Abstract
BackgroundAmorphophallus is a genus of perennial plants widely distributed in the tropics or subtropics of West Africa and South Asia. Its corms contain a high level of water-soluble glucomannan; therefore, it has long been used as a medicinal herb and food source. Genetic studies of Amorphophallus have been hindered by a lack of genetic markers. A large number of molecular markers are required for genetic diversity study and improving disease resistance in Amorphophallus. Here, we report large scale of transcriptome sequencing of two species: Amorphophallus konjac and Amorphophallus bulbifer using deep sequencing technology, and microsatellite (SSR) markers were identified based on these transcriptome sequences.ResultscDNAs of A. konjac and A. bulbifer were sequenced using Illumina HiSeq™ 2000 sequencing technology. A total of 135,822 non-redundant unigenes were assembled from about 9.66 gigabases, and 19,596 SSRs were identified in 16,027 non-redundant unigenes. Di-nucleotide SSRs were the most abundant motif (61.6%), followed by tri- (30.3%), tetra- (5.6%), penta- (1.5%), and hexa-nucleotides (1%) repeats. The top di- and tri-nucleotide repeat motifs included AG/CT (45.2%) and AGG/CCT (7.1%), respectively. A total of 10,754 primer pairs were designed for marker development. Of these, 320 primers were synthesized and used for validation of amplification and assessment of polymorphisms in 25 individual plants. The total of 275 primer pairs yielded PCR amplification products, of which 205 were polymorphic. The number of alleles ranged from 2 to 14 and the polymorphism information content valued ranged from 0.10 to 0.90. Genetic diversity analysis was done using 177 highly polymorphic SSR markers. A phenogram based on Jaccard’s similarity coefficients was constructed, which showed a distinct cluster of 25 Amorphophallus individuals.ConclusionA total of 10,754 SSR markers have been identified in Amorphophallus using transcriptome sequencing. One hundred and seventy-seven polymorphic markers were successfully validated in 25 individuals. The large number of genetic markers developed in the present study should contribute greatly to research into genetic diversity and germplasm characterization in Amorphophallus.
Highlights
Amorphophallus is a genus of perennial plants widely distributed in the tropics or subtropics of West Africa and South Asia
The genus includes more than 170 species, which are mainly distributed in the tropics or subtropics of West Africa and South Asia [3]
Of Amorphophallus transcriptome data from Illumina sequencing After stringent quality assessment and data filtering, Illumina HiSeqTM 2000 sequencing produced 54,986,020 reads for A. konjac and 52,334,098 reads for A. bulbifer with 92.27% Q20 bases and 93.48% Q20 bases, respectively (Table 1)
Summary
Amorphophallus is a genus of perennial plants widely distributed in the tropics or subtropics of West Africa and South Asia. Its corms contain a high level of water-soluble glucomannan; it has long been used as a medicinal herb and food source. A large number of molecular markers are required for genetic diversity study and improving disease resistance in Amorphophallus. The diploid species A. konjac [6] is widely planted in China, its corm contained high-level glucomannan and possessed wide adaptability, but poor disease resistance and low coefficient of propagation [7]; in the contrast, A. bulbifer is a triploid (2n = 39) [6] with high disease resistance, characteristics of multi-seeding relay growth and high coefficient of propagation [8]. As far as we know, gene discovery and molecular breeding of new Amorphophallus economic cultivars have not been initiated because of the lack of genetic and genomic information
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