Construction of a high-density genetic linkage map and identification of gene controlling resistance to cucumber mosaic virus in Luffa cylindrica (L.) Roem. based on specific length amplified fragment sequencing

2020 ◽  
Vol 47 (8) ◽  
pp. 5831-5841
Author(s):  
Lina Lou ◽  
Xiaojun Su ◽  
Xiaohong Liu ◽  
Zhe Liu
Agronomy ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 114 ◽  
Author(s):  
Xiaoxia Yu ◽  
Mingfei Zhang ◽  
Zhuo Yu ◽  
Dongsheng Yang ◽  
Jingwei Li ◽  
...  

Specific length amplified fragment sequencing (SLAF-seq) is a recently developed high-resolution strategy for the discovery of large-scale de novo genotyping of single nucleotide polymorphism (SNP) markers. In the present research, in order to facilitate genome-guided breeding in potato, this strategy was used to develop a large number of SNP markers and construct a high-density genetic linkage map for tetraploid potato. The genomic DNA extracted from 106 F1 individuals derived from a cross between two tetraploid potato varieties YSP-4 × MIN-021 and their parents was used for high-throughput sequencing and SLAF library construction. A total of 556.71 Gb data, which contained 2269.98 million pair-end reads, were obtained after preprocessing. According to bioinformatics analysis, a total of 838,604 SLAF labels were developed, with an average sequencing depth of 26.14-fold for parents and 15.36-fold for offspring of each SLAF, respectively. In total, 113,473 polymorphic SLAFs were obtained, from which 7638 SLAFs were successfully classified into four segregation patterns. After filtering, a total of 7329 SNP markers were detected for genetic map construction. The final integrated linkage map of tetraploid potato included 3001 SNP markers on 12 linkage groups, and covered 1415.88 cM, with an average distance of 0.47 cM between adjacent markers. To our knowledge, the integrated map described herein has the best coverage of the potato genome and the highest marker density for tetraploid potato. This work provides a foundation for further quantitative trait loci (QTL) location, map-based gene cloning of important traits and marker-assisted selection (MAS) of potato.


2019 ◽  
Author(s):  
Lina Lou ◽  
Zhe Liu ◽  
Xiaohong Liu ◽  
Xiaojun Su

Abstract Background: Luffa cylindrica L. is an economically important vegetable crop that is consumed globally. Cucumber mosaic virus (CMV) is an important virus affecting Luffa spp. No specific high-density maps have been constructed owing to a lack of efficient markers. Furthermore, no genes or quantitative trait loci (QTLs) are reportedly responsible for CMV resistance in Luffa spp. The development of next-generation sequencing has enabled discovery of single nucleotide polymorphisms and high-throughput genotyping of large populations. Results: A total of 271.01 Mb pair-end reads were generated. The average sequencing depth was 86.19× in both maternal and parental lines, and 14.57× in each F 2 individual. When filtering low-depth specific locus amplified fragment (SLAF) tags, 100,077 high-quality SLAFs were detected, and 7,405 of them were polymorphic. Finally, 3,701 of the polymorphic markers were selected for genetic map construction, and 13 linkage groups were generated. The map spanned 1,518.56 cM with an average distance of 0.41 cM between adjacent markers. Our results also revealed that CMV resistance was regulated by QTLs. Based on the newly constructed high-density map, two loci located on chromosome 1 (100.072 ~ 100.457 cM) and 4 (42.475 ~ 44.398 cM) were identified to regulate CMV resistance in L. cylindrica . A gag-polypeptide of LTR copia-type retrotransposon was predicted as the candidate gene responsible for CMV resistance in L. cylindrica . Conclusions: A high-density linkage map of L. cylindrica was constructed using SLAF. QTL mapping based on CMV disease phenotypes of F 2 led to the identification of two QTL on chromosome 1 and 4, respectively. Kompetitive allele-specific PCR analysis of 60 F 2 individuals, which gave rise to F 2:3 individuals, was carried out. We found that the QTL on chromosome 1 was associated with CMV resistance. Mapping of CMV QTL combined with the transcriptomic sequence alignment identified a gag-polypeptide of LTR copia-type retrotransposon as the most likely causal gene.


2019 ◽  
Author(s):  
Lina Lou ◽  
Zhe Liu ◽  
Xiaohong Liu ◽  
Xiaojun Su

Abstract Background: Luffa cylindrica L. is an economically important vegetable crop that is consumed globally. Cucumber mosaic virus (CMV) is an important virus affecting Luffa spp. No specific high-density maps have been constructed owing to a lack of efficient markers. Furthermore, no genes or quantitative trait loci (QTLs) are reportedly responsible for CMV resistance in Luffa spp. The development of next-generation sequencing has enabled discovery of single nucleotide polymorphisms and high-throughput genotyping of large populations. Results: A total of 271.01 Mb pair-end reads were generated. The average sequencing depth was 86.19× in both maternal and parental lines, and 14.57× in each F 2 individual. When filtering low-depth specific locus amplified fragment (SLAF) tags, 100,077 high-quality SLAFs were detected, and 7,405 of them were polymorphic. Finally, 3,701 of the polymorphic markers were selected for genetic map construction, and 13 linkage groups were generated. The map spanned 1,518.56 cM with an average distance of 0.41 cM between adjacent markers. Our results also revealed that CMV resistance was regulated by QTLs. Based on the newly constructed high-density map, two loci located on chromosome 1 (100.072 ~ 100.457 cM) and 4 (42.475 ~ 44.398 cM) were identified to regulate CMV resistance in L. cylindrica . A gag-polypeptide of LTR copia-type retrotransposon was predicted as the candidate gene responsible for CMV resistance in L. cylindrica . Conclusions: A high-density linkage map of L. cylindrica was constructed using SLAF. QTL mapping based on CMV disease phenotypes of F 2 led to the identification of two QTL on chromosome 1 and 4, respectively. Kompetitive allele-specific PCR analysis of 60 F 2 individuals, which gave rise to F 2:3 individuals, was carried out. We found that the QTL on chromosome 1 was associated with CMV resistance. Mapping of CMV QTL combined with the transcriptomic sequence alignment identified a gag-polypeptide of LTR copia-type retrotransposon as the most likely causal gene.


2020 ◽  
Vol 66 (3) ◽  
pp. 161-173 ◽  
Author(s):  
Xiaoxia Yu ◽  
Yanhong Ma ◽  
Zhiyan Jiang ◽  
Yue Shi ◽  
Dongsheng Yang ◽  
...  

BMC Genomics ◽  
2020 ◽  
Vol 21 (1) ◽  
Author(s):  
Guosong Zhang ◽  
Jie Li ◽  
Jiajia Zhang ◽  
Xia Liang ◽  
Tao Wang ◽  
...  

Abstract Background A high-density genetic linkage map is essential for QTL fine mapping, comparative genome analysis, identification of candidate genes and marker-assisted selection in aquaculture species. Pelteobagrus vachelli is a very popular commercial species in Asia. However, some specific characters hindered achievement of the traditional selective breeding based on phenotypes, such as lack of large-scale genomic resource and short of markers tightly associated with growth, sex determination and hypoxia tolerance related traits. Results By making use of 5059 ddRAD markers in P. vachelli, a high-resolution genetic linkage map was successfully constructed. The map’ length was 4047.01 cM by using an interval of 0.11 cm, which is an average marker standard. Comparative genome mapping revealed that a high proportion (83.2%) of markers with a one-to-one correspondence were observed between P. vachelli and P. fulvidraco. Based on the genetic map, 8 significant genome-wide QTLs for 4 weight, 1 body proportion, 2 sex determination, and 1 hypoxia tolerance related traits were detected on 4 LGs. Some SNPs from these significant genome-wide QTLs were observably associated with these phenotypic traits in other individuals by Kompetitive Allele Specific PCR. In addition, two candidate genes for weight, Sipa1 and HSD11B2, were differentially expressed between fast-, medium- and slow-growing P. vachelli. Sema7a, associated with hypoxia tolerance, was induced after hypoxia exposure and reoxygenation. Conclusions We mapped a set of suggestive and significant QTLs as well as candidate genes for 12 growth, 1 sex determination and 1 hypoxia tolerance related traits based on a high-density genetic linkage map by making use of SNP markers for P. fulvidraco. Our results have offered a valuable method about the much more efficient production of all-male, fast growth and hypoxia tolerance P. vachelli for the aquaculture industry.


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