryza minuta |
| CN103229674A (en) * | 2013-05-21 | 2013-08-07 | 江苏胜田农业科技发展有限公司 | Method for simply and rapidly identifying resistance of pyricularia grisea |
| CN104381125A (en) * | 2014-11-19 | 2015-03-04 | 长江大学 | Authentication method of rice blast disease resistant breeding material |
| CN105104048A (en) * | 2015-09-28 | 2015-12-02 | 江苏丘陵地区镇江农业科学研究所 | Method for inducing, identifying and screening rice blast resistance breeding materials |
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