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Pollen, the male gametophyte of seed plants, is extremely sensitive to UV light, which may prevent fertilization. As a result, strategies to improve plant resistance to solar ultraviolet (UV) radiation are required. The tardigrade damage suppressor protein (Dsup) is a putative DNA-binding protein that enables tardigrades to tolerate harsh environmental conditions, including UV radiation, and was therefore considered as a candidate for reducing the effects of UV exposure on pollen. Tobacco pollen was genetically engineered to express Dsup and then exposed to UV-B radiation to determine the effectiveness of the protein in increasing pollen resistance. To establish the preventive role of Dsup against UV-B stress, we carried out extensive investigations into pollen viability, germination rate, pollen tube length, male germ unit position, callose plug development, marker protein content, and antioxidant capacity. The results indicated that UV-B stress has a significant negative impact on both pollen grain and pollen tube growth. However, Dsup expression increased the antioxidant levels and reversed some of the UV-B-induced changes to pollen, restoring the proper distance between the tip and the last callose plug formed, as well as pollen tube length, tubulin, and HSP70 levels. Therefore, the expression of heterologous Dsup in pollen may provide the plant male gametophyte with enhanced responses to UV-B stress and protection against harmful environmental radiation. [ABSTRACT FROM AUTHOR]

Fig types such as "Smyrna" and "San Pedro" require pollination (called caprification in fig cultivation) to produce a commercial crop, based on the crop and pollination characteristics of figs. Caprification is the process of hanging caprifig (male fig) fruits on female fig trees to ensure the transfer of pollen from the female fig to the caprifig by a wasp (Blastophaga psenes) that lives within the caprifig. It is necessary to extend the caprification period by using caprifig genotypes that ripen at different times, as female fig fruits ripen gradually. However, as caprifigs may not be continuously available for pollinating female figs, storing suitable caprifigs is necessary. The aim of this study was to assess changes in Blastophaga psenes, the duration of Blastophaga's exit, and the viability of pollen from caprifigs of different genotypes (16 08 05, 16 08 09, 16 08 10, 16 09 10, and 16 ZF 08) stored for caprification. These stored caprifig genotypes were subsequently used for pollination three times at 8-day intervals, after which their impact on the set and quality of the edible fig fruits was evaluated. According to the average data, at the end of storage, the least B. psenes loss was obtained from the 16 08 05 (61.03%) genotype, and the highest was obtained from the 16 09 10 (67.00%) genotype. Pollen germination tended to increase with the storage of caprifig fruits, but this increase was not linear. After storage, the 16 08 09 and 16 09 10 genotypes exhibited greater pollen germination. The highest fruit set and quality were obtained when the 16 08 09 and 16 09 10 genotypes were used as pollen sources. Furthermore, since the 16 08 10 genotype is the latest ripening caprifig genotype, it has been determined that it can pollinate late-ripening "Bursa Siyahı" fruits. Principal component and path analysis demonstrated that pollen viability and germination rate were crucial in selecting caprifig genotypes for fruit set and quality. [ABSTRACT FROM AUTHOR]

Rising temperature affects agricultural production, causing food insecurity. Thus, the objective of this study was to evaluate how increased temperature influences pollen viability, photosynthetic and enzymatic responses, and their consequences on the final yield of cowpea cultivars. The cultivars BRS Acauã, BRS Guariba, BRS Gurguéia, and BRS Pajeú were used, kept in growth chambers under two temperature regimes: 24.8–30.8–37.8 °C and 20–26–33 °C. The cultivars BRS Acauã, BRS Guariba, and BRS Pajeú showed prolonged flowering and greater flower abortion, at 23.58%, 34.71%, and 25.55%, respectively, under an increase of 4.8 °C in temperature. This increase also reduced the viability of BRS Acauã and BRS Pajeú pollen by 34 and 7%, respectively. Heating increased stomatal opening and transpiration but reduced chlorophyll content. The enzymatic response varied according to cultivars and temperature. Changes in photosynthetic and enzymatic activities contribute to reducing pollen viability and productivity. BRS Acauã was the most affected, with an 82% reduction in the number of seeds and a 70% reduction in production. BRS Gurguéia maintained its production, even with an increase of 4.8 °C, and can be selected as a cultivar with the potential to tolerate high temperatures as it maintained pollen viability, with less flower abortion, with the synchrony of physiological and biochemical responses and, consequently, greater production. [ABSTRACT FROM AUTHOR]

Pollen cells require large amounts of sugars from the anther to support their development, which is critical for plant sexual reproduction and crop yield. Sugars Will Eventually be Exported Transporters (SWEETs) have been shown to play an important role in the apoplasmic unloading of sugars from anther tissues into symplasmically isolated developing pollen cells and thereby affect the sugar supply for pollen development. However, among the 17 CsSWEET genes identified in the cucumber (Cucumis sativus L.) genome, the CsSWEET gene involved in this process has not been identified. Here, a member of the SWEET gene family, CsSWEET5a, was identified and characterized. The quantitative real-time PCR and β-glucuronidase expression analysis revealed that CsSWEET5a is highly expressed in the anthers and pollen cells of male cucumber flowers from the microsporocyte stage (stage 9) to the mature pollen stage (stage 12). Its subcellular localization indicated that the CsSWEET5a protein is localized to the plasma membrane. The heterologous expression assays in yeast demonstrated that CsSWEET5a encodes a hexose transporter that can complement both glucose and fructose transport deficiencies. CsSWEET5a can significantly rescue the pollen viability and fertility of atsweet8 mutant Arabidopsis plants. The possible role of CsSWEET5a in supplying hexose to developing pollen cells via the apoplast is also discussed. [ABSTRACT FROM AUTHOR]

Wheat pollen, which is characterized by its short lifespan, exhibits rapid germination after anthesis. The preservation of wheat pollen is contingent upon environmental factors including temperature, relative humidity, light, and wind. The aim is to explicate the process for efficiently storing wheat pollen, particularly with regard to breeding. The short longevity of wheat pollen grains renders it impractical to conduct tests for pollen viability and in vitro germination on a large scale. Herein, the impact of storage temperatures and duration was assessed on pollen viability and in vitro germination in order to optimize storage conditions for preserving pollen viability. Pollen grains from 50 diverse spring wheat genotypes, each with three replicates, were harvested and stored at temperatures of 22 °C, −20 °C, and 4 °C. Subsequently, pollen viability and in vitro germination rates were determined after storage for 1, 3, and 6 days. The results revealed that storage temperatures, durations, genotypes, and their interactions had a statistically significant impact on both pollen viability and in vitro germination. Notably, when pollen was kept at 22 °C, almost all genotypes exhibited a loss of pollen viability and in vitro germination after 1, 3, and 6 days of storage. Likewise, storage at −20 °C failed to extend pollen germination. However, at a storage temperature of 4 °C, the pollen of 36 wheat genotypes exhibited a range of 6–14% for in vitro pollen germination and even remained viable for 6 days. The ANOVA revealed a substantial variation in grain number per spike between wheat genotypes, thereby highlighting the significant influence of genetic variations on grain yield. Moreover, a slight positive association between the viability of wheat pollen and the number of grains was found in the current study, suggesting that a variety of factors affect the number of grains produced. Simple linear regression analysis further revealed a significant negative correlation between pollen viability, in vitro germination, and storage time and temperature. In conclusion, our findings underscore that 4 °C is the optimal temperature for preserving pollen viability and in vitro pollen germination in spring wheat for up to 6 days. The results of the present study suggests that the pollen viability of wheat is dependent on genotype, storage temperature, and storage duration. Thus, the 36 wheat genotypes identified during the present work could be efficiently maintained at 4 °C for short-term storage (6 days) and could be further used for genetic and breeding purposes. [ABSTRACT FROM AUTHOR]

Lily (Lilium spp.) is a popular ornamental plant. Traditional genetic transformation methods have low efficiency in lily, thus development of a high-efficiency genetic transformation system is important. In this study, a novel transient transformation method involving pollen magnetofection was established and optimized pollen viability, and exogenous gene expression in magnetofected pollen and that of different germplasm were assessed. The highest germination percentage of Lilium regale pollen was 85.73% in medium containing 100 g/L sucrose, 61.5 mg/L H3BO3, and 91.5 mg/L CaCl2. A 1:4 ratio of nanomagnetic beads to DNA plasmid and transformation time of 0.5 h realized the highest transformation efficiency (88.32%). The GFP activity in transformed pollen averaged 69.66%, while that of the control pollen was 0.00%. In contrast to the control, transgenic seedlings obtained by pollination with magnetofected pollen showed strong positive GUS activity with 56.34% transformation efficiency. Among the lily germplasm tested, 'Sweet Surrender' and L. leucanthum had the highest transformation efficiency (85.80% and 54.47%), whereas L. davidii var. willmottiae was not successfully transformed. Transformation efficiency was positively correlated with pollen equatorial diameter and negatively correlated with polar axis/equatorial diameter ratio. The results suggest that pollen magnetofection-mediated transformation can be applied in Lilium but might have species or cultivar specificity. [ABSTRACT FROM AUTHOR]

Both drought and high temperature can influence the antioxidant metabolism of crop reproductive organs in different ways, affecting the fertility of reproductive organs and yield formation. However, the combined effects of drought stress and high temperature on the crop reproductive physiology have not yet been widely considered. In order to broaden our understanding of this mechanism of influence, a pond experiment was conducted using a cotton variety Yuzaomian 9110 divided into four treatment groups: control (CK), drought stress (DS), high temperature (HT), and drought stress coupled with high temperature (DS+HT). Results showed a significant negative correlation between pollen viability and superoxide anion (O2−) content, as well as hydrogen peroxide (H2O2). Compared with CK, DS did not alter O2− content in anthers, but HT treatment resulted in higher anther O2−. Compared with single-stress groups, DS+HT further promoted the formation of O2− in anthers, leading to more malondialdehyde in anthers. Moreover, a higher H2O2 content in anthers was found in DS and HT than in CK. DS+HT did not show altered H2O2 content relative to HT treatment, although its H2O2 was higher than in DS. Further analyses of the antioxidant enzyme system showed that DS had no significant effect on superoxide dismutase gene (GhCu/ZnSOD) expression, but HT and DS+HT significantly downregulated its expression. The expression of GhCu/ZnSOD was lower under DS+HT than HT, which might be why O2− content was not altered under DS treatment compared with CK and was higher in DS+HT than HT. DS and HT significantly downregulated the expression of the peroxidase gene (GhPOD) and catalase gene (GhCAT), which should be the main reason for the larger accumulation of H2O2 under drought stress and high-temperature conditions. Compared with single-stress groups, DS+HT had lower expression of GhCAT, resulting in a larger H2O2 content. Regarding the ascorbic acid–glutathione (AsA–GSH) cycle, DS and HT significantly downregulated the expression of monodehydroascorbate reductase gene (GhMDHAR) to hinder the production of AsA and upregulated the expression of ascorbate oxidase gene (GhAAO) to promote the oxidation of AsA, which was theoretically detrimental to AsA accumulation. However, HT downregulated the expression of the ascorbate peroxidase gene (GhAPX), hindering the reduction of H2O2 by AsA, which was the reason for AsA and H2O2 accumulation. Moreover, DS also significantly upregulated the expression of the dehydroascorbate reductase gene (GhDHAR2) to enhance the reduction of dehydroascorbate to form AsA, leading to a higher content of AsA under DS than HT. The combined stress significantly downregulated the expression of GhAAO to inhibit the oxidation of AsA but significantly upregulated the expression of GhMDHAR and GhDHAR2, promoting the AsA production, and downregulated the expression of GhAPX, hindering the reduction of H2O2 by AsA. All these resulted in increased AsA content under combined stresses. In addition, HT significantly downregulated the glutathione reductase gene (GhGR) expression, hindering the reduction of oxidized glutathione (GSSG), which led to the reduction of GSH. However, DS and DS+HT significantly downregulated the glutathione peroxidase gene (GhGPX) expression, resulting in the accumulation of GSH. Overall, compared with single-stress treatments, the effects of DS+HT on cotton pollen fertility and peroxide accumulation were more significant. The effects of DS+HT on the antioxidant enzyme system were mainly caused by high temperature, while the mechanism of abnormal accumulation of AsA and GSH caused by DS+HT was different from those of single-stress groups. [ABSTRACT FROM AUTHOR]

Ogura cytoplasmic male sterility (CMS) is one of the important methods for hybrid seed production in cruciferous crops. The lack of a restorer of fertility gene (Rfo) in Brassica rapa L. restricts the development and utilization of its germplasm resources. In this research, Brassica napus with the Rfo gene was used to restore the fertility of Ogura CMS B. rapa with the golden heart trait. Through the distant cross of two B. rapa and four B. napus, six interspecific hybrid combinations received F1 seeds. The six combinations were different in seed receiving. By morphological observation and molecular marker-assisted selection (MAS), in F1, individuals containing the Rfo gene all appeared fertile, while those without it remained male-sterile. The pollen viability of the fertile individuals was measured, and the fertile lines of the six interspecific hybrid combinations were different (40.68–80.49%). Three individuals (containing both Rfo and GOLDEN genes) with the highest pollen vitality (≥60%) were backcrossed with fertile cytoplasmic B. rapa, resulting in a total of 800 plants. Based on the MAS, a total of 144 plants with GOLDEN but no Rfo were screened (18%). Moreover, through morphological investigation, one individual with normal cytoplasm, stable fertility but without the restoring gene Rfo, the GOLDEN gene, and morphological characteristics similar to those of B. rapa was obtained. These results increased the diversity of B. rapa germplasm and provided a new method for the utilization of CMS germplasm in Brassica crops. [ABSTRACT FROM AUTHOR]

The hormonal imbalances, including abscisic acid (ABA) and brassinosteroid (BR) levels, caused by salinity constitute a key factor in hindering spikelet development in rice and in reducing rice yield. However, the effects of ABA and BRs on spikelet development in plants subjected to salinity stress have been explored to only a limited extent. In this research, the effect of ABA and BRs on rice growth characteristics and the development of spikelets under different salinity levels were investigated. The rice seedlings were subjected to three different salt stress levels: 0.0875 dS m−1 (Control, CK), low salt stress (1.878 dS m−1, LS), and heavy salt stress (4.09 dS m−1, HS). Additionally, independent (ABA or BR) and combined (ABA+BR) exogenous treatments of ABA (at 0 and 25 μM concentration) and BR (at 0 and 5 μM concentration) onto the rice seedlings were performed. The results showed that the exogenous application of ABA, BRs, and ABA+BRs triggered changes in physiological and agronomic characteristics, including photosynthesis rate (Pn), SPAD value, pollen viability, 1000-grain weight (g), and rice grain yield per plant. In addition, spikelet sterility under different salt stress levels (CK, LS, and HS) was decreased significantly through the use of both the single phytohormone and the cocktail, as compared to the controls. The outcome of this study reveals new insights about rice spikelet development in plants subjected to salt stress and the effects on this of ABA and BR. Additionally, it provides information on the use of plant hormones to improve rice yield under salt stress and on the enhancement of effective utilization of salt-affected soils. [ABSTRACT FROM AUTHOR]

Unlike animals that can escape threats, plants must endure and adapt to biotic and abiotic stresses in their surroundings. One such condition, cold stress, impairs the normal growth and development of plants, in which most phases of reproductive development are particularly susceptible to external low temperature. Exposed to uncomfortably low temperature at the reproductive stage, meiosis, tapetal programmed cell death (PCD), pollen viability, and fertilization are disrupted, resulting in plant sterility. Of them, cold-induced tapetal dysfunction is the main cause of pollen sterility by blocking nutrition supplements for microspore development and altering their timely PCD. Further evidence has indicated that the homeostatic imbalances of hormones, including abscisic acid (ABA) and gibberellic acid (GA), and sugars have occurred in the cold-treated anthers. Among them, cold stress gives rise to the accumulation of ABA and the decrease of active GA in anthers to affect tapetal development and represses the transport of sugar to microspores. Therefore, plants have evolved lots of mechanisms to alleviate the damage of external cold stress to reproductive development by mainly regulating phytohormone levels and sugar metabolism. Herein, we discuss the physiological and metabolic effects of low temperature on male reproductive development and the underlying mechanisms from the perspective of molecular biology. A deep understanding of cold stress response mechanisms in anther development will provide noteworthy references for cold-tolerant crop breeding and crop production under cold stress. [ABSTRACT FROM AUTHOR]

Fixing the genomic composition and multiplication through true potato seed (TPS) is an important challenge in autotetraploid potato. Disrupted meiotic cDNA (DMC1) is a meiotic gene that plays a central role in DNA recombination through crossing over in meiosis. Using the Arabidopsis DMC1 (AtDMC1) gene sequence, we retrieved Solanum tuberosum DMC1(StDMC1) from the diploid potato genome, and subsequently, sense and antisense regions of the StDMC1 gene were amplified in potato cv. Kufri Jyoti. The sense and antisense fragments were confirmed by Sanger-sequencing and cloned in the pRI101 vector. Agrobacterium-mediated transformation of the RNAi construct resulted in 44% transformation efficiency, and a total of 137 mutant lines were obtained. These mutant lines were further validated through pollen viability testing, and selected lines were used for gene expression analysis. The acetocarmine-based pollen staining showed reduced pollen viability ranging from 14 to 21% in four DMC1 mutant lines (DMC4-37, DMC4-41, DMC6-20, and DMC6-21), as compared to the Kufri Jyoti control plants, which on average exhibited 78% pollen viability. The phenotypic data was supported by the reduced expression of the StDMC1 gene in these four mutant lines compared to the control Kufri Jyoti. The results confirmed the generation of StDMC1 knockdown lines. This is the first report of StDMC1 mutant line generation in tetraploid potatoes and will be a step forward in generating non-recombinant mutants through sexual reproduction in potatoes. [ABSTRACT FROM AUTHOR]

Titanium dioxide nanoparticles (nTiO2) are widely used as fertilizers in agricultural production because they promote photosynthesis and strong adhesion. Low pollination and fertilization due to rainy weather during the litchi plant's flowering phase result in poor fruit quality and output. nTiO2 would affect litchi during the flowering and fruiting stages. This study considers how nTiO2 affects litchi's fruit quality and pollen viability during the flowering stage. The effects of nTiO2 treatment on pollen vigor, yield, and fruit quality were investigated. nTiO2 effectively improved the pollen germination rate and pollen tube length of litchi male flowers. The germination rate reached 22.31 ± 1.70%, and the pollen tube reached 237.66 μm in the 450 mg/L reagent-treated group. Spraying with 150 mg/L of nTiO2 increased the germination rate of pollen by 2.67% and 3.67% for two types of male flowers (M1 and M2) of anthesis, respectively. After nTiO2 spraying, the fruit set rates of 'Guiwei' and 'Nomici' were 46.68% and 30.33%, respectively, higher than those of the boric acid treatment group and the control group. The edibility rate, titration calculation, and vitamin C of nTiO2 treatment were significantly higher than those of the control. The nTiO2-treated litchi fruit was more vividly colored. Meanwhile, the adhesion of nTiO2 to leaves was effectively optimized by using ATP and BCS to form nTiO2 carriers and configuring nTiO2 complex reagents. These results set the foundation for future applications of titanium dioxide nanoparticles as fertilizers for agriculture and guide their application to flowers and fruits. [ABSTRACT FROM AUTHOR]

Pollen grains, the male gametophytes for reproduction in higher plants, are vulnerable to various stresses that lead to loss of viability and eventually crop yield. A conventional method for assessing pollen viability is manual counting after staining, which is laborious and hinders high-throughput screening. We developed an automatic detection tool (PollenDetect) to distinguish viable and nonviable pollen based on the YOLOv5 neural network, which is adjusted to adapt to the small target detection task. Compared with manual work, PollenDetect significantly reduced detection time (from approximately 3 min to 1 s for each image). Meanwhile, PollenDetect can maintain high detection accuracy. When PollenDetect was tested on cotton pollen viability, 99% accuracy was achieved. Furthermore, the results obtained using PollenDetect show that high temperature weakened cotton pollen viability, which is highly similar to the pollen viability results obtained using 2,3,5-triphenyltetrazolium formazan quantification. PollenDetect is an open-source software that can be further trained to count different types of pollen for research purposes. Thus, PollenDetect is a rapid and accurate system for recognizing pollen viability status, and is important for screening stress-resistant crop varieties for the identification of pollen viability and stress resistance genes during genetic breeding research. [ABSTRACT FROM AUTHOR]

Oil tea (Camellia oleifera Abel.) is one of the most important woody edible oil tree species in China, with intraspecific polyploid. In order to study the variation in pollen size and vigor of C. oleifera at ploidy level, four ploidy covers a total of 32 types of Camellia pollens as the material for the experiment. The results showed that the pollen sizes of diploid, tetraploid, hexaploidy, and octaploid were positively correlated with the ploidy level. Pollen viability of C. oleifera was determined by fluorescein diacetate (FDA) dye solution staining and medium containing 10% sucrose, 0.01% boric acid, and 1% agar germination in vitro, which indicated that the pollen viability and germination rate of the hexaploid were relatively high among the four ploidy levels, at 79.69% and 71.78% respectively. The pollen vigor of diploid NR-3, tetraploid DP43, hexaploid CJ-12, and octoploid YNYC-1 was higher than that of other materials with the same ploidy level. Knowledge of different ploidy pollen sizes and pollen viability provides basic information for formulating pollen breeding plans and pollination methods of C. oleifera. [ABSTRACT FROM AUTHOR]

The use of biotechnology for the genetic improvement of wheat (Triticum aestivum L.) has been hampered by its recalcitrance to standard transformation and regeneration protocols. Male gametes present a potentially useful option for introducing gene edits via clustered regularly interspaced short palindromic repeats (CRISPR). However, the utility of male gametes for introducing genetic improvements would be dependent on the retention of viability after treatment to introduce the CRISPR components. We have studied wheat pollen morphology and its viability in a range of germination media to identify conditions that optimize the viability of in vitro hydrated pollen. The size, shape, and aperture from seven different wheat genotypes were compared using scanning electron microscope (SEM). The SEM results revealed that the pollen of all of the wheat genotypes examined in this study were monoporate; however, a significant variation in the size of the mature pollen grains was observed. The hydrated pollen of the wheat genotypes remained viable for up to five hours at 20 ± 2 °C. Of all of the germination media tested, the medium containing 5% sucrose, 10% PEG4000, 100 mg/L boric acid, 200 mg/L calcium nitrate, 100 mg/L potassium nitrate, and 100 mg/L magnesium sulphate at pH 6.5 achieved the highest percentage of pollen germination after 5 h of hydration. Impedance Flow Cytometry (IFC) provided similar results to the in vitro germination study. This work elucidates important factors that can form the basis for a pollen-based non-genetically modified system for gene editing in wheat. [ABSTRACT FROM AUTHOR]

The development of varieties with strong tolerance is one of the important strategies to diminish the negative impact of chilling stress during heading on the spikelet fertility and yield formation of late-season rice. However, whether such genetic improvement has been made in inbred late rice lines in China is not clear. In the present study, three late-season inbred rice varieties, Xiangwanxian2 (XWX2, released in 1988), Xiangwanxian8 (XWX8, released in 1998) and Xiangwanxian17 (XWX17, released in 2008) were subjected to moderate (20 °C) and extreme (17 °C) chilling stress during heading, and the grain yield components and flowering-related traits of the three varieties in response to different temperature were investigated. The results showed that the newly released inbred late rice variety XWX17, demonstrated better chilling tolerance during heading than the early released varieties with respect to higher grain filling percentage. The improved grain filling percentage in XWX17 might be the results of increased spikelet fertility, which was attributed to the increase in pollen viability, anther dehiscence length and anther volume. In addition, the SPAD value and the chlorophyll a content of the flag leaf can be used as indicators to predict the rice spikelet fertility when suffering from chilling stress during heading. The present study provides evidence that the genetic approach has been made to improve the chilling tolerance of inbred late rice lines during heading; however, further research is needed to explore the physiological and molecular mechanism underlying the relationship between leaf characteristics and function with rice spikelet fertility. [ABSTRACT FROM AUTHOR]

Pollen fertility plays an important role in the application of heterosis in wheat (Triticum aestivum L.). However, the key genes and mechanisms underlying pollen abortion in K-type male sterility remain unclear. TAA1a is an essential gene for pollen development in wheat. Here, we explored the mechanism involved in its transcriptional regulation during pollen development, focusing on a 1315-bp promoter region. Several cis-acting elements were identified in the TAA1a promoter, including binding motifs for Arabidopsis thaliana AtAMS and AtMYB103 (CANNTG and CCAACC, respectively). Evolutionary analysis indicated that TaTDRL and TaMYB103 were the T. aestivum homologs of AtAMS and AtMYB103, respectively, and encoded nucleus-localized transcription factors containing 557 and 352 amino acids, respectively. TaTDRL and TaMYB103 were specifically expressed in wheat anthers, and their expression levels were highest in the early uninucleate stage; this expression pattern was consistent with that of TAA1a. Meanwhile, we found that TaTDRL and TaMYB03 directly interacted, as evidenced by yeast two-hybrid and bimolecular fluorescence complementation assays, while yeast one-hybrid and dual-luciferase assays revealed that both TaTDRL and TaMYB103 could bind the TAA1a promoter and synergistically increase its transcriptional activity. Furthermore, TaTDRL-EAR and TaMYB103-EAR transgenic Arabidopsis plants displayed abnormal microspore morphology, reduced pollen viability, and lowered seed setting rates. Additionally, the expression of AtMS2, a TAA1a homolog, was significantly lower in the two repressor lines than in the corresponding overexpression lines or WT plants. In summary, we identified a potential transcriptional regulatory mechanism associated with wheat pollen development. [ABSTRACT FROM AUTHOR]

Reproductive-stage heat stress (RSHS) poses a major constraint to cereal crop production by damaging main plant reproductive structures and hampering reproductive processes, including pollen and stigma viability, pollination, fertilization, grain setting and grain filling. Despite this well-recognized fact, research on crop heat stress (HS) is relatively recent compared to other abiotic stresses, such as drought and salinity, and in particular, RSHS studies in cereals are considerably few in comparison with seedling-stage and vegetative-stage-centered studies. Meanwhile, climate change-exacerbated HS, independently or synergistically with drought, will have huge implications on crop performance and future global food security. Fortunately, due to their sedentary nature, crop plants have evolved complex and diverse transient and long-term mechanisms to perceive, transduce, respond and adapt to HS at the molecular, cell, physiological and whole plant levels. Therefore, uncovering the molecular and physiological mechanisms governing plant response and tolerance to RSHS facilitates the designing of effective strategies to improve HS tolerance in cereal crops. In this review, we update our understanding of several aspects of RSHS in cereals, particularly impacts on physiological processes and yield; HS signal perception and transduction; and transcriptional regulation by heat shock factors and heat stress-responsive genes. We also discuss the epigenetic, post-translational modification and HS memory mechanisms modulating plant HS tolerance. Moreover, we offer a critical set of strategies (encompassing genomics and plant breeding, transgenesis, omics and agronomy) that could accelerate the development of RSHS-resilient cereal crop cultivars. We underline that a judicious combination of all of these strategies offers the best foot forward in RSHS tolerance improvement in cereals. Further, we highlight critical shortcomings to RSHS tolerance investigations in cereals and propositions for their circumvention, as well as some knowledge gaps, which should guide future research priorities. Overall, our review furthers our understanding of HS tolerance in plants and supports the rational designing of RSHS-tolerant cereal crop cultivars for the warming climate. [ABSTRACT FROM AUTHOR]

Roses are one of the most highly produced and purchased ornamental plants worldwide. Procurement and preservation of pollen is essential for the production of diverse rose varieties. In this study, we analyzed pollen management conditions, such as the pollen collection stage, drying time, and storage temperature, to determine optimal conditions for rose pollen management. Pollens were stored under different conditions and the pollen vitality and germination rate were investigated through an optical microscope. The vitality of pollen was an essential factor for rose breeding and depended on the storage conditions. Collecting pollen in the seventh flowering stage resulted in a relatively higher pollen yield. Drying the flower for 5 h after the anther opened improved pollen germination. The germination rate of freshly collected pollen was similar to that of pollen stored at temperatures between −20 °C and −72 °C for up to 30 days, indicating the efficacy of pollen storage at sub-zero temperatures. Since the rate of fruiting increases when pollination is performed three times, considering the time and cost of breeding, it is appropriate to pollinate three times to increase the number of seeds. This study provides an efficient pollen management method to collect and store pollen for breeding. [ABSTRACT FROM AUTHOR]

Meiosis is the least explored stage for thermotolerance in wheat. We evaluated the impact of 5 d of moderate transient daily heat stress during meiosis in the main stem spike on physiological and grain yield traits in 30 diverse wheat cultivars which vary widely in heat tolerance and sensitivity. We found that a moderate heat stress event during meiosis in the main stem spike had lasting impacts on plant growth and reproduction in heat-sensitive, but not heat-tolerant, wheat cultivars. Heat-tolerant cultivars maintained grain yield, grain number and individual grain weight in the main stem spike and also total plant grain yield and biomass in the heat stress treatment relative to the control. Heat-sensitive cultivars responded to heat stress by producing fewer and smaller grains per spikelet on the main stem, fewer tillers, lower biomass and lower total plant grain yield in the high temperature treatment relative to the control. Heat-sensitive cultivars produced higher flag leaf chlorophyll content in the high temperature treatment relative to the control than heat-tolerant cultivars during the first 3 d of heat treatment. There was small reduction in pollen viability from 98% to 96% following heat stress during meiosis which was unrelated to heat tolerance or sensitivity. Moderate transient heat stress during meiosis did not greatly reduce the production of viable male gametes, but had long-lasting negative impacts on fertilization and subsequent seed production in heat-sensitive cultivars. [ABSTRACT FROM AUTHOR]

Cultivated yam (D. rotundata) is a staple tuber crop in West Africa whose sexual reproduction control remains largely unknown despite its importance for plant breeding programs. In this paper, we compared self-pollination, intracultivar cross-pollination and intercultivar cross-pollination in three monoecious cultivars (Amoula, Heapala and Yassi). Results showed that pollen viability (49%) and stigma receptivity (40%) were similar in monoecious and dioecious plants, suggesting that autogamy could occur in monoecious plants. However, fruit and seed sets were significantly lower after self-pollination compared to cross-pollination. Overall, autogamy reached 11% and pollen lability was almost zero (<1%). The low percentage of pollen grains germinating on the stigma (37%) and pollen tubes reaching the ovules (25%) after self-pollination partly explained the low seed set. Strong inbreeding depression was observed after self-pollination and almost all fruits and about 75% of the seeds resulting from self-pollination showed malformations. Seed germination was also 20 times lower after self-pollination compared to cross-pollination. Sexual reproduction remained low in D. rotundata even after cross-pollination as fruit and seed set did not exceed 18% and 13% respectively. Moreover, comparison between intracultivar cross-pollination and self-pollination revealed intravarietal genetic diversity inside the analyzed yam cultivars. Overall, our results showed that D. rotundata has a very low tolerance to autogamy in monoecious cultivars and has developed pre- and postzygotic mechanisms to limit selfing. [ABSTRACT FROM AUTHOR]

Heat stress frequently imposes a strong negative impact on vegetative and reproductive development of plants leading to severe yield losses. Wheat, a major temperate crop, is more prone to suffer from increased temperatures than most other major crops. With heat waves becoming more intense and frequent, as a consequence of global warming, a decrease in wheat yield is highly expected. Here, we examined the impact of a short-term (48 h) heat stress on wheat imposed during reproduction at the pollen mitosis stage both, at the physiological and molecular level. We analyzed two sets of summer wheat germplasms from Australia (Kukri, Drysdale, Gladius, and RAC875) and Europe (Epos, Cornetto, Granny, and Chamsin). Heat stress strongly affected gas exchange parameters leading to reduced photosynthetic and transpiration rates in the European cultivars. These effects were less pronounced in Australian cultivars. Pollen viability was also reduced in all European cultivars. At the transcriptional level, the largest group of heat shock factor genes (type A HSFs), which trigger molecular responses as a result of environmental stimuli, showed small variations in gene expression levels in Australian wheat cultivars. In contrast, HSFs in European cultivars, including Epos and Granny, were strongly downregulated and partly even silenced, while the high-yielding variety Chamsin displayed a strong upregulation of type A HSFs. In conclusion, Australian cultivars are well adapted to moderate heat stress compared to European summer wheat. The latter strongly react after heat stress application by downregulating photosynthesis and transpiration rates as well as differentially regulating HSFs gene expression pattern. [ABSTRACT FROM AUTHOR]

N-linked glycosylation is one of the key post-translational modifications. α1,3-Fucosyltransferase (OsFucT) is responsible for transferring α1,3-linked fucose residues to the glycoprotein N-glycan in plants. We characterized an Osfuct mutant that displayed pleiotropic developmental defects, such as impaired anther and pollen development, diminished growth, shorter plant height, fewer tillers, and shorter panicle length and internodes under field conditions. In addition, the anthers were curved, the pollen grains were shriveled, and pollen viability and pollen number per anther decreased dramatically in the mutant. Matrix-assisted laser desorption/ionization time-of-flight analyses of the N-glycans revealed that α1,3-fucose was lacking in the N-glycan structure of the mutant. Mutant complementation revealed that the phenotype was caused by loss of Osfuct function. Transcriptome profiling also showed that several genes essential for plant developmental processes were significantly altered in the mutant, including protein kinases, transcription factors, genes involved in metabolism, genes related to protein synthesis, and hypothetical proteins. Moreover, the mutant exhibited sensitivity to an increased concentration of salt. This study facilitates a further understanding of the function of genes mediating N-glycan modification and anther and pollen development in rice. [ABSTRACT FROM AUTHOR]

Increasing use of iron oxide nanoparticles in medicine and environmental remediation has led to concerns regarding exposure of these nanoparticles to the public. However, limited studies are available to evaluate their effects on the environment, in particular on plants and food crops. Here, we investigated the effects of positive (PC) and negative (NC) charged iron oxide (Fe2O3) nanoparticles (IONPs) on the physiology and reproductive capacity of Arabidopsis thaliana at concentrations of 3 and 25 mg/L. The 3 mg/L treated plants did not show evident effects on seeding and root length. However, the 25 mg/L treatment resulted in reduced seedling (positive-20% and negative-3.6%) and root (positive-48% and negative-negligible) length. Interestingly, treatment with polyethylenimine (PEI; IONP-PC coating) also resulted in reduced root length (39%) but no change was observed with polyacrylic acid (PAA; IONP-NC coating) treatment alone. However, treatment with IONPs at 3 mg/L did lead to an almost 5% increase in aborted pollen, a 2%-6% reduction in pollen viability and up to an 11% reduction in seed yield depending on the number of treatments. Interestingly, the treated plants did not show any observable phenotypic changes in overall size or general plant structure, indicating that environmental nanoparticle contamination could go dangerously unnoticed. [ABSTRACT FROM AUTHOR]

Owing to the wide variation in their morphological characteristics across diverse geographies, the identification and classification of plants in the Nymphaea genus are challenging. Therefore, the present study investigated the pollen morphological characteristics of hardy water lilies (N. 'Rose Arey', N. 'Perry's Fire Opal', and N. 'Peter Slocum'), their relationship with species classification and system evolution, and their cross-compatibility with three strains of Nymphaeahybrid (NH-1, NH-2, and NH-3), a tropical waterlily. Pollen of the hardy water lilies was single-grained, oblate, and 18.31–20.47 × 32.51–37.64 μm. The pollen apertures were of the ring-groove type, and the pollen exine ornamentation was rod- and tumour-shaped. Pollen grains of different species differed in size, the obviousness of tumour ornamentation, and the size and density of rod-like ornamentation; their germination rates also differed significantly. Viabilities of N. 'Rose Arey' and N. 'Peter Slocum' pollen were the highest and lowest, respectively. According to the artificial pollination results, all hybrid combinations except three (NH-1 × N. 'Peter Slocum', NH-2 × N. 'Peter Slocum', and NH-3 × N. 'Peter Slocum') bore seeds; combinations with NH-2 as the female parent and N. 'Rose Arey' as the male parent had the highest seed-setting rates. [ABSTRACT FROM AUTHOR]

High summer temperatures (day/night: 40 °C/30 °C) are known to cause poor fruiting, reduced fruit/seed weight, and delayed fruit development in the "Da Hong" red-fleshed pitaya (Hylocereus polyrhizus); however, the mechanisms of these effects are unknown. This study examined how high temperature (HT, 40 °C/30 °C) affects stamen and pistil fertility through pollination combinations and fruit set in "Da Hong" pitaya and evaluated photosystem II (PSII) efficiency in yellowed shoots (cladodes). The in-vitro pollen germination rate was significantly higher at optimal temperature (OT, 30 °C/20 °C) than at HT, which was also associated with a reduced flower length, poor fruit set, small fruit size, and low number of seeds after self-pollination. Meanwhile, fruit set and fruit development were remarkably improved by using the pollens taken from plants grown at OT and moderate temperature (MH, 35 °C/25 °C) after mutual cross pollination, indicating that the reduction in seed number caused by incomplete fertilization due to inferior pollen viability was the main cause of poor fruiting under HT rather than pistil or photoinhibition, although HT treatment was linked to reductions of PSII efficiency at both the adaxial and abaxial ends of shoots. The results suggested that pollen viability was most affected under HT stress; thus, yielding remarkably reduced fruiting. [ABSTRACT FROM AUTHOR]

In Brassica, interspecific hybridisation plays an important role in the formation of allopolyploid cultivars. In this study, the ploidy of F1 and F2 generations resulting from interspecific hybridisation between B. oleracea inbred lines of head cabbage (B. oleracea L. var. capitata) (2n = 18) and kale (B. oleracea L. var. acephala) (2n = 18) with inbred lines of rapeseed (B. napus L.) (2n = 38) was examined by flow cytometry analysis and chromosome observation. Furthermore, the effect of meiotic polyploidisation on selected phenotypic and anatomical traits was assessed. The F1 hybrids of head cabbage × rapeseed (S3) and kale × rapeseed crosses (S20) were allotriploids with 2n = 28 chromosomes, and nuclear DNA amounts of 1.97 (S3) and 1.99 pg (S20). These values were intermediate between B. oleracea and B. napus. In interspecific hybrids of the F2 generation, which were derived after self-pollination of F1 hybrids (FS3, FS20) or by open crosses between F1 generation hybrids (FC320, FC230), the chromosome numbers were similar 2n = 56 or 2n = 55, whereas the genome sizes varied between 3.81 (FS20) and 3.95 pg 2C (FC230). Allohexaploid F2 hybrids had many superior agronomic traits compared to parental B. napus and B. oleracea lines and triploid F1 hybrids. In the generative stage, they were characterised by larger flowers and flower elements, such as anthers and lateral nectaries. F2 hybrids were male and female fertile. The pollen viability of F2 hybrids was comparable to parental genotypes and varied from 75.38% (FS3) to 88.24% (FC320), whereas in triploids of F1 hybrids only 6.76% (S3) and 13.46% (S20) of pollen grains were fertile. Interspecific hybrids of the F2 generation derived by open crosses between plants of the F1 generation (FC320, FC230) had a better ability to set seed than F2 hybrids generated from the self-pollination of F1 hybrids. In the vegetative stage, F2 plants had bigger and thicker leaves, larger stomata, and significantly thicker layers of palisade and spongy mesophyll than triploids of the F1 generation and parental lines of B. oleracea and B. napus. The allohexaploid F2 hybrids analysed in this study can be used as innovative germplasm resources for further breeding new vegetable Brassica crops at the hexaploid level. [ABSTRACT FROM AUTHOR]

Olea europaea L. is a crop typical of the Mediterranean area that has an important role in economy, society, and culture of this region. Climate change is expected to have significant impact on this crop, which is typically adapted to certain pedo-climatic characteristics of restricted geographic areas. In this scenario, the aim of this study was to evaluate the time-course response of pollen viability to different combinations of temperature and humidity. The study was performed comparing flowering time and pollen functionality of O. europaea from twelve cultivars growing at the same site belonging to the Campania olive collection in Italy. Pollen was incubated at 12 °C, 22 °C, and 36 °C in combination with 50% RH or 100% RH treatments for 5 days. The results highlighted that a drastic loss of pollen viability occurs when pollen is subjected to a combination of high humidity and high temperature, whereas 50% RH had less impact on pollen thermotolerance, because most cultivars preserved a high pollen viability over time. In the ongoing climate change scenario, it is critical to assess the effect of increasing temperatures on sensitive reproductive traits such as pollen viability to predict possible reduction in crop yield. Moreover, the results highlighted that the effect of temperature increase on pollen thermotolerance should be evaluated in combination with other environmental factors such as humidity conditions. The screening of olive cultivars based on pollen thermotolerance is critical in the ongoing climate change scenario, especially considering that the economic value of this species relies on successful fertilization and embryo development, and also that production cycle of Olea europaea can be longer than a hundred years. [ABSTRACT FROM AUTHOR]

Simple Summary: The genus Ranunculus (buttercup) includes over 600 species, some of which are endangered, e.g., Illyrian Buttercup. Knowledge of the reproductive biology of such species may be crucial for conservation action. For this purpose, six species with different reproduction modes (nonclonal reproducing sexually by seeds only, clonal propagating by seeds and additionally vegetatively and apomictic) were observed. Selected features related to the efficiency of sexual reproduction were described: pollen viability, number of fruit set, seed viability and germination. It has been shown that in clonal species, which include the Illyrian Buttercup, the efficiency of sexual reproduction is lower compared to nonclonal species. The results will support conservation action taken for this species. Generative processes have been evaluated in six European buttercup species in order to verify the hypothesis that the reproduction efficiency of clonal species is lower than that of nonclonal ones. The study covered common species (Ficaria verna, Ranunculus auricomus, R. bulbosus, R. cassubicus, R. lanuginosus) and the endangered R. illyricus. The following properties have been assessed: pollen viability (staining method), pollen grain germination and the pollen-tube elongation in pistil tissues (fluorescence microscopy), seed formation efficiency, seed viability (tetrazolium test) and germination ability by introducing factors interrupting dormancy (low temperature and gibberellin application). Additionally, the pistil morphology was documented for R. bulbosus, R. illyricus and R. cassubicus using SEM techniques. It was demonstrated that the reproductive efficiency, expressed as the production of viable seeds able to germinate, was significantly higher in the species reproducing sexually (especially in R. lanuginosus) compared to the clonal ones. However, the complexity observed leads to separation of an additional group (cluster) of apomictic species: R. auricomus and R. cassubicus, distinguished by the lowest pollen viability and a low ability of the seeds to germinate. In the vegetatively reproducing R. illyricus, the seed formation efficiency was just 13.2% despite the having highest number of pistils in its flowers. The developed seeds of this species observed in our experiment were viable, but in general effective methods to stimulate their germination have not been proposed yet. Here, the first comparative study concerning the biology of sexual reproduction of R. illyricus is presented in the context of its decreasing distribution in natural habitats. [ABSTRACT FROM AUTHOR]

Among many challenges in chrysanthemum cross-breeding, the access to viable pollen for hybridization of cultivars distant in location and different in flowering time is required. Low pollen viability along with incompatibility are mainly responsible for low seed set in modern chrysanthemum cultivars. The aim of the study was to test various temperatures and periods of pollen storage of Chrysanthemum × morifolium in order to elaborate the method of chrysanthemum pollen preservation for cross-breeding purposes. In the first experiment, in vitro pollen germination of four cultivars was investigated following storage at 20 °C, 4 °C, −20 °C, and −80 °C, for one, four, and eight weeks. The second experiment focused on in vivo seed set after one week pollen treatment with 20 °C, 4 °C, −20 °C, and −80 °C (three pollen donor cultivars tested). Pollen in vitro germinability, as well as seed set efficiency, was generally low and cultivar dependent. Independent of the period of storage, stored pollen germinability was lower (5.30–6.63%) than fresh pollen (8.15%). Incubation of pollen in −80 °C significantly increased pollen germinability (9.80%), as well as seed set efficiency in comparison to control (19.28% and 10.21%, respectively) provided the cultivars are compatible. Among cultivars, the highest germinability of pollen was found in 'Brda' and 'Donna' (8.2% and 8.23%, respectively), while 'Bydgoszczanka' showed the lowest germinability (2.97%). There were also pollen genotype dependent effects in in vivo seed set efficiency, which was highest in 'Brda' (17.57%) and much lower in 'Jutrzenka' and 'Polka' (1.34% and 0.39%, respectively), which contributed to the incompatibility of crossed cultivars rather than pollen viability. [ABSTRACT FROM AUTHOR]

Abiotic stress, particularly heat stress, affects various parts of the cotton plant and ultimately impacts the seed cotton yield. Different portions of a single cotton plant of a cultivar exhibit variable responses to stress during reproductive and vegetative phases. To test this hypothesis, physiological and morphological traits related to heat stress were observed for two flowering positions in 13 genotypes of upland cotton. These genotypes were sown in field conditions in triplicate following a randomized complete block design. Data were collected for pollen germination, pollen viability, cell membrane thermostability, chlorophyll content, boll weight, and boll retention for both the top and bottom branches of each genotype. The collected data were analyzed for the identification of variability within and between genotypes for these two flowering positions. Tukey's test was applied to estimate the significance of differences between genotypes and positions within each genotype. Results showed that the two positions within the same plant statistically varied from each other. The bottom branches of the genotypes performed significantly better for all traits measured except boll weight. The genotype AA-933 performed best for pollen germination and boll retention, while CYTO-608 exhibited maximum pollen viability in both the bottom and top flower positions compared with other genotypes. Overall, MNH-1016 and CIM-602 showed better cell membrane thermostability and chlorophyll content, respectively. This intra-plant variability can be further exploited in breeding programs to enhance the stress tolerance capabilities of the resulting varieties. [ABSTRACT FROM AUTHOR]

Global warming leads to a progressive rise in environmental temperature. Plants, as sessile organisms, are threatened by these changes; the male gametophyte is extremely sensitive to high temperature and its ability to preserve its physiological status under heat stress is known as acquired thermotolerance. This latter can be achieved by exposing plant to a sub-lethal temperature (priming) or to a progressive increase in temperature. The present research aims to investigate the effects of heat priming on the functioning of tobacco pollen grains. In addition to evaluating basic physiological parameters (e.g., pollen viability, germination and pollen tube length), several aspects related to a correct pollen functioning were considered. Calcium (Ca2+) level, reactive oxygen species (ROS) and related antioxidant systems were investigated, also to the organization of actin filaments and cytoskeletal protein such as tubulin (including tyrosinated and acetylated isoforms) and actin. We also focused on sucrose synthase (Sus), a key metabolic enzyme and on the content of main soluble sugars, including UDP-glucose. Results here obtained showed that a pre-exposure to sub-lethal temperatures can positively enhance pollen performance by altering its metabolism. This can have a considerable impact, especially from the point of view of breeding strategies aimed at improving crop species. [ABSTRACT FROM AUTHOR]

Heat tolerance is important for the sustainable production of many crops, including chile pepper. Tolerance to high temperature is complex and involves various component traits, with pollen viability being among the most important. in vitro pollen assays for heat tolerance have been widely used in chile pepper; however, associations between the pollen treatment and pollination have not been widely explored. The objectives of this study were to validate the utility of in vitro heat stress pollen characterization through in vivo pollination during summer and winter seasons and to evaluate the cross-compatibility among wild and domesticated species to initiate introgression population development. Seven entries of wild and domestic Capsicum species grown during the summer and winter seasons were used to evaluate pollination success rate. Pollen was either used directly or treated at 38 °C for four hours before making reciprocal self- and cross-pollination among all the entries. Significant associations between in vitro pollen treatment and pollination success rate during summer and winter seasons were identified. Heat treatment was a greater contributor to variability than the growing environment, which validates previous reports on the usefulness of studying pollen in vitro in selection for heat tolerance. Accessions of the wild progenitor C. annuum var glabriusculum, PBC 1969 and PBC 1970, were identified as a potential heat-tolerant source for use in breeding and future research. This work provides a basis for future research in exploring additional heat tolerance components as well as for the development of phenotyping assays for pollen or other floral traits. [ABSTRACT FROM AUTHOR]

Gradually increasing temperatures at global and local scales are causing heat stress for cool and summer-season food legumes, such as lentil (Lens culinaris Medik.), which is highly susceptible to heat stress, especially during its reproductive stages of development. Hence, suitable strategies are needed to develop heat tolerance in this legume. In the present study, we tested the effectiveness of heat priming (HPr; 6 h at 35 °C) the lentil seeds and a foliar treatment of γ-aminobutyric acid (GABA; 1 mM; applied twice at different times), singly or in combination (HPr+GABA), under heat stress (32/20 °C) in two heat-tolerant (HT; IG2507, IG3263) and two heat-sensitive (HS; IG2821, IG2849) genotypes to mitigate heat stress. The three treatments significantly reduced heat injury to leaves and flowers, particularly when applied in combination, including leaf damage assessed as membrane injury, cellular oxidizing ability, leaf water status, and stomatal conductance. The combined HPr+GABA treatment significantly improved the photosynthetic function, measured as photosynthetic efficiency, chlorophyll concentration, and sucrose synthesis; and significantly reduced the oxidative damage, which was associated with a marked up-regulation in the activities of enzymatic antioxidants. The combined treatment also facilitated the synthesis of osmolytes, such as proline and glycine betaine, by upregulating the expression of their biosynthesizing enzymes (pyrroline-5-carboxylate synthase; betaine aldehyde dehydrogenase) under heat stress. The HPr+GABA treatment caused a considerable enhancement in endogenous levels of GABA in leaves, more so in the two heat-sensitive genotypes. The reproductive function, measured as germination and viability of pollen grains, receptivity of stigma, and viability of ovules, was significantly improved with combined treatment, resulting in enhanced pod number (21–23% in HT and 35–38% in HS genotypes, compared to heat stress alone) and seed yield per plant (22–24% in HT and 37–40% in HS genotypes, in comparison to heat stress alone). The combined treatment (HPr+GABA) was more effective and pronounced in heat-sensitive than heat-tolerant genotypes for all the traits tested. This study offers a potential solution for tackling and protecting heat stress injury in lentil plants. [ABSTRACT FROM AUTHOR]

Our study attempted to elucidate the significance of floral and pollen traits of the highly nutritious tropical trees Moringa oleifera and Moringa peregrina for their reproductive success under arid conditions. We found that the pollen grains of both species were immersed in a pollenkitt that constituted ~60% of the pollen. Successful pollination was achieved by large bees inserting the pollen into a narrow stylar tube. We found that, upon removal of the pollenkitt, approximately 65% fewer pollen grains penetrated the stylar tube for both species. The pollenkitt protected against heat and desiccation, while removing the pollenkitt resulted in significantly reduced levels of the viability of pollen grains, especially in M. oleifera, and significantly reduced levels of germinability in both species. The stylar tube provided high protection for pollen grains against heat and desiccation even when the pollenkitt was removed. Chemical analysis of pollenkitts of the two species revealed a waxy blend of 21 hydrocarbon compounds, in which n-alkanes constituted >90% of the compounds and their identity corresponded to known plant and animal hydrocarbons, associated with protection against heat and water stress. We concluded that, under arid conditions, the reproductive success of both Moringa species is potentially enhanced by their unique floral and pollen traits. This supports the prospect of cultivating M. oleifera and Moringa peregrina as food crops in arid regions across the globe. [ABSTRACT FROM AUTHOR]

Simple Summary: On wheat polyploidisation, the major meiotic gene ZIP4, duplicated and diverged, such that tetraploid and hexaploid wheat each carry three and four copies of ZIP4, respectively. Surprisingly, this study demonstrates that, in hexaploid wheat, despite the presence of the other three ZIP4 copies, the duplicated ZIP4 copy is required to prevent major abnormalities during meiosis. Although there is greater disruption of subsequent male rather than female fertility, the duplicated ZIP4 copy preserves up to 50% of the grain number. High wheat fertility is important since it is consumed by over 4.5 billion people on the planet, of whom 2.5 billion are dependent on it. This study highlights the potentially extraordinary value of the wheat ZIP4 duplication, mandating further studies to unravel the complexity of the ZIP4 phenotype in this global crop. Although most flowering plants are polyploid, little is known of how the meiotic process evolves after polyploidisation to stabilise and preserve fertility. On wheat polyploidisation, the major meiotic gene ZIP4 on chromosome 3B duplicated onto 5B and diverged (TaZIP4-B2). TaZIP4-B2 was recently shown to promote homologous pairing, synapsis and crossover, and suppress homoeologous crossover. We therefore suspected that these meiotic stabilising effects could be important for preserving wheat fertility. A CRISPR Tazip4-B2 mutant was exploited to assess the contribution of the 5B duplicated ZIP4 copy in maintaining pollen viability and grain setting. Analysis demonstrated abnormalities in 56% of meiocytes in the Tazip4-B2 mutant, with micronuclei in 50% of tetrads, reduced size in 48% of pollen grains and a near 50% reduction in grain number. Further studies showed that most of the reduced grain number occurred when Tazip4-B2 mutant plants were pollinated with the less viable Tazip4-B2 mutant pollen rather than with wild type pollen, suggesting that the stabilising effect of TaZIP4-B2 on meiosis has a greater consequence in subsequent male, rather than female gametogenesis. These studies reveal the extraordinary value of the wheat chromosome 5B TaZIP4-B2 duplication to agriculture and human nutrition. Future studies should further investigate the role of TaZIP4-B2 on female fertility and assess whether different TaZIP4-B2 alleles exhibit variable effects on meiotic stabilisation and/or resistance to temperature change. [ABSTRACT FROM AUTHOR]

In the era of technology, nanotechnology has been introduced as a new window for agriculture. However, no attention has been paid to the effect of cerium dioxide nanoparticles (nCeO2) on the reproductive stage of plant development to evaluate their toxicity and safety. To address this important topic, bean plants (Phaseolus vulgaris L.) treated aerially with nCeO2 suspension at 250–2000 mg L−1 were cultivated until flowering and seed production in the greenhouse condition. Microscopy analysis was carried out on sectioned anthers and ovules at different developmental stages. The pollen's mother cell development in nCeO2 treatments was normal at early stages, the same as control plants. However, the results indicated that pollen grains underwent serious structural damages, including chromosome separation abnormality at anaphase I, pollen wall defect, and pollen grain malformations in nCeO2-treated plants at the highest concentration, which resulted in pollen abortion and yield losses. On the ovule side, the progression of development only at the highest concentration was modified in the two-nucleated embryo sac stage, probably due to apoptosis in nuclei. Nevertheless, the findings confirmed the more pronounced vulnerability of male reproductive development under nCeO2 exposure than female development. The higher concentration decreased seed productivity, including seed set in either pods or whole plant (13% and 18% compared to control, respectively). The data suggested the potential application of nCeO2 at optimal dosages as a plant productivity ameliorative. However, a higher dosage is considered as an eco-environmental hazard. To our best knowledge, this is the first study analyzing reproductive plant response upon exposure to nCeO2. [ABSTRACT FROM AUTHOR]

Gac is valued for the nutritious aril surrounding its seed. When pollinators are limited or when flower sex expression is female-biased, hand pollination is necessary. Here, female flowers were hand pollinated with male flowers or pollen stored for up to 84 days at 4 or −20 °C, and fruit set and some qualities of mature fruit were evaluated. Cool storage reduced pollen viability (germinability and pollen tube length) and compromised fruit set (10–87%) compared with fresh pollen (97%). Fruit weight was also reduced at least by 8%, and oil concentration in aril by 40%. However, the lycopene and β-carotene concentrations in aril were largely uncompromised, and some fruits were of a marketable weight (>1.2 kg) and quality. Cool storage is a low-cost method for the short-term storage of Gac pollen. However, methods for drying pollen to an inactive state need investigation for a storage protocol, and for improvements in fruit set and fruit physicochemical qualities using hand pollination. [ABSTRACT FROM AUTHOR]

Interspecific hybridization between B. oleracea inbred lines of head cabbage, Brussels sprouts, kale and B. taurica and inbred lines of rapeseed (B. napus L.) were performed aiming at the development of the new sources of genetic variability of vegetable Brassicas. Using conventional crossings and the embryo-rescue techniques the following interspecific hybrids were developed: 11 genotypes of F1 generation, 18 genotypes of F2 and F1 × F2 generations (produced after self- and cross-pollination of interspecific F1 hybrids), 10 plants of the BC1 generation (resulted from crossing head cabbage cytoplasmic male-sterile lines with interspecific hybrids of the F2 and F1 generations) and 8 plants of BC1 × (F1 × F2). No viable seeds of the BC2 generation (B. oleracea) were obtained due to the strong incompatibility and high mortality of embryos. The morphological characteristics during the vegetative and generative stages, pollen characteristics, seed development and propagation, nuclear DNA contents and genome compositions of interspecific hybrids were analyzed. All the interspecific F1 hybrids were male-fertile with a majority of undeveloped and malformed pollen grains. They showed intermediate values for morphological traits and nuclear DNA contents and had nearly triploid chromosomal numbers (27 to 29) compared with parental lines. The F2 generation had a doubled nuclear DNA content, with 52 and 56 chromosomes, indicating their allohexaploid nature. F2 hybrids were characterized by a high heterosis of morphological characteristics, viable pollen and good seed development. F1 × F2 hybrids were male-fertile with a diversified DNA content and intermediate pollen viability. BC1 plants were male-sterile with an intermediate nuclear DNA content between the F2 and head cabbage, having 28 to 38 chromosomes. Plants of the BC1 × (F1 × F2) generation were in majority male-fertile with 38–46 chromosomes, high seed set, high heterosis and intermediate values for morphological traits. The obtained interspecific hybrids are valuable as new germplasm for improving Brassica-breeding programs. [ABSTRACT FROM AUTHOR]

Products containing graphene-related materials (GRMs) are becoming quite common, raising concerns for environmental safety. GRMs have varying effects on plants, but their impact on the sexual reproduction process is largely unknown. In this study, the effects of few-layer graphene (FLG) and a similarly layered phyllosilicate, muscovite mica (MICA), were tested in vivo on the reproductive structures, i.e., pollen and stigma, of Cucurbita pepo L. ssp. pepo 'greyzini' (summer squash, zucchini). Pollen was exposed to FLG or MICA, after careful physical-chemical characterization, at concentrations of 0.5 and 2 mg of nanomaterial (NM) per g of pollen for up to six hours. Following this, pollen viability was tested. Stigmas were exposed to FLG or MICA for three hours and then analyzed by environmental scanning electron microscopy to verify possible alterations to their surface. Stigmas were then hand-pollinated to verify the effects of the two NMs on pollen adhesion and in vivo pollen germination. FLG and MICA altered neither pollen viability nor the stigmatic surface. However, both NMs equivalently decreased pollen adhesion and in vivo germination compared with untreated stigmas. These effects deserve further attention as they could impact on production of fruits and seeds. Importantly, it was shown that FLG is as safe as a naturally occurring nanomaterial. [ABSTRACT FROM AUTHOR]

In the present study, we analyzed the main parameters related with the potential fertilization ability of two grapevine varieties, Godello and Mencía, during the years 2017 and 2018. The research was carried out in two vineyards of the Galician winegrowing Designation of Origin areas of Ribeiro and Ribeira Sacra. Ten vines of each variety were selected for bunch and flower counting, pollen calculations, pollen viability studies by means of aceto-carmine (AC) stain and 2, 3, 5-triphenyl tetrazolium chloride (TTC) methods, and the determination of their germination rate. In all vineyards the 50% fruitset was reached, except for Godello in Cenlle during 2017. The mean coulure value was higher for Godello (40.5%) than for Mencía (31%). Analyzing the pollen production per plant and airborne pollen levels, we observed important discordances between them, which can be due to the influence of weather conditions and be related with self-pollination processes. We found important differences on pollen viability depending on the applied method and variety, with higher values for the AC method than the TTC for both varieties in all study plots, and higher values for Mencía variety than Godello. Regarding germination rates, we observed a marked reduction in 2017 with respect to 2018, in all study sites and for both varieties. The analyzed parameters were useful to explain the different productive abilities of Godello and Mencía varieties in the two studied bioclimatic regions of Ribeiro and Ribeira Sacra. [ABSTRACT FROM AUTHOR]

The seeds of the dioecious shrub jojoba (Simmondsia chinensis (Link) Schneider) yield a liquid wax that is in high demand for the cosmetics industry. While elite female cultivars of this species are currently clonally propagated, male plants are grown from seed, resulting in large variations in both the flowering period and the pollen viability, and hence large variation in yields. We characterized the existing male plant material in a local plantation as a platform for future selection of elite male cultivars that would produce sufficient amounts of viable pollen throughout the extended flowering period of the female cultivars. Using as a guide the number of viable pollen grains per 1-m branch, defined here as the calculated effective pollen productivity (EPP), we identified plants with an elevated EPP that flower concurrently with the female cultivars. [ABSTRACT FROM AUTHOR]

Heat stress is a major limiting factor for crop productivity. Tomato is highly sensitive to heat stress, which can result in a total yield loss. To adapt to current and future heat stress, there is a dire need to develop heat tolerant cultivars. Here, we review recent attempts to improve screening for heat tolerance and to exploit genetic and genomic resources in tomatoes. We provide key factors related to phenotyping environments and traits (morphological, physiological, and metabolic) to be considered to identify and breed thermo-tolerant genotypes. There is significant variability in tomato germplasm that can be harnessed to breed for thermo-tolerance. Based on our review, we propose that the use of advanced backcross populations and chromosome segments substitution lines is the best means to exploit variability for heat tolerance in non-cultivated tomato species. We applied a meta quantitative trait loci (MQTL) analysis on data from four mapping experiments to co-localize QTL associated with heat tolerance traits (e.g., pollen viability, number of pollen, number of flowers, style protrusion, style length). The analysis revealed 13 MQTL of which 11 were composed of a cluster of QTL. Overall, there was a reduction of about 1.5-fold in the confidence interval (CI) of the MQTL (31.82 cM) compared to the average CI of individual QTL (47.4 cM). This confidence interval is still large and additional mapping resolution approaches such as association mapping and multi-parent linkage mapping are needed. Further investigations are required to decipher the genetic architecture of heat tolerance surrogate traits in tomatoes. Genomic selection and new breeding techniques including genome editing and speed breeding hold promise to fast-track development of improved heat tolerance and other farmer- and consumer-preferred traits in tomatoes. [ABSTRACT FROM AUTHOR]

The sensitivity of sorghum to pre-flowering cold stress, resulting in reduced pollen viability and poor seed set, is a major constraint for expanding growing areas into higher altitudes and latitudes. Nevertheless, compared to juvenile cold tolerance, reproductive cold tolerance in sorghum has received much less attention so far, and very little is known about its inheritance in F1-hybrids. We have composed a representative factorial (n = 49 experimental F1-hybrids) for a comprehensive study on heterosis and combining ability for crucial tolerance traits as spikelet fertility (panicle harvest index), seed yield and pollen viability, using field trials in stress- and control environments in Germany and Mexico as well as climate chamber experiments. Our results indicate a heterotic and rather dominant inheritance of reproductive cold tolerance in sorghum, with strong effects of female general combining ability (GCA) on F1-hybrid performance in our material. These findings, together with the comparatively low contribution of specific combining ability (SCA) effects and high heritability estimates, suggest that robust and efficient enhancement of reproductive cold tolerance is feasible via hybrid breeding. [ABSTRACT FROM AUTHOR]