Long-read sequencing and de novo assembly of the Luffa cylindrica (L.) Roem. genome.

Sponge gourd (Luffa cylindrica (L.) Roem.) or luffa is a diploid herbaceous plant with 26 chromosomes (2n = 26) and belongs to the family Cucurbitaceae. To address the limited knowledge of the genome of Luffa species, the chromosome-level genome of L. cylindrica was assembled and analysed using PacBio long reads and Hi-C data. We combined Hi-C data with a draft genome assembly to generate chromosome-length scaffolds. Thirteen scaffolds corresponding to the 13 chromosomes were assembled from 1,156 contigs to a final size of 669 Mb with a contig N50 size of 5 Mb and a scaffold N50 size of 53 Mb. After removing redundant sequences, 416.31 Mb (62.18% of the genome) of repeat sequences was detected. Subsequently, 31,661 protein-coding genes with an average of 5.69 exons per gene were identified in the L. cylindrica genome using de novo methods, transcriptome data and homologue-based approaches. In addition, 27,552 protein-coding genes (87.02%) were annotated in five databases. According to the phylogenetic analysis, L. cylindrica is closely related to Cucurbita and Cucumis species and diverged from their common ancestor ~28.6-67.1 million years ago. Genome collinearity analysis was performed in Cucurbita moschata, Cucumis sativus and L. cylindrica, and it demonstrated a high degree of conserved gene order in these three species. The completeness of the genome will provide high-quality genomic knowledge on breeding and reveal genetic variation in L. cylindrica.

The establishment of the species-delimits and varietal-identities of the cultivated germplasm of Luffa acutangula and Luffa aegyptiaca in Sri Lanka using morphometric, organoleptic and phylogenetic approaches.

Luffa acutangula and L. aegyptiaca are two vegetable species commonly found in South and South East Asia. L. acutangula is widely grown; however, L. aegyptiaca is considered as an underutilized crop. The species delimits, phylogenetic positions, and the varietal identities of L. acutangula and L. aegyptiaca in Sri Lanka are not known. Thus, in the present study, we aimed to establish the species delimits and varietal identities of L. acutangula and L. aegyptiaca varieties grown in Sri Lanka using morphometric, phylogenetic and organoleptic assessments. We assessed five varieties of L. acutangula and three varieties of L. aegyptiaca. The vegetative and reproductive data were collected for the morphometric analysis and DNA sequence polymorphism of the makers rbcL, trnH-psbA and ITS for the phylogenetic analysis. We also conducted an organoleptic assessment based on taste parameters; aroma, bitterness, color, texture, and overall preference using the dishes prepared according to the most common Sri Lankan recipe for Luffa. The variation of the vegetative and reproductive traits grouped L. acutangula varieties into two distinct clusters. The trnH-psbA polymorphism provided the basis for the species delimits of L. acutangula and L. aegyptiaca. The rbcL and ITS polymorphisms provided the basis for the identities of the varieties of L. aegyptiaca and L. acutangula respectively. In the phylogeny, the L. acutangula varieties of Sri Lanka formed a unique clade and the L. aegyptiaca varieties formed a reciprocal monophyletic group in comparison to worldwide L. aegyptiaca reported. The taste parameters aroma, texture, color, and overall preference were significantly different among the Luffa varieties. The L. aegyptiaca varieties received lower preference in the organoleptic assessment. The present study sets the species delimits, phylogenetic positions and the varietal identities of the cultivated germplasm of Luffa and revealed the distinct morphological and organoleptic properties of each variety.

Genome Survey Sequencing of Luffa Cylindrica L. and Microsatellite High Resolution Melting (SSR-HRM) Analysis for Genetic Relationship of Luffa Genotypes.

Luffa cylindrica (L.) Roem. is an economically important vegetable crop in China. However, the genomic information on this species is currently unknown. In this study, for the first time, a genome survey of L. cylindrica was carried out using next-generation sequencing (NGS) technology. In total, 43.40 Gb sequence data of L. cylindrica, about 54.94× coverage of the estimated genome size of 789.97 Mb, were obtained from HiSeq 2500 sequencing, in which the guanine plus cytosine (GC) content was calculated to be 37.90%. The heterozygosity of genome sequences was only 0.24%. In total, 1,913,731 contigs (>200 bp) with 525 bp N50 length and 1,410,117 scaffolds (>200 bp) with 885.01 Mb total length were obtained. From the initial assembled L. cylindrica genome, 431,234 microsatellites (SSRs) (≥5 repeats) were identified. The motif types of SSR repeats included 62.88% di-nucleotide, 31.03% tri-nucleotide, 4.59% tetra-nucleotide, 0.96% penta-nucleotide and 0.54% hexa-nucleotide. Eighty genomic SSR markers were developed, and 51/80 primers could be used in both "Zheda 23" and "Zheda 83". Nineteen SSRs were used to investigate the genetic diversity among 32 accessions through SSR-HRM analysis. The unweighted pair group method analysis (UPGMA) dendrogram tree was built by calculating the SSR-HRM raw data. SSR-HRM could be effectively used for genotype relationship analysis of Luffa species.

[Biological characters of the different varieties for Luffa cylindrica].

The yield of Luffa cylindrica cultivated in Taiyuan of Shanxi Province, which introduced from Zhejiang Province, was obviously high than that of local variety. The paper reported the biological characters of the two varieties.