Modern pollen-vegetation relationships along altitudinal vegetation belts in the Leigong Mountain of southwestern China

Understanding the distribution,dispersal,and correlation of modern pollen with vegetation in mountainous regions is essential for establishing accurate modern analogs for fossil pollen records.This study,conducted in Leigong Mountain on the YunnanGuizhou Plateau of southwestern China,involved the collection of 35 surface soil samples from diverse vegetation communities along an elevational gradient ranging from 1210 to 1875 meters.The results reveal a close correspondence between modern pollen assemblages and vegetation zones.Principal Component Analysis(PCA)results indicate that pollen assemblages can effectively distinguish between subtropical montane evergreen broad-leaved forest(SEBF)and subtropical montane deciduous broadleaved forest(SDBF).However,both SEBF and SDBF show significant overlap with subtropical montane evergreen-deciduous broad-leaved mixed forest(SEMF).Detrended Correspondence Analysis(DCA)results clearly distinguish the three vegetation zones,and the first axis of DCA shows a significant positive correlation with elevation(p<0.01,R=0.48).Discriminant Analysis(DA)successfully assigns 94.4%of the modern pollen samples to their respective vegetation zones.Pollen taxa such as Impatiens,Astertype,and Rosaceae exhibit significant indicative capabilities for the SEBF zone,effectively distinguishing this vegetation zone from others.Pinus and Alnus display overrepresentation in the Leigong Mountain region,while Quercus(D,deciduous-type)and Poaceae exhibit high representation in the SEBF zone.In the SEBF zone,both pollen diversity and richness are the lowest.Our study reveals the complex relationship between the richness and diversity of pollen and vegetation.The diversity and richness of tree and shrub pollen are found to be lower than those of the corresponding plants.The pollen-vegetation relationship elucidated in this study serves as a critical reference for reconstructing ancient environments from fossil pollen retrieved in this region.

Enhancer of Shoot Regeneration 2(ESR2)regulates pollen maturation and vitality in watermelon(Citrullus lanatus)

Watermelon(Citrullus lanatus)holds global significance as a fruit with high economic and nutritional value.Exploring the regulatory network of watermelon male reproductive development is crucial for developing male sterile materials and facilitating cross-breeding.Despite its importance,there is a lack of research on the regulation mechanism of male reproductive development in watermelon.In this study,we identified that ClESR2,a VIIIb subclass member in the APETALA2/Ethylene Responsive Factor(AP2/ERF)superfamily,was a key factor in pollen development.RNA insitu hybridization confirmed significantClESR2 expression in the tapetum and pollen during the later stage of anther development.The pollens of transgenic plants showed major defects in morphology and vitality at the late development stage.The RNA-seq and protein interaction assay confirmed that ClESR2 regulates pollen morphology and fertility by interacting with key genes involved in pollen development at both transcriptional and protein levels.These suggest that Enhancer of Shoot Regeneration 2(ESR2)plays an important role in pollen maturation and vitality.This study helps understand the male reproductive development of watermelon,providing a theoretical foundation for developing male sterile materials.

High temperature treatment induced production of unreduced 2n pollen in Camellia oleifera

Unreduced gametes through chromosome doubling play a major role in the process of plant polyploidization.Our previous work confirmed that Camellia oleifera can produce natural 2n pollen,and it is possible to induce the 2n pollen formation by high temperature treatment.This study focused on the optimization of the 2n pollen induction technique and the mechanisms of high temperature-induced2n pollen formation in C.oleifera.We found that the optimal protocol for inducing 2n pollen via high temperature was to perform 45℃with4 h at the prophaseⅠstage of the pollen mother cells(PMCs).Meanwhile,high temperature significantly decreased the yield and fertility of2n pollen.Through the observation of meiosis,abnormal chromosome and cytological behaviour was discovered under high-temperature treatment,and we confirmed that the formation of 2n pollen is caused by abnormal cell plate.Based on weighted gene co-expression network analysis,fifteen hub genes related to cell cycle control were identified.After male flower buds were exposed to heat shock,polygalacturonase gene(CoPGX3)was significantly upregulated.We inferred that high temperature causes the CoPGX3 gene to be overexpressed and that CoPGX3 is redistributed into the cytosol where it degrades cytoplasmic pectin,which leads to an abnormal cell plate.Furthermore,abnormal cytokinesis resulted in the formation of dyads and triads,and PMCs divided to produce 2n pollen.Our findings provide new insights into the mechanism of 2n pollen induced by high temperature in a woody plant and lay a foundation for further ploidy breeding of C.oleifera.

Comparative Study on Floral Organs and Pollen Characteristics of Different Varieties of Sorrel

The floral organ morphology,pollen morphology,quantity,and viability of Rumex,Rumex hanus F1,Rumex hanus 1,and Rumex hanus 2 were compared and analyzed.The results showed that the longitudinal diameter of petals was not significantly different among different varieties of sorrel,and the transverse diameter and corolla diameter showed differences to varying degrees.They all had six stamens and three stigmas.The pollen shape of the four varieties was prolate,and the polar view was trilobated and round.They all had three germinal furrows,and the outer wall of the pollen was decorated with small cavities and spiny patterns,with warty protrusions.The equatorial axis length of pollen of different varieties of sorrel was significantly different(P<0.05).There were no significant differences in pollen amount among different varieties.There were six anthers in a single flower,featuring anther dehiscence.The pollen viability of Rumex hanus 1 was significantly lower than that of the other three varieties(P<0.05).Therefore,through the analysis of the floral organs and morphological characteristics of anthers of sorrel,the varieties of sorrel can be effectively distinguished and identified.

BRASSICA DE-EXINED POLLEN AS A NEW EXPERIMENTAL SYSTEM FOR STUDYING POLLEN GERMINATION

Exine deprived pollen only coated with inline has been isolated and artificially germinated in Brassica L. In view of the lack of exine and germ furrows, the de-exined pollen is an interesting experimental system for studying pollen germination. This report focuses on the microscopic and ultrastructural changes in the aspects of polarization, predetermination of the germination site and new wall synthesis during the course of isolation and germination of the de-exined pollen. It was shown that the de-exined pollen was already in an active state and had established its polarity and germination sites prior to exine detachment. The germination sites were still localized at the region of the previous germ furrows even after exine detachment. The new wall deposited at the germination sites appeared to have an important morphogenetic role in setting a limit to the size of pollen tube diameter . These results support our assumption that the de-exined pollen may have an extensive application in the research of pollen biology.

GAMYB transcription factor LoMYB65 from lily plays a vital role in pollen development

Lily(Lilium spp.)is an important horticultural crop,but its use is limited due to serious pollen contamination problems.There are many studies on pollen development in model plants,but few on flower crops such as lilies.Gibberellin(GA)is a large class of hormones and plays an important role in plant vegetative growth and reproductive development.GAMYB is a group of the R2R3-MYB family upregulated by gibberellin,and plays an important role in anther development.Here,we isolated a novel GAMYB,named LoMYB65,from lily,which was closely related to the AtMYB65 and AtMYB33 in Arabidopsis.Fluorescence quantitative PCR results showed that LoMYB65 was mainly expressed in lily anthers.LoMYB65 could be activated by 288μmol·L^(-1)GA3treatment and the LoMYB65 protein was located in the nucleus and cytoplasm,and had transactivation in yeast and tobacco leaf cells.The conserved motif within 226 amino acids of the C-terminal of LoMYB65 contributed to its transactivation.Overexpression of LoMYB65 caused dwarf phenotype,unnormal tapetum development,less seeds of siliques in transgenic Arabidopsis plants,the transgenic plants showed partly male sterile.Simultaneously,silencing of LoMYB65 with VIGS(Virus Induced Gene Silencing)in lily anthers caused unnormal pollen development and reduced the pollen amount.Overexpression of LoMYB65 in Arabidopsis and silencing of LoMYB65 in lily resulted in decreased pollen counts,so we speculate that LoMYB65 may be dose-dependent.Overall,these findings suggest that LoMYB65 may play an important role in anther development and pollen formation in lily.LoMYB65 may provide a useful candidate gene for pollenless breeding of lily.

Effect of wall-disruption on nutrient composition and in vitro digestion of camellia and lotus bee pollens

The nutrient digestion,absorption and biological activity of bee pollen may be limited due to the complex pollen wall.Here,the effect of superfine grinding technology on the release of nutrients from bee pollen were investigated,and their antioxidant activities and in vitro digestion were explored in this study.Results showed that the content of nutrients in bee pollen increased after wall disruption.Among them,fat content increased by 22.55%-8.31%,protein content increased by 0.54%-4.91%,starch content increased by 36.31%-48.64%,soluble sugar content increased by 20.57%-29.67%,total phenolic acid content increased by 11.73%-86.98%and total flavonoids content increased by 14.29%-24.79%.At the same time,the antioxidant activity increased by 14.84%-46.00%.Furthermore,the active components such as phenolic compounds in the wall-disruption bee pollen were more readily to be released during the in vitro digestion,and easier to be absorbed because of their higher bioaccessibility.Antioxidant activities during in vitro digestion were also improved in walldisruption bee pollen.These findings provide evidence that bee pollen wall disruption was suggested,thus,it is more conducive to exerting the value of bee pollen in functional foods.

Effect of different drying methods on the amino acids,α-dicarbonyls and volatile compounds of rape bee pollen

The significant demand for high quality food has motivated us to adopt appropriate processing methods to improve the food nutritional quality and flavors.In this study,the effects of five drying methods,namely,pulsed vacuum drying(PVD),freeze drying(FD),infrared drying(IRD),hot-air drying(HAD)and sun drying(SD)on free amino acids(FAAs),α-dicarbonyl compounds(α-DCs)and volatile compounds(VOCs)in rape bee pollen(RBP)were determined.The results showed that FD significantly released the essential amino acids(EAAs)compared with fresh samples while SD caused the highest loss.Glucosone was the dominantα-DCs in RBP and the highest loss was observed after PVD.Aldehydes were the dominant volatiles of RBP and SD samples contained more new volatile substances(especially aldehydes)than the other four drying methods.Comprehensively,FD and PVD would be potential methods to effectively reduce the quality deterioration of RBP in the drying process.

Morphological and cytological assessments reveal pollen degradation causes pollen abortion in cotton cytoplasmic male sterility lines

Background Understanding the mechanism of male sterility is crucial for producing hybrid seeds and developing sterile germplasm resources.However,only a few cytoplasmic male sterility(CMS)lines of cotton have been produced due to several challenges,like inadequate variation of agronomic traits,incomplete sterility,weak resilience of restorer lines,and difficulty in combining strong dominance.Therefore,the morphological and cytological identification of CMS in cotton will facilitate hybrid breeding.Results Two F_(2) segregating populations of cotton were constructed from cytoplasmic male sterile lines(HaA and 01A,maternal)and restorer lines(HaR and 26R,paternal).Genetic analysis of these populations revealed a segregation ratio of 3:1 for fertile to sterile plants.Phenotypic analysis indicated no significant differences in traits of flower bud development between sterile and fertile plants.However,sterile plants exhibited smaller floral organs,shortened filament lengths,and anther atrophy on the flowering day in comparison with the fertile plants.When performed scanning electron microscopy(SEM),the two F_(2) populations revealed morphological variations in the anther epidermis.Cellular analysis showed no significant differences in pollen development before pollen maturation.Interestingly,between the pollen maturation and flowering stages,the tapetum layer of sterile plants degenerated prematurely,resulting in abnormal pollen grains and gradual pollen degradation.Conclusion The results of this study suggest that fertility-restoring genes are controlled by a single dominant gene.Sterile plants exhibit distinctive floral morphology,which is characterized by stamen atrophy and abnormal anthers.Pollen abortion occurs between pollen maturity and flowering,indicating that premature tapetum degradation may be the primary cause of pollen abortion.Overall,our study provides a theoretical basis for utilizing CMS in hybrid breeding and in-depth investigation of the dominant configuration of cotton hybrid combinations,mechanisms of sterility,and the role of sterile and restorer genes.

Allergen degradation of bee pollen by lactic acid bacteria fermentation and its alleviatory effects on allergic reactions in BALB/c mice

Food allergy as a global health problem threatens food industry.Bee pollen(BP)is a typical food with allergenic potentials,although it performs various nutritional/pharmacological functions to humans.In this study,lactic acid bacteria(LAB)were used to ferment Brassica napus BP for alleviating its allergenicity.Four novel allergens(glutaredoxin,oleosin-B2,catalase and lipase)were identified with significant decreases in LAB-fermented BP(FBP)than natural BP by proteomics.Meanwhile,metabolomics analysis showed significant increases of 28 characteristic oligopeptides and amino acids in FBP versus BP,indicating the degradation of LAB on allergens.Moreover,FBP showed alleviatory effects in BALB/c mice,which relieved pathological symptoms and lowered production of allergic mediators.Microbial high-throughput sequencing analysis showed that FBP could regulate gut microbiota and metabolism to strengthen immunity,which were closely correlated with the alleviation of allergic reactivity.These findings could contribute to the development and utilization of hypoallergenic BP products.

DIA-based proteome profiling with PRM verification reveals the involvement of ER-associated protein processing in pollen abortion in Ogura CMS cabbage

Ogura cytoplasmic male sterility(Ogura CMS)is extensively applied in hybrid seed production in cruciferous crops.However,the posttranscriptional molecular basis of Ogura CMS in cruciferous crops remains elusive.Here,a data-independent acquisition-based proteomic approach coupled with a parallel reaction monitoring-based targeted proteomic assay was used to analyze the proteome dynamics of Ogura CMS cabbage line RM and its maintainer line RF during floral bud development to obtain insights into the mechanism underlying Ogura CMS in cruciferous crops.A total of 9162 proteins corresponding to 61464 peptides were identified in RM and RF floral buds.The proteomic fluctuation of RM was weaker than that of RF.Differences in protein expression between RM and RF gradually enlarged with floral bud development.Fifteen continually up-regulated and eight continually down-regulated proteins were found in RM relative to RF throughout floral bud development.Differentially expressed proteins between RM and RF during floral bud development were implicated in the endoplasmic reticulum(ER)-associated protein processing pathway,in which most of them exhibited down-regulated expression in RM.These data suggest that ER-associated protein processing may be involved in pollen abortion in Ogura CMS cabbage by inhibiting the expression of critical factors.Our findings not only deepen the understanding of the molecular mechanisms of Ogura CMS in cruciferous crops but also provide better guidance for applying Ogura CMS in the hybrid breeding of cruciferous crops.

Treatments of Pollen with Different Solvents Affect the Seed Setting Rate of Rice(Oryza sativa L.)

This study aimed to investigate the effects of treatments of rice pollen with different solvents on the seed setting rate.Rice pollen was treated with ultrapure water,0.5%dimethyl sulfoxide,ethanol(1%,5%,and 8%),and 0.5%dimethyl sulfoxide+8%ethanol,and the pollen without treatment was taken as the control.Then,crossing was conducted with the treated rice plants as the male parents and plants subjected to emasculation by warm water as the female parents.The seed setting rates of the male and female parents were then recorded.The results showed that the treatments with different solvents significantly decreased the seed setting rates of both male and female parents.The treatment with 1%ethanol resulted in the highest seed setting rates for both male and female parents,while the treatment with 0.5%dimethyl sulfoxide resulted in the lowest seed setting rates for both parents.Although these solvents affected the seed setting rate of rice,they served as intermediate solvents to dissolve chemicals used in the induction of rice haploids.This study layed a technical foundation for subsequent chemical induction of rice haploids.

Influence of Plant Growth Regulators on Pollen Germination and Growth of Xinjiang Apricots

[Objective] This study aimed to investigate the influence of different plant growth regulators on apricot pollen germination and pollen tube growth. [Method] Pollens of six kinds of Xinjiang apricots were cultured in solid media supplemented with five plant growth regulators （GA3 , NAA, 2, 4-D, 6-BA, IAA）. Then the rate of pollen germination and the length of pollen tube were respectively measured. [Result] In a certain concentration range, GA3 most significantly promoted the pollen germination and the pollen tube growth of Shushanggan, Kalayulvke, Dayoujia, Yiliakeyulvke and Kabakehuanna; NNA had the strongest improvement function on Kumaiti’s pollen germination and pollen tube growth. [Conclusion] All the five plant growth regulators promoted the pollen germination and the pollen tube growth of apricots at low concentration but inhibited them at high concentration.

Modern pollen assemblages of the forest communities and their relationships with vegetation and climate in northern China

53 pollen traps and surface samples were collected in order to detect the characteristics of pollen assemblages and their relationships with vegetation and climate in 16 forest communities located in 10 mountains in northern China. The results show that 72% of the pollen taxa (80 taxa) are the same between the traps and the surface samples. The dominant taxa in the plant communities are consistent with the main pollen taxa in the pollen assemblages at the same sites. In Pinus plant communities, both Pinus pollen influx and concentration are higher, indicating the high pollen productivity and good pollen preservation ability of Pinus. In Picea and Abies plant communities, Picea and Abies have lower pollen influxes but higher concentrations, suggesting their low pollen productivities but better pollen preservation abilities. In Betula and Quercus plant communities, Betula and Quercus have higher pollen influxes but lower concentrations, revealing their high pollen productivities but poor pollen preservation abilities. The study of relationships between pollen and vegetation with discriminant analysis shows that pollen assemblages from both trap and surface samples can reflect the characteristics of different communities and distinguish different ecological areas, but surface samples can reflect the dominant components of communities much better than the traps. The study on correlations between pollen assemblages and climate with DCCA reveals that significant correlations exist between pollen assemblages and mean temperature of the coldest month (r = 0.84 for trap samples, r = 0.72 for surface samples), and then annual mean precipitation (r = 0.73 for trap samples, r = 0.71 for surface samples).

Pollen Viability in Three Xinjiang Hawthorn Species

[Objective] The aim was to explore the most efficient medium for pollen germination and suitable pollen storage temperature for three different hawthorn species in Xinjiang, and to lay foundation for the development and utilization of wild hawthorn resources. [Method] A randomized block design was adopted to select the most suitable concentrations of sucrose and boric acid in the medium for pollen germination under in vitro culture. The vitality of the pollens stored at different tem- peratures for six mouths was measured once every 10 d using the optimum medi- um selected in above steps. [Result] The most efficient medium for Altai hawthorn pollen germination was 1% agar ＋ 0.05% boric acid ＋ 15% sucrose, and its germi- nation rate in this medium was 30.8%; the most suitable medium for crabapple hawthorn pollen germination was 1% agar ＋ 0.01% boric acid ＋ 15% sucrose, and its germination rate in this medium was 58.7%; and the highest germination rate of Junggar hawthorn pollen was found in the medium containing 1% agar ＋ 0.01% boric acid ＋ 20% sucrose, and the germination rate reached 67.2%. There were extremely significant differences （P〈0.01） in germination rate between these treat- ments and others in each species. The pollen vitality of Altai hawthorn, crabapple hawthorn and Junggar hawthorn were lost completely after they were stored at room temperature for 40, 60 and 70 d; at 4-5 ℃ for 90, 130, and 140 d; or at -18 ℃ for 130, 160 and 170 d. Therefore, -18 ℃ was found the most efficient storage temperature for the three hawthorn species＇ pollen among the three different stor- age temperatures. [Conclusion] A certain concentration of sucrose and boric acid can promote pollen germination of the three hawthorn species. Under different stor- age temperatures, the pollen vitality of the three hawthorn species can maintain for a longer period at -18 ℃ than at 4-5 ℃, and for the shortest period at room tem- perature.

Plant Pollen Grains:A Move Towards Green Drug and Vaccine Delivery Systems

Pollen grains and plant spores have emerged as innovative biomaterials for various applications such as drug/vaccine delivery,catalyst support,and the removal of heavy metals.The natural microcapsules comprising spore shells and pollen grain are designed for protecting the genetic materials of plants from exterior impairments.Two layers make up the shell,the outer layer(exine)that comprised largely of sporopollenin,and the inner layer(intine)that built chiefly of cellulose.These microcapsule shells,namely hollow sporopollenin exine capsules have some salient features such as homogeneity in size,non-toxic nature,resilience to both alkalis and acids,and the potential to withstand at elevated temperatures;they have displayed promising potential for the microencapsulation and the controlled drug delivery/release.The important attribute of mucoadhesion to intestinal tissues can prolong the interaction of sporopollenin with the intestinal mucosa directing to an augmented effectiveness of nutraceutical or drug delivery.Here,current trends and prospects related to the application of plant pollen grains for the delivery of vaccines and drugs and vaccine are discussed.

Vitality and storage condition of Syringa pollen

The fresh pollen vitality, the effect of different storage conditions on the pollen vitality, and the difference of vitality among the species of seven species of Syringa were determined in Shenyang, China. The results indicated that the pollen vi-tality (81.5%) of Syringa villosa was the highest among the seven tested species, followed by S. microphylla and S. meyeri, and that of S. oblata var. affinis was the lowest. The low temperature was the best condition for storage of pollen of Syringa, and the most proper temperature for the storage was 0-2 癈. The storability of S. microphylla was the best of all, and it could be stored over 60 days at the temperature of 0-2 癈, next was S. villosa and S. meyeri.

Effects of Low Energy Argon Ion Implantation on Maize Pollen Germination and the Cytosolic Ca^(2+) Distribution during Pollen Germination Process

To investigate the effect of low energy ion implantation on maize pollen germination and cytosolic Ca2+ distribution during pollen germination process, the argon ion (Ar+) with energy of 30 keV, dose of 0.78 ×1015-13×1015 ion/cm2 was implanted into maize pollen by irradiation, and the germination of pollen and cytosolic Ca2+ distribution during pollen germination process of the Ar+ implanted pollen were studied. The results showed that when been irradiated with Ar+ with dose of 5.2×1015 ion/cm2, the germination rate of maize pollen increased remarkably, while implantation of ions with dose exceeding 5.2×1015 ion/cm2 sharply decreased the germination rate of maize pollen. Meanwhile, tracing of esterified Ca2+ fluorescence probe fluo-3 AM for intact pollen showed that variation of cytosolic Ca2+ concentration was consistent with the change of pollen fertility. The dynamics of cytosolic Ca2+ concentration caused by low energy ion implantation may be concluded as the initial effects of pollen germination.

Mapping of S-b Locus for F_1 Pollen Sterility in Cultivated Rice Using PCR Based Markers

In cultivated rice ( Oryza sativa L.), F-1 pollen sterility is controlled by at least 6 loci of the F, pollen sterility genes. To map S-b, one of loci, rice variety Taichung 65 (T65) carrying S-b(j)/S-b(j) and its near-isogenic line TIST2 carrying S-b(i)/S-b(i) were used to develop the mapping population. One hundred and fifty-eight microsatellite markers were selected to survey T65 and TISL2. RM13 on chromosome 5 was found to be polymorphic between them. Cosegregation indicated that RM13 was closely linked with locus S-b. Eleven RFLP markers were selected on the corresponding region from the genetic map of Rice Genome Research Program (RGP) of Japan to convert into sequence-tagged site (STS) markers. Amplicon length polymorphism (ALP) was carried out, but none of them was found to be polymorphic between T65 and TISL2. Then PCR-based RFLP (PBR) was done using six 4-nucleotide recognizing restriction endonucleases. Polymorphism was detected when PCR products of R830STS and R2213SSTS were digested with Taq I. Genetic analysis indicated that the distance between locus S-b and markers, R830STS, RM13 and R2213SSTS were 3.3 cM (centi-Morgan), 5.2 cM and 5.5 cM, respectively. These PCR-based markers could be directly used in marker-assisted selection. The technical system combining genetic mapping and PCR-based marker-assisted selection will facilitate the development of molecular breeding.

Pollen Assemblage Features of Modern Water Samples from the Shiyang River Drainage, Arid Region of China

Pollen analysis of 30 modem water samples from the Shivang River, an internal river system located between the Tengger and Badain Jaran deserts, Northwest China was carried out to examine the river's capacity to carry pollen and spores, and to assess the contribution of the water-borne pollen to pollen assemblages in lake sediments at the end of the river system. Results indicate the pollen assemblages in water samples consist of both local and upland pollen. Percentages of upland pollen reach 30% - 60%, and pollen assemblages in water samples do not indicate the nature of local vegetation at the sampling sites. Fluvial currents have the capacity to transport large quantities of pollen long distances, and the contribution of this fluvial transported pollen is relatively high, For example, percentages of Picea Dietr. pollen in water samples at sampling sites 130 km and 145 km away from Picea forests reach 16.5% and 7.7%, respectively. Fluvial pollen transport occurs primarily during flood periods, and pollen concentrations from the flood samples are 17.1 - 12.5 times those from normal fluvial flow. Reservoirs affect pollen transportation since pollen is deposited at reservoir inlets and pollen concentrations are much reduced at reservoir outlets. Human activity can thus change natural features of pollen transportation and deposition. The main factors influencing pollen concentrations and assemblages are sampling time, sampling location, and rainfall intensity.