• Title/Summary/Keyword: marker-assisted selection (MAS)

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Identification of Coupling and Repulsion Phase DNA Marker Associated With an Allele of a Gene Conferring Host Plant Resistance to Pigeonpea sterility mosaic virus (PPSMV) in Pigeonpea (Cajanus cajan L. Millsp.)

  • Daspute, Abhijit;Fakrudin, B.
    • The Plant Pathology Journal
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    • v.31 no.1
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    • pp.33-40
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    • 2015
  • Pigeonpea Sterility Mosaic Disease (PSMD) is an important foliar disease caused by Pigeonpea sterility mosaic virus (PPSMV) which is transmitted by eriophyid mites (Aceria cajani Channabasavanna). In present study, a F2 mapping population comprising 325 individuals was developed by crossing PSMD susceptible genotype (Gullyal white) and PSMD resistant genotype (BSMR 736). We identified a set of 32 out of 300 short decamer random DNA markers that showed polymorphism between Gullyal white and BSMR 736 parents. Among them, eleven DNA markers showed polymorphism including coupling and repulsion phase type of polymorphism across the parents. Bulked Segregant Analysis (BSA), revealed that the DNA marker, IABTPPN7, produced a single coupling phase marker (IABTPPN $7_{414}$) and a repulsion phase marker (IABTPPN $7_{983}$) co-segregating with PSMD reaction. Screening of 325 F2 population using IABTPPN7 revealed that the repulsion phase marker, IABTPPN $7_{983}$, was co-segregating with the PSMD responsive SV1 at a distance of 23.9 cM for Bidar PPSMV isolate. On the other hand, the coupling phase marker IABTPPN $7_{414}$ did not show any linkage with PSMD resistance. Additionally, single marker analysis both IABTPPN $7_{983}$ (P<0.0001) and IABTPPN $7_{414}$ (P<0.0001) recorded a significant association with the PSMD resistance and explained a phenotypic variance of 31 and 36% respectively in $F_2$ population. The repulsion phase marker, IABTPPN7983, could be of use in Marker-Assisted Selection (MAS) in the PPSMV resistance breeding programmes of pigeonpea.

Construction of an Integrated Pepper Map Using RFLP, SSR, CAPS, AFLP, WRKY, rRAMP, and BAC End Sequences

  • Lee, Heung-Ryul;Bae, Ik-Hyun;Park, Soung-Woo;Kim, Hyoun-Joung;Min, Woong-Ki;Han, Jung-Heon;Kim, Ki-Taek;Kim, Byung-Dong
    • Molecules and Cells
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    • v.27 no.1
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    • pp.21-37
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    • 2009
  • Map-based cloning to find genes of interest, marker-assisted selection (MAS), and marker-assisted breeding (MAB) all require good genetic maps with high reproducible markers. For map construction as well as chromosome assignment, development of single copy PCR-based markers and map integration process are necessary. In this study, the 132 markers (57 STS from BAC-end sequences, 13 STS from RFLP, and 62 SSR) were newly developed as single copy type PCR-based markers. They were used together with 1830 markers previously developed in our lab to construct an integrated map with the Joinmap 3.0 program. This integrated map contained 169 SSR, 354 RFLP, 23 STS from BAC-end sequences, 6 STS from RFLP, 152 AFLP, 51 WRKY, and 99 rRAMP markers on 12 chromosomes. The integrated map contained four genetic maps of two interspecific (Capsicum annuum 'TF68' and C. chinense 'Habanero') and two intraspecific (C. annuum 'CM334' and C. annuum 'Chilsungcho') populations of peppers. This constructed integrated map consisted of 805 markers (map distance of 1858 cM) in interspecific populations and 745 markers (map distance of 1892 cM) in intraspecific populations. The used pepper STS were first developed from end sequences of BAC clones from Capsicum annuum 'CM334'. This integrated map will provide useful information for construction of future pepper genetic maps and for assignment of linkage groups to pepper chromosomes.

Genetic analysis of clubroot resistance in Chinese cabbage using single spore isolate of Plasmodiophora brassicae and development of RAPD marker linked to its resistance gene

  • Cho, Kwang-Soo;Hong, Su-Young;Han, Young-Han;Yoon, Bong-Kyeong;Ryu, Seoung-Ryeol;Woo, Jong-Gyu
    • Journal of Crop Science and Biotechnology
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    • v.11 no.2
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    • pp.101-106
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    • 2008
  • To identify inheritance of clubroot disease resistance genes in Chinese cabbage, seedling tests of $BC_1P_1,\;BC_1P_2$, and $F_2$ populations derived from $F_1$ hybrid(var. CR Saerona) using single spore isolate(race 4 identified with William's differential host) from Plasmodiophora brassciae were conducted. Resistance(R) and susceptible(S) plants segregated to 1:0 in backcross to the resistant parent. The $F_2$ population segregated in a 3(R):1(S) ratio. This result implied that the resistance of clubroot disease is controlled by a single dominant gene to the race 4 of P. brassicae in CR Saerona. To develop DNA markers linked to clubroot resistance genes, 185 plants of CR Saerona among $F_2$ populations were used. A total of 300 arbitrary decamer was applied to $F_2$ population using BSARAPD(Bulked segregant analysis-Randomly amplified polymorphic DNA). One RAPD marker linked to clubroot resistance gene in CR Saerona($OPJ_{1100}$) was identified. This marker was 3.1 cM in distance from resistance gene in $F_2$ population. This marker may be useful for a marker-assisted selection(MAS) and gene pyramiding of the clubroot disease resistant gene in Chinese cabbage breeding programs.

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Development of a SNP marker set related to crown gall disease in grapevines by a genome wide association study

  • Kim, Dae-Gyu;Jang, Hyun A;Lim, Dong Jun;Hur, Youn Young;Lee, Kyo-Sang;Min, Jiyoung;Oh, Sang-Keun
    • Korean Journal of Agricultural Science
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    • v.47 no.3
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    • pp.693-705
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    • 2020
  • Grapes (Vitis spp. L.) are the third most produced fruit in the world. Crown gall disease caused by Agrobacterium vitis forms galls in the stems of the grapevines and reduces the vitality of the fruit trees, resulting in reduced yields. This pathogen has occurred in vineyards worldwide and caused serious economic losses. It is a soil-borne disease, so Agrobacterium vitis can survive for several years in vineyards and is difficult to control. Additionally, since there is no effective chemical control method, the most effective control method is the breeding of resistant varieties. To make the resistant variety, marker-assisted selection (MAS) enables fast breeding with low cost. In this study, we applied a genome-wide association study (GWAS), by combining phenotyping and genotyping-by-sequencing (GBS), for the development of a single nucleotide polymorphism (SNP) marker set related to crown gall disease using 350 grapevine varieties. As a result of the GBS based genotyping analysis, about 58,635 SNPs were obtained. In addition, the phenotypic analysis showed 35.2% resistance, 73% moderate susceptibility and 16.4% highly susceptibility. Moreover, after confirmation, two genes (VvARF4 and VvATL6-like) were shown to be related to crown gall disease based on the results of GWAS analysis, using the phenotypic data, and GBS. High-resolution melting analysis (HRMA) was performed using the Luna® Universal Probe with real-time PCR to distinguish the melting peaks of the resistant and susceptible varieties. Our data show that these SNP markers are expected to be helpful in evaluating resistance against grapevine crown gall disease and in breeding.

Mapping of RFLP Markers Linked to Bacterial Blight Resistant Genes (Xa-1, Xa-3) in Rice (벼 흰잎마름병 저항성 유전자(Xa-1, Xa-3)연관 RFLP 마커 탐색)

  • 강현중;김현순;남정권;이영태;이승엽;김석동
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.6
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    • pp.419-423
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    • 2003
  • Bacterial blight caused by Xantomonas oryzae pv. oryzae is one of the most serious diseases of rice especially in southern area of Korea. Three races, $\textrm{K}_1$, $\textrm{K}_2$ and $\textrm{K}_3$, are the most dominant species. lo improve rice breeding efficiency using marker assisted selection, some RFLP markers were surveyed for polymorphism between resistant and susceptible to $\textrm{K}_1$ and $\textrm{K}_3$. And, 127 doubled-haploid (DH) lines derived from Milyang121/HRl1650-1-4-2 and 131 DH lines derived from Milyang123/HR10624-AC5 were evaluated to bacterial blight ($\textrm{K}_1$ and $\textrm{K}_3$). Milyang121 and HR10624-AC5 have Xa-1, resistant to $\textrm{K}_1$ race, and Milyang123 has Xa-3, resistant to $\textrm{K}_1$ and $\textrm{K}_3$ race. Three markers, RZ590, RZ536 and RG303, showing polymorphism between parents and resistance gene, Xa-1 and Xa-3, were analysed in the two combinations of DH lines. The segregation pattern of resistant DH population of Milyang123/HR10624-AC5 to susceptible showed 3:1 and 1:1 in $\textrm{K}_1$ and $\textrm{K}_3$ race. In three RFLP markers, RZ590 was linked to Xa-1 on chromosome 4, and RZ536 and RG303 were linked to Xa-3 on chromosome 11. The map distance between Xa-1 and RZ590 was 3.1cM on chromosome 4, and Xa-3 and RZ536/RG303 were 7.6/16.0cM on chromosome 11, respectively. The results of RFLP mapping will be useful for the selection and pyramiding of bacterial blight resistant genes.

Characterization and Utilization of the Clubroot Resistant Genes in Chinese Cabbage (Brassica rapa L.)

  • Hatakeyama, Katsunori
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.33-33
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    • 2015
  • Clubroot disease is the major threat to the production of Chinese cabbage (Brassica rapa L.) in Japan. Although the breeding of the clubtoot resistant (CR) cultivars is one of the most efficient ways to control this disease, the CR cultivars do not always have effects due to the breakdown of resistance. Therefore, it is necessary to develop the breeding strategy to accumulate multiple CR genes in a single cultivar effectively. We have identified two incomplete dominant CR loci, Crr1 and Crr2, which are originated from the European CR turnip Siloga. To investigate the effectiveness of marker-assisted selection (MAS) for CR breeding, the inbred line with Crr1 and Crr2 was crossed with parental lines of the existing CR $F_1$ cultivar of Chinese cabbage, followed by 5 times of MAS and backcrossing. The $F_1$ derived from a cross between the resulting parental lines improved the clubroot resistance as expected and had the same morphological characters as the original $F_1$ cultivar. We have shown that the Crr1 locus comprised two loci: Crr1a, which by itself conferred resistance to the mild isolate; and Crr1b, which had a minor effect, but was not required for Crr1a-mediated resistance. Further genetic analysis suggested that Crr1b was necessary to acquire resistance to the more virulent isolate in combination with Crr2. Molecular characterization of Crr1a encoding TIR-NB-LRR class of R protein revealed that there were at least 4 alleles in Japanese CR cultivars of Chinese cabbage. PCR analysis with Crr1a-specific markers demonstrated that the functional alleles were predicted to be present in European CR turnips, Debra and 77b besides Siloga, whereas rarely in Japanese CR cultivars, indicating that Crr1a is an useful source to improve the resistance of Chinese cabbage cultivars.

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Characterization of the Lsi1 Homologs in Cucurbita moschata and C. ficifolia for Breeding of Stock Cultivars Used for Bloomless Cucumber Production

  • Jung, Jaemin;Kim, Joonyup;Jin, Bingkui;Choi, Youngmi;Hong, Chang Oh;Lee, Hyun Ho;Choi, Youngwhan;Kang, Jumsoon;Park, Younghoon
    • Horticultural Science & Technology
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    • v.35 no.3
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    • pp.333-343
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    • 2017
  • Bloomless cucumber fruits are commercially produced by grafting onto the pumpkin stocks (Cucurbita moschata) to restricted silicon ($SiO_2$) absorption. Inhibition of silicon absorption in bloomless stocks is conferred by a mutant allele of the CmLsi1 homologous to Lsi1 in rice. In this study, we characterized the Lsi1 homologs in pumpkin (C. moschata) and its cold-tolerant wild relative C. ficifolia ('Heukjong') in order to develop a DNA marker for selecting a bloomless trait and to establish the molecular basis for breeding bloomless stock cultivars of C. ficifolia. A Cleaved amplified polymorphic sequence (CAPS) marker (CM1-CAPS) was designed based on a non-sysnonymous single nucleotide polymorphism (SNP, C>T) of the CmLsi1 mutant-type allele, and its applicability for Marker-assisted selection (MAS) was confirmed by evaluating three bloom and five bloomless pumpkin stock cultivars. Quantitative RT-PCR of the CmLsi1 for these stock cultivers implied that expression level of the CmLsi1 gene does not appear to be associated with the bloom/bloomless trait and may differ depending on plant species and tissues. A full length cDNA of the Lsi1 homolog [named CfLsi1($B^+$)] of 'Heukjong' (C. ficifolia), was cloned and sequence comparison between CmLsi1($B^+$) and CfLsi1($B^+$) revealed that there exists total 24 SNPs, of which three were non-synonymous. Phylogenetic analysis of CfLsi1($B^+$) and Lsi1 homologs further revealed that CfLsi1($B^+$) is closesly related to Nodulin 26-like intrinsic proteins (NIPs) and most similar to CpNIP1 of C. pepo than C. moschata.

Improvement of cultural efficiency using DNA markers in anther and seed culture of rice (DNA marker를 이용한 벼의 조직배양 효율개선)

  • Kim, Hong-Jib;Kim, Tae-Heon;Sohn, Jae-Keun
    • Current Research on Agriculture and Life Sciences
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    • v.27
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    • pp.21-28
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    • 2009
  • The purpose of this study was to improve the culturability of 'IR 36', a indica type rice cultivar using DNA marker associated with the ability of plant regeneration in anther and seed culture. The culturability of 6 rice cultivars and 2 indica/japonica lines ('MGRI 036', 'MGRI 079') were investigated in anther and seed culture. The culturability of 3 japonica rice cultivars were much higher than tongil and indica rice cultivars, and 'MGRI 036' and 'MGRI 079' has high culturability with 20% regenerability, also. 34 $BC_2F_4$ 4 lines were selected by marker screening using RZ400 among 90 $BC_2F_4$ lines derived from a cross between 'MGRI 079' and 'IR 36'. The frequency of callus formation of 10 $BC_2F_4$ lines were higher than 'IR 36' in anther culture among selected 34 $BC_2F_4$ lines. The ability of plant regeneration of 10 lines were higher than 'IR 36' in the seed culture among selected 34 $BC_2F_4$ lines. A promising line, $BC_2F_4$-28, was selected to have better culturability in the anther and seed culture among selected 34 $BC_2F_4$ lines. The heading date and grain shape of the $BC_2F_4$-28 was similar to 'IR 36'. Using the RZ400 DNA marker associated with the culturability will be useful method for improving of indica rice culticvar's culturability in rice breeding program.

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Development of Selectable Marker of High Oleate Trait in Peanut (Arachis hypogaea L.) (땅콩에서 고 올레인산 형질관련 분자마커의 선발)

  • Yang, Kiwoung;Pae, Suk-Bok;Park, Chang-Hwan;Lee, Myoung Hee;Jung, Chan-Sik;Son, Jeong-Hee;Park, Keum-Yong
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.507-514
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    • 2010
  • Peanut(Arachis hypogaea L.) is one of the major oilseed crops. The peanut oil consists of palmitic, oleic and linoleic acids, which are present at levels of 10%, 36-67% and 15-43%, respectively. High oleate mutant of peanut F435 contains 80% oleate and as little as 2% linoleate in seed oil. Previous study indicated that delta 12 fatty acid desaturase is a major enzyme controlling the oleate content in seeds of oilseed crops. F435 sequence alignment of their coding regions disclosed that an extra A(adenine) was inserted at the position +2,823 bp of delta 12 fatty acid desaturase gene. This study was to develop molecular marker (SNP marker) co-segregating with the high oleate trait. Chopyeong ${\times}$ F435 $F_2$ 41 population were investigated using molecular marker and fatty acid assay (NIR and gas chromatography). Finally, this marker segregates Chopyeong type 26 lines, heterotype 9 lines and F435 type 6 lines. These results in our study suggested that SNP marker conform fatty acid assay.

Identification of DNA Markers Related to Resistance to Herbicide Containing Mesotrione in Tongil Type Rice (통일형 벼에서 메소트리온계 제초제 저항성 연관 DNA marker 탐색)

  • Lee, Ji-Yoon;Cho, Jun-Hyeon;Lee, Jong-Hee;Cho, Su-Min;Kwon, Young-Ho;Park, Dong-Soo;Song, You-Chun;Ko, Jong-Min
    • Korean Journal of Breeding Science
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    • v.50 no.4
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    • pp.387-395
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    • 2018
  • This study was conducted to identify DNA markers related to resistance to herbicide containing mesotrione in Tongil type rice. Two Tongil type elite lines; Milyang154 and Suweon382, showed resistance to mesotrione, whereas the others were susceptible at 20 days after mesotrione application, and severe growth inhibition was observed in the remaining 13 lines. As a result of analysis of mesotrione resistance using 190 $F_2$ populations derived from a cross of Hanareum2 (susceptible) and Milyang154 (resistant), the mesotrione resistance locus was shown to be a single dominant gene with a 3:1 segregation ratio ($X^2=1.19$, P=0.31). To identify a DNA marker closely linked to the mesotrione resistance gene, bulked segregant analysis (BSA) was adopted. The DNA marker RM3501 was identified on chromosome 2 with a recombinant value of 0.53 to the mesotrione resistance gene. Mst1(t) was located between SSR (simple sequence repeat) markers RM3501 and RM324 with a physical map distance of 10.2 Mb-11.4 Mb on chromosome 2. The band pattern of agarose gel electrophoresis of the SSR marker RM3501 showed the same segregation pattern with respect to mesotrione treatment in 20 Tongil type varieties and a $BC_2F_2$ segregation population derived from a cross between Unkwang (resistant) and Hanareum2 (susceptible). Thus, the RM3501 DNA marker could be used in breeding programs for Marker Assisted Selection in mesotrione resistant rice breeding.