Genetic Diversity Analysis of Indonesian Pigmented Rice Accessions Based on Functional Molecular Markers

Pigmented rice is functional staple food that becomes popular because of its anthocyanin content which is beneficial for health. Studies on the diversity of the local variety of Indonesian pigmented rice accessions have been carried out, but are still limited to one region of germplasm origin. This study aimed to analyze the genetic diversity of local varieties of pigmented rice collections of the IAARD-ICABIOGRAD Gene Bank. A total of 93 pigmented rice accessions from 16 provinces in Indonesia were analyzed using 15 functional molecular markers of SSR, STS, and indel. The total alleles detected were 115 with an average per locus of genetic diversity value of 0.71. There were five markers with PIC values >0.75, i.e. RM167, RM223, R8M33, R10M10, and GBSS1. The accessions were divided into two main groups based on their pericarp color. It is necessary to analyze the physicochemical content of the local rice accessions to complement the existing diversity information and identify potential pigmented rice accessions with high palatability.

Transcriptome wide SSR discovery cross-taxa transferability and development of marker database for studying genetic diversity population structure of Lilium species

Lily belongs to family liliaceae, which mainly propagates vegetatively. Therefore, sufficient number of polymorphic, informative, and functional molecular markers are essential for studying a wide range of genetic parameters in Lilium species. We attempted to develop, characterize and design SSR (simple sequence repeat) markers using online genetic resources for analyzing genetic diversity and population structure of Lilium species. We found di-nucleotide repeat motif were more frequent (4684) within 0.14 gb (giga bases) transcriptome than other repeats, of which was two times higher than tetra-repeat motifs. Frequency of di-(AG/CT), tri-(AGG/CTT), tetra-(AAAT), penta-(AGAGG), and hexa-(AGAGGG) repeats was 34.9%, 7.0%, 0.4%, 0.3%, and 0.2%, respectively. A total of 3607 non-redundant SSR primer pairs was designed based on the sequences of CDS, 5′-UTR and 3′-UTR region covering 34%, 14%, 23%, respectively. Among them, a sub set of primers (245 SSR) was validated using polymerase chain reaction (PCR) amplification, of which 167 primers gave expected PCR amplicon and 101 primers showed polymorphism. Each locus contained 2 to 12 alleles on average 0.82 PIC (polymorphic information content) value. A total of 87 lily accessions was subjected to genetic diversity analysis using polymorphic SSRs and found to separate into seven groups with 0.73 to 0.79 heterozygosity. Our data on large scale SSR based genetic diversity and population structure analysis may help to accelerate the breeding programs of lily through utilizing different genomes, understanding genetics and characterizing germplasm with efficient manner.

Molecular analysis of five rice blast resistant genes in landraces from Myanmar and Laos

In order to understand rice blast resistance gene patterns in rice landraces from Myanmar and Laos, we analysed the Pita, Pib, Pikh, Pi9 and Pi5 genes in 80 landraces through the use of functional molecular markers and functional fragment sequencing. These landraces were separated into four haplotypes (H1–H4) based upon three mutations identified in the Pita sequence. All four haplotypes were detected among landraces from Myanmar, whereas only the H1 and H2 haplotypes were detected among landraces from Laos. We additionally grouped landraces carrying 0–4 resistance genes into 18 genotypes based upon their resistance-susceptibility patterns and found that 16 of these genotypes were detected among Myanmar landraces at relative frequencies ranging from 2.50–12.50%, while 10 were detected among landraces from Laos at relative frequencies of 2.50–42.50%. The ‘Pib( + ) pikh(−) pi9(−) pi5(−) pita(−)’ genotype was found to be dominant, accounting for 12.5 and 42.5% of landraces from Myanmar and Laos, respectively. The Pib, Pikh, Pi5 and Pita genes were detected in 52.50, 32.50, 37.50 and 30.00% of landraces from Myanmar respectively, whereas they were detected in 52.50, 12.50, 30.00 and 17.50% of landraces from Laos, respectively. The Pi9 gene was rarely detected among analysed landraces and was particularly rare in those from Laos. These findings suggest that rice landraces from Myanmar are more genetically diverse than those from Laos, with clear differences in blast resistance gene patterns between landraces from these two countries.

Isolation and Molecular Marker Detection of Badh2 Gene from Aromatic Rice Germplasm Resources in Southern Henan

The production of aroma in aromatic rice is due to the increase of 2-acetyl-1-pyrroline (2-AP) precursor substances caused by the functional deletion of Badh2 gene on chromosome 8, and the accumulation of 2AP makes rice produce aroma. In this study, Badh2 gene was isolated and cloned from 18 representative aromatic rice cultivars in Southern Henan, and the bioinformatics analysis of Badh2 gene was carried out. Meanwhile, seven functional molecular markers developed by Badh2 gene were used to detect and analyze Badh2 gene in 18 aromatic rice varieties from Southern Henan. The results showed that the coding region of Badh2 gene was 1509 bp in length. It contained 15 exons and 14 introns, and encoded 503 amino acids. There are many types of variation of the Badh2 gene in the 18 aromatic rice varieties. According to the variation of Badh2 gene, the tested aromatic rice varieties could be divided into three groups, among which Xinxianggeng 1, Xiangnuo 25, Heixiangdao 193 and Xiangbao 2 were concentrated in group Ⅰ, while the other 14 kinds of aromatic rice were concentrated in group II. Seven functional molecular markers of Badh2 gene were used to detect different varieties mutation types in exon 2, exon 4~5, exon 7 and exon 13 of Badh2 gene. No aromatic rice varieties with different mutation types were found in promoter region, exon 12 and exon 14 of Badh2 gene. Therefore, our results provide important information for understanding the genetic basis of fragrant genes in aromatic rice germplasm resources in Southern Henan and breeding new varieties of high-quality aromatic rice using molecular marker-assisted selection.

De novo transcriptome assembly of Vitis flexuosa grapevines inoculated with Elsinoe ampelina

In this study, the transcriptome of Vitis flexuosa leaves inoculated with Elsinoe ampelina was analysed to identify useful genes and elucidate their function and differential expression patterns through assembly and annotation gene ontology of data from sequencing short reads on the Illumina platform. We assembled ~121 million high-quality trimmed reads using Velvet and Oases with optimal parameters into a non-redundant set of 70,899 transcripts ( ≥ 200 bp in length). The transcripts exhibited an average length of 1138 bp and a N50 length of 1695 bp, with the largest contig length being 9623 bp. Functional categorization revealed the conservation of genes involved in various molecular functions, including protein binding (21.1%) and oxidoreductase activity (11.7%), in V. flexuosa. The V. flexuosa transcript set generated in this study will serve as a resource for gene discovery and development of functional molecular markers.