Vol. 13(12), pp. 1314-1322, 19 March, 2014
DOI: 10.5897/AJB2013.13021
ISSN 1684-5315
Copyright © 2014 African Journal of Biotechnology
Author(s) retain the copyright of this article
http://www.academicjournals.org/AJB

Full Length Research Paper

Identification and utility of sequence related amplified

polymorphism (SRAP) markers linked to bacterial wilt
resistance genes in potato
Zhi Yanping, Li Hui, Zhang Hairui, and Gang Gao*
School of Life Science, Shanxi Normal University, Linfen041004, Shanxi, China.
Received 11 July, 2013; Accepted 26 February, 2014

Bacterial wilt caused by Ralstonia solanacearum is one of the most economically important diseases
affecting potato (Solanum tuberosum). It is necessary to develop more molecular markers for potential
use in potato genetic research. A highly resistant primitive cultivated species Solanum phureja was
employed to generate a F1 mapping population to perform the bulked segregant analysis (BSA) for
screening and identifying of sequence related amplified polymorphism (SRAP) markers linked to the
potato resistance to bacterial wilt. A linkage map containing 23 DNA markers distributed on three
linkage groups, and covering a genetic distance of 111 cm with an average distance of 5.8 cm between
two markers was developed. Two SRAP markers, Me2em5 linked in repulsion phase and Me2em2 in
coupling phase, flanked the resistance genes at genetic distances of 3.5 and 3.7 cm distance,
respectively. These markers and two others were used for early seedling selection in a BC1 population.
The results show that this marker system could be used in marker assisted selection (MAS) breeding
program.

Key words: Sequence related amplified polymorphism (SRAP) marker, potato, bacterial wilt.

INTRODUCTION

Bacterial wilt caused by Ralstonia solanacearum is one of borne vascular parasite is known to vary considerably
the most widely spread and very destructive plant according to climate, farming practices, soil type and
disease, causing enormous economic losses. The vas- geographic locations (Guidot et al., 2007). Control of wilt
cular pathogen enter and colonize the plant vascular diseases is also complicated by the scarcity of sources of
(xylem) system, disrupting water transport, and causing disease-resistant host germplasm, and the soil and vas-
the characteristic symptoms of wilting, and often vascular cular habitats of the pathogen (Hayward, 1991; Esposito
discoloration, and death of aerial tissues (He, 1983; Hayward, et al., 2008). Despite decades of interests in the patho-
1991). The bacterial parasite cause diseases on over 400 logy, epidemiology and control of bacterial wilt, very little
plant species, including many crops such as potato, is known about the sources and resistance of the disease
tomato and eggplant. The severity of attacks by this soil- and about the genetic or molecular mechanisms under-

*Corresponding author. E-mail: [email protected].

Author(s) agree that this article remain permanently open access under the terms of the Creative Commons Attribution License 4.0
International License
 Yanping et al. 1315

lying host plant resistance (Qu, 1996). Some nuclear controlling wilt diseases.
SSR alleles derived from the wild species S. chacoense
appeared to be related to bacterial wilt resistance (Chen MATERIALS AND METHODS
et al.,2013).
Plant materials
The mode of inheritance of resistance in some tetra-
ploid primitively cultivated species is not yet clearly One genotype ED13, derived from the cross hybridization between
known. Therefore, it is difficult to select bacterial wilt 772102.37 (resistant) and USW7589.2 (susceptible) collected both
resistant clones or varieties in potato breeding. Although, from the Department of Plant Breeding, Wageningen University, the
Netherlands was used as the resistant parent. The susceptible
method of biological control could be used in some situa- parent was derived from USW5337.3 × Solanum phureja (provided
tions to combat the pathogens, development of resistant by Institute of Vegetables and Flowers, Chinese Academy of
cultivars is usually the best agronomic solution. Disease Agricultural Sciences, China). A segregating F1 population (Figure
resistance breeding has traditionally been done by 4A) composed of 230 seedlings from hybrid true potato seed (TPS)
phenotypic selection. Efficiency of phenotypic selec-tion was used for mapping the SRAP markers. Another segregating
is reduced by variability in the pathogen, infection and BC1 population were generated by first backcrossing a CE26
(susceptible, derived from a cross USW 5337.3 × 772102.37) with
disease development. Molecular markers tightly lin-ked to the parent 772102.37 for assessing the utility of candidate SRAP
the resistance genes can eliminate these sources of markers in identifying bacterial wilt-resistant BC1 plants (Figure 4B).
phenotypic variation to enable more efficient breeding
strategies in potato improvement (Li et al., 2013; Virupaksh Wounded root plus soil inoculation
et al., 2012). Marker-assisted selection can be helpful to
bacterial wilt resistance breeding. We have identified Inocula for potato plants at a concentration of 1 × 108 cfu/ml were
made from cultures grown on BPG plates at 28°C for 48 h using
some molecular markers like AFLP and RAPD makers SMM without agar and were incubated at 28°C for 4 h. Potato
that linked to potato resistance, these markers were found plants with six to eight fully expanded leaves were inoculated by
not to be effective to screen a large population containing pouring 50 ml of bacterial suspension into the soil around the base
up to 200 true potato seeds in the actual application (Gao of the stem with wounding the roots with a knife. 15 potato seed-
et al., 2002, 2005). To accelerate introgression of the lings were inoculated with the strain PO41 of the bacterium (provi-
resistance into cultivated lines, it would be desirable to ded by the Institute of Plant Protection, Chinese Academy of
Agricultural Sciences, China). The virulent, wild colony type of R.
have more and better molecular markers to perform solanacearum cultures was selected on 2, 3, 5-tetrazolium chloride
selection of those possible resistance genes. medium (TZC) and used for inoculation. Inoculated plants were
Sequence-related amplified polymorphism (SRAP) cultivated at 28°C under a 16/8 h (light/dark) photoperiod in a
technology has been recognized as a new and useful growth chamber and were not watered five days before and after
molecular marker system for mapping and gene tagging inoculation. Control plants were mock-inoculated with sterile water.
Each treated plant was rated daily for disease for 21 days after
in many crops plants (Valdez-Ojeda et al., 2008; Cao et inoculation. Symptoms were scored daily on a 0 to 5 disease index,
al., 2011; Niu et al., 2011; Guo et al., 2012; Deng et al., 2013). where 0 indicates no disease, 1 indicates 1 to 10% of leaves wilted,
They are also useful in positional cloning of genes and in 2 indicates 10 to 25% of leaves wilted, 3 indicates 25 to 50% of lea-
elucidating the genetic mode of complex traits that do not ves wilted, 4 indicates 50 to 75% of leaves wilted, and 5 indi-cates
display Mendelian segregation (Levi, 2002). Marker-facili- 75 to 100% of leaves wilted. Each experiment contained 16 plants
tated selection would be particularly effective for pyramid- per treatment, and experiments were repeated at least three times.
Leave tissues of the susceptible were sampled 4 to 7 dpi into liquid
ding more resistance genes to provide effective and nitrogen for DNA extraction, while the resistant samples were
potentially stable resistance to disease (Martin et al., collected 8 to 14 dpi due to slower disease development in this
1991). resistant host.
The objectives of this study were to determine the
feasibility of using SRAP makers to develop a molecular Sequence-related amplified polymorphism analysis
linkage map of the S. tuberosum, and to identify marker
DNA was isolated according to Ducreux et al. (2008). Bulk segre-
loci associated with bacterial wilt. Both of these gant analysis (BSA) (Michelmore et al., 1991) was used to identify
objectives were met, and we identified the SRAP markers SRAP markers linked to the bacterial wilt resistance gene. For BSA,
that proved useful in selecting for the gene conferring resistant and susceptible DNA bulks were composed of 10 most
bacterial wilt resistance in a BC1 population. A longer- resistant (disease index 0) plants and most susceptible (disease
term goal of this project will be to use closely linked index 5), respectively. SRAP analysis was conducted according to
molecular markers to introduce these resistant loci into previously established protocols with minor modifications (Li et al.,
2001). The PCR reaction was set up in a final volume of 20 µL
potato cultivars as a potential strategy to control potato containing 50 ng of DNA, 5.0 pmoL of primer, 200 mM dNTPs, 1.5
bacterial wilt disease in the field. In this study, the set of mM MgCl2, and 0.5U of Taq polymerase (Sangon Biotech Co. Ltd.,
markers provide an impactful marker combination for use Shanghai, China) in 1× Taq buffer. The PCR program included an
in a marker assisted selection (MAS) breeding program initial denaturing at 94°C for 3 min followed by 8 cycles of 94°C for
to identify genotypes containing resistance from original 30 s, 37°C for 30 s, and at 72°C for 90 s and then 35 cycles of 94°C
potato cultivars. Taken together, this study provides and for 30 s, 48°C for 30 s, and 72°C for 90 s with a final extension of
72°C for 10 min. PCR products were separated using 8% denatu-
sheds light into potential directions for development of ring polyacryamide gel electrophoresis and visualized by fast silver
novel management strategies for molecular breeding to staining (Bassam et al., 1991).
1316 Afr. J. Biotechnol.

Table 1. The primer sequences of SRAP used in this study.

Forward primer Reverse primer

me1 TGAGTCCAAACCGGATA em1 GACTGCGTACGAATTAAT
me2 TGAGTCCAAACCGGAGC em2 GACTGCGTACGAATTTGC
me3 TGAGTCCAAACCGGAAT em3 GACTGCGTACGAATTGAC
me4 TGAGTCCAAACCGGACC em4 GACTGCGTACGAATTTGA
me5 TGAGTCCAAACCGGAAG em5 GACTGCGTACGAATTCGA
me6 TGAGTCCAAACCGGTAA em6 GACTGCGTACGAATTAAC
me7 TGAGTCCAAACCGGACG em7 GACTGCGTACGAATTGCA
em8 GACTGCGTACGAATTCAA

The primer combinations (Table 1) that generated polymorphic RESULTS

bands between the bulks were tested on the bulked individuals to
eliminate false-positive markers.
Polymorphism and screening of SRAP markers

Linkage analysis of markers 100 primer combinations were first tested for selective
amplification of DNA fragments from the resistant and
Several markers obtained previously were employed before the susceptible bulks to screening the polymorphism in the
construction of linkage map to increase reliability; those include
three AFLP, one RAPD markers. Preliminary screening for candi-
diploid mapping population. Prior to linkage analysis
date seedling was carried out on the two populations using these segregation rates, SRAP fragments that ranged in size of
markers. As expected, more than 80% of the individual genotype 75 to 500 bp were scored for marker. Fifty-six (56) of the
was detected to contain all or one relevant marker DNA fragment. polymorphic SRAP primer combinations amplified
Based on the preliminary selection, Linkage analysis was perfor- inconsistent band patterns per line. This inconsistency
med with the JoinMap software (Van Ooijen and Vooripps, 2001). may have been the results of residual heterozygosity or
Prior to linkage analysis segregation rates, SRAP fragments that
ranged in size of 75 to 500 bp were scored for marker. Presence or
the amplification of similar sequences in two separate ge-
absence of each polymorphic fragment was coded as “1” and “0”, nomic regions. It may also result from the use of poly-
where “1” indicated the presence of a specific allele, and “0” indi- acrylamide gels, which have a higher resolution power
cated its absence. For each segregating marker, a chi-square test than most agarose gels. A total of 314 unambiguous
was performed to fit for deviation from the 1:1 expected segregation bands were amplified by the 38 of 54 SRAP primer com-
ratio in a testcross or BC1 population under the ‘locus genotypic binations, of which 187 bands were polymorphic (59.55%)
frequency’ command. Markers were sorted on the basis of the chi-
squares test with a P -value of <0.05; and skewed markers were and ranged in size from 50 to 1000 bp. The number of
excluded from the analysis. All markers were analyzed for linkage, polymorphic fragments for each SRAP primer combina-
and recombination fractions were converted into map distances tion varied from 2 to 15, with an average of 6.0 fragments
(centimorgans) while employing the Kosambi mapping function. per primer combination. Polymorphisms were amplified
Logarithm of the odds ratio for linkage (LOD) scores of 2.0 to 6.0 with 38 primer pairs (67.8%) resulting in 146 polymorphic
were used for grouping of markers, followed by high threshold LOD
scores of 7.0 to 10.0 for final mapping of markers in each linkage
bands between the resistant and susceptible parental
group. Loci showing weak or suspect linkages were removed from genotypes. Based on these results, primer pairs that ge-
the analysis. nerated polymorphic bands were tested on the resistant
and susceptible bulks.
Assessing the utility of a marker set in identifying bacterial
SRAP bands present in one pool and absent in the
wilt-resistant BC1 plants other were regarded as candidate markers linked to
bacterial wilt resistance. SRAP fragments that ranged in
Forty-one (41) BC1 plants were generated by first backcrossing size of 75 to 500 bp were scored for marker. A relatively
bacterial wilt-susceptible CE26 genotype with the recurrent parent small number of these primer combinations (4 of 54 pairs;
772102.37. The marker set identified here were used to select for 7.41%) were not suitable for the mapping experiment
those progeny that presumably contain the corresponding DNA
allele conferring bacterial-wilt resistance, and then three times strict within the tested population because of the lack of
phenotypic identification (the disease index was scored for wilt polymorphism in size of 75 to 500 bp. Hence, out of the
symptoms as described earlier) were employed to exanimate their original 56 primer pairs, only 30 combinations were used.
identical degree. Spearman correlation coefficient was used to However, three (5.56%) of these combinations were
analyze the correlation between the molecular markers and the skewed from the expected 1:1 segregation ratio and had
disease parameters statistical software package S-Plus Version 6.
Probabilities <0.01 were considered as significant. All p values were
to be excluded from the final linkage analysis. Twenty-
based on two-sided tests, and the differences were considered seven (27) primer combinations produced 86 SRAP
statistically significant when the p value was ≤ 0.05. All molecular candidate markers that were used in the next linkage
detection and phenotypic identification were repeated three times. analysis.
 Yanping et al. 1317

Identification of SRAP markers linkage map all, as there are some medium-resistant or medium-sus-
ceptible genotypes in the population detected by phenol-
In order to obtain more closely linked markers and to type identification. As expected, the band types in the gel
avoid any possible mapping errors, the stringent linkage were present reproducibly in the BC1 population (Figure
analysis criteria used with the JoinMap analysis (Van 3).
Ooijen et al., 2001) resulted in linkage groups. We com-
pared for each primer combination, all primer pairs that
generated polymorphic bands were tested on the resis- Practical utility of the markers
tant and susceptible bulks. SRAP bands present in one In previous studies, several markers including AFLP,
pool and absent in the other were regarded as candidate RAPD and SSR markers were described to link with bac-
markers linked to bacterial wilt resistance. These SRAP terial wilt resistance loci, but did not determine their use-
markers were uniquely present in one of the donor pa- fulness well in MAS programs. Here, we tested 41 BC1
rents and in the F1 individual genotypes. Although, a total plants inoculated with the pathogen for the presence or
of eighty-six SRAP markers were suitable for the map- absence of the new SRAP markers. Four markers were
ping analysis using the first filial generation population, used to select for those progeny that presumably contain
23 (26.7%) of these markers were skewed from the the corresponding DNA allele conferring bacterial-wilt
expected 1:1 segregation ratio and had to be excluded resistance following three times strict phenotypic identi-
from the final linkage analysis. Sixty-three (63) SRAP fication (Figure 3). As shown in Figure 3A, of the 17
markers were used in the final linkage analysis. Five plants whose disease index were rated as 0 (highly resis-
markers (5.8%) did not show strong linkage to our aim, tant), 11 were detected to contain all four markers as
and they were excluded from the analysis. A total of 58 homozygous genotypes, six plants were heterozygous for
SRAP markers were analyzed for linkage. Of these, 23 the four markers, in which five plants were detected to
could be mapped with high confidence on the linkage contain three markers, while one plant contained only
map, four markers were clustered on linkage group 1, one SRAP marker. Of the 15 plants whose disease inde-
and seven markers were on linkage group 2 and twelve xes were rated as 5 (highly susceptible), 14 were detec-
on linkage group 3 (Figures 1 and 2). ted to contain the repulsion marker. These results sug-
gest that there is at least one major locus in the four
Markers linked in coupling or repulsion phase marker alleles, because there were one or two markers
detected in other nine plants whose disease indexes
Among the twelve polymorphic bands on linkage group 3, showed mildly resistant or mildly susceptible. These results
we found four markers tightly linked to the resistance were consistent with the expectation.
locus, two of them linked in coupling phase and the
others linked in repulsion phase. The primer combina- DISCUSSION
tions me2em5 and me5em2 (Table 1) were respectively
detected, a band polymorphic between the resistant bulk Although, SRAP markers are feasible to generate poly-
and the susceptible bulk, since the marker band named morphic (Levi et al., 2006; Poczai et al., 2013), a rela-
as Me2em5 and Me5em2 was present in all of the tively small number of these primer combinations were
susceptible plants in the bulks. They were screened out not suitable for the mapping experiment within the tested
from the F1 population segregating for bacterial wilt population because of the lack of polymorphism in size of
resistance. When the phenotype identification was perfor- 75 to 500 bp. Hence, out of the original 56 primer pairs,
med, this marker also co-segregated with disease reac- only 30 combinations were used. However, three (5.56%)
tion in the same fashion. And the linkage was further of these combinations were skewed from the expected
tested in the BC1 plants. The band was present in most 1:1 segregation ratio and had to be excluded from the
of the susceptible genotypes and absent in the resistant final linkage analysis. These SRAP markers proved to be
ones. These results indicated a tight linkage between the enough efficient and reliable in the mapping analysis in
marker me1em2 and the dominant susceptibility allele in this study. We initially used bulk segregant analysis (BSA)
the repulsion phase. The markers linked in repulsion strategy (Michelmore et al., 1991) to identify SRAPs. BSA
phase exhibited similar electrophoretic images (Figures 2 has been widely used in many crop species for detecting
and 3); While the primer combinations me2em2 and markers linked to genes conferring disease resistance
me5em1 detected approximately two close-migrating (Hyten et al., 2009) and is a powerful method for identi-
concomitant polymorphic bands between the resistant fying molecular markers that show association with a
and susceptible individuals (Figure 2). These double- gene of interest or a specific region of the genome (Ren
banded marker linked in coupling phase were designated et al., 2012; Salinas et al., 2013).
Me2em2 and Me5em1, respectively. Segregation distortion is widespread in plant popula-
The two polymorphic bands appeared in most of the tions and is a common feature of plant genetic linkage
resistant genotypes of the F1 population, and of which maps; it is frequent in progeny derived from interspecific
this proportion basically fits the expected 1:1 ratio, after crosses and distortion tends to increase with increasing
1318 Afr. J. Biotechnol.

Figure 1. Genetic linkage groups of F1 population constructed with SRAP markers using the JoinMap
software. Distances in centiMorgans are indicated to the left and marker names to the right followed
by the fragment size in nucleotide bases. Me2em5 and Me2em2 flanked the resistance gene locus
(Rbw) at genetic distances of 3.5 and 3.7 cm distance, respectively.

numbers of meioses (Zhang et al., 2012). disadvantage to its respective gametes or zygotes (Lyttle,
In this study, 16.8% of the total loci showed segre- 1991; Buckler et al., 1999). Both biological factors and
gation distortion (P < 0.05), which is larger than other technical problems potentially contribute to segregation
reports on cotton, wheat and Aegilops tauschii (Faris et distortion. Integration of distorted segregation markers in
al., 1998; Lin et al., 2005; Guo et al., 2007; Kumar et al., linkage construction possibly lead to untrue distance bet-
2007; Yu et al., 2007). Skewed segregation frequently ween the adjacent markers in linkage groups (Weber et
occurs in populations derived from interspecific crosses al., 2003; Lu et al., 2012). Therefore, in order to increase
and may be influenced by many factors, such as the accuracy of the genetic map constructed, the distortion
differential genes controlling the reproduction processes, segregation markers were ignored in this study. It has
meiotic drive controlling unique structural features and been proven that highly skewed markers may contribute
genetic properties rendering selective advantage or to overestimation of recombination frequency and to
 Yanping et al. 1319

Figure 2. Samples of markers detected by primer combination Me2em2, Me5em1, Me2em5 and Me5em2 in the resistant
and susceptible DNA bulks. Lanes: 1 to 10, most resistant (disease index 0) genotypes. 11 to 20, most susceptible
(disease index 5) genotypes. Standard size markers are given on left side.

Figure 3. Assessment of the utility of 4 markers in a segregating BC1 population. A) Spearman correlation coefficient analysis for degree of
coincidence between the molecular markers and the disease parameters. For disease index, 0 = no disease, 1 = 1 to 10% of leaves wilted, 2
= 10 to 25% of leaves wilted, 3 = 25 to 50% of leaves wilted, 4 = 50 to 75% of leaves wilted, and 5 = 75 to 100% of leaves wilted, or plant
death 21 dpi; B) Distribution of markers genotype data from bacterial wilt resistant polymorphism and phenotype data from potato BC1
population derived from the bacterial wilt-susceptible CE26 genotype with the recurrent parent 772102.37. The results of one of three
independent experiments were shown; C) Electrophoresis patterns of polymerase chain reaction-amplified with genomic DNA of 20
genotypes. M, molecular marker showed by arrows followed by base number; Lanes 1 to 10, F1 plants; lanes 11 to 20, susceptible F1 plants.
Primer combinations are given on left side.
1320 Afr. J. Biotechnol.

A P195198.13* × W1 P225696.1* × W42

W482 × USW5279.14 VH34211* × USW5337.3 [C]

772102.37 × USW7589.2 [D] USW5337.3× 772102.37 [E]

ED13 × S.phurejia CE26 × 772102.37

Mapping population Testing population  

Figure 4. Genetic background of the plant materials used in this study. A) The mapping population
composed of 230 genotypes; B) The testing population composed of 47 genotypes. *Pedigree from S.
phurejia and S. vernei involved (Qu, 1996).

loose linkages between markers while they may cause notypic identification and molecular marker detection. It
the merging of two linkage groups (Saliba-Colombani et was not surprising that two moderately susceptible geno-
al., 2000). Thus, in this study, the skewed markers had to types were detected to contain a molecular marker. Simi-
be excluded from the mapping analysis. lar results were confirmed in others of similar experi-
Five markers (5.8%) did not show strong linkage to our ments in eggplant (Lebeau et al., 2013). Using spearman
aim, and they were excluded from the analysis. Conver- correlation for analysis we found that there was not very
sely, they may cause the merging of two linkage groups. strong correlation existed between the SRAP markers (4
Although, this phenomenon was manifested in others primer recombination) and bacterial wilt disease scores.
(Levi et al., 2006; Saliba-Colombani et al., 2000) and our On the interpretation of the results, we inferred that the
results, we speculate that these molecular markers in symptom and disease scores were not significantly corre-
group 1 or 2 may represent additional resistance genes; lated with the numbers of the SRAP markers used here.
they are more likely to link within other different chromo- In general, the correlation between symptom scores and
somes. There may be of different origin derived from gene functional status measures should be stronger than
those remoter ancestors. This result suggested that the the correlation between disease scores and gene number
bacterial wilt resistance may be controlled by quantitative measures. After all, the other markers in linkage groups 1
trait loci, or there may be more loci exist in potato ge- and 2 were not used for correlation analysis. It is worth
nome. A recent report on resistance to R. solanacearum paying more attentions to that those markers should be-
in eggplant (Lebeau et al., 2013) seems to be able to come the object of special consideration in MAS breeding
support this speculation. It is worthy that a more in-depth program because the contribution from the gene loci
genetic analysis of bacterial wilt resistance in potato, linked with the markers is not inconsiderable.
especially in tetrapoid potato, needs to be considered. It has been demonstrated that SRAP markers have
The markers Me2em2, Me5em1, Me2em5 and Me5em2 good coverage of the genome and was able to rapidly
found associated with the locus can be used readily for detect markers linked to the resistance gene (Guo et al.,
marker assisted selection, helping to introgress the 2012; Lu et al., 2012; Zhao et al., 2012). The results in
recessive resistance allele of this gene into cultivated this study suggest that the resistance to bacterial wilt is
lines. Although, these are not codominant markers, the not simply inherited, but possibly controlled by a series of
linkage in coupling phase of Me2em2, Me5em1 and in genes. Here, we identified the SRAP markers that proved
repulsion phase of marker Me2em5 and Me5em2 to the useful in selecting for the gene conferring bacterial wilt
resistance allele, makes it possible to identify almost all resistance in a BC1 potato population. The set of markers
R. solanacearum isolates. provide a robust marker combination for use in MAS
Use of these markers might circumvent in many cases breeding program to identify genotypes containing the
progeny testing of resistant plants, thereby reducing in relative allele conferring bacterial wilt resistance in potato
half the time required to develop bacterial wilt resistant cultivars.
lines. Since the BC1 plants generated did not contain the
same level of resistance found in the mapping population, Conclusion
as the smaller population was more likely to exhibit a
larger segregation distortion and different genetic recom- In this study, based on an F1 segregating generation, we
bination; it was possible that a certain deviation of the identified the SRAP markers that proved useful in
matching degree occurred between the symptomatic phe- selecting for the gene conferring bacterial wilt resistance
 Yanping et al. 1321

in a BC1 potato population. The set of markers provide a Guo HL, Xuan JP, Liu JX, Zhang YM, Zheng YQ (2012). Association of
molecular markers with cold tolerance and green period in
robust marker combination for use in MAS breeding pro-
zoysiagrass (Zoysia Willd). Breed. Sci. 62:320-327.
gram to identify genotypes containing the relative allele Guo WZ, Cai CP, Wang CB, Han ZG, Song XL, Wang K, Niu XW,
conferring bacterial wilt resistance in potato cultivars. Wang C, Lu KY, Shi B, Zhang TZ (2007). A microsatellite-based,
gene-rich linkage map reveals genome structure, function and
evolution in Gossypium. Genetics.176:527-541.
Conflict of Interests Hayward AC (1991). Biology and epidemiology of bacterial wilt caused
by Pseudomonas solanacearum. Annu. Rev. Phytopathol. 29:65-87.
He LY, Sequeira L, Kelmen A (1983). Characterization of strains of
The author(s) have not declared any conflict of interests.
Pseudomonas solanacearum from China. Plant. Dis. 67:1357-1361.
Hyten DL, Smith JR, Frederick RD, Tucker ML, Song Q, Cregan PB
(2009). Bulked segregant analysis using the Golden Gate assay to
ACKNOWLEDGEMENTS locate the Rpp3 locus that confers resistance to soybean rust in
soybean. Crop. Sci. 49:265–271.
Kumar S, Gill BS, Faris JD (2007). Identification and characterization of
This work was supported by a high-tech industrialization
segregation distortion loci along chromosome 5B in tetraploid wheat.
project of the Department of Education of Shanxi Pro- Mol. Genet. Genomics. 278:187-196.
vince (2010011) and a program of the Science and Lebeau A, Gouy M, Daunay MC, Wicker E, Chiroleu F, Prior P, Frary A,
Technology Agency of Shanxi Province (2012011032-2) Dintinger J (2013). Genetic mapping of a major dominant gene for
resistance to Ralstonia solanacearum in eggplant. Theor. Appl.
and a project of National Natural Science Fund (31260344).
Genet. 126(1):143-158.
The authors thank the membership of the potato breeding Levi A, Thomas E, Joobeur T, Zhang X, Davis A (2002). A genetic
group of Institute of Vegetables and flowers, Chinese linkage map for watermelon derived from a testcross population:
Academy of Agricultural Sciences, and Dr. Yuan Yuxiang (Citrullus lanatus var citroides x C. lanatus var lanatus) x Citrullus
colocynthis. Theor. Appl. Genet. 105:555-563.
from Institute of Horticulture, Henan Academy of Agricul-
Levi A, Thomas CE, Trebitsh T, Salman A, King J, Karalius J, Newman
tural Sciences, for their technical assistance and helpful M, Reddy OUK, Xu Y, Zhang X (2006). An extended linkage map for
suggestions. watermelon based on SRAP, AFLP, SSR, ISSR, and RAPD markers.
J. Am. Soc. Hortic. Sci. 131:393-402.
Li G, Quiros CF (2001). Sequence-related amplified polymorphism
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