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Petalized anther abortion is an important characteristic of male sterility in plants. The male sterile plants (HB-21) evincing petalized anther abortion previously discovered in a clone population of the Camellia oleifera cultivar Huashuo by our research group were selected as the experimental material in this study. Using plant microscopy and anatomic methods and given the correspondence between external morphology and internal structure, we studied the anatomic characteristics of petalized anther abortion (with a fertile plant as the control group) in various stages, from flower bud differentiation to anther maturity, in hopes of providing a theoretical basis for research on and applications of male sterile C. oleifera plants, a new method for the selection of male sterile C. oleifera cultivars, and improvements in the yield and quality of C. oleifera. In this study, the development of anthers in C. oleifera was divided into 14 stages. Petalized anther abortion in male sterile plants was mainly initiated in the second stage (the stage of sporogenous cells). Either the petalized upper anther parts did not form pollen sacs, or the entire anthers did not form pollen sacs. The lower parts of some anthers could form deformed pollen sacs and develop, and these anthers could be roughly divided into two types: fully and partially petalized anthers. Abnormal callose and the premature degradation of the tapetum occurred in the pollen sacs formed by partially petalized anthers during the development process, resulting in the absence of inclusions in the pollen grains formed. Small quantities of mature pollen grains withered inward from the germinal furrows, exhibiting obvious abortion characteristics. The relative in vitro germination rate of the pollen produced by the partially petalized anthers of sterile plants was 11.20%, and the relative activity of triphenyltetrazolium chloride was 3.24%, while the fully petalized anthers did not generate pollen grains. Either the petalized anthers in male sterile plants did not produce pollen, or the vitality of the small amounts of pollen produced by sterile plants was very low compared with that of fertile plants. Such male sterile plants could be used to select correct clones and have good prospects for application in production.
This study aimed to investigate the flowering biological characteristics, floral organ characteristics, and pollen morphology of Camellia weiningensis Y.K. Li. These features of adult C. weiningensis plants were observed via light microscopy and scanning electron microscopy (SEM). Pollen viability and stigma receptivity were detected using 2,3,5-triphenyltetrazole chloride (TTC) staining and the benzidine–hydrogen peroxide reaction method. C. weiningensis is monoecious, with alternate leaves and glabrous branchlets. Its flowering period lasts 2 to 4 months, and the flowering time of individual plants lasts ≈50 days, with the peak flowering period from the end of February to the middle of March. It is a “centralized flowering” plant that attracts a large number of pollinators. Individual flowers are open for 12 to 13 days, mostly between 1230 and 1630 hr, and include four to six sepals, six to eight petals, ≈106 stamens, an outer ring of ≈24.6-mm-long stamens, an inner ring of ≈13.4-mm-long stamens, one pistil, and nine to 12 ovules. The flowers are light pink. The style is two- to three-lobed and 16.6 mm long, showing a curly “Y” shape. The contact surface of the style is covered with papillary cells and displays abundant secretory fluid and a full shape, facilitating pollen adhesion. The pollen is rhombohedral cone-shaped, and there are germ pores (tremoids). The groove of the germ pore is slender and extends to the two poles (nearly reaching the two poles). The pollen is spherical in equatorial view and trilobate in polar view. The pollen vitality was highest at the full flowering stage, and the stigma receptivity was greatest on days 2 to 3 of flowering. The best concentration of sucrose medium for pollen germination was 100 g/L. The number of pollen grains per anther was ≈2173, and the pollen-to-ovule ratio was 23,034:1. C. weiningensis is cross-pollinated. Seventy-two hours after cross-pollination, the pollen tube reached the base, and a small part entered the ovary. The time when the pollen tube reached the base after pollination was later than that in commonly grown Camellia oleifera. The results of this study might lay an important foundation for the flowering management, pollination time selection, and cross-breeding of C. weiningensis.
MicroRNAs (miRNAs) related to phytohormone signal transduction and self-incompatibility may play an important role in the xenia effect. However, associated research in this area is still lacking in rabbiteye blueberry (Vaccinium ashei). In this study, we identified miRNAs, predicted their target genes, performed functional enrichment analysis of the target genes, and screened for miRNAs related to phytohormone signaling and self-incompatibility. A total of 491 miRNAs were identified, of which 27 and 67 known miRNAs as well as 274 and 416 new miRNAs were found in the rabbiteye blueberry cultivars Brightwell and Premier, respectively. Compared with ‘Premier’, 31 miRNAs were upregulated and 62 miRNAs were downregulated in ‘Brightwell’. Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analysis indicated that the 4985 target genes predicted were involved in biosynthesis of amino acids, plant–pathogen interaction, and spliceosome pathways. A total of 10, one, one, five, two, five, and two candidate miRNAs related to auxin, cytokinin, gibberellin, abscisic acid, ethylene, brassinosteroid, and salicylic acid signaling, respectively, in rabbiteye blueberry pollen were identified. Further analysis indicated that novel_miR_49 was a candidate miRNA related to self-incompatibility, and their target gene was maker-VaccDscaff21-snap-gene-21.37. In addition, the KEGG enrichment analysis of the target genes of novel_miR_49 showed that they were involved in the ribosome, aminoacyl-tRNA biosynthesis, and glycosylphosphatidylinositol-anchor biosynthesis pathways. The results revealed that the microRNAs of rabbiteye blueberry pollen regulated to phytohormone signal transduction and self-incompatibility signal transduction based on related to auxin, cytokinin, gibberellin, abscisic acid, ethylene, brassinosteroid, and salicylic acid signaling. Results suggest that more research of the effects of miRNAs on regulation of hormone signal transduction and self-incompatibility is necessary for elucidating the molecular mechanism of the xenia effect.
The application of plant growth regulators (PGRs), such as abscisic acid (ABA), putrescine (Put), and 2,4-epibrassinolide (EBR), has been shown to enhance a plant's resistance to various abiotic stresses. However, the protective effects of these PGRs on tomato (Solanum lycopersicum) seedlings under suboptimal temperature stress have not yet been evaluated. We also do not know the most effective method of application of PGRs for various tomato cultivars. We studied the effects of three rates of exogenous ABA, Put, or EBR in limiting damage from suboptimal temperature stress on two tomato cultivars, Zhongshu6 (considered sensitive to suboptimal temperatures) and SANTIAM (considered tolerant to suboptimal temperatures). Results showed that application of these PGRs at appropriate concentrations could effectively reduce the decline in the net photosynthetic rate (Pn) and the chlorophyll (Chl) content in leaves caused by suboptimal temperature stress in both ‘Zhongshu6’ and ‘SANTIAM’ and could promote an increase in organic osmolyte (proline and soluble sugar) contents and root 2,3,5-triphenyltetrazolium chloride (TTC)-reducing activity for ‘Zhongshu6’. However, these effects were inferior on ‘SANTIAM’. For both cultivars, the best treatment concentrations are 1 mm ABA, 0.1 mm Put, or 0.02 μM EBR. Results indicate that in tomato production, exogenous application of ABA, Put, or EBR at appropriate concentrations can effectively limit damage from suboptimal temperature stress.
Baby primrose (Primula forbesii) is a newly cultivated and valuable ornamental plant with great market potential for both indoor and landscape use. As a container plant, baby primrose has long, weak flower stalks that can easily lodge, resulting in poor-quality plants, especially during transportation. To control plant height and subsequently prevent flower peduncle lodging, we investigated the effects of two plant growth regulators (PGRs), chlormequat chloride (CCC) at 0, 250, 500, or 750 ppm and uniconazole (UNI) at 25, 50, or 75 ppm on growth, development, and flowering of two cultivars of baby primrose, Fragrant Luolan and Red Star. Plant growth regulators at the proposed concentrations were applied twice throughout the experiment. Both PGRs significantly suppressed plant height in both cultivars, with a 16% to 27% reduction by CCC and 50% to 59% by UNI compared with untreated plants. Among CCC-treated groups, plants were shortest when CCC was applied at 500 ppm; plant height was suppressed more when treated with UNI. In both cultivars, UNI significantly suppressed the first, second, and third peduncle lengths. Furthermore, CCC affected peduncle length, but to a lesser extent than UNI. Plant growth regulator applications generally had little effect on flower characteristics of baby primrose. Neither PGRs influenced the inflorescence number and flower size; however, PGRs did increase the number of floral whorls and suppressed pedicel length of ‘Red Star’. New leaf growth was suppressed by both PGRs. In addition, peduncle cell length and width were both significantly suppressed by PGR applications. We concluded that two foliar applications of UNI at 25 ppm comprised the most effective method of controlling baby primrose plant height while maintaining desirable flower traits at a relatively low production cost. Results of this study provide guidance for techniques that can be used to effectively control the plant height of potted baby primrose for commercial greenhouse production.
The Himalayan yew, Taxus wallichiana Zucc., is an endangered species with a scatted distribution in the Eastern Himalayas and southwestern China. In the present study, 10 microsatellite markers from the genome of T. wallichiana were developed using the protocol of fast isolation by amplified fragment length polymorphism of sequences containing repeats (FIASCO). Polymorphism of each locus was assessed in 28 samples from four wild populations of the Himalayan yew. The allele number of the microsatellites ranged from two to five with an average of 2.9 per allele. The observed and expected heterozygosity varied from 0.00 to 1.00 and from 0.3818 to 0.7552, respectively. Cross-species amplification in another two yew species showed eight of them holding promise for sister species. Two of the 10 loci (TG126 and TC49) significantly deviated from Hardy-Weinberg expectations. No significant linkage disequilibrium was detected between the comparisons of these loci. These polymorphic microsatellite markers would be useful tools for population genetics studies and assessing genetic variations to establish conservation strategy of this endangered species.
Ascorbic acid (AsA) is a major antioxidant and redox buffer in plants. Dehydroascorbate reductase (DHAR; EC 1.8.5.1) catalyzes the conversion of dehydroascorbate (DHA) to AsA and is crucial for AsA regeneration. In this study, we developed transgenic tomato plants that overexpressed PbDHAR2 to investigate whether PbDHAR2 could limit the deleterious effects of salt and chilling stresses. These transgenic plants contained significantly higher AsA levels than the wild-type (WT) plants. Overexpression of PbDHAR2 increased the expression of the AsA-glutathione (GSH) cycle genes in transgenic lines under salt and chilling stresses. In addition, the transgenic lines subjected to salt and chilling stresses showed higher levels of antioxidant enzyme activity, lower malondialdehyde (MDA) levels, and higher chlorophyll contents than the WT. Thus, our results demonstrate that the regulation of PbDHAR2 during AsA regeneration contributes to enhanced salt and chilling tolerance in tomato.
To assess the genetic diversity among lotus (Nelumbo) accessions and evaluate the correlation between genetic variation and morphological classification, we sampled 138 accessions: two of N. lutea, 112 of N. nucifera, 17 of hybrids between N. nucifera and N. lutea, and seven Japanese cultivars. The 11 selected combinations of amplified fragment length polymorphism (AFLP) primers produced 138 polymorphic loci, and the percentage of polymorphism was 28.7%. The unweighted pair group method with arithmetic mean (UPGMA) dendrogram clustered all the accessions into two groups: Group I comprised N. lutea and its hybrids with N. nucifera; Group II included N. nucifera and its hybrids with N. lutea and Japanese cultivars. Population structure analysis identified four main clusters: N. lutea clustered mainly in C1, whereas N. nucifera clustered in C2, C3, and C4, which was consistent with the UPGMA and principal coordinate analysis results. The Japanese cultivars were related more closely to N. nucifera (genetic similarity coefficient = 0.74) than to N. lutea (0.46); hence, the Japanese cultivars can be classified as N. nucifera. Moreover, rhizome lotuses formed a separate subclade, whereas seed lotuses were interspersed among flower lotuses, which demonstrated that rhizome lotuses were distinct from flower and seed lotuses. Plant size, flower color, and other morphological criteria used commonly to classify lotuses were correlated with genetic variation to a certain extent but not sufficiently for accurate classification. It appears that it is necessary to use both DNA markers and morphological characteristics to classify lotus species and cultivars.
The appropriate timing of bolting and flowering is one of the keys to the reproductive success of Isatis indigotica. Several flowering regulatory pathways have been reported in plant species, but we know little about flowering regulatory in I. indigotica. In the present study, we performed RNA-seq and annotated I. indigotica transcriptome using RNA from five tissues (leaves, roots, flowers, fruit, and stems). Illumina sequencing generated 149,907,857 high-quality clean reads and 124,508 unigenes were assembled from the sequenced reads. Of these unigenes, 88,064 were functionally annotated by BLAST searches against the public protein databases. Functional classification and annotation assigned 55,991 and 23,072 unigenes to 52 gene ontology (GO) terms and 25 clusters of orthologous group (COG) categories, respectively. A total of 19,927 unigenes were assigned to 124 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, and 80 candidate genes related to plant circadian rhythm were identified. We also identified a number of differentially expressed genes (DEG) and 91 potential bolting and flowering-related genes from the RNA-seq data. This study is the first to identify bolting and flowering-related genes based on transcriptome sequencing and assembly in I. indigotica. The results provide foundations for the exploration of flowering pathways in I. indigotica and investigations of the molecular mechanisms of bolting and flowering in Brassicaceae plants.
Ultraviolet-A (UV-A) is the main component of UV radiation in nature. However, its role on plant growth, to a large extent, remains unknown. In this study, tomato (Solanum lycopersicum ‘Beijing Cherry Tomato’) seedlings were cultivated in an controlled environment in which UV-A radiation was provided by UV-A fluorescent lamps (λmax = 369 nm) with a fluence rate of 2.28 W·m−2. The photoperiod of UV-A radiation was 0, 4, 8, and 16 hours, which corresponds to control, UV-A4, UV-A8, and UV-A16 treatments, respectively. The photosynthetic photon flux density (PPFD) was 220 μmol·m−2·s−1, which was provided by light-emitting diodes (LEDs) with a blue/red light ratio of 1:9, the photoperiod of PPFD was 16 hours. We showed that supplementing 8 and 16 hours of UV-A to visible radiation (400–700 nm) stimulated plant biomass production by 29% and 33%, respectively, compared with that of control. This resulted mainly from larger leaves (i.e., 22% and 31% in 8 and 16 hours UV-A, respectively), which facilitated light capture. Supplemental UV-A also enhanced photosynthetic capacity, as indicated by greater net photosynthesis rates in response to CO2 under saturating PPFD. Furthermore, the greatest stomatal conductance (g S) value was observed in UV-A16, followed by UV-A8, which correlated with the greater stomatal density in the corresponding treatments. Moreover, supplemental UV-A did not induce any stress, as the maximum quantum efficiency of photosynthetic system II (PSII) (F v/F m) remained ≈0.82 in all treatments. Similarly, chlorophyll content and leaf mass area (LMA) were also unaffected by UV-A radiation. Taken together, we conclude that supplementing reasonable levels of UV-A to visible radiation stimulates growth of indoor cultivated tomato seedlings.