Deep transcriptome sequencing allows for the acquisition of large-scale microsatellite information, and it is especially useful for genetic diversity analysis and mapping in plants without reference genome sequences. In this study, a total of 14,004 simple sequence repeats (SSRs) were mined from 10,511 unigenes screening of 63,608 nonredundant transcriptome unigenes in loquat (Eriobotrya japonica) with a frequency of 22 SSR loci distributed over 100 unigenes. Dinucleotide and trinucleotide repeat SSRs were dominant, accounting for 20.62%, and 42.1% of the total, respectively. Seventy primer pairs were designed from partial SSRs and used for polymerase chain reaction (PCR) amplification. Of these primer pairs, 54 exhibited amplification and 33 were polymorphic. The number of alleles at these loci ranged from two to 17, and the polymorphism information content values ranged from 0.24 to 0.89. We tested the transferability of 33 SSR polymorphic primer pairs in apple and pear, and the transferability rates in these two species were 90.9% and 87.9%, respectively. A high level of marker polymorphism was observed in apple [Malus ×domestica (66.7%)], whereas a low level was observed in pear [Pyrus sp. (51.5%)]. In addition, the PCR products from seven SSR primer pairs were selected for sequence analysis, and 89.2% of the fragments were found to contain SSRs. SSR motifs were conserved among loquat, apple, and pear. According to our sequencing results for real SSR loci, ≈12,490 SSR loci were present in these loquat unigenes. The cluster dendrogram showed a distinct separation into different groups for these three species, indicating that these SSR markers were useful in the evaluation of genetic relationships and diversity between and within the species of Maloideae in the Rosaceae. The results of our identified SSRs should be useful for genetic linkage map construction, quantitative trait locus mapping, and molecular marker-assisted breeding of loquat and related species.
Xiaoying Li, Hongxia Xu, Jianjun Feng and Junwei Chen
Kunkun Li, Weiqi Dong, Yun Zhao, Hongxia Xu, Junwei Chen and Changjie Xu
Loquat (Eriobotrya japonica Lindl.) is an economically important subtropical fruit tree, originating and commercially produced mainly in China and Japan. The aseptic seed germination of 13 loquat cultivars, including 9 white-fleshed cultivars and 4 red-fleshed ones, was studied, and the relationship between the germination rate and the content of endogenous ABA in seeds was explored. The germination rate and the seedling height at 80 days after sowing of white-fleshed cultivars were generally higher than that of red-fleshed ones. The ABA content in seeds was generally lower in white-fleshed cultivars, and the ABA content negatively correlated with the germination rate at 80 days after sowing. A moderate detrimental effect of 75% ethanol disinfection on aseptic germination of loquat seeds was observed, especially, for the cultivar Dahongpao, and the germination rate and seedling height were reduced by around half and one-third, respectively. The addition of plant preservative mixture (PPM) to the germination medium at a final concentration of 25 mg·L−1 effectively controlled contamination when the 75% ethanol disinfection procedure was omitted.