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  • Author or Editor: Li Xu x
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Paeonia ostii ‘Feng Dan’ is an economically important, multipurpose woody plant in terms of its medical, ornamental, and oil values; however, there is a noticeable contradiction between the increasing demands and the lack of excellent germplasm resources because of traditional breeding and propagation approaches. In vitro embryo culture is an attractive option for this issue. This study presents a protocol for in vitro immature embryo culture in P. ostii ‘Feng Dan’, which involves two steps: 1) immature seeds at 30 days after anthesis (DAA) (cellularization stage of endosperm, proembryo stage) or after being cultured in vitro for cotyledon embryo formation (upward micropyle with placenta was the best inoculation method with the highest ratio of seed with cotyledon embryo of 66.67%); and 2) seedling establishment was realized within 7 months via embryo (at 40 DAA or after) germination, shoot induction, rooting, and acclimatization. The multiplication potential was increased with embryo maturity. This protocol provides an available reference for embryo rescue and propagation of tree peony and will be beneficial to shortening the breeding cycle.

Open Access
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Anthers contain starch and neutral lipids, which have key roles in microspore ontogeny and gametophyte development. In this study, we observed the dynamic changes in starch and neutral lipids in the anther developmental processes of castor (Ricinus communis) by cytochemical methods. Starch grains and neutral lipids presented a regular dynamic distribution during anther development. In young anthers, some neutral lipids accumulated in sporogenous cells, whereas neutral lipids disappeared with microspore growth. At the late microspore stage, starch grains began to accumulate in microspores, and the starch content of bicellular pollen significantly increased after microspore mitosis. At anthesis, starch grains and neutral lipids accumulated in the mature pollen grains. Visible changes occurred in anther wall cells. The epidermis, middle layer, and tapetum were degenerated, and only a single layer of endothecium remained at anthesis. The dynamic variation of starch grains and neutral lipids in tapetal cells was consistent with the changes in microspores and pollen during anther development. All these findings demonstrated that tapetal cells directly interacted with the developing gametophytes. The tapetal cells play an important role in supplying nutritional substances for microspore absorption. Moreover, the endothecium protects the pollen and contributes to anther dehiscence. The results of this study provide a foundation for the further research on sexual reproduction in angiosperms.

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An improved three-stage protocol for plant regeneration via somatic embryogenesis of the horticulturally important plant Iris germanica L. was developed using shoot apex segments as explants. At the first stage of the experiment, 60% of callus was obtained from shoot apex segments of I. germanica on Murashige and Skoog’s (MS) medium supplemented with 4.52 μm 2,4-dichloropheoxyacetic acid (2,4-D) and 0.44 μm 6-benzyladenine (6-BA). When nonembryogenic calli were subcultured on MS medium with 11.31 μm 2,4-D and 0.44 μm 6-BA, maximum frequency of embryogenic callus (66.0%) was obtained. At the second stage, the treatment of 9% (w/v) sucrose resulted in the optimum somatic embryo (SE) formation (70.0%). More than 90.0% of SEs germinated with bipolar structure and regenerated into plantlets on plant growth regulator-(PGR)free MS medium during the third stage. Regenerated plantlets were successfully acclimatized in greenhouse environment with little somaclonal variation. Histological study showed that somatic embryogenesis stages were asynchronous and SEs developed from the surface and inner tissue of embryogenic calli.

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Twenty-four representative melon varieties and six parental cultivars were examined in this study. Among 159 pairs of simple sequence repeat (SSR) primers, 18 SSR core primers with rich polymorphic information, a large number of genotypes, and the ability to distinguish different melon varieties were selected. A total of 113 genotypes were detected among the 30 experimental materials, with an average of 6.28 genotypes for each pair of primers. The polymorphic information content was on average 0.6807, ranging from 0.5618 to 0.7885. Specific bands of the primers for the 30 experimental materials were analyzed, and by combining different primer loci, all 30 varieties were identified. Unique barcodes for molecular identity cards for the 30 experimental materials were established using the fingerprints formed with this SSR marker system. Each variety has a unique identity card that can be applied for the registration of the newly bred varieties, the protection of breeders’ rights, and the authenticity of breeds after promulgation of the new Seed Law of the People’s Republic of China.

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A static experiment in greenhouse was conducted to investigate the growth of three grasses in high and medium eutrophic water and the effects of their removal on ammonia nitrogen (NH4 +-N), nitrate nitrogen (NO3 -N), total nitrogen (TN), total phosphorus (TP), and the chemical oxygen demand (COD), and compared with cattail (Typha angustifolia). The results showed that 1) the removal efficiency of NH4 +-N, NO3 -N, TN, TP, and COD treated by the four plants in eutrophic water were significantly higher than that in non-plant water. With the extension of treatment time, the concentrations of NH4 +-N, NO3 -N, TN, TP, and COD in the eutrophic water decreased first and then tend to be stable. 2) Cynodon dactylon ‘Tifton 85’ (C. dactylon ‘Tifton 85’), Cortaderia selloana ‘Pumila’ (C. selloana ‘Pumila’) and T. angustifolia absorbed more than 95.7% and 88.6% of TN and TP in eutrophic water, and accumulate more than 89.5% and 82.0% in plants, respectively. However, the ratio of Cortaderia selloana ‘Silver Comet’ (C. selloana ‘Silver Comet’) was significantly lower. 3) The high abilities of these three plants to purify eutrophic water may be directly related to their rapid growth. 4) The comprehensive purification ability of the four plants to eutrophic was significantly different, in the order of C. dactylon ‘Tifton 85’ > C. selloana ‘Pumila’ ≈ T. angustifolia > C. selloana ‘Silver Comet’. These results indicated that C. dactylon ‘Tifton 85’ and C. selloana ‘Pumila’ can be used as alternative plants to T. angustifolia for the purification of eutrophic water. The results of this study can provide new materials and ideas for phytoremediation.

Open Access

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.

Free access
Authors: , , , and

The investigation of hybridization processes and embryogenesis of heterozygote is an effective approach for early hybrids’ identification, which could provide reliable information for successful crossbreeding. In this study, we reported the whole hybridization processes of the direct cross and reciprocal cross between Michelia yunnanensis Franch. ex Finet et Gagnep. and Michelia crassipes Law using fluorescence microscopy after aniline blue staining, with the pollen germination on stigmas, pollen tube growth in styles, and subsequent extension into the embryo sac as well as the double fertilization processes are documented in detail. The M. yunnanensis × M. crassipes combination displayed considerable cross-compatibility, and the heterozygote embryogenesis was further observed with an approach of modified cryosectioning technique. Besides, the whole formation processes of hybrid seeds from artificial pollination to maturation were successfully observed. However, in the reciprocal cross, we found incompatibility between pollen grains of M. yunnanensis and stigmas of M. crassipes for the reason of hysteretic identification, as well as the abnormal callose deposition which belongs to the prefertilization barriers. This is the first study in which the complete and clear hybridization processes in Michelia were reported. We inferred that unilateral incompatibility of M. crassipes detected in this study may also exist in some other Michelia species. In artificial hybridization practices, we suggest some special treatments for overcoming prefertilization barrier should be taken when treating M. crassipes as the maternal parent.

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Soil water deficit impacts cold acclimation and freezing tolerance in creeping bentgrass (Agrostis stolonifera L.), but the mechanisms underlying have not been well understood. The objectives of this study were to investigate the effects of deficit irrigation before and during cold acclimation on osmoprotectants, antioxidant metabolism, and freezing tolerance in creeping bentgrass. The grass was subjected to three-soil moisture levels: well-watered [100% container capacity (CC)], deficit irrigation induced-mild drought stress (60% CC), and severe drought stress (30% CC) for 35 days including 14 days at 24/20 °C (day/night) and then 21 days under cold acclimation treatment (2 °C) in growth chambers. Leaf proline and total soluble sugar (TSS) levels were higher in the grass under mild drought stress relative to that under severe drought stress. Superoxide (O2 −·), hydrogen peroxide (H2O2), and malondialdehyde (MDA) content were higher in the grass under severe drought relative to that under well-watered and mild drought stress at day 35. Mild drought stress increased catalase (CAT) and guaiacol peroxidase (POD) activity, induced new isoforms and increased band intensities of superoxide dismutase (SOD), CAT, and POD during cold acclimation (days 14 to 35). No differences in osmoprotectants, antioxidant metabolism, and freezing tolerance were found between mild drought and well-watered treatments. The results of this study suggest deficit irrigation-induced mild drought stress in late fall and winter could induce accumulation of osmoprotectants and improve antioxidant metabolism, and freezing tolerance, but severe drought stress could reduce freezing tolerance of creeping bentgrass in the region with limited precipitation.

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Thirteen Chinese cabbage (Brassica rapa) hybrid cultivars and 26 parental inbred lines were used as experimental materials to screen for primers producing hybrid and parental complementary bands and for primers with high polymorphism information contents and low genotype frequencies. A total of 18 pairs of core primers were designed to identify the purity of Chinese cabbage. There was no significant difference in the purity percentage measured between different loci of the same strain. The fingerprint obtained by the amplification of each locus could be used to identify purity to obtain an authentic purity percentage. Curve mapping and significance analyses were conducted using the purity percentage of eight different seed samples and confirmed a sampling seed number of 96. The results of the purity test were verified by comparison with the grow-out test (GOT) using molecular markers. In conclusion, the simple sequence repeat (SSR) detection system could be used for the rapid identification of the purity of the tested Chinese cabbage hybrids.

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