• Title/Summary/Keyword: recombinant inbred lines

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Identification of QTLs Associated with Physiological Nitrogen Use Efficiency in Rice

  • Cho, Young-Il;Jiang, Wenzhu;Chin, Joong-Hyoun;Piao, Zhongze;Cho, Yong-Gu;McCouch, Susan R.;Koh, Hee-Jong
    • Molecules and Cells
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    • v.23 no.1
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    • pp.72-79
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    • 2007
  • Demand for low-input sustainable crop cultivation is increasing to meet the need for environment-friendly agriculture. Consequently, developing genotypes with high nutrient use efficiency is one of the major objectives of crop breeding programs. This study was conducted to identify QTLs for traits associated with physiological nitrogen use efficiency (PNUE). A recombinant inbred population (DT-RILs) between Dasanbyeo (a tongil type rice, derived from an indica ${\times}$ japonica cross and similar to indica in its genetic make-up) and TR22183 (a Chinese japonica variety) consisting of 166 $F_8$ lines was developed and used for mapping. A frame map of 1,409 cM containing 113 SSR and 103 STS markers with an average interval of 6.5 cM between adjacent marker loci was constructed using the DT-RILs. The RILs were cultivated in ordinary-N ($N-P_2O_5-K_2O=100-80-80kg/ha$) and low-N ($N-P_2O_5-K_2O=50-80-80kg/ha$) (100 kg/ha) conditions. PNUE was positively correlated with the harvest index and grain yield in both conditions. Twenty single QTLs (S-QTLs) and 58 pairs of epistatic loci (E-QTLs) were identified for the nitrogen concentration of grain, nitrogen concentration of straw, nitrogen content of shoot, harvest index, grain yield, straw yield and PNUE in both conditions. The phenotypic variance explained by these S-QTLs and E-QTLs ranged from 11.1 to 44.3% and from 16.0% to 63.6%, respectively. The total phenotypic variance explained by all the QTLs for each trait ranged from 35.8% to 71.3%, showing that the expression of PNUE and related characters depends signify- cantly upon genetic factors. Both S-QTLs and E-QTLs may be useful for marker-assisted selection (MAS) to develop higher PNUE genotypes.

Quantitative Trait Locus Mapping and Candidate Gene Analysis for Plant Architecture Traits Using Whole Genome Re-Sequencing in Rice

  • Lim, Jung-Hyun;Yang, Hyun-Jung;Jung, Ki-Hong;Yoo, Soo-Cheul;Paek, Nam-Chon
    • Molecules and Cells
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    • v.37 no.2
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    • pp.149-160
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    • 2014
  • Plant breeders have focused on improving plant architecture as an effective means to increase crop yield. Here, we identify the main-effect quantitative trait loci (QTLs) for plant shape-related traits in rice (Oryza sativa) and find candidate genes by applying whole genome re-sequencing of two parental cultivars using next-generation sequencing. To identify QTLs influencing plant shape, we analyzed six traits: plant height, tiller number, panicle diameter, panicle length, flag leaf length, and flag leaf width. We performed QTL analysis with 178 $F_7$ recombinant inbred lines (RILs) from a cross of japonica rice line 'SNU-SG1' and indica rice line 'Milyang23'. Using 131 molecular markers, including 28 insertion/deletion markers, we identified 11 main- and 16 minor-effect QTLs for the six traits with a threshold LOD value > 2.8. Our sequence analysis identified fifty-four candidate genes for the main-effect QTLs. By further comparison of coding sequences and meta-expression profiles between japonica and indica rice varieties, we finally chose 15 strong candidate genes for the 11 main-effect QTLs. Our study shows that the whole-genome sequence data substantially enhanced the efficiency of polymorphic marker development for QTL fine-mapping and the identification of possible candidate genes. This yields useful genetic resources for breeding high-yielding rice cultivars with improved plant architecture.

Development of Molecular Markers for Xanthomonas axonopodis Resistance in Soybean

  • Kim Ki-Seung;Van Kyujung;Kim Moon Young;Lee Suk-Ha
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.5
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    • pp.429-433
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    • 2004
  • A single recessive gene, rxp, controls the bacterial leaf pustule (BLP) resistance in soybean and in our previous article, it has been mapped on linkage group (LG) D2 of molecular genetic map of soybean. A total of 130 recombinant inbred lines (RILs) from a cross between BLP-resistant SS2-2 and BLP-susceptible Jangyeobkong were used to identify molecular markers linked to rxp. Fifteen simple sequence repeat (SSR) markers on LG D2 were screened to construct a genetic map of rxp locus. Only four SSR markers, Satt135, Satt372, Satt448, and Satt486, showed parental polymorphisms. Using these markers, genetic scaffold map was constructed covering 26.2cM. Based on the single analysis of variance, Satt372 among these four SSR markers was the most significantly associated with the resistance to BLP. To develop new amplified fragment length polymorphism (AFLP) marker linked to the resistance gene, bulked segregant analysis (BSA) was employed. Resistance and susceptible bulks were made by pooling equal amount of genomic DNAs from ten of each in the segregating population. A total of 192 primer combinations were used to identify specific bands to the resistance, selecting three putative AFLP markers. These AFLP markers produced the fragment present in SS2-2 and the resistant bulk, and not in Jangyeobkong and the susceptible bulk. Linkage analysis revealed that McctEact97 $(P=0.0004,\;R^2=14.67\%)$ was more significant than Satt372, previously reported as the most closely linked marker.

Identification of SNPs tightly linked to the QTL for pod shattering in soybean[Glycine max (L.) Merr.]

  • Kim, Kyung-Ryun;Kim, Kyung Hye;Go, Hong Min;Lee, Ju Seok;Moon, Jung-Kyung;Ha, Bo-Keun;Jeong, Soon-Chun;Kim, Namshin;Kang, Sungtaeg
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.146-146
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    • 2017
  • The pod shattering or dehiscence is essential for the propagation of pod-bearing plant species in the wild, but it causes significant yield losses during harvest of domesticated crop plants. Identifying novel molecular makers, which are linked to seed-shattering genes, is needed to employ the molecular marker-assisted selection for efficiently developing shattering-resistant soybean varieties. In this study, a genetic linkage map was constructed using 115 recombinant inbred lines (RILs) developed from crosses between the pod shattering susceptible variety, Keunol, and resistant variety, Sinpaldal. A 180 K Axiom(R) SoyaSNPs data and pod shattering data from two environments in 2001 and 2015 were used to identify quantitative trait loci (QTL) for pod shattering. A major QTL was identified between two flanking single nucleotide polymorphism (SNP) markers, AX-90320801 and AX-90306327 on chromosome 16 with 1.3 cM interval, 857 kb of physical range. In sequence, genotype distribution analysis was conducted using extreme phenotype RILs. This could narrow down the QTL down to 153 kb on the physical map and was designated as qPDH1-KS with 6 annotated gene models. All exons within qPDH1-KS were sequenced and the 6 polymorphic SNPs affecting the amino acid sequence were identified. To develop universally available molecular markers, 38 Korean soybean cultivars were investigated by the association study using the 6 identified SNPs. Only two SNPswere strongly associated with the pod shattering. These two identified SNPs will help to identify the pod shattering responsible gene and to develop pod shattering-resistant soybean plants using marker-assisted selection.

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Positional mapping for foxglove aphid resistance with 180k SNP array in soybean [Glycine max (L.) Merr.]

  • Park, Sumin;Kim, Kyung Hye;Go, Hong Min;Lee, Ju Seok;Jung, Jin Kyo;Bilyeu, Kristin D.;Lee, Jeong-Dong;Kan, Sungtaeg
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.145-145
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    • 2017
  • Foxglove aphid, Aulacorthum solani (Kaltenbach), is a Hemipteran insect that infected a wide variety of plants worldwide and caused serious yield losses in crops. The objective of this study was to identify the putative genes to foxglove aphid resistance in wild soybean, PI 366121 (Glycine soja Sieb. and Zucc.). One hundred and forty-one F4:8 recombinant inbred lines developed from a cross between susceptible variety, Williams 82 and foxglove aphid resistance wild soybean, PI 366121 were used. The two type of resistance response, antibiosis and antixenosis resistance were evaluated through choice and no-choice test, graded by the degree of total plant damage and primary infestation leaf damage; a genome-wide molecular linkage map was constructed with 29,898 single-nucleotide polymorphism markers utilizing a Axiom(R) 180K soyaSNP array. Using inclusive composite interval mapping analysis for foxglove aphid resistance, one major candidate QTL on chromosome 7 was identified. The major QTL on chromosome 7 showed both antixenosis and antibiosis resistance responses. The newly identified major QTL was consistent with previously reported QTL, Raso2, which showed around 5 times narrow down interval range with 8 candidate genes. Furthermore, total 1,115 soybean varieties including Glycine soja and Glycine max were exposed to germplasm screening, and 31 varieties, which showed significant antibiosis type foxglove aphid resistance were identified. This result could be useful in breeding for new foxglove aphid resistant soybean cultivars and developing novel insecticides.

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Identification of QTLs controlling somatic embryogenesis using RI population of cultivar ${\times}$ weedy soybean

  • Choi, Pilson;Mano, Yoshiro;Ishikawa, Atsuko;Odashima, Masashi;Umezawa, Taishi;Fujimura, Tatsuhito;Takahata, Yoshihito;Komatsuda, Takao
    • Plant Biotechnology Reports
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    • v.4 no.1
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    • pp.23-27
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    • 2010
  • Quantitative trait loci (QTLs) controlling ability of somatic embryogenesis were identified in soybean. A frame map with 204-point markers was developed using an RI population consisting of 117 $F_{11}$ lines derived from a cross between cultivar 'Keburi' and a weedy soybean 'Masshokutou Kou 502'. The parents differed greatly in their abilities of somatic embryogenesis using immature cotyledons as explants. The ability of somatic embryogenesis was evaluated in five different experiments: the $F_{11}$ (evaluated in 1998) and $F_{15}$ (2002) generations cultured on basal media supplemented with $40\;mg\;l^{-1}$ 2,4-D (2,4-D1998 and 2,4-D2002), $F_{14}$ (2001) generation on medium with $40\;mg\;l^{-1}$ 2,4-D and high sucrose concentration [2,4-D2001 ($30\;g\;l^{-1}$ sucrose)], and the $F_{11}$ (1998) and $F_{12}$ (1999) generations on medium with $10\;mg\;l^{-1}$ NAA (NAA1998 and NAA1999). The RILs showed wide and continuous variations in each of the five experiments. In the composite interval mapping analysis, 2 QTLs were found in group 8 (D1b + W, LOD = 5.42, $r^2$ = 37.5) in the experiment of 2,4-D1998 and in group 6 (C2, LOD = 6.03, $r^2$ = 26.0) in the experiment of 2,4-D2001 (high concentration sucrose). In both QTLs, alleles of 'Masshokutou Kou 502' with high ability of somatic embryogenesis contributed to the QTLs. For the other three experiments, no QTL was detected in the criteria of LOD >3.0, suggesting the presence of minor genes.

Inheritance of Agronomic Traits and Their Interrelationship in Mungbean(Vigna radiata(L.) Wilczek)

  • Sriphadet, Sukhumaporn;Lambrides, Christopher J.;Srinives, Peerasak
    • Journal of Crop Science and Biotechnology
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    • v.10 no.4
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    • pp.249-256
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    • 2007
  • A study was conducted to observe the variation and inheritance of agronomic traits and their interrelationship in mungbean. The objective of the study was to compare agronomic traits and hardseed percentage of 268 recombinant inbred lines(RILs) developed from the cross between wild Vigna subspecies sublobata "ACC 41" with the mungbean cultivar "Berken". The RIL population and their parents were evaluated under controlled conditions in a glass house at the University of Queensland, Brisbane, Australia. The results showed significant differences among the RILs and among the parents in all traits under study. Berken had a longer flowering date and a higher seed weight per plant, but less total leaf number and pod number per plant than ACC 41. A germination test between papers revealed that ACC 41 was 100% hard-seeded and did not germinate at all, while Berken germinated up to 100%. Their RILs distributed well between 0 to 100% hardseed. Upon scarification, all hardseed germinated within seven days. Narrowsense heritability estimates of total leave number, hardseedness, pod length, and pod width were highly heritable at 89.9, 98.9, 93.7, and 93.2%, respectively. The heritability of seed weight per plant and number of seeds per plant were lower at 63.1 and 58.4%, respectively. Seed weight per plant showed positive transgressive segregation when compared with ACC 41 and a positive correlation with 100 seed weight. While the number of seeds per pod showed a negative transgressive segregation when compared with Berken and a negative correlation with pod length and pod width. The RILs gave a 1:1 segregation ratio in leaflet shape, growth habit, and growth pattern, indicating that these traits were controlled by a single dominant gene.

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Identification of QTLs Associated with Resistance to Riptortus clavatus Thunberg (Heteroptera: Alydidae) in Soybean (Glycine max L. Merr.)

  • Li, Wenxin;Van, Kyujung;Zheng, Da-Hao;Liu, Weixian;Lee, Yeong-Ho;Lee, Sue-Yeon;Lee, Joon-Ho;Lee, Suk-Ha
    • Journal of Crop Science and Biotechnology
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    • v.11 no.4
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    • pp.243-248
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    • 2008
  • The bean bug Riptortus clavatus Thunberg (Heteroptera: Alydidae) is an important pest, causing serious yield loss in soybean. But the information on mechanism of resistance to R. clavatus is limited. The objective of this study was to identify QTLs for R. clavatus resistance using simple sequence repeat (SSR) markers in a soybean population of recombinant inbred lines (RILs) developed from the cross PI 171451 ${\times}$ Hwaeomputkong. A genetic map from this population was constructed with a total of 136 SSR markers covering 1073.9 cM on 20 linkage groups (LGs). With 126 $F_5$ RILs, two independent QTLs for resistance to R. clavatus were mapped on LGs B1 and C2. The amount of phenotypic variation explained by these QTLs ranged from 12 to 16%. PI 171451 showed an escape response to R. clavatus. Under feeding conditions, 14.4% of RILs showed greater resistance to R. clavatus than the resistant parent. The resistance to R. clavatus in soybean from PI 171451 was incomplete and quantitatively inherited and the QTLs for resistance to R. clavatus detected in the RIL population were not significantly affected by epistatic interactions.

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Resistance Potential of Bread Wheat Genotypes Against Yellow Rust Disease Under Egyptian Climate

  • Mahmoud, Amer F.;Hassan, Mohamed I.;Amein, Karam A.
    • The Plant Pathology Journal
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    • v.31 no.4
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    • pp.402-413
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    • 2015
  • Yellow rust (stripe rust), caused by Puccinia striiformis f. sp. tritici, is one of the most destructive foliar diseases of wheat in Egypt and worldwide. In order to identify wheat genotypes resistant to yellow rust and develop molecular markers associated with the resistance, fifty F8 recombinant inbred lines (RILs) derived from a cross between resistant and susceptible bread wheat landraces were obtained. Artificial infection of Puccinia striiformis was performed under greenhouse conditions during two growing seasons and relative resistance index (RRI) was calculated. Two Egyptian bread wheat cultivars i.e. Giza-168 (resistant) and Sakha-69 (susceptible) were also evaluated. RRI values of two-year trial showed that 10 RILs responded with RRI value >6 <9 with an average of 7.29, which exceeded the Egyptian bread wheat cultivar Giza-168 (5.58). Thirty three RILs were included among the acceptable range having RRI value >2 <6. However, only 7 RILs showed RRI value <2. Five RILs expressed hypersensitive type of resistance (R) against the pathogen and showed the lowest Average Coefficient of Infection (ACI). Bulked segregant analysis (BSA) with eight simple sequence repeat (SSR), eight sequence-related amplified polymorphism (SRAP) and sixteen random amplified polymorphic DNA (RAPD) markers revealed that three SSR, three SRAP and six RAPD markers were found to be associated with the resistance to yellow rust. However, further molecular analyses would be performed to confirm markers associated with the resistance and suitable for marker-assisted selection. Resistant RILs identified in the study could be efficiently used to improve the resistance to yellow rust in wheat.

Progress and Prospect of Rice Biotechnology in Korea

  • Tae Young, Chung
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 1997.06a
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    • pp.23-49
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    • 1997
  • This is a progress report of rice biotechnology including development of gene transformation system, gene cloning and molecular mapping in rice. The scope of the research was focused on the connection between conventional breeding and biotech-researches. Plant transformation via Agrobacterium or particle bombardment was developed to introduce one or several genes to recommended rice cultivars. Two chimeric genes containing a maize ribosome inactivating protein gene (RIP) and a gerbicide resistant gene (bar) were introduced to Nipponbare, a Japonica cultivar, and transmitted to Korean cultivars. The homozygous progenies of herbicide resistant transgenic plant showed good fertility and agronomic characters. To explore the genetic resourses in rice, over 8,000 cDNA clones from immature rice seed have been isolated and sequenced. About 13% of clones were identified as enzymes related to metabolic pathway. Among them, twenty clones have high homology with genes encoding enzymes in the photorespiratory carbon cycle reaction. Up to now about 100 clones were fully sequenced and registered at EMBL and GenBank. For the mapping of quantitative tarits loci (QTL) and eternal recombinant inbred population with 164 F13 lines (MGRI) was developed from a cross between Milyang 23 and Gihobyeo, Korean rice cultivars. After construction of fully saturated RFLP and AFLP map, quantitative traits using MGRI population were analyzed and integrated into the molecular map. Eighty seven loci were determined with 27 QTL characters including yield and yield components on rice chromosomes. Map based cloning was also tried to isolate semi-dwarf (sd-1) gene in rice. A DNA probe, RG 109, the most tightly linked to sd-1 gene was used to screen from bacterial artifical chromosome (BAC) libraries and five over lapping clones presumably containing sd-1 gene were isolated. Rice genetic database including results of biotech reasearch and classical genetics is provided at Korea Rice Genome Server which is accessible with world wide web (www) browser. The server provides rice cDNA sequences and map informations linked with phenotypic images.