Pollen Number and Ribosome Gene Expression Altered in a Genome-Editing Mutant of REDUCED POLLEN NUMBER1 Gene

The number of pollen grains varies within and between species. However, little is known about the molecular basis of this quantitative trait, in contrast with the many studies available on cell differentiation in the stamen. Recently, the first gene responsible for pollen number variation, REDUCED POLLEN NUMBER1 (RDP1), was isolated by genome-wide association studies of Arabidopsis thaliana and exhibited the signature of natural selection. This gene encodes a homolog of yeast Mrt4 (mRNA turnover4), which is an assembly factor of the large ribosomal subunit. However, no further data were available to link ribosome function to pollen development. Here, we characterized the RDP1 gene using the standard A. thaliana accession Col-0. The frameshift mutant, rdp1-3 generated by CRISPR/Cas9 revealed the pleiotropic effect of RDP1 in flowering, thus demonstrating that this gene is required for a broad range of processes other than pollen development. We found that the natural Col-0 allele conferred a reduced pollen number against the Bor-4 allele, as assessed using the quantitative complementation test, which is more sensitive than transgenic experiments. Together with a historical recombination event in Col-0, which was identified by sequence alignment, these results suggest that the coding sequence of RDP1 is the candidate region responsible for the natural phenotypic variation. To elucidate the biological processes in which RDP1 is involved, we conducted a transcriptome analysis. We found that genes responsible for ribosomal large subunit assembly/biogenesis were enriched among the differentially regulated genes, which supported the hypothesis that ribosome biogenesis is disturbed in the rdp1-3 mutant. Among the pollen-development genes, three key genes encoding basic helix-loop-helix (bHLH) transcription factors (ABORTED MICROSPORES (AMS), bHLH010, and bHLH089), as well as direct downstream genes of AMS, were downregulated in the rdp1-3 mutant. In summary, our results suggest a specialized function of ribosomes in pollen development through RDP1, which harbors natural variants under selection.

SYP72 interacts with the mechanosensitive channel MSL8 to protect pollen from hypoosmotic shock during hydration

In flowering plants, hydration of desiccated pollen grains on stigma is a prerequisite for pollen germination, during which pollen increase markedly in volume through water uptake, requiring them to survive hypoosmotic shock to maintain cellular integrity. However, the mechanisms behind the adaptation of pollen to this hypoosmotic challenge are largely unknown. Here, we identify the Qc-SNARE protein SYP72, which is specifically expressed in male gametophytes, as a critical regulator of pollen survival upon hypoosmotic shock during hydration. SYP72 interacts with the MSCS-LIKE 8 (MSL8) and is required for its localization to the plasma membrane. Intraspecies and interspecies genetic complementation experiments reveal that SYP72 paralogs and orthologs from green algae to angiosperms display conserved molecular functions and rescue the defects of Arabidopsis syp72 mutant pollen facing hypoosmotic shock following hydration. Our findings demonstrate a critical role for SYP72 in pollen resistance to hypoosmotic shock through the MSL8 cascade during pollen hydration.

Apple Autotetraploids—Phenotypic Characterisation and Response to Drought Stress

Polyploidization is an important source of variability for plant breeding. Polyploids are often characterised by increased resistance to biotic and abiotic stresses. Since drought and pathogen attack are the main threats to apple cultivation, obtaining new sources of resistance is an important issue for apple breeding. The newly obtained autotetraploid clones of apple cv. ‘Redchief’ showed superior resistance to fire blight. The aim of the presented research was the in-depth phenotypic characterisation of ‘Redchief’ tetraploids and assessment of their response to drought at the physiological and genetic level. The growth of own-rooted five-year-old trees of ‘Redchief’ tetraploids was poor compared with diploids; all growth parameters—the number and length of current season shoots, the total length of current season shoots per tree and the cross-section area of the trunk—were reduced in tetraploid clones. Grafting on M9 rootstock improved the growth characteristics of ‘Redchief’ tetraploids. Compared with diploid plants, the leaves of tetraploids were thicker, with altered shape, higher chlorophyll content, and larger stomata, but the stomatal density decreased. The leaf anatomical structure of tetraploids was changed, the adaxial and abaxial epidermis and both types of mesophyll were significantly thicker than in diploids. Moreover, the pollen grains of tetraploids were larger, but their viability and germination were reduced. Under conditions of limited water supply, the reduction in growth parameters was smaller and the physiological parameters were higher in the ‘Redchief’ tetraploid clone 4x-25 than in diploid plants. The expression of APX gene was higher in tetraploids than in diploids 15 days after drought stress induction. The results suggest the enhanced drought tolerance of the studied ‘Redchief’ autotetraploid clone compared with its diploid counterpart.

Ephedra aurea (Ephedraceae), a new species from Sicily

Ephedra aurea, a new species occurring in Sicily, is described and illustrated. Taxonomically, it shows close relationships with Ephedra nebrodensis mainly for the small size with internodes scabrous-wrinkled not fragile at the nodes, but relevant differences regarding the size and shape of twigs, leaves, male and female cones, ovules, seeds, fruiting strobili and pollen grains allow to distinguish very well the two species. Its ecology, conservation status and relationships with other allied Mediterranean species are discussed.

Micromorphology of endemic Centaurea glaberrima subsp. divergens (Asteraceae)

In this study, the micromorphology of the vegetative and reproductive structures of the endemic Centaurea glaberrima Tausch subsp. divergens (Vis.) Hayek (Asteraceae), using scanning electron microscope (SEM), is presented for the first time. Uniseriate whip-like non-glandular and biseriate glandular trichomes are found on the surface of all aboveground parts (stem, leaves, peduncles, involucral bract). On the adaxial leaf epidermis ribbed thickenings (striation pattern) of outer periclinal cell walls, slightly curved anticlinal cell walls and anomocytic stomata are noticed. Rugose abaxial surface with thorny protuberances of the involucral bract is documented. Corolla is glabrous with longitudinally parallel epidermal cells with distinct straight outline. Isopolar, radially symmetric and tricolporate microechinate pollen grains are seen. Short stylar hairs, without cuticular striations, are present along the outer sides of the style, while the inner sides (abaxial surface) constitute the papillate stigmatic surface. Microcharacters found in cypsela are as follows: slightly ribbed body; rotund base; lateral and concave insertion; short, unicellular curly acute trichomes; smooth epidermis; fine-sulcate ornamentation; rod shaped epidermal cells with short, obtuse end walls and straight anticlinal walls; poorly developed minutely dentate pericarp rim; and dimorphic pappus with bristles of different length and morphology, with pinnules restricted to the margins of the bristles. The results obtained contribute to knowledge about the micromorphology of the studied endemic species and provide features for its better identification. The taxonomic significance of the analyzed characters is discussed. Some well defined microcharacters of the studied species might have taxonomic value

Fossil Leaves of Meliosma (Sabiaceae) With Associated Pollen and a Eupodid Mite From the Eocene of Maoming Basin, South China

A first occurrence of the genus Meliosma (Sabiaceae) is reported from the upper Eocene of the Maoming Basin of South China. This fossil is one of the oldest reliable records of the genus within its modern center of diversity. Fossil leaves are assigned to a new species, Meliosma eosinica sp. nov. based on leaf morphology and epidermal characters. The leaf epidermal anatomy of fossil Meliosma is illustrated for the first time. We also provide the first SEM observation of pollen grains associated with Meliosma. This study also documents an occurrence of mites within the leaf domatia previously unknown from the fossil record. We presume that the studied mite belongs to the superfamily Eupodoidea (Arthropoda), and probably the family Eupodidae, which comprises very small soft-bodied cosmopolitan mites occupying a wide range of terrestrial habitats. Additionally, we analyze the damage types on the fossil leaves of Meliosma. They exhibit exclusively external foliage feeding damage caused by arthropods and traces of probable fungal infection. A review of currently known fossil occurrences of leaves, fruits, and wood of Meliosma provides evidence for the geological and geographical distribution of the genus.

Viability of pollen grains and stigma receptivity in Desert Rose

Adenium obesum (Forssk.) Roem. & Schult., popularly known as desert rose, has become a valuable ornamental plant. In floriculture, the production of hybrids is prioritized. Hence, knowledge on floral biology and sexual reproduction of the target species is fundamental. The objectives of this study were: (1) to test sucrose concentrations and temperatures for in vitro germination of A. obesum pollen grains; (2) to identify the effect of temperature on the viability of A. obesum pollen grains; and (3) to evaluate the viability of pollen grains and stigma receptivity in pre-anthesis, at flower opening, and 72 h post-flower opening in three accessions of A. obesum. A significant relationship between temperatures and sucrose concentrations was observed in the in vitro germination test. The highest percentage of in vitro germination of pollen grains, 39.81%, was observed at an estimated temperature of 26.05 °C. Desert rose accessions maintained in biochemical oxygen demand (BOD) chambers at 30 °C during a 16-h light photoperiod showed faster flowering, and temperatures ≥ 25 °C induced pollen grain viability percentages above 69%. Temperature is one of the most important abiotic factors, influencing mainly in pollen germination, pollen tube growing and in efficiency fertilization. The ICA-wd accession stood out and can be considered a pollen donor in artificial pollination. The stigmas of flowers were receptive from a day before flower opening until three days after. The two parameters presented above, stigma receptivity and pollen viability, allow inferences about the appropriate time for successful pollination and subsequent fertilization in desert roses.

Evaluation of Native and Nonnative Ornamentals as Pollinator Plants in Florida: II. Floral Resource Value

Consumer demand for novel, visually attractive ornamentals has often overshadowed the functional value plants may provide for flower-visiting insects. As native and nonnative species are hybridized for form, color, flowering, and disease resistance, it is important to assess whether some of these alterations influence plant nutrient quality for foraging insect pollinators. A study was conducted to ascertain the resource value of ornamental cultivars compared with their native congeners. The nectar volume and pollen quantity, viability, and protein content of 10 species of popular herbaceous flowering plants, commonly advertised as pollinator-friendly, were evaluated in northcentral Florida. Each genus encompassed a native and nonnative species, apart from pentas. Native species included blanket flower (Gaillardia pulchella), lanceleaf coreopsis (Coreopsis lanceolata), pineland lantana (Lantana depressa), and scarlet sage (Salvia coccinea). Nonnative species included Barbican™ yellow-red ring blanket flower (Gaillardia aristata ‘Gaiz005’), Bloomify™ rose lantana (Lantana camara ‘UF-1011-2’), mysty salvia (Salvia longispicata × farinacea ‘Balsalmysty’), Lucky Star® dark red pentas (Pentas lanceolata ‘PAS1231189’), ruby glow pentas (Pentas lanceolata ‘Ruby glow’) and UpTick™ Gold & Bronze coreopsis (Coreopsis × ‘Baluptgonz’). Floral rewards differed significantly across species. The native scarlet sage exhibited the largest nectar volume per flower in the summer (2.13 ± 0.17 µL), followed by the nonnative mysty salvia (1.26 ± 0.17 µL). In the fall, ruby glow pentas exhibited the largest nectar volume per flower (1.09 ± 0.17 µL) compared with all other ornamentals. The composite flowers of the native and nonnative blanket flower and coreopsis species had the lowest nectar volume per flower regardless of sampling date. Likewise, ruby glow pentas displayed the highest quantity of pollen grains (96.29 ± 0.12) per sample, followed by Lucky star pentas (52.33 ± 0.12), and Barbican blanket flower (50.98 ± 0.12). Pollen viability was similarly high (92% to 98%) among all species, apart from Bloomify rose lantana (20%) and pineland lantana (48%). Pollen protein content was highest in Uptick coreopsis (11.378 ± 1.860 μg/mg dry weight) and Lucky star pentas (10.656 ± 3.726 μg/mg dry weight), followed by lanceleaf coreopsis (7.918 ± 1.793 μg/mg dry weight). These results largely showed that the nonnative ornamentals selected provided resource-rich floral rewards, comparable to native congeners. Still, care should be taken in making similar assessments of other modern floral types.