Effect of Crown Layers on Reproductive Effort and Success in Andromonoecious Aesculus indica (Wall. ex Camb.) Hook (Sapindaceae) in a Temperate Forest of Garhwal Himalaya.

The andromonoecy is an unusual sex expression in trees in which an individual plant bears both functionally staminate and hermaphrodite flowers on the inflorescences. This study aims to investigate the effect of crown layers on the floral biology and reproductive effort of Aesculus indica (Wall. ex Camb.) Hook. The results revealed that the peak period of anthesis was between 06:00 and 08:00 h of the day. Male flower production was predominantly higher as compared to the perfect flowers on the inflorescences. There was no significant variation between total pollen production in staminate and perfect flowers. Features like protogyny and inter-level asynchrony promote xenogamy; however, intra-level asynchrony results in geitonogamy. Controlled pollination treatments revealed the existence of self-incompatibility in flowers. Pollination syndromes in flowers support ambophily. A trend of consistent improvement in reproductive success from lower canopy layers to upper crown layers in the analyzed trees was recorded. The crown layers have a significant impact on flower production, fruit, and seed set. An increase in male flower production due to the increment in the crown is a mechanism of reproductive assurance as a pollen donor and pollinator recipient and also due to the differential cost of expenditure of reproduction in crown layers. Andromonoecy in A. indica promotes self-incompatibility, and there was a tapering trend of reproductive success in the crown layers.

Comparative floral development in male and female plants of Palmer amaranth (Amaranthus palmeri).

PREMISE: Characterizing the developmental processes in the transition from hermaphroditism to unisexuality is crucial for understanding floral evolution. Amaranthus palmeri, one of the most devastating weeds in the United States, is an emerging model system for studying a dioecious breeding system and understanding the biological traits of this invasive weed. The objectives of this study were to characterize phases of flower development in A. palmeri and compare organogenesis of flower development in female and male plants. METHODS: Flower buds from male and female plants were dissected for light microscopy. Segments of male and female inflorescences at different stages of development were cut longitudinally and visualized using scanning electron microscopy. RESULTS: Pistillate flowers have two to three styles, one ovary with one ovule, and five obtuse tepals. Staminate flowers have five stamens with five acute tepals. Floral development was classified into 10 stages. The distinction between the two flower types became apparent at stage four by the formation of stamen primordia in staminate flowers, which developed female and male reproductive organs initially, as contrasted to pistillate flowers, which produced carpel primordia only. In staminate flowers, the putative carpel primordia changed little in size and remained undeveloped. CONCLUSIONS: Timing of inappropriate organ termination varies across the two sexes in A. palmeri. Our study suggests that the evolution of A. palmeri from a cosexual ancestral state to complete dioecy is still in progress since males exhibited transient hermaphroditism and females produced strictly pistillate flowers.

Transcriptomic landscape of staminate catkins development during overwintering process in Betula platyphylla.

Betula platyphylla, belonging to the cold-specialized lineage Betulaceae, exhibits a unique reproductive strategy where staminate catkins emerge in the first summer and undergo an overwintering process, culminating in flowering in the following year. However, the underlying regulatory mechanism remains unclear. In this study, we investigated the male germline development of B. platyphylla in four distinct stages: microsporocytes in Oct. (S1), uninuclear microspores from Dec. (S2) to Mar. of the following year (S3), and bicellular microspores in Apr. (S4). We performed RNA sequencing on mature pollen and the four stages of staminate catkins. Using weighted gene co-expression network analysis (WGCNA), we identified five highly correlated gene modules with distinct expression profiles. These modules exhibited strong correlations with sugar metabolism, cell cycle, flowering, and cell wall dynamics, highlighting their dynamic roles during male germline developmental stages. During the overwintering process, we observed that the expression of transcription factors such as BpDUO1 and BpAMS at the appropriate developmental stages, suggests their significant roles in male germline development. The expression patterns of BpFLC and BpFT suggest their potential involvement in temperature perception during male reproductive development. These findings offer valuable insights into the reproductive success of plants adapting to cold environments.

Geniposidic Acid from Eucommia ulmoides Oliver Staminate Flower Tea Mitigates Cellular Oxidative Stress via Activating AKT/NRF2 Signaling

Eucommia ulmoides Oliver staminate flower (ESF) tea enjoys a good reputation in folk medicine and displays multiple bioactivities, such as antioxidant and antifatigue properties. However, the underlying biological mechanisms remain largely unknown. In this study, we aimed to investigate whether ESF tea can mitigate cellular oxidative stress. Crude ethyl alcohol extract and its three subfractions prepared by sequential extraction with chloroform, n-butyl alcohol and residual water were prepared from ESF tea. The results of antioxidant activity tests in vitro manifested n-butyl alcohol fraction (n-BUF) showed the strongest antioxidant capacity (DPPH: IC50 = 24.45 ± 0.74 µg/mL, ABTS: IC50 = 17.25 ± 0.04 µg/mL). Moreover, all subfractions of ESF tea, especially the n-BUF, exhibited an obvious capacity to scavenge the reactive oxygen species (ROS) and stimulate the NRF2 antioxidative response in human keratinocytes HaCaT treated by H2O2. Using ultra-high-performance liquid chromatography, we identified geniposidic acid (GPA) as the most abundant component in ESF tea extract. Furthermore, it was found that GPA relieved oxidative stress in H2O2-induced HaCaT cells by activating the Akt/Nrf2/OGG1 pathway. Our findings indicated that ESF tea may be a source of natural antioxidants to protect against skin cell oxidative damage and deserves further development and utilization.

Floral Quality Characterization in Olive Progenies from Reciprocal Crosses.

Despite the importance of flowering for fruit formation, it has been considered very little in breeding programs involving fruit species, including olives. We evaluated the principal morphological flower-quality components in the olive cultivars, 'Arbequina' and 'Picual', and in the progenies of their crosses. Wide ranges of variation were obtained for all the inflorescence traits and ovary tissue sizes. An analysis of variance indicated that the residual error was the main contributor to the inflorescence traits, except for the number of perfect flowers, underlining the need to evaluate adequate numbers of inflorescences for accurate measurements of these traits. However, the high repeatability obtained for the inflorescence traits suggests that simple evaluation procedures could be accurate enough for genotype characterization. The average values for 'Arbequina' were in the upper range for all the traits; the opposite occurred for 'Picual', and the values for most of the progenies were intermediate. No significant differences between the maternal and paternal effect on inheritance were found. Some interesting transgressive segregants showed a higher flower number, greater ovary and mesocarp size, or percentage of ovaries with all four fully developed ovules. The correlations among the parameters may have reflected a relatively consistent distribution of the ovaries' structural components and a close relationship between the ovaries and their mesocarp growth.

Phytochemical constituents, chemotaxonomic significance and anti-arthritic effect of Eucommia ulmoides Oliver staminate flowers.

Phytochemical investigation of Eucommia ulmoides Oliver staminate flowers provided twenty-seven compounds, including 11 triterpenes (1-11), 4 fatty acids (12-15), 1 coumarin (16), 1 chromone (17), 7 flavonoids (18-24) and 3 cyclopeptide alkaloids (25-27). Compounds 4-6, 8-9, 11-15, 17 and 25-27 were reported for the first time in the genus Eucommia. Their structures were determined by nuclear magnetic resonance, mass spectroscopy, optical rotation, circular dichroism spectroscopy analyses and comparison with literature data. The anti-arthritic activity of these compounds was also evaluated in this study. The result showed 1, 2, 4, 9 and 14 could significantly inhibit proliferation of fibroblast-like synovial cells from rats with IC50 values ranging from 11.83 to 50.12 µM. Furthermore, the chemotaxonomic significance of the isolated compounds has also been discussed. 12 and 25-27 might possess chemotaxonomic value for staminate flowers.

Both small and large plants are likely to produce staminate (male) flowers in a hermaphrodite lily.

Resource allocation to female and male function may vary among individual plants in species with variable sex expression. Size-dependent sex allocation has been proposed in hermaphrodites, in which female-biased allocation may increase with plant size. In many hermaphrodites with large floral displays, however, later-produced flowers tend to be functionally male. This paradoxical relationship between female and male function and plant size remains poorly understood. The subalpine lily Lilium lankongense has individuals of three sexual types: males with only staminate flowers, hermaphrodites with only perfect flowers, and andromonoecious plants with both perfect and staminate flowers. Here we tested theoretical predictions of size-dependent sex allocation in L. lankongense by measuring plant height and flower number of individuals of each sex at five field sites in the mountainous region of Shangri-La, southwestern China. To investigate variation in phenotypic gender, we identified sex expression of 457 individuals one year later. Our investigation showed that male plants, which usually produced one flower, were significantly smaller than andromonoecious and hermaphrodite plants. In addition, the total flower numbers of andromonoecious and hermaphrodite plants increased significantly with plant size. Large individuals were more likely to produce terminal staminate flowers, as there were more flowers in andromonoecious than in hermaphrodite individuals. Non-flowered plants were significantly smaller than flowering ones. Perfect flowers had significantly larger petals and pistils than staminate flowers, but they did not differ in dry weight of stamens. Our findings indicate that when plants are small, the less costly sex is favored, consistent with the 'size-advantage hypothesis'. When plants are large, both female and male investments change isometrically, as later-produced flowers tend to be functionally male.

Selection and Validation of Appropriate Reference Genes for Quantitative Real-Time PCR Normalization in Staminate and Perfect Flowers of Andromonoecious Taihangia rupestris.

Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) is the most commonly used and powerful method for gene expression analysis due to its high sensitivity, specificity, and high throughput, and the accuracy of this approach depends on the stability of reference genes used for normalization. Taihangia rupestris Yu and Li (Rosaceae), an andromonoecious plant, produces both bisexual flowers and unisexual male flowers within the same individual. Using qRT-PCR technique, investigation of the gene expression profiling in staminate and perfect flowers would improve our understanding of the molecular mechanism in regulation of flower formation and sex differentiation in andromonoecious T. rupestris. To accurate normalize the gene expression level in Taihangia flower, 16 candidate reference genes, including 10 traditional housekeeping genes, and 6 newly stable genes, were selected based on transcriptome sequence data and previous studies. The expressions of these genes were assessed by qRT-PCR analysis in 51 samples, including 30 staminate and perfect flower samples across developmental stages and 21 different floral tissue samples from mature flowers. By using geNorm, NormFinder, BestKeeper, and comprehensive RefFinder algorithms, ADF3 combined with UFD1 were identified as the optimal reference genes for staminate flowers, while the combination of HIS3/ADF3 was the most accurate reference genes for perfect floral samples. For floral tissues, HIS3, UFD1, and TMP50 were the most suitable reference genes. Furthermore, two target genes, TruPI, and TruFBP24, involved in floral organ identity were selected to validate the most and least stable reference genes in staminate flowers, perfect flowers, and different floral tissues, indicating that the use of inappropriate reference genes for normalization will lead to the adverse results. The reference genes identified in this study will improve the accuracy of qRT-PCR quantification of target gene expression in andromonoecious T. rupestris flowers, and will facilitate the functional genomics studies on flower development and sex differentiation in the future.

Larval feeding behaviour affects the impact of staminate flower production on the suitability of balsam fir trees for spruce budworm.

Laboratory rearing of spruce budworm, Choristoneura fumiferana, in conjunction with field rearing indicated that the feeding behaviour of the larvae, which is affected by the insect population density, significantly influenced the impact of balsam fir, Abies balsamea, staminate flowering on spruce budworm biology. At low budworm density, the production of pollen in the midcrown of host trees reduced the insect development time by 5 days without affecting pupal weight, fecundity and survival. However, at high budworm density, the small amount of current-year foliage produced by flowering branches forced old larvae (sixth instar) either to feed on 1-year-old foliage (backfeeding) or to move from the midcrown to the lower crown section where staminate flowers are absent and more current-year foliage is available. When old larvae fed on old foliage, they exhibited reduced pupal weight and fecundity without losing the advange in development time that they obtained from feeding on pollen during their early stages of development. On the other hand, when old larvae moved to the lower crown section, they avoided the negative effects of backfeeding but lost the advantage in development time that was gained from feeding on pollen. Results from this study indicated that the production of staminate flowers by balsam fir trees could have opposite effects on spruce budworm population dynamics depending upon the insect population density when flowering occurs.