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.
Lilium davidii var. unicolor Salisb is a cultivar of Lilium (Liliaceae) with important edible and ornamental characteristic. The application and production of Lilium davidii var. unicolor Salisb were still facing large problems because of its several disadvantages such as narrow range of adaptability, small annual growth increment, and low fertility. To achieve broader environmental adaptability and obtain a more nutritious germplasm, we used colchicine and oryzalin to induce chromosome doubling via the soaking method. Tissue culture bulbs were treated with colchicine at 0.03%, 0.05%, or 0.08% for 32, 40, or 48 hours or with oryzalin at 0.002%, 0.005%, 0.008%, or 0.01% for 3, 6, 9, 12, or 24 hours before being transferred to a differentiating medium. The results showed that colchicine treatment resulted in the highest induction rate when applied at 0.05% for 48 hours, whereas oryzalin treatment produced fewer tetraploid plants. The chromosome number of induced plants with small stoma density and longer guard cells is twice than that of the diploid. The plants were identified as tetraploid. In this study, a new germplasm of Lilium davidii var. unicolor Salisb was innovative and showed novel genetic characteristic.
To explore differences in sucrose metabolism between peach fruit subjected to chilling stress (5 °C) and nonchilling stress (10 °C), sucrose concentration as well as the activities and gene expression levels for enzymes associated with sucrose metabolism were compared. Fruits stored at 5 °C accumulated higher concentrations of H2O2 and developed severe chilling injury (CI) compared with fruit kept at 10 °C. Activities and gene expression levels for enzymes related to sucrose metabolism, such as acid invertase (AI), neutral invertase (NI), sucrose synthase (SS), and sucrose phosphate synthase (SPS) were higher in fruit stored at 5 °C than at 10 °C throughout or late in storage. A sharp increase in net sucrose cleavage activity dramatically decreased sucrose concentration and increased reducing sugars at 5 °C. The sucrose concentration at 10 °C increased over the first 21 days and then declined slightly, and was higher than in fruit at 5 °C throughout storage. The increase in net sucrose cleavage activity at 5 °C is contrary to the expectation that biochemical reactions ordinarily proceed more rapidly with increasing temperature. We conclude that chilling stress stimulates the activities and transcription levels of enzymes involved in sucrose metabolism, resulting in increased sucrose cleavage.
Chinese bayberry (Morella rubra) is an economically important subtropical evergreen fruit crop native to China and other Asian countries. For facilitating cultivar discrimination and genetic diversity analysis, a total of 38 high-quality and highly polymorphic expressed sequence tags-simple sequence repeat (EST-SSR) markers, with little or no polymerase chain reaction (PCR) stutter bands, including 21 screened from those obtained previously and 17 newly developed markers, were developed. The average number of alleles (Na) per locus was 5.6, and polymorphism information content varied from 0.34 to 0.86, with a mean value of 0.57. With these markers, all 42 Chinese bayberry accessions analyzed were successfully discriminated and the phylogenetic relationship between accessions was revealed. The accessions can be separated into two groups with six subgroups. The grouping of four main cultivars in three subgroups and 12 white-fruited accessions, each with little or no anthocyanin accumulation in ripe fruit, into five subgroups suggested the preservation of broad diversity among cultivated populations. These EST-SSR markers and the findings obtained in this study can assist the discrimination of cultivars and lines and contribute to genetic and breeding studies in Chinese bayberry.
Jinyan (Actinidia eriantha × A. chinensis) is one of the gold-fleshed kiwifruit cultivars currently being promoted in south China. However, its fruit dry matter is usually less than 16%, which seriously affects fruit quality including taste and flavor. This causes a financial loss to growers: not only are the prices paid for the fruit low because of their bad reputation for quality, but some orchards have been removed. Improvement of fruit quality is essential. In this study, a method is described for squeezing and twisting flowering shoots before flowering and removing the distal vegetative parts of flowering shoots after fruit set. The effects on fruit quality were determined. The dry matter of fruit was increased by 6.6%. Fruit size also increased as did the chlorophyll a content and the chlorophyll:carotenoid ratio. The significantly increased fruit dry matter, resulting in significant increases in fruit soluble solids concentrations (P < 0.01), thereby possibly improving fruit taste. Fruit weight, fruit length, and carotenoid and ascorbic acid concentrations were significantly enhanced in comparison with controls (P < 0.01), increasing by 20%, 7%, 12%, and 19%, respectively. However, there was no significant difference in soluble sugar concentrations, titratable acid concentrations, and the reduced chlorophyll b concentrations. This research provides a practical method to increase fruit dry matter, and hence a way to allow fruit quality to reach commercial requirements for cultivars such as Jinyan, which under previous management systems had significant shortcomings in fruit flavor and taste.