• Title/Summary/Keyword: Backcross breeding

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Application of genomics into rice breeding

  • Ando, Ikuo
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.13-13
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    • 2017
  • By the progress of genome sequencing, infrastructures for marker-assisted breeding (MAB) of rice came to be established. Fine mapping and gene isolation have been conducted using the breeding materials derived from natural variations and artificial mutants. Such genetic analysis by the genome-wide dense markers provided us the knowledge about the many genes controlling important traits. We identified several genes or quantitative trait loci (QTL) for heading date, blast resistance, eating quality, high-temperature stress tolerance, and so on. NILs of each gene controlling heading date contribute to elongate the rice harvest period. Determination of precise gene location of blast resistance gene pi21, allowed us to overcome linkage drag, co-introduction of undesirable eating quality. We could also breed the first practical rice cultivar in Japan with a brown planthopper resistance gene bph11 in the genetic back-ground of an elite cultivar. Discovery of major and minor QTLs for good eating quality allowed us to fine-tune of eating quality according to the rice planting area or usage of rice grain. Many rice cultivars have bred efficiently by MAB for several traits, or by marker-assisted backcross breeding through chromosome segment substitution lines (CSSLs) using genetically diverse accessions. We are also systematically supporting the crop breeding of other sectors by MAB or by providing resources such as CSSLs. It is possible to pyramid many genes for important traits by using MAB, but is still difficult to improve the yielding ability. We are performing a Genomic Selection (GS) for improvement of rice biomass and grain yield. We are also trying to apply the genome editing technology for high yield rice breeding.

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$\alpha$- and $\beta$-Amylase Isozyme Expresser Native Proteins in Tropical Silkworm Bombyx mori L.

  • Chattopadhyay, G.K.;Verma, A.K.;Sengupta, A.K.;Das, S.K.;Urs, S.Raje
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.2
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    • pp.189-194
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    • 2004
  • Amylase isozyme based three multivoltine viz., N+p, Np, N+ $p^{cho}$ and two bivoltine-D6+p, D6p syngenic lines (Syn. L) were developed from germplasm (GP) stocks Nistari (N) and D6 respectively. haemolymph isozyme pattern at pH 7.0 and 8.5 depicted a total 11 number (Am $y_{1 to 6}$ at pH 7.0 and Am $y^{l to 5}$ at pH 8.5) of native proteins (NP) of various sizes are amylase isozyme expressers. Among eleven NPs, two NPs of 770 kDa (Am $y^{6}$ at pH 7.0) and 376 kDa (Am $y^3$ at pH 8.5) are $\alpha$-amylase expressers and remaining NPs of 370, 364, 350, 329 and 274 kDa at pH 7.0 and 206, 292, 416, 725 kDa at pH 8.5 are $\beta$-amylase expressers. Accordingly, digestive juice amylase isozyme pattern at aforesaid pH also depicted a total number of 10 NPs (Am $y^{1 to 5}$) at each pH 7.0 and 8.5 are amylase expressers of which NP of 387 kDa (Am $y^4$ at pH 7.0) and 780 kDa (Am $y^{5}$ at pH 8.5) are a-amylase expresser. Remaining NPs of 338,297 & 216 kDa at pH 7.0 and 370, 341, 329 &302 kDa at pH 8.5 are $\beta$-amylase expresser. Recurrent backcross lines (RBL) viz., N+pRBL and NpRBL were developed through introgression of high shell weight character (a multigenic trait) to be used further for congenic line (Con. L) development and to understand any association with introgressed character. Isozyme pattern in haemolymph of RBLs depicted only one $\alpha$-amylase of 770 kDa at pH 7.0 and 376 kDa at pH 8.0 with three and four respective $\beta$-amylase bands but in bivoltine lines numbers of $\beta$-amylase bands vary between 1 to 2 at aforesaid pH. Variability was also observed in digestive juice of multivolitine and its RBLs but bivoltine lines express null activity at both pH except appearance of one very week $\alpha$-amylase band D6+p at pH 8.5. Overall study suggests that not a single NP at both pH is common for expression of any band of amylase isozyme i.e., a totally different set of proteins are the amylase isozyme expresser at specific pH and no molecular factor of amylase is associated in developed RBLs which showed improvement on survival, single cocoon shell weight (SCSW) and single filament length over receptor parents.s.s.s.

Breeding of Bivoltine Breeds of Bombyx mori L Suitable for Variable Climatic Conditions of the Tropics

  • Moorthy, S. M.;Das, S. K.;Kar, N. B.;Urs, S. Raje
    • International Journal of Industrial Entomology and Biomaterials
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    • v.14 no.2
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    • pp.99-105
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    • 2007
  • The success of rearing with presently available conventional bivoltine is unpredictable in some seasons of the tropical regions due to highly fluctuating adverse climatic conditions. Thus, in order to popularize bivoltine breeds in tropical parts of India, it is very much essential to have a bivoltine breed(s), which can give stable cocoon crop under variable environments. With this objective a breeding programme was undertaken to improve the survival trait in bivoltine silkworm by introducing multivoltine genes into bivoltine through back crossing. Resultant bivoltine lines showed significantly higher survival in compared to the receptor (Bivoltine) parent and control bivoltine breed. Esterase isozyme analysis revealed similar banding pattern in the developed bivoltine and in the donor multivoltine, which predicts the introgression of multivoltine character into evolved bivoltine.

QTL Mapping of Agronomic Traits in an Advanced Backcross Population from a Cross between Oryza sativa L. cv. Milyang 23 and O. glaberrima

  • Kang, Ju-Won;Suh, Jung-Pil;Kim, Dong-Min;Oh, Chang-Sik;Oh, Ji-Min;Ahn, Sang-Nag
    • Korean Journal of Breeding Science
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    • v.40 no.3
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    • pp.243-249
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    • 2008
  • In the previous study, 141 $BC_3F_2$ lines from a cross between the Oryza sativa cv. Milyang 23 and O. glaberrima were used to identify favorable wild QTL alleles for yield component traits. In this study, we carried out QTL analysis of four grain morphology as well as four yield component traits using 141 $BC_3F_5$ lines from the same cross and compared QTLs detected in two different generations. The mean number of O. glaberrima segments in the 141 $BC_3F_5$ lines ranged from 1 to 13 with 2.69 and 5.71 of the average means of homozygous and heterozygous segments, respectively. There was a three-fold difference in the number of QTLs detected for four traits commonly evaluated in two generations (seven QTLs in the $BC_3F_5$ vs 21 in the $BC_3F_2$ population). The percentages of the phenotypic variance explained by QTLs in the BC3F5 population were similar to or less than those in the $BC_3F_2$ population. This is probably due to the difference in the genetic composition of two populations and the environmental effects. The locations of the QTLs commonly detected in both generations were in good agreement except for one QTL for spikelets per panicle. The yield QTL, yd3 was colocalized with the spikelets per panicle, spp3. Yield increase at this locus is due to the increase in spikelets per panicle, because both traits were associated with increase in spikelets per panicle and yield due to the presence of an O. glaberrima allele. Clusters of QTLs for grain morphology traits were observed in two chromosome regions. One cluster harboring five QTLs near SSR markers RM106 and RM263 was detected on chromosome 2. This population would serve as a foundation for development of the introgression line population from a cross between Milyang 23 and O. glaberrima.

Near-Isogenic Lines for Genes Conferring Hypersensitive Resistance to Bacterial Spot in Chili Pepper

  • Kim, Byung-Soo;Kim, Young-Chun;Shin, Kwang-Sik;Kim, Jeong-Hoon
    • The Plant Pathology Journal
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    • v.23 no.3
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    • pp.155-160
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    • 2007
  • In order to develop chili pepper bacterial spot resistant cultivars and near-isogenic lines (NILs) to prompt the molecular mapping of the resistance gene, we have run backcross breeding program since 1994. Two resistance genes against Xanthomonas axonopodis pv. vesicatoria Bs2 from Fla. XVR 3-25 and Bs3 from our breeding line 25-11-3-2, were introduced into a land race, Chilseongcho (abbreviated to Chilseong hereafter) with good fruit guality. We report here the testing of $BC_4F_3\;to\;BC_4F_5$. We found that $BC_4F_5$ lines of the crosses were homozygous with respect to the respective genes of introduction. The lines, in which Bs2 gene was introduced, were hypersensitively resistant to both race 1 and race 3 of X. axonopodis pv. vesicatoria, whereas, those in which Bs3 was introduced were resistant to race 1.

Recurrent parent genome (RPG) recovery analysis in a marker-assisted backcross breeding based on the genotyping-by-sequencing in tomato (Solanum lycopersicum L.) (토마토 MABC 육종에서 GBS(genotyping-by-sequencing)에 의한 RPG(recurrent parent genome) 회복률 분석)

  • Kim, Jong Hee;Jung, Yu Jin;Seo, Hoon Kyo;Kim, Myong-Kwon;Nou, Ill-Sup;Kang, Kwon Kyoo
    • Journal of Plant Biotechnology
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    • v.46 no.3
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    • pp.165-171
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    • 2019
  • Marker-assisted backcrossing (MABC) is useful for selecting an offspring with a highly recovered genetic background for a recurrent parent at early generation to various crops. Moreover, marker-assisted backcrossing (MABC) along with marker-assisted selection (MAS) contributes immensely to overcome the main limitation of the conventional breeding and it accelerates recurrent parent genome (RPG) recovery. In this study, we were employed to incorporate rin gene(s) from the donor parent T13-1084, into the genetic background of HK13-1151, a popular high-yielding tomato elite inbred line that is a pink color fruit, in order to develop a rin HK13-1084 improved line. The recurrent parent genome recovery was analyzed in early generations of backcrossing using SNP markers obtained from genotyping-by-sequencing analysis. From the $BC_1F_1$ and $BC_2F_1$ plants, 3,086 and 4868 polymorphic SNP markers were obtained via GBS analysis, respectively. These markers were present in all twelve chromosomes. The background analysis revealed that the extent of RPG recovery ranged from 56.7% to 84.5% and from 87.8% to 97.8% in $BC_1F_1$ and $BC_2F_1$ generations, respectively. In this study, No 5-1 with 97.8% RPG recovery rate among $BC_2F_1$ plants was similar to HK13-1151 strain in the fruit shape. Therefore, the selected plants were fixed in $BC_2F_2$ generation through selfing. MAS allowed identification of the plants that are more similar to the recurrent parent for the loci evaluated in the backcross generations. MABC can greatly reduce breeding time as compared to the conventional backcross breeding. For instance, MABC approach greatly shortened breeding time in tomato.

Development and Application of Speed Vernalization System for Practical Speed Breeding in Wheat (Triticum aestivum L.)

  • Jin-Kyung Cha;Hyunjin Park;Youngho Kwon;So-Myeong Lee;Dongjin Shin;Jong-Hee Lee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.20-20
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    • 2022
  • A traditional wheat breeding program needs more than 12-13 years to develop a new cultivar. In recent years, 'Speed breeding (SB)' system, which uses extremely extended photoperiod (22 h), enabled up to 4-6 generations of spring wheat per year. However, since almost 70% of wheat cultivars are winter type, and over 95% of total cultivation area is for winter wheat in Korea, optimized vernalization treatment was essential for improving the SB system. Several vernalization temperatures and durations were tested with various genotypes, and the 4 weeks of 8-10 ℃ vernalization treatment was the most effective to develop 4 generations per year, for both spring and winter type wheat cultivars. This 'Speed vernalization (SV)' system followed by SB, allowed developing a new F6 recombinant inbred lines (RILs) within 2 years. Among the 184 RILs, which derived from a cross between Jokoyung and Joongmo2008, two outstanding lines were selected for yield trial test, and then named Milyang52 and Milyang53. Compared to the traditional wheat breeding program, over 60% of the time was saved to develop these two lines. Marker-assisted selection and backcross were also combined with the SV system. YW3215-2B-1 (Jokoyung*3/Gamet), which has similar agronomic traits with Jokyoung and the same Glu-B1 allele with Garnet, was developed within 2.5 years. Thus, the SV system combined with molecular breeding technology would help breeders to make a new cultivar with less time and high efficiency.

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Development of an SNP set for marker-assisted breeding based on the genotyping-by-sequencing of elite inbred lines in watermelon (수박 엘리트 계통의 GBS를 통한 마커이용 육종용 SNP 마커 개발)

  • Lee, Junewoo;Son, Beunggu;Choi, Youngwhan;Kang, Jumsoon;Lee, Youngjae;Je, Byoung Il;Park, Younghoon
    • Journal of Plant Biotechnology
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    • v.45 no.3
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    • pp.242-249
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    • 2018
  • This study was conducted to develop an SNP set that can be useful for marker-assisted breeding (MAB) in watermelon (Citrullus. lanatus L) using Genotyping-by-sequencing (GBS) analysis of 20 commercial elite watermelon inbreds. The result of GBS showed that 77% of approximately 1.1 billion raw reads were mapped on the watermelon genome with an average mapping region of about 4,000 Kb, which indicated genome coverage of 2.3%. After the filtering process, a total of 2,670 SNPs with an average depth of 31.57 and the PIC (Polymorphic Information Content) value of 0.1~0.38 for 20 elite inbreds were obtained. Among those SNPs, 55 SNPs (5 SNPs per chromosome that are equally distributed on each chromosome) were selected. For the understanding genetic relationship of 20 elite inbreds, PCA (Principal Component Analysis) was carried out with 55 SNPs, which resulted in the classification of inbreds into 4 groups based on PC1 (52%) and PC2 (11%), thus causing differentiation between the inbreds. A similar classification pattern for PCA was observed from hierarchical clustering analysis. The SNP set developed in this study has the potential for application to cultivar identification, F1 seed purity test, and marker-assisted backcross (MABC) not only for 20 elite inbreds but also for diverse resources for watermelon breeding.

Marker Assisted Selection of Brown Planthopper Resistance and Development of Multi-Resistance to Insect and Diseases in Rice (Oryza sativa L.) (DNA 마커를 이용한 벼멸구 저항성 선발 및 복합내병충성 벼 계통 육성)

  • Lee, Jong-Hee;Yeo, Un-Sang;Cho, Jun-Hyun;Lee, Ji-Yoon;Song, You-Chun;Shin, Mun-Sik;Kang, Hang-Won;Sohn, Jae-Keun
    • Korean Journal of Breeding Science
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    • v.43 no.5
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    • pp.413-421
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    • 2011
  • The main objective of this study was to develop the multi-resistance lines to insects(brown planthopper; BPH, rice green leafhopper; GRH) and disease(blast; BL, bacterial blight; BB and rice stripe virus disease;RSV) with good grain quality and plant type by combining conventional breeding and marker assisted selection(MAS) and to eliminate the linkage drag effects between Bph1 gene and culm length, we conducted MAS of Bph1 gene in advanced backcross and double cross progenies. 'Nampyeong', 'Junam' and 'Milyang220' were used as the parent in this study. 'Milyang220' was used as the donor of brown planthopper resistance gene Bph1 with tall culm length. Two backcross progenies were developed using two recipients 'Nampyeong' carrying GRH resistance gene Grh3(t) with good grain appearance and 'Junam' harboring bacterial blight resistance gene Xa3 with short culm length. Two $BC_1$ generations were resulted from the backcrossing of the $F_1$ plants with recurrent parents 'Nampyeong' and 'Junam'. The second rounds of backcrossing($BC_2$) were derived from the cross of selected resistant $BC_1F_1$ plants based on heterozygous genotype of RM28493 linked to Bph1 gene. The double crossed population was constructed from the cross of between each heterozygous $BC_2F_1$ plants at RM28493 locus of '$Nampyeong^*3$ / Milyang220' and '$Junam^*3$ / Milyang220'., The homozygous alleles in Bph1 gene were selected using co-dominant DNA marker RM28493 in double crossed population. Eighty-five lines with multi-resistance to BL, BB, RSV, GRH and BPH were selected by bio-assay and MAS in generation of double crossing. The culm length, head rice ratio and yield of the selected multi resistance lines was ranged from 71 to 88 cm, from 51 to 93%, from 449 to 629 kg/10a. respectively. We can select a promising multi resistance line similar with 'Nampyeong' of major agronomic traits such as culm legnth, head rice ratio and yield. It was designated as Milyang265. Finally this study was developed the multi resistant varieties against to insects and diseases with the good grain quality 'Milyang265' by the advanced backcross and double cross combining MAS and it can be used as genetic resources of multi-resistance to insect and diseases in rice breeding programs.

Identification of a Rice Gene (Bph 1) Conferring Resistance to Brown Planthopper (Nilaparvata lugens Stal) Using STS Markers

  • Kim, Suk-Man;Sohn, Jae-Keun
    • Molecules and Cells
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    • v.20 no.1
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    • pp.30-34
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    • 2005
  • This study was carried out to identify a high-resolution marker for a gene conferring resistance to brown planthopper (BPH) biotype 1, using japonica type resistant lines. Bulked segregant analyses were conducted using 520 RAPD primers to identify RAPD fragments linked to the BPH resistance gene. Eleven RAPDs were shown to be polymorphic amplicons between resistant and susceptible progeny. One of these primers, OPE 18, which amplified a 923 bp band tightly linked to resistance, was converted into a sequence-tagged-site (STS) marker. The STS marker, BpE18-3, was easily detectable as a dominant band with tight linkage (3.9cM) to Bph1. It promises to be useful as a marker for assisted selection of resistant progeny in backcross breeding programs to introgress the resistance gene into elite japonica cultivars.