AFLP targeting of the 1-cM region conferring the vrs1 gene for six-rowed spike in barley, Hordeum vulgare L.

Genome ◽  
2004 ◽  
Vol 47 (6) ◽  
pp. 1122-1129 ◽  
Author(s):  
Congfen He ◽  
Badraldin Ebrahim Sayed-Tabatabaei ◽  
Takao Komatsuda
Keyword(s):  
Hordeum Vulgare ◽  
Fragment Length ◽  
Positional Cloning ◽  
Aflp Markers ◽  
Near Isogenic Lines ◽  
Hordeum Vulgare L ◽  
Plant Materials ◽  
Multiple Alleles ◽  
Isogenic Lines ◽  
Sequence Tagged Sites

Spike morphology is a key characteristic in the study of barley domestication, yield, and use. Multiple alleles at the vrs1 locus control the development and fertility of the lateral spikelets of barley. We developed five amplified fragment length polymorphism (AFLP) markers tightly linked to the vrs1 locus using well-characterized near-isogenic lines as plant materials. The AFLP markers were integrated into three different maps, in which 'Azumamugi' was used as the maternal parent. Of the three maps, Hordeum vulgare L. 'Azumamugi' × H. vulgare 'Golden Promise' showed recombination of the AFLP markers and the vrs1 locus (closest, 0.05 cM), providing the best mapping population for positional cloning of alleles at the vrs1 locus. Conversion of AFLP bands into polymorphic sequence-tagged sites (STSs) is necessary for further high-throughput genotype scoring and for bacterial artificial chromosome (BAC) library screening. We cloned and sequenced the five AFLP bands and synthesized primer pairs. PCR amplification generated DNAs of the same size from all four parental lines for each marker. Restriction endonuclease treatment of e40m36-1110/AccIII, e34m13-260/Psp1406I, e52m32-270/FokI, and e31m26-520/MnlI revealed fragment length polymorphisms between 'Azumamugi' and all the two-rowed parents. Allelism between the AFLPs and corresponding STS markers was confirmed genetically, indicating the usefulness of the STSs as genetic markers.Key words: positional cloning, codominance, near-isogenic lines, high-resolution maps, STSs.

Construction of a genetic map of barley (Hordeum vulgare L.) cross 'Azumamugi' × 'Kanto Nakate Gold' using a simple and efficient amplified fragment-length polymorphism system

Genome ◽  
2001 ◽  
Vol 44 (2) ◽  
pp. 284-292 ◽  
Author(s):  
Y Mano ◽  
S Kawasaki ◽  
F Takaiwa ◽  
T Komatsuda
Keyword(s):  
Hordeum Vulgare ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Recombinant Inbred ◽  
Fragment Length Polymorphism ◽  
Recombinant Inbred Lines ◽  
Amplified Fragment Length Polymorphism ◽  
Buffer System ◽  
Hordeum Vulgare L ◽  
Sequence Tagged Sites

We have devised a simple and efficient amplified fragment-length polymorphism (AFLP) system consisting of small slab gels, a discontinuous buffer system, and silver staining. Using this system, a single worker developed a barley map with 227 polymorphic fragments in 2 months. As a mapping population, 99 recombinant inbred lines of barley cultivars 'Azumamugi' × 'Kanto Nakate Gold' were used. Most of the 227 AFLP fragments showed a Mendelian segregation ratio of 1:1, and all were assigned to the seven barley chromosomes. Thus, these fragments are useful as molecular markers. They were integrated with 40 previously characterized sequence-tagged sites, 3 isozymes, and 2 morphological markers to construct an integrated map. The resulting map covered 925.6 cM with 272 markers (detecting 150 loci) at an average interval of 6.5 cM/locus. This system greatly simplifies map construction.Key words: amplified fragment length polymorphism (AFLP), Hordeum vulgare, linkage map, recombinant inbred line (RIL), sequence-tagged site (STS).

Transfer and mapping of the shoot-differentiation locus Shd1 in barley chromosome 2

Genome ◽  
1995 ◽  
Vol 38 (5) ◽  
pp. 1009-1014 ◽  
Author(s):  
T. Komatsuda ◽  
F. Taguchi-Shiobara ◽  
S. Oka ◽  
F. Takaiwa ◽  
T. Annaka ◽  
...  
Keyword(s):  
Hordeum Vulgare ◽  
Immature Embryo ◽  
Gene Products ◽  
Chromosome 2 ◽  
Near Isogenic Lines ◽  
Hordeum Vulgare L ◽  
Isogenic Lines ◽  
Shoot Differentiation ◽  
Backcross Breeding ◽  
Positive Effect

The locus Shd1, which we previously mapped to the long arm of chromosome 2 of Hordeum vulgare L., controls the differentiation of shoots from immature barley embryo callus. The locus has major effects and its action explains more than 65% of the total genetic variance in the shoot-differentiation rate. The allele of cultivar Kanto Nakate Gold designated Shd1K has a significant positive effect on the shoot-differentiation rate, whereas Shd1A of cultivar Azumamugi does not promote shoot differentiation. To identify gene products and characterize the function of Shd1, a set of near-isogenic lines is essential. In this study we produced BC5F1 plants by repeated backcrossing of 'Azumamugi' to F1 plants ('Azumamugi' × 'Kanto Nakate Gold'). The BC5F1 plants were examined for their RFLP genotype and for the shoot-differentiation ability of immature embryo-derived callus. The results indicated that the Shd1 locus was located in a chromosomal region between MWG2081 and MWG503 that flanks the MWG801, cMWG699, v (ear type), and MWG865 loci. Shd1K from 'Kanto Nakate Gold' functions effectively in the genetic background of 'Azumamugi', an indication that backcross breeding is possible for production of near-isogenic lines that would be very suitable for tissue culture.Key words: Hordeum vulgare L., shoot-differentiation ability, immature embryo callus, backcross breeding, RFLPs.

Identification of restriction fragment length polymorphism and random amplified polymorphic DNA markers linked to downy mildew resistance genes in lettuce, using near-isogenic lines

Genome ◽  
1991 ◽  
Vol 34 (6) ◽  
pp. 1021-1027 ◽  
Author(s):  
Ilan Paran ◽  
Richard Kesseli ◽  
Richard Michelmore
Keyword(s):  
Restriction Fragment Length Polymorphism ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Rapd Markers ◽  
Fragment Length Polymorphism ◽  
Random Amplified Polymorphic Dna ◽  
Near Isogenic Lines ◽  
Isogenic Lines ◽  
Restriction Fragment Length

Near-isogenic lines were used to identify restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) markers linked to genes for resistance to downy mildew (Dm) in lettuce. Two pairs of near-isogenic lines that differed for Dm1 plus Dm3 and one pair of near-isogenic lines that differed for Dm11 were used as sources of DNA. Over 500 cDNAs and 212 arbitrary 10-mer oligonucleotide primers were screened for their ability to detect polymorphism between the near-isogenic lines. Four RFLP markers and four RAPD markers were identified as linked to the Dm1 and Dm3 region. Dm1 and Dm3 are members of a cluster of seven Dm genes. Marker CL922 was absolutely linked to Dm15 and Dm16, which are part of this cluster. Six RAPD markers were identified as linked to the Dm11 region. The use of RAPD markers allowed us to increase the density of markers in the two Dm regions in a short time. These regions were previously only sparsely populated with RFLP markers. The rapid screening and identification of tightly linked markers to the target genes demonstrated the potential of RAPD markers for saturating genetic maps.Key words: lettuce, downy mildew, near-isogenic lines, disease resistance, restriction fragment length polymorphism, random amplified polymorphic DNA.

scholarly journals Understanding Mechanisms of Salinity Tolerance in Barley by Proteomic and Biochemical Analysis of Near-Isogenic Lines

2020 ◽  
Vol 21 (4) ◽  
pp. 1516 ◽  
Author(s):  
Juan Zhu ◽  
Yun Fan ◽  
Sergey Shabala ◽  
Chengdao Li ◽  
Chao Lv ◽  
...  
Keyword(s):  
Salinity Tolerance ◽  
Crop Production ◽  
Biochemical Analysis ◽  
Mass Spectrometry Analysis ◽  
Near Isogenic Lines ◽  
Ros Scavenging ◽  
Hordeum Vulgare L ◽  
Two Dimensional Gel Electrophoresis ◽  
Spectrometry Analysis ◽  
Isogenic Lines

Salt stress is one of the major environmental factors impairing crop production. In our previous study, we identified a major QTL for salinity tolerance on chromosome 2H on barley (Hordeum vulgare L.). For further investigation of the mechanisms responsible for this QTL, two pairs of near-isogenic lines (NILs) differing in this QTL were developed. Sensitive NILs (N33 and N53) showed more severe damage after exposure to 300 mM NaCl than tolerant ones (T46 and T66). Both tolerant NILs maintained significantly lower Na+ content in leaves and much higher K+ content in the roots than sensitive lines under salt conditions, thus indicating the presence of a more optimal Na+/K+ ratio in plant tissues. Salinity stress caused significant accumulation of H2O2, MDA, and proline in salinity-sensitive NILs, and a greater enhancement in antioxidant enzymatic activities at one specific time or tissues in tolerant lines. One pair of NILs (N33 and T46) were used for proteomic studies using two-dimensional gel electrophoresis. A total of 53 and 51 differentially expressed proteins were identified through tandem mass spectrometry analysis in the leaves and roots, respectively. Proteins which are associated with photosynthesis, reactive oxygen species (ROS) scavenging, and ATP synthase were found to be specifically upregulated in the tolerant NIL. Proteins identified in this study can serve as a useful resource with which to explore novel candidate genes for salinity tolerance in barley.

YIELD COMPONENT DEVELOPMENT IN FOUR ISOGENIC BARLEY LINES DIFFERING IN AWN LENGTH

1974 ◽  
Vol 54 (2) ◽  
pp. 315-322 ◽  
Author(s):  
D. G. FARIS
Keyword(s):  
Hordeum Vulgare ◽  
Yield Components ◽  
Early Ontogeny ◽  
Yield Component ◽  
High Yield ◽  
Hordeum Vulgare L ◽  
Breeding Programs ◽  
Isogenic Lines ◽  
Component Development ◽  
Northern Alberta

Four backcross-derived isogenic lines of Atlas barley (Hordeum vulgare L.) differing in awn length (Full-, Half-, Quarter-awned and Awnless) were grown in the field for 3 yr in northern Alberta. On the average the highest yielding line was Half-awned outyielding (P < 0.10) Awnless by 5%. Weight per seed was linearly related to awn length with a regression of 0.33 mg/cm of awn. Examination of other components of yield showed that increased awn length was associated with reduced number of spikes and florets per plant. The results suggested that the magnitude of these yield components was partially dependent on competition for nutrients during the early ontogeny of the spike between the awns at the base and the developing florets at the tip of the spike. The long daylength was suggested as a factor which might put Full-awned at a disadvantage at Beaverlodge despite high yield levels there. Application of the findings to barley breeding programs is proposed.

Restriction fragment length polymorphisms linked to genes for resistance to crown rust (Puccinia coronata) in near-isogenic lines of hexaploid oat (Avena sativa)

Genome ◽  
1994 ◽  
Vol 37 (5) ◽  
pp. 823-831 ◽  
Author(s):  
A. L. Bush ◽  
R. P. Wise ◽  
P. J. Rayapati ◽  
M. Lee
Keyword(s):  
Restriction Fragment ◽  
Fragment Length ◽  
Avena Sativa ◽  
Crown Rust ◽  
Puccinia Coronata ◽  
Recurrent Parent ◽  
Near Isogenic Lines ◽  
Isogenic Lines ◽  
Length Polymorphisms ◽  
Restriction Fragment Length

Crown rust, perhaps the most important fungal disease of oat, is caused by Puccinia coronata. An examination of near-isogenic lines (NILs) of hexaploid oat (Avena sativa) was conducted to identify markers linked to genes for resistance to crown rust. These lines were created such that a unique resistance gene is present in each of the two recurrent parent backgrounds. The six NILs of the current study, X434-II, X466-I, and Y345 (recurrent parent C237-89) and D486, D494, and D526 (recurrent parent Lang), thus provide a pair of lines to study each of three resistance genes. Restriction fragment length polymorphisms and resistance loci were mapped using BC1F2 populations. Three markers were found linked to a locus for resistance to crown rust race 203, the closest at 1.9 cM in line D494 and 3.8 cM in line X466-I. In lines D526 and Y345 a marker was placed 1.0 and 1.9 cM, respectively, from the locus conferring resistance to crown rust race 345, and in D486 and X434-II a marker mapped at 8.0 and 10.2 cM from the locus for resistance to rust race 264B.Key words: genetics, disease resistance, monocots, near-isogenic lines.

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