CN103014153A - Anti-ustilaginoidea virens major gene and molecular marker thereof - Google Patents
Anti-ustilaginoidea virens major gene and molecular marker thereof Download PDFInfo
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- CN103014153A CN103014153A CN2012104755622A CN201210475562A CN103014153A CN 103014153 A CN103014153 A CN 103014153A CN 2012104755622 A CN2012104755622 A CN 2012104755622A CN 201210475562 A CN201210475562 A CN 201210475562A CN 103014153 A CN103014153 A CN 103014153A
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Abstract
The invention relates to a molecular marker of a new anti-ustilaginoidea virens gene of rice, belonging to the field of rice breeding for disease resistance. According to the invention, each strain of Teqing background backcross introgression lines obtained by the cross and backcross between a susceptible parent Teqing and a disease-resistant parent Lemont is subjected to evaluation of disease resistance and molecular genetic linkage analysis and positioned to the anti-ustilaginoidea virens gene qFSR10.5 on the 10th chromosome of rice; and the molecular marker linked with the qFSR10.5 is RM258. By checking the RM258 banding pattern data and screening the ustilaginoidea virens resistance controlled by the qFSR10.5 resistance allele, the breeding efficiency of the anti-ustilaginoidea virens rice variety can be improved.
Description
Technical field
The present invention relates to the molecule marker of the new gene qFSR10.5 of Rice Resistance false smut, belong to paddy disease-resistant breeding field, be applicable to molecule marker carries out false smut to said gene assisted selection and pyramiding breeding.
Background technology
False smut (Ustilaginoidea virens (Cooke) Tak.) is a kind of important rice fungus diseases evil, and germ is transformed into the rice curve with the grain of rice, a large amount of powderies of surface coverage, blackish green chlamydospore.Since the eighties in 20th century, along with hybrid rice, Xian round-grained rice are handed over the popularization of the high-yield variety of breeding and the application of corresponding well stimulation, false smut rises to Major Diseases in China by Minor diseases, spreads all at present each main paddy fields.False smut reduces rice yield and rice quality significantly.Fall ill the serious time, the production loss rate reaches 30-50%; And the rice that processes is mixed with a large amount of chlamydospores, and chlamydospore contains Antitubulin, and is totally unfavorable to HUMAN HEALTH, and the edible cereal that pollutes of domestic animal can cause poisoning (Nakamura, 1994; Koiso, 1994).
The pests occurrence rule of Japan's early start research fifties in last century false smut, the investigator generally believes that false smut is monocyclic disease both at home and abroad at present, and to the Spatial distribution of false smut, the relation that weather, cultivation condition, varietal character and false smut occur has been carried out many investigation.Yet because false smut is fringe section disease, ustilaginoidea virens is complicated the life history in addition, and up to the present, false smut infects with regularty of epidemic and still imperfectly understands.
For a long time, the false smut control is always to spray chemical pesticide as main.False smut is fringe section disease, and is without any symptom, late if symptom to be found is prevented and treated again at premorbid.Therefore, the colony of monitoring pathogenic bacteria is dynamic, in good time dispenser is effectively to prevent and treat false smut and realize the prerequisite that the chemistry security decrement is used, but not yet sets up the popular monitoring method of false smut both at home and abroad so far.
Cultivating and plant disease-resistant quality is that controlling plant diseases is most economical effectively, also one of environment amenable measure.A large amount of field investigation and tests show, there is significant difference (Zhou et al. in sickness rate to false smut between the different rice varieties, 2010), Li Yusheng etc. think that there is obvious major gene effect (Li Yusheng etc., 2010) in paddy rice to the resistance of false smut.Fujita in 1989 etc. have reported the potted plant artificial inoculation technology of carrying out false smut that adopts, impel the plant morbidity owing to needing the condition of low temperature and high relative humidity after the inoculation, thereby simultaneously to a large amount of strains inoculate be difficult to the operation, the false smut localization of disease resistance genes is difficult to carry out always.
In recent years, the investigator begins successively by field natural occurrence or field provocative inoculation method location false smut resistant gene.2002, Xu Jianlong etc. adopt the near isogene introgressive line of 266 blue or green backgrounds of spy, in conjunction with 2 years field false smut nature qualification results, on the 10th and the 12nd karyomit(e), respectively navigate to 1 anti-false smut gene, its allelotrope that strengthens resistance is from parent Lemont, and the disease-resistant gene Qfsr10 and the mark AK10150 that wherein are positioned on the 10th karyomit(e) are chain; The high point rice that 157 familys of the employing such as Li Yusheng form/IR28 RIL, adopt and cause the false smut Inoculation Method, detected 7 disease-resistant genes in 2 years, wherein qFsr10 and qFsr11 two are detected every year, qFsr10 is positioned at RM6691-RM5620 interval on the karyomit(e), and qFsr11 is positioned at RM229-RM254 interval on the karyomit(e).Present patent application person is used to come from the introgressive line colony that backcrosses that disease-resistant parent Lemont and special 213 the blue or green strains of Susceptible parent consist of, by continuous 2 years each strains of natural induction in fields inoculated identification resistance to false smut, find 2 new resistance site qFSR10.2 and qFSR10.5 at the 10th karyomit(e), wherein qFSR10.2 is positioned in RM222-RM216 than the minizone, but may with control rice ear sprouting period gene linkage.QFSR10.5 is positioned in the RM271-RM258 interval, with the closely linked molecule marker of qFSR10.5 be that RM258 is expected to marker assisted selection and the pyramiding breeding for this gene.
Summary of the invention
The object of the present invention is to provide Rice Resistance false smut gene locus, and provide the molecule marking method of gene locus, by detecting the molecule marker chain with false smut resistant gene site, can predict rice plant to the resistance of false smut, thereby for the false smut resistance breeding provides may.
Purpose of the present invention is achieved through the following technical solutions:
Utilize the introgressive line that backcrosses of the special blue or green special blue or green background that makes up of disease-resistant parent Lemont and Susceptible parent, by linkage analysis, navigate to the new gene qFSR10.5 of false smut resistance (Fig. 1) at the 10th karyomit(e).
QFSR10.5 and mark RM258 close linkage, RM258 forward primer sequence is: 5 '-tgctgtatgtagctcgcacc-3 '; The reverse primer sequence is: 5 '-tg gcctttaaagctgtcgc-3 '.With this primer amplification Lemont genomic dna, can amplify 1 fragment (Fig. 2) about 140bp.
Invention provides the molecule marking method of above-mentioned anti-false smut gene locus.Take the kind Lemont that carries false smut resistant gene qFSR10.5 as donor parents, hybridize and selfing take the good rice varieties of economical character as recurrent parent, in segregating generation is selected, carry out molecular marker assisted selection: namely with the RM258 labeled primer colony's individual plant and parent's genomic dna is carried out pcr amplification, amplified production is carried out polyacrylamide gel electrophoresis, judge by amplification anti-false smut gene locus whether contained in rice strain.Fig. 2 is that RM258 with chain qFSR10.5 is at F
2Marker genetype in the colony by the electrophoresis banding pattern of each individual plant electrophoresis banding pattern and donor parents in the comparison colony, can be determined the individual plant that target site isozygotys.
The present invention compared with prior art has the following advantages and effect:
(1) discovery of qFSR10.5 and location can be used for the seed selection work of Rice Varieties for Resistance false smut.By molecular mark, this gene and other disease-resistant gene can be carried out polymerization.
(2) the method can fast and effeciently be selected anti-false smut strain, reduces Breeding Scale, improves breeding efficiency.
Figure of description
Fig. 1 rice varieties Lemont anti-false smut gene locus qFSR10.5 and Linkage mapl
The molecule marker RM258 of Fig. 2 q FSR10.5 is to donor parents Lemont and F
2The amplification collection of illustrative plates of individual plant
*The expression homozygous plants
Embodiment
Below in conjunction with the implementation example, further set forth the present invention.Following example only is used for explanation the present invention and is not used in the claimed scope of restriction the present invention.In the situation that do not deviate from essence of the present invention, to the replacement of the inventive method, step or receptor parent, all belong to scope of the present invention.Experimental technique in the following embodiment if no special instructions, is ordinary method.
1, target group and Resistance Identification
The special blue or green background introgressive line colony that backcrosses is comprised of 213 strains, comprises 133 BC
2F
5With 80 BC
3F
4Strain.Special green grass or young crops is the Indica Rice kind of China's spread, and Lemont is the good quality and high output japonica rice variety that southern US is promoted.
At Beijing and Shenyang false smut Prevalent district, select the serious field of false smut morbidity to plant the special blue or green background introgressive line colony that backcrosses, in the autumn upper one year of test, to collect the rice curve and evenly spread the field, rice curve addition is 20g/m
2Randomized complete-block design is adopted in test, single this plantation, and each strain kind 1 row 10 strain, distance between rows and hills is 20cm * 15cm, three repetitions.When between the sword-like leave auricle, reaching 5cm, will isolate around the experimental plot with the plastic cloth that 2m is high, top sunshade net shading, the 1hr that sprays water noon every day is to increase humidity and to reduce temperature.After the rice grain maturation, investigate the infected seed rate (morbidity grain number accounts for the per-cent of total number) of middle 6 strains of each strain as the disease index of each strain.Disease index such as the table 1 of parent and colony.
The false smut infected seed rate of the special green grass or young crops of table 1, Lemont and the introgressive line colony that backcrosses thereof
*,
*Represent respectively the significance of difference of P＜0.01 and 0.001 level.
2, QTL location
SSR Linkage mapl (the covering gene group 1677cM that utilizes introgressive line to make up, mean distance between adjacent two marks is 10.5cM) (Xu JL, Lafitte YM, Gao YM, Fu BY, Torres R, Li ZK (2005) QTLs for drought escape and tolerance identified in a set of random introgression lines of rice.Theor Appl Genet111:1642-1650), detect the QTL that affects false smut infected seed rate in conjunction with SAS PROC GLM One-way ANOVA program, with the threshold value of 0.05≤P≤0.005 conspicuous level as the choice main effect QTL, when 1 QTL and two or more marks are chain, with the highest mark of F value as listing with the chain mark of QTL.
According to the test-results of colony in Shenyang and Pekinese, detect altogether 10 anti-false smut QTL from the introgressive line offspring, two stable genes that are positioned on the 10th karyomit(e) are expressed, and all can detect in two places, reduce the allelotrope of infected seed rate all from Lemont (table 2).Wherein, disease-resistant gene qFSR10.2 is positioned at the RM222-RM216 interval, contiguous RM216, but should there be the control gene at heading stage in the interval; QFSR10.5 between RM271-RM258, contiguous RM258, interval in no-fix to the gene relevant with heading stage, be convenient to utilization in the breeding.
Two main effect sites that table 2 navigates in special green grass or young crops/Lemont backcrosses introgressive line colony and linked marker thereof
The underscore mark represents the adjacent marker near QTL; Additive effect represents the effect that produces after receptor allele is by the donor allelic substitution, i.e. the effect that produces after being substituted by Lemont of special blue or green allelotrope.
3, qFSR10.5 molecular marker assisted selection
Obtain F with the special green grass or young crops of Susceptible parent and disease-resistant parent Lemont hybridization, selfing
2Colony.Spy green grass or young crops, Lemont and F2 colony thereof are planted in false smut grave illness field, manual-induced false smut.In the autumn upper one year of test, to collect the rice curve and evenly spread the field, rice curve addition is 20g/m
2Every row kind 8 strains, the parent repeats for three times.Plant respectively the special blue or green plant of 6 row around the residential quarter as guard rows.When between plant sword-like leave auricle, reaching 5cm, will isolate around the experimental plot with the plastic cloth that 2m is high, top sunshade net shading, the 1hr that sprays water noon every day is to increase humidity and to reduce temperature.At the special green grass or young crops of clip in tillering phase, Lemont and F
2The blade of individual plant extracts DNA, adopts molecule marker RM258 to increase, and analyzes the amplification collection of illustrative plates of each plant.After the rice grain maturation, investigate the infected seed rate (morbidity grain number accounts for the per-cent of total number) of each plant as the disease index of plant.
Amplification collection of illustrative plates (Fig. 2) shows F
2Plant is 3 kinds of banding patterns of performance after RM258 amplification, namely with respectively with parent Lemont and special blue or green identical banding pattern, and heterozygosis banding pattern.The disease resistance investigation result is respectively 4.30 ± 0.19% and 2.16 ± 0.05% for special infected seed rate blue or green and Lemont; F
2The infected seed rate of homozygous plants and heterozygous plant be respectively 2.98 ± 0.13% and 3.87 ± 0.39%, RM258 can be used for molecular marker assisted selection.
Claims (2)
1. the method for the new gene molecule marker of Rice Resistance false smut, it is characterized in that backcrossing with special green grass or young crops/Lemont, to import be mapping population, according to the result of Disease Resistance Identification and the linkage analysis between the SSR flag data, obtain and the closely linked molecule marker RM258 of the new gene qFSR10.5 of anti-false smut at paddy rice the 10th karyomit(e).
According to right 1 that obtain with the introgressive line that backcrosses with special green grass or young crops/Lemont in the application of the closely linked molecule marker RM258 of the new gene qFSR10.5 of anti-false smut in the anti-false smut rice varieties of seed selection on the 10th karyomit(e), it is characterized in that in the filial generation with susceptible variety and disease-resistant variety Lemont, by the banding pattern data of RM258 mark, the false smut resistance that screening is controlled by the qFSR10.5 resistance allele can improve the breeding efficiency of anti-false smut rice varieties.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106086196A (en) * | 2016-06-29 | 2016-11-09 | 无锡南理工科技发展有限公司 | A kind of rice green smut Resistance detecting method |
| CN112257925A (en) * | 2020-10-22 | 2021-01-22 | 安徽省农村综合经济信息中心（安徽省农业气象中心） | Rice false smut prediction method based on rainfall and temperature |
| CN113016603A (en) * | 2021-03-09 | 2021-06-25 | 广西壮族自治区农业科学院 | Molecular breeding method of primary high-quality long-grain broad-spectrum rice blast-resistant normal recovery line |
| CN117305506A (en) * | 2023-11-07 | 2023-12-29 | 四川省农业科学院植物保护研究所 | Accurate rice false smut resistance gene identification method |
| CN117587158A (en) * | 2023-11-07 | 2024-02-23 | 四川省农业科学院植物保护研究所 | Tightly-linked SSR molecular marker of rice false smut resistance QTL and application thereof |
| CN117587158B (en) * | 2023-11-07 | 2024-07-05 | 四川省农业科学院植物保护研究所 | Tightly-linked SSR molecular marker of rice false smut resistance QTL and application thereof |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106086196A (en) * | 2016-06-29 | 2016-11-09 | 无锡南理工科技发展有限公司 | A kind of rice green smut Resistance detecting method |
| CN106086196B (en) * | 2016-06-29 | 2019-09-20 | 青岛欧易生物科技有限公司 | A kind of rice green smut Resistance detecting method |
| CN112257925A (en) * | 2020-10-22 | 2021-01-22 | 安徽省农村综合经济信息中心（安徽省农业气象中心） | Rice false smut prediction method based on rainfall and temperature |
| CN113016603A (en) * | 2021-03-09 | 2021-06-25 | 广西壮族自治区农业科学院 | Molecular breeding method of primary high-quality long-grain broad-spectrum rice blast-resistant normal recovery line |
| CN113016603B (en) * | 2021-03-09 | 2022-03-11 | 广西壮族自治区农业科学院 | Molecular breeding method of primary high-quality long-grain broad-spectrum rice blast-resistant normal recovery line |
| CN117305506A (en) * | 2023-11-07 | 2023-12-29 | 四川省农业科学院植物保护研究所 | Accurate rice false smut resistance gene identification method |
| CN117587158A (en) * | 2023-11-07 | 2024-02-23 | 四川省农业科学院植物保护研究所 | Tightly-linked SSR molecular marker of rice false smut resistance QTL and application thereof |
| CN117305506B (en) * | 2023-11-07 | 2024-05-28 | 四川省农业科学院植物保护研究所 | Accurate rice false smut resistance gene identification method |
| CN117587158B (en) * | 2023-11-07 | 2024-07-05 | 四川省农业科学院植物保护研究所 | Tightly-linked SSR molecular marker of rice false smut resistance QTL and application thereof |
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