Genome survey sequencing and characterization of simple sequence repeat (SSR) markers in Platostoma palustre (Blume) A.J.Paton (Chinese mesona)

Platostoma palustre (Blume) A.J.Paton is an annual herbaceous persistent plant of the Labiatae family. However, there is a lack of genomic data for this plant, which severely restricts its genetic improvement. In this study, we performed genome survey sequencing of P. palustre and developed simple sequence repeat (SSR) markers based on the resulting sequence. K-mer analysis revealed that the assembled genome size was approximately 1.21 Gb. A total of 15,498 SSR motifs were identified and characterized in this study; among them, dinucleotide, and hexanucleotide repeats had the highest and lowest, respectively. Among the dinucleotide repeat motifs, AT/TA repeat motifs were the most abundant, and GC/CG repeat motifs were rather rare, accounting for 44.28% and 0.63%, respectively. Genetic similarity coefficient analysis by the UPMGA methods clustered 12 clones, of P. palustre and related species into two subgroups. These results provide helpful information for further research on P. palustre resources and variety improvements.

Genome-wide development of lncRNA-derived-SSR markers for Dongxiang wild rice (Oryza rufipogon Griff.)

Dongxiang wild rice (Oryza rufipogon Griff.) (DXWR) is the northernmost distributed wild rice found in the world. Similar to other populations of O. rufipogon, DXWR contains a large number of agronomically valuable genes, which makes it a natural gene pool for rice breeding. Molecular markers, especially simple repeat sequence (SSR) markers, play important roles in plant breeding. Although a large number of SSR markers have been developed, most of them are derived from the genome coding sequences, rarely from non-coding sequences. Meanwhile, long non-coding RNAs (lncRNAs), which are derived from the transcription of non-coding sequences, play vital roles in plant growth, development and stress responses. In our previous study, we obtained 1655 lncRNA transcripts from DXWR using strand-specific RNA sequencing. In this study, 1878 SSR loci were detected from the lncRNA sequences of DXWR, and 1258 lncRNA-derived-SSR markers were developed on the genome-wide scale. To verify the validity and applicability of these markers, 72 pairs of primers were randomly selected to test 44 rice accessions. The results showed that 42 (58.33%) pairs of primers have abundant polymorphism among these rice materials; the polymorphism information content values ranged from 0.04 to 0.87 with an average of 0.50; the genetic diversity index of SSR loci varied from 0.04 to 0.88 with an average of 0.56; and the number of alleles per marker ranged from 2 to 11 with an average of 4.36. Thus, we concluded that these lncRNA-derived-SSR markers are a very useful source for future basic and applied research.

Identification of Lens Cultivars in Market by Molecular Tools: DNA Barcoding and SSRs

Substitution of plant cultivars of high commercial value with a cheaper, lower-quality one is a common fraud committed against consumers and producers. Since it is one of the most widely grown legumes, lentil (Lens culinaris Medik.) are a suitable for such frauds. This study aimed to identify lentil cultivars which are registered and authorized in the market in Turkey by using current molecular methods. For this purpose, 26 lentil cultivars were analyzed for 15 SSR markers and 2 DNA barcode loci (trnH-psbA and matK). A high allele diversity was observed by 12 scorable SSR markers, and the average number of alleles was determined to be 16. One of the important findings was the presence of “cultivar-specific alleles” that can be used to identify each cultivar in the lentil market in Turkey. At least one “cultivar-specific allele” was obtained for each cultivar. The lentil cultivars were analyzed in terms of 2 DNA barcode regions as trnH-psbA and matK. Sequences that could identify 14 of the 26 cultivars were obtained. While the rate of the intra-species variation for the trnH-psbA region was observed to be low, a higher rate was found for matK. Nevertheless, it was observed that intra-species discrimination can be made more effective when both loci are used together. We expect that the results of this study, especially the cultivar-specific SSR alleles and DNA barcoding sequence data may be used routinely to identify on production and packaged products that are commercially available in markets.

Evaluation of yellow rust resistance in backcross populations of wheat

Aim: To screen wheat populations derived from cross DBW17 × WH1105 for loci imparting yellow rust resistance and selection of plants using polymorphic SSRs. Methodology: The study for yellow rust resistance was carried out on two populations, i.e., BC1F2 and BC2F2. Stress was provided by planting infector rows between the blocks and by artificial inoculation using a mixture of races 46S102, 47S103 and 78S84 of stripe rust pathogen. DNA isolated from young leaves was checked for the presence of yellow rust resistance genes using gene specific primers. Results: Fifteen primers were found to be polymorphic among parents DBW17 and WH1105. Fifteen polymorphic SSR markers were dispersed over the wheat genome (AABBDD), with allele range 2-5. These polymorphic SSR markers were used to produce molecular diversity among progeny lines. Cluster analysis of parents and both the populations, showed that two parents were diverse genetically and in both backcrosses progeny lines resembled their respective recurrent parent. Single marker analysis using data revealed that primers on nine chromosomes were associated with grain yield per plant, other yield attributes and yellow rust resistance in both populations. Interpretation: This study showed that a linked marker like Xgwm582 could be a promising tool for breeding wheat with enhanced tolerance to yellow rust resistance. However, growth rates and biomass production provide reliable criteria for assessing the degree of yellow rust resistance and the ability of a plant to withstand it.

Genome-wide identification and characterization of functionally relevant microsatellite markers from transcription factor genes of Tea (Camellia sinensis (L.) O. Kuntze)

Tea, being one of the most popular beverages requires large set of molecular markers for genetic improvement of quality, yield and stress tolerance. Identification of functionally relevant microsatellite or simple sequence repeat (SSR) marker resources from regulatory “Transcription factor (TF) genes” can be potential targets to expedite molecular breeding efforts. In current study, 2776 transcripts encoding TFs harbouring 3687 SSR loci yielding 1843 flanking markers were identified from traits specific transcriptome resource of 20 popular tea cultivars. Of these, 689 functionally relevant SSR markers were successfully validated and assigned to 15 chromosomes (Chr) of CSS genome. Interestingly, 589 polymorphic markers including 403 core-set of TF-SSR markers amplified 2864 alleles in key TF families (bHLH, WRKY, MYB-related, C2H2, ERF, C3H, NAC, FAR1, MYB and G2-like). Their significant network interactions with key genes corresponding to aroma, quality and stress tolerance suggests their potential implications in traits dissection. Furthermore, single amino acid repeat reiteration in CDS revealed presence of favoured and hydrophobic amino acids. Successful deployment of markers for genetic diversity characterization of 135 popular tea cultivars and segregation in bi-parental population suggests their wider utility in high-throughput genotyping studies in tea.