Sensitivity of sorghum pollen and pistil to high-temperature stress.
ABSTRACT: High temperature (HT) decreases seed set percentage in sorghum (Sorghum bicolor [L.] Moench). The relative sensitivity of pollen and particularly pistil and the mechanistic response that induces tolerance or susceptibility to HT are not well known and hence are the major objectives of this research. The male sterile (ATx399) and fertile (RTx430) lines were exposed to 30/20 °C (optimum temperature), 36/26 °C (HT1 ), and 39/29 °C (HT2 ) from the start of booting to seed set in a controlled environment. Similarly, in the field, HT stress was imposed using heat tents. HT stress decreased pollen germination. Relatively high levels of reactive oxygen species and decreased antioxidant enzyme activity and phospholipid unsaturation were observed in pollen compared to pistil under HT. The severe cell organelle damage was observed in pollen and pistil at 36/26 and 39/29 °C, respectively. The seed set percentage was higher in HT-stressed pistil pollinated with optimum-temperature pollen. Direct and reciprocal crosses indicate that pollen was more sensitive with larger decreases in seed set percentage than pistil under HT stress. The negative impact was greater in pollen than pistil at lower temperatures. Overall, pollen was more sensitive than pistil to HT stress because it is more susceptible to oxidative damage than pistil.
Project description:The global climate change is leading to increased frequency of heatwaves with crops getting exposed to extreme temperature events. Such temperature spikes during the reproductive stage of plant development can harm crop fertility and productivity. Here we report the response of short-term heat stress events on the pollen and pistil tissues in a commercially grown cultivar of <i>Brassica napus</i>. Our data reveals that short-term temperature spikes not only affect pollen fitness but also impair the ability of the pistil to support pollen germination and pollen tube growth and that the heat stress sensitivity of pistil can have severe consequences for seed set and yield. Comparative transcriptome profiling of non-stressed and heat-stressed (40°C for 30 min) pollen and pistil (stigma + style) highlighted the underlying cellular mechanisms involved in heat stress response in these reproductive tissues. In pollen, cell wall organization and cellular transport-related genes possibly regulate pollen fitness under heat stress while the heat stress-induced repression of transcription factor encoding transcripts is a feature of the pistil response. Overall, high temperature altered the expression of genes involved in protein processing, regulation of transcription, pollen-pistil interactions, and misregulation of cellular organization, transport, and metabolism. Our results show that short episodes of high-temperature exposure in <i>B. napus</i> modulate key regulatory pathways disrupted reproductive processes, ultimately translating to yield loss. Further investigations on the genes and networks identified in the present study pave a way toward genetic improvement of the thermotolerance and reproductive performance of <i>B. napus</i> varieties.
Project description:Understanding the adaptive changes in wheat pollen lipidome under high temperature (HT) stress is critical to improving seed set and developing HT tolerant wheat varieties. We measured 89 pollen lipid species under optimum and high day and/or night temperatures using electrospray ionization-tandem mass spectrometry in wheat plants. The pollen lipidome had a distinct composition compared with that of leaves. Unlike in leaves, 34:3 and 36:6 species dominated the composition of extraplastidic phospholipids in pollen under optimum and HT conditions. The most HT-responsive lipids were extraplastidic phospholipids, phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol, phosphatidic acid, and phosphatidylserine. The unsaturation levels of the extraplastidic phospholipids decreased through the decreases in the levels of 18:3 and increases in the levels of 16:0, 18:0, 18:1, and 18:2 acyl chains. PC and PE were negatively correlated. Higher PC:PE at HT indicated possible PE-to-PC conversion, lower PE formation, or increased PE degradation, relative to PC. Correlation analysis revealed lipids experiencing coordinated metabolism under HT and confirmed the HT responsiveness of extraplastidic phospholipids. Comparison of the present results on wheat pollen with results of our previous research on wheat leaves suggests that similar lipid changes contribute to HT adaptation in both leaves and pollen, though the lipidomes have inherently distinct compositions.
Project description:Most chrysanthemum cultivars are self-incompatible, so it is very difficult to create pure lines that are important in chrysanthemum breeding and theoretical studies. In our previous study, we obtained a self-compatible chrysanthemum cultivar and its self-pollinated seed set was 56.50%. It was interesting that the seed set of its ten progenies ranged from 0% to 37.23%. Examination of the factors causing the differences in the seed set will lead to an improved understanding of chrysanthemum self-incompatibility, and provide valuable information for creating pure lines. Pollen morphology, pollen germination percentage, pistil receptivity and embryo development were investigated using the in vitro culture method, the paraffin section technique, scanning electron microscopy and transmission electron microscopy. Moreover, RNA sequencing and bioinformatics were applied to analyzing the transcriptomic profiles of mature stigmas and anthers. It was found that the self-pollinated seed set of "Q10-33-1?","Q10-33-1?","Q10-33-1?" and "Q10-33-1?" were 37.23%, 26.77%, 7.97% and 0%, respectively. The differences in fertility among four progenies were mainly attributable to differences in pollen germination percentage and pistil receptivity. Failure of the seed set in "Q10-33-1?" was possibly due to self-incompatibility. In the transcriptomic files, 22 potential stigma S genes and 8 potential pollen S genes were found out.
Project description:Members of the P4 subfamily of P-type ATPases are thought to create and maintain lipid asymmetry in biological membranes by flipping specific lipids between membrane leaflets. In Arabidopsis, 7 of the 12 Aminophospholipid ATPase (ALA) family members are expressed in pollen. Here we show that double knockout of ALA6 and ALA7 (ala6/7) results in siliques with a ~2-fold reduction in seed set with a high frequency of empty seed positions near the bottom. Seed set was reduced to near zero when plants were grown under a hot/cold temperature stress. Reciprocal crosses indicate that the ala6/7 reproductive deficiencies are due to a defect related to pollen transmission. In-vitro growth assays provide evidence that ala6/7 pollen tubes are short and slow, with ~2-fold reductions in both maximal growth rate and overall length relative to wild-type. Outcrosses show that when ala6/7 pollen are in competition with wild-type pollen, they have a near 0% success rate in fertilizing ovules near the bottom of the pistil, consistent with ala6/7 pollen having short and slow growth defects. The ala6/7 phenotypes were rescued by the expression of either an ALA6-YFP or GFP-ALA6 fusion protein, which showed localization to both the plasma membrane and highly-mobile endomembrane structures. A mass spectrometry analysis of mature pollen grains revealed significant differences between ala6/7 and wild-type, both in the relative abundance of lipid classes and in the average number of double bonds present in acyl side chains. A change in the properties of the ala6/7 plasma membrane was also indicated by a ~10-fold reduction of labeling by lipophilic FM-dyes relative to wild-type. Together, these results indicate that ALA6 and ALA7 provide redundant activities that function to directly or indirectly change the distribution and abundance of lipids in pollen, and support a model in which ALA6 and ALA7 are critical for pollen fitness under normal and temperature-stress conditions.
Project description:<h4>Background</h4>Decreased spikelet fertility is often responsible for reduction in grain yield in rice (Oryza sativa L.). In this study, two varieties with different levels of heat tolerance, Liangyoupeijiu (LYPJ, heat susceptible) and Shanyou63 (SY63, heat tolerant) were subjected to two temperature treatments for 28 days during the panicle initiation stage in temperature/relative humidity-controlled greenhouses: high temperature (HT; 37/27 °C; day/night) and control temperature (CK; 31/27 °C; day/night) to investigate changes in anther development under HT during panicle initiation and their relationship with spikelet fertility.<h4>Results</h4>HT significantly decreased the grain yield of LYPJ by decreasing the number of spikelets per panicle and seed setting percentage. In addition, HT produced minor adverse effects in SY63. The decreased spikelet fertility was primarily attributed to decreased pollen viability and anther dehiscence, as well as poor pollen shedding of the anthers of LYPJ under HT. HT resulted in abnormal anther development (fewer vacuolated microspores, un-degraded tapetum, unevenly distributed Ubisch bodies) and malformation of pollen (obscure outline of the pollen exine with a collapsed bacula, disordered tectum, and no nexine of the pollen walls, uneven sporopollenin deposition on the surface of pollen grains) in LYPJ, which may have lowered pollen viability. Additionally, HT produced a compact knitted anther cuticle structure of the epidermis, an un-degraded septum, a thickened anther wall, unevenly distributed Ubisch bodies, and inhibition of the confluent locule, and these malformed structures may be partially responsible for the decreased anther dehiscence rate and reduced pollen shedding of the anthers in LYPJ. In contrast, the anther wall and pollen development of SY63 were not substantially changed under HT.<h4>Conclusions</h4>Our results suggest that disturbed anther walls and pollen development are responsible for the reduced spikelet fertility and grain yield of the tested heat susceptible variety, and noninvasive anthers and pollen formation in response to HT were associated with improved heat tolerance.
Project description:Successive stamen movement directly controls pollen presentation schedules through sequential stamen maturation and changes the extent of herkogamy by altering the positions of sexual organs. However, the implications of such movements in terms of pollination are not well understood. Pollen presentation theory predicts that staggered pollen presentation should be favoured when plants are subject to diminishing returns on pollen transfer. Herkogamy on the other hand, has been interpreted as an adaptive trait that reduces sexual interference in hermaphrodite flowers. In this study, we conducted floral manipulations to determine the function of successive stamen movement in pollen transfer. By artificially manipulating the flowers to present two anthers simultaneously in the floral centre, we attempted to investigate whether changes in the anther presentation strategy affect pollen removal, deposition and the efficiency of pollinators. Compared with the natural treatment, the pollen transfer efficiency of halictid bees decreased significantly when the flowers were manipulated to present two anthers simultaneously. Although the presentation of two anthers simultaneously led to a similar pollen removal rate, there was a significant reduction in pollen deposition on neighbouring stigmas. To evaluate the effect of movement herkogamy on pollen export and deposition and seed set, the flowers were manipulated with or without the movement of stamen bending out from the floral centre. Pollen export decreased significantly when the central anther was moved away from the pistil, and pollen deposition and seed set declined significantly when the five spent anthers were retained on the pistil. Our study provides good support for the pollen presentation theory and provides direct experimental evidence that successive stamen movement could increase pollen transfer efficiency by sequential stamen maturation. Moreover, movement herkogamy promotes pollen export, deposition and seed set, and could therefore be regarded as an effective mechanism to reduce interference between male and female functions.
Project description:BACKGROUND:Pollen tube elongation in the pistil is a key step for pollination success in plants, and auxins play an important role in this process. However, the function of auxins in pollen tube elongation in the pistil of rice under heat stress has seldom been previously reported. RESULTS:Two rice genotypes differing in heat tolerance were subjected to heat stress of 40 °C for 2 h after flowering. A sharp decrease in spikelet fertility was found in the Nipponbare (NPB) and its mutant High temperature susceptible (HTS) under heat stress, but the stress-induced spikelet sterility was reversed by 1-naphthaleneacetic acid (NAA), especially the HTS. Under heat stress, the pollen tubes of NPB were visible in ovule, while those of HTS were invisible. However, we found the pollen tubes in ovule when sprayed with NAA. During this process, a significant increase in indole-3-acetic acid (IAA) and reactive oxygen species (ROS) levels was found in the pistil of heat-stressed NPB, while in heat-stressed HTS they were obviously decreased. Additionally, the peroxidase (POD) activity in pistil of NPB was significantly decreased by heat stress, whereas there was no difference between the heat-stressed and non-heat-stressed pistils of HTS. CONCLUSION:It was concluded that the enhancement of heat tolerance in plants by NAA was achieved through the increase of the levels of auxins, which prevented the inhibition of pollen tube elongation in pistil, and the crosstalk between auxins and ROS, which might be involved in this process. In addition, POD might be a negative mediator in pollen tube elongation under heat stress due to its ability to scavenge ROS and degrade auxin.
Project description:Although self-incompatibility (SI) in plants has been studied extensively, far less is known about interspecific reproductive barriers. One interspecific barrier, known as unilateral incongruity or incompatibility (UI), occurs when species display unidirectional compatibility in interspecific crosses. In the wild tomato species Solanum pennellii, both SI and self-compatible (SC) populations express UI when crossed with domesticated tomato, offering a useful model system to dissect the molecular mechanisms involved in reproductive barriers. In this study, the timing of reproductive barrier establishment during pistil development was determined in SI and SC accessions of S. pennellii using a semi-in vivo system to track pollen-tube growth in developing styles. Both SI and UI barriers were absent in styles 5 days prior to flower opening, but were established by 2 days before flower opening, with partial barriers detected during a transition period 3-4 days before flower opening. The developmental expression dynamics of known SI factors, S-RNases and HT proteins, was also examined. The accumulation of HT-A protein coincided temporally and spatially with UI barriers in developing pistils. Proteomic analysis of stigma/styles from key developmental stages showed a switch in protein profiles from cell-division-associated proteins in immature stigma/styles to a set of proteins in mature stigma/styles that included S-RNases, HT-A protein and proteins associated with cell-wall loosening and defense responses, which could be involved in pollen-pistil interactions. Other prominent proteins in mature stigma/styles were those involved in lipid metabolism, consistent with the accumulation of lipid-rich material during pistil maturation.
Project description:Phylogenetic analysis was conducted on 9 kDa non-specific lipid transfer protein (nsLTP) genes from nine plant species. Each of the five classified types in angiosperms exhibited eight conserved cysteine patterns. The most abundant nsLTP genes fell into the type I category, which was particularly enriched in a grass-specific lineage of clade I.1. Six pairs of tandem copies of nsLTP genes on the distal region of rice chromosomes 11 and 12 were well-preserved under concerted evolution, which was not observed in sorghum. The transgenic promoter-reporter assay revealed that both rice and sorghum nsLTP genes of type I displayed a relatively conserved expression feature in the epidermis of growing tissue, supporting its functional roles in cutin synthesis or defence against phytopathogens. For type I, the frequent expression in the stigma and seed are indicative of functional involvement in pistil-pollen interactions and seed development. By way of contrast, several type V genes were observed, mainly in the vascular bundle of the rosette as well as the young shoots, which might be related with vascular tissue differentiation or defence signalling. Compared with sorghum, the highly redundant tissue-specific expression pattern among members of rice nsLTP genes in clade I.1 suggests that concerted evolution via gene conversion favours the preservation of crucial expression motifs via the homogenization of proximal promoter sequences under high selection constraints. However, extensive regulatory subfunctionalization might also have occurred under relative low selection constraints, resulting in functional divergence at the expression level.
Project description:Elevated [CO<sub>2</sub>] (e[CO<sub>2</sub>]) environments have been predicted to improve rice yields under future climate. However, a concomitant rise in temperature could negate e[CO<sub>2</sub>] impact on plants, presenting a serious challenge for crop improvement. High temperature (HT) stress tolerant NL-44 and high yielding basmati Pusa 1121 rice cultivars, were exposed to e[CO<sub>2</sub>] (from panicle initiation to maturity) and a combination of e[CO<sub>2</sub>] + HT (from heading to maturity) using field based open top chambers. Elevated [CO<sub>2</sub>] significantly increased photosynthesis, seed-set, panicle weight and grain weight across both cultivars, more prominently with Pusa 1121. Conversely, e[CO<sub>2</sub>] + HT during flowering and early grain filling significantly reduced seed-set and 1000 grain weight, respectively. Averaged across both the cultivars, grain yield was reduced by 18 to 29%. Despite highly positive response with e[CO<sub>2</sub>], Pusa 1121 exposure to e[CO<sub>2</sub>] + HT led to significant reduction in seed-set and sink starch metabolism enzymatic activity. Interestingly, NL-44 maintained higher seed-set and resilience with starch metabolism enzymes under e[CO<sub>2</sub>] + HT exposure. Developing rice cultivars with higher [CO<sub>2</sub>] responsiveness incorporated with increased tolerance to high temperatures during flowering and grain filling using donors such as NL-44, will minimize the negative impact of heat stress and increase global food productivity, benefiting from [CO<sub>2</sub>] rich environments.