scholarly journals 049 Development of Molecular Markers Linked to High Pigment (hp) and Dark Green (dg) Loci in Tomato

HortScience ◽  
1999 ◽  
Vol 34 (3) ◽  
pp. 449D-449
Author(s):  
Yiping Zhang ◽  
John R. Stommel

The carotenoids have an important influence on tomato fruit quality and enhance the fruit contribution to human nutrition. Expression of the high pigment (hp) locus in tomato results in increased total carotenoids and increased efficiency of utilization of the polyenes. A similar mutant, dark green (dg), contains higher level of chlorophyll in immature fruit and results in darker red pigmentation, both externally and internally in ripe fruit. Random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) analyses were performed using two pairs of near isogenic lines (NILs) designed to be isogenic at the hp and dg loci. Sixty-four AFLP primer pairs and more than 1000 RAPD 10-mer primers were screened for polymorphism between each pair of the NILs. One RAPD marker was identified to be linked to the hp gene, and two AFLP primer pairs showed polymorphic fragments which distinguished the dg NILs. The markers identified in this study will be converted to allele specific SCAR (sequence characterized amplified region) markers, which are more useful in marker-assisted selection breeding programs.

2017 ◽  
Vol 142 (6) ◽  
pp. 444-453
Author(s):  
Matthew R. Mattia ◽  
John W. Scott

Tomato (Solanum lycopersicum) breeders have observed that plants with uniform green-shouldered fruit are less prone to yellow shoulder (YS) than are plants with (dark) green-shouldered tomatoes and thus have selected for tomato cultivars with uniform green-shouldered fruit. However, a recent publication reported that a cultivar with green-shouldered fruit had significantly higher soluble solids content (SSC) than an isogenic cultivar with uniform green-shouldered fruit and postulated that selection of uniform green shoulder has negatively affected tomato flavor and processing quality. Lines with dark green (u+), uniform green (u), uniform gray-green (ug), apple green (uAg), medium green, and pale green (uPg) immature fruit colors were crossed in all combinations to produce F1 plants that were self-pollinated to produce F2 seed. Parents, F1, and F2s were planted in the field in completely randomized block designs over two seasons. Plants were visually phenotyped for immature fruit color, and fruit from each plant were selected to measure shoulder and base color with a colorimeter. Ripening fruit were harvested to measure the incidence of YS, and SSC was measured on ripe fruit from each plant with a refractometer. In the spring season, fruit from F2 plants with green-shouldered fruit had significantly higher YS incidence than all phenotypes with uniform fruit colors. In the fall, phenotypes with medium-green shoulders were also tested, and these had greater YS than all other phenotypes except green shoulder. YS incidence for green shoulder was not significantly greater than that in the other phenotypes. Fla. 7956, the apple green parent, had 0% YS and appeared to be resistant. Higher SSC was observed in the spring season than in the fall season. However, in both seasons, when comparisons were made between phenotypes that segregated in the F2s, the SSC of green-shouldered phenotypes was not significantly higher than that of other phenotypes. Plants with apple green fruit tended to be higher in SSC in the fall, but this may relate to the dark green foliage of apple green plants and not just the fruit color. A hypothesis that stress may relate to reported SSC increases because of u+ is discussed.


2002 ◽  
Vol 92 (3) ◽  
pp. 237-244 ◽  
Author(s):  
Fernando M. Alves-Santos ◽  
Brisa Ramos ◽  
M. Asunción García-Sánchez ◽  
Arturo P. Eslava ◽  
José María Díaz-Mínguez

We have characterized strains of Fusarium oxysporum from common bean fields in Spain that were nonpathogenic on common bean, as well as F. oxysporum strains (F. oxysporum f. sp. phaseoli) pathogenic to common bean by random amplified polymorphic DNA (RAPD) analysis. We identified a RAPD marker (RAPD 4.12) specific for the highly virulent pathogenic strains of the seven races of F. oxysporum f. sp. phaseoli. Sequence analysis of RAPD 4.12 allowed the design of oligonucleotides that amplify a 609-bp sequence characterized amplified region (SCAR) marker (SCAR-B310A280). Under controlled environmental and greenhouse conditions, detection of the pathogen by polymerase chain reaction was 100% successful in root samples of infected but still symptomless plants and in stem samples of plants with disease severity of ≥4 in the Centro Internacional de Agricultura Tropical (CIAT; Cali, Colombia) scale. The diagnostic procedure can be completed in 5 h and allows the detection of all known races of the pathogen in plant samples at early stages of the disease with no visible symptoms.


1994 ◽  
Vol 119 (1) ◽  
pp. 122-125 ◽  
Author(s):  
Scott D. Haley ◽  
Phillip N. Miklas ◽  
Lucia Afanador ◽  
James D. Kelly

The objective of this study was to evaluate the degree of RAPD marker variability between and within commercially productive market classes representative of the Andean and Middle American gene pools of common bean (Phaseolus vulgaris L.). Six sets of near-isogenic lines were screened with oligonucleotide primers in the polymerase chain reaction-based RAPD assay. Simultaneous analyses with at least three sets of lines enabled us to score RAPD markers between the two major gene pools, races within the same gene pool, and different genotypes of the same race (within race). A “three-tiered” pattern of polymorphism was observed: between gene pools> between races> within races. The overall level of polymorphism between the Andean and Middle American gene pools was 83.4%. The overall level of polymorphism between races within the same gene pool was similar for Andean races (60.4%) and Middle American races (61.7%). The level of polymorphism between related commercial navy bean lines was 39.2% and between related commercial snap bean lines was 53.6 %. The inherent simplicity and efficiency of RAPD analyses, coupled with the number of polymorphisms detectable between related commercial genotypes, should facilitate the construction of RAPD-based genetic linkage maps in the context of populations representative of most bean breeding programs.


HortScience ◽  
1998 ◽  
Vol 33 (1) ◽  
pp. 140-142 ◽  
Author(s):  
Jean-Guy Parent ◽  
Danièle Pagé

Five polymorphic random amplified polymorphic DNA (RAPD) markers for 13 red raspberry (Rubus idaeus L.) and two purple raspberry (R. idaeus L. × R. occidentalis L.) cultivars were cloned and their termini sequenced. Sequence-specific 24-mer primer pairs were synthesized as extended RAPD primers and used in sequence characterized amplified region (SCAR) DNA analysis. All primer pairs generated polymorphic SCAR markers of the original RAPD marker sizes and length variants. Markers from four of the primer pairs could be easily scored and were adequate to identify the raspberry cultivars of the certification program of the province of Quebec.


2001 ◽  
Vol 91 (4) ◽  
pp. 377-382 ◽  
Author(s):  
Jianhua Xu ◽  
Takashi Narabu ◽  
Takayuki Mizukubo ◽  
Tadaaki Hibi

Root-knot nematodes of the genus Meloidogyne are economically important pathogens of a wide range of crops. The tomato resistance gene Mi typically confers resistance to the three major species, M. incognita, M. javanica, and M. arenaria. However, virulent populations completely overcoming the Mi resistance still occur. In an attempt to develop molecular markers for virulence against Mi and gain insights into the genetic relationships among virulent populations of different species and origins, random amplified polymorphic DNA (RAPD) analyses of laboratory-selected virulent, field virulent, and avirulent populations of M. incognita, M. javanica, and M. arenaria were carried out. A RAPD marker, specific for selected virulent populations, was identified, and subsequently, converted to a sequence characterized amplified region (SCAR). Sequence characterization of the SCAR locus showed that alleles from laboratory- and field-selected virulent populations were highly similar to each other and clearly different from alleles from natural virulent and avirulent populations. This result suggests that the genetic mechanism for virulence against Mi may be similar among selected virulent populations of the three Meloidogyne spp., but different between selected and natural virulent populations. Based on the nucleotide polymorphisms at the SCAR locus, codominant and dominant polymerase chain reaction-based markers were developed enabling rapid diagnosis of selected virulent genotypes in M. incognita, M. javanica, and M. arenaria.


HortScience ◽  
2007 ◽  
Vol 42 (1) ◽  
pp. 11-15 ◽  
Author(s):  
Aoxue Wang ◽  
Fanjuan Meng ◽  
Xiangyang Xu ◽  
Yong Wang ◽  
Jingfu Li

Leaf mold, caused by the fungus Cladosporium fulvum, is a serious disease of tomato. In the current study, the main physiological races of C. fulvum collected from three northeastern provinces of China were identified using a set of identification hosts. The results showed that the prevalent pathogenic physiological races were 1.2.3, 1.3, 3, 1.2.3.4, and 1.2.4. F1, F2, and BC1 tomato plants were obtained by crossing C. fulvum-resistant cultivar 03748 carrying the Cf-6 gene and susceptible cultivar 03036. Three 10-mer oligonucleotide random amplified polymorphic DNA (RAPD) primers and two simple sequence repeat (SSR) primers were selected for the further molecular marking analysis after 210 RAPD primers and 50 SSR primers were screened using the bulked segregate analysis method. The polymorphic DNA bands were amplified among parents, 10 F1 plants, 184 F2 plants including 145 resistant plants and 39 sensitive plants using three RAPD primers and two SSR primers so that three RAPD molecular markers and two SSR molecular markers linked to the Cf-6 loci were identified. Three RAPD markers were linked to the Cf-6 resistant locus separated with 8.7 cM, 20.3 cM, and 33.4 cM. Also, one RAPD codominant marker S374619/559 was found. The locations of the two SSR markers were 12.6 cM and 9.7 cM away from the Cf-6 locus. After cloning and sequencing two specific DNA fragments closely connected to the Cf-6 resistant and susceptible alleles respectively, in the RAPD codominant marker S374619/559 and one codominant sequence characterized amplified region marker S674619/559 was converted from RAPD marker S374619/559. In the RAPD marker S374619/559, the length difference of two specific fragments, 619-bp fragment and 559-bp fragment, is the result of one insertion (60 bp) in the 619-bp fragment. These markers will facilitate the selection of resistant tomato germplasm containing the Cf-6 gene and cloning of Cf-6 to breed new C. fulvum resistant tomato cultivars.


2002 ◽  
Vol 127 (4) ◽  
pp. 602-607 ◽  
Author(s):  
Melissa T. McClendon ◽  
Debra A. Inglis ◽  
Kevin E. McPhee ◽  
Clarice J. Coyne

Dry pea (Pisum sativum L.) production in many areas of the world may be severely diminished by soil inhabiting pathogens such as Fusarium oxysporum f. sp. pisi race 1, the causal organism of fusarium wilt race 1. Our objective was to identify closely linked marker(s) to the fusarium wilt race 1 resistance gene (Fw) that could be used for marker assisted selection in applied pea breeding programs. Eighty recombinant inbred lines (RILs) from the cross of Green Arrow (resistant) and PI 179449 (susceptible) were developed through single-seed descent, and screened for disease reaction in race 1 infested field soil and the greenhouse using single-isolate inoculum. The RILs segregated 38 resistant and 42 susceptible fitting the expected 1:1 segregation ratio for a single dominant gene (χ2 = 0.200). Bulk segregant analysis (BSA) was used to screen 64 amplified fragment length polymorphism (AFLP) primer pairs and previously mapped random amplified polymorphic DNA (RAPD) primers to identify candidate markers. Eight AFLP primer pairs and 15 RAPD primers were used to screen the RIL mapping population and generate a linkage map. One AFLP marker, ACG:CAT_222, was within 1.4 cM of the Fw gene. Two other markers, AFLP marker ACC:CTG_159 at 2.6 cM linked to the susceptible allele, and RAPD marker Y15_1050 at 4.6 cM linked to the resistant allele, were also identified. The probability of correctly identifying resistant lines to fusarium wilt race 1, with DNA marker ACG:CAT_222, is 96% percent. These markers will be useful for marker assisted breeding in applied pea breeding programs.


2004 ◽  
Vol 94 (4) ◽  
pp. 320-325 ◽  
Author(s):  
Richard C. Larsen ◽  
Phillip N. Miklas

A random amplified polymorphic DNA (RAPD) marker directly linked (0.0 cM) with a resistance gene was identified in a snap bean recombinant inbred population (Moncayo × Primo) consisting of 94 F5:7 recombinant inbred lines that had uniform segregation for disease reaction to Beet curly top virus (BCTV) across three field locations. Resistance was conditioned by a single dominant allele tentatively designated Bct. Seven hundred and fifty decamer primers were screened to obtain the linked RAPD marker that was then converted to a sequence characterized amplified region (SCAR) marker SAS8.1550. The SCAR mapped within a cluster of resistance genes on linkage group B7 of the core map. A survey of 103 BCTV-resistant and -susceptible snap and dry bean genotypes was conducted using SAS8.1550. Results showed that the SCAR would be highly useful for marker-assisted selection of Bct in snap and dry bean originating from the Andean gene pool. Marker-assisted selection for Bct will expedite the development of BCTV-resistant cultivars and minimize the need for cumbersome pathogen tests.


2016 ◽  
Vol 97 (6) ◽  
pp. 1307-1315 ◽  
Author(s):  
Elangovan Dilipan ◽  
Jutta Papenbrock ◽  
Thirunavakkarasu Thangaradjou

In India 14 seagrass species can be found with monospecific genera (Enhalus, ThalassiaandSyringodium),Cymodoceawith two species andHalophilaandHalodulerepresented by more than two taxonomically complex species. Considering this, the present study was made to understand the level and pattern of genetic variability among these species collected from Tamilnadu coast, India. Random amplified polymorphic DNA (RAPD) analysis was used to evaluate the level of polymorphism existing between the species. Out of the 12 primers tested, 10 primers amplified 415 DNA fragments with an average of 41.5 fragments per primer. Of the total 415 amplified fragments only 123 (29.7%) were monomorphic and the remaining 292 (70.3%) were polymorphic for Indian seagrass species. Among the 10 primers used four are identified as the key primers capable of distinguishing all the Indian seagrasses with a high degree of polymorphism and bringing representative polymorphic alleles in all the tested seagrasses. From the present investigation, this study shows that the RAPD marker technique can be used not only as a tool to analyse genetic diversity but also to resolve the taxonomic uncertainties existing in the Indian seagrasses. The efficiency of these primers in bringing out the genetic polymorphism or homogeneity among different populations of theHalophilaandHalodulecomplex still has to be tested before recommending these primers as an identification tool for Indian seagrasses.


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