Transcriptome and Physiological Analysis of Rapeseed Tolerance to Post-Flowering Temperature Increase.

Climate-change-induced temperature fluctuations pose a significant threat to crop production, particularly in the Southern Hemisphere. This study investigates the transcriptome and physiological responses of rapeseed to post-flowering temperature increases, providing valuable insights into the molecular mechanisms underlying rapeseed tolerance to heat stress. Two rapeseed genotypes, Lumen and Solar, were assessed under control and heat stress conditions in field experiments conducted in Valdivia, Chile. Results showed that seed yield and seed number were negatively affected by heat stress, with genotype-specific responses. Lumen exhibited an average of 9.3% seed yield reduction, whereas Solar showed a 28.7% reduction. RNA-seq analysis of siliques and seeds revealed tissue-specific responses to heat stress, with siliques being more sensitive to temperature stress. Hierarchical clustering analysis identified distinct gene clusters reflecting different aspects of heat stress adaptation in siliques, with a role for protein folding in maintaining silique development and seed quality under high-temperature conditions. In seeds, three distinct patterns of heat-responsive gene expression were observed, with genes involved in protein folding and response to heat showing genotype-specific expression. Gene coexpression network analysis revealed major modules for rapeseed yield and quality, as well as the trade-off between seed number and seed weight. Overall, this study contributes to understanding the molecular mechanisms underlying rapeseed tolerance to heat stress and can inform crop improvement strategies targeting yield optimization under changing environmental conditions.

Overcoming the trade-off between grain weight and number in wheat by the ectopic expression of expansin in developing seeds leads to increased yield potential.

Wheat is the most widely grown crop globally, providing 20% of all human calories and protein. Achieving step changes in genetic yield potential is crucial to ensure food security, but efforts are thwarted by an apparent trade-off between grain size and number. Expansins are proteins that play important roles in plant growth by enhancing stress relaxation in the cell wall, which constrains cell expansion. Here, we describe how targeted overexpression of an α-expansin in early developing wheat seeds leads to a significant increase in grain size without a negative effect on grain number, resulting in a yield boost under field conditions. The best-performing transgenic line yielded 12.3% higher average grain weight than the control, and this translated to an increase in grain yield of 11.3% in field experiments using an agronomically appropriate plant density. This targeted transgenic approach provides an opportunity to overcome a common bottleneck to yield improvement across many crops.

Transcriptome Analysis of Seed Weight Plasticity in Brassica napus.

A critical barrier to improving crop yield is the trade-off between seed weight (SW) and seed number (SN), which has been commonly reported in several crops, including Brassica napus. Despite the agronomic relevance of this issue, the molecular factors involved in the interaction between SW and SN are largely unknown in crops. In this work, we performed a detailed transcriptomic analysis of 48 seed samples obtained from two rapeseed spring genotypes subjected to different source-sink (S-S) ratios in order to examine the relationship between SW and SN under different field conditions. A multifactorial analysis of the RNA-seq data was used to identify a group of 1014 genes exclusively regulated by the S-S ratio. We found that a reduction in the S-S ratio during seed filling induces the expression of genes involved in sucrose transport, seed weight, and stress responses. Moreover, we identified five co-expression modules that are positively correlated with SW and negatively correlated with SN. Interestingly, one of these modules was significantly enriched in transcription factors (TFs). Furthermore, our network analysis predicted several NAC TFs as major hubs underlying SW and SN compensation. Taken together, our study provides novel insights into the molecular factors associated with the SW-SN relationship in rapeseed and identifies TFs as potential targets when improving crop yield.

Pericarp growth dynamics associate with final grain weight in wheat under contrasting plant densities and increased night temperature.

BACKGROUND AND AIMS: The pericarp weight comprises <17 % of wheat grain weight at harvest. The pericarp supports the hydration and nutrition of both the embryo and endosperm during early grain filling. However, studies of the pericarp and its association with final grain weight have been scarce. This research studied the growth dynamics of wheat pericarp from anthesis onwards and its relationship to final grain weight under contrasting plant densities and night warming. METHODS: Two spring wheat cultivars contrasting in kernel weight (Bacanora and Kambara) were sown in field conditions during seasons 2012-13 and 2014-15. Both genotypes were grown under contrasting plant density (control, 370 plants m-2; and low plant density, 44 plants m-2) and night temperatures, i.e. at ambient and increased (>6 °C) temperature for short periods before and after anthesis. From anthesis onward, grains were harvested every 3 or 4 d. Grain samples were measured and the pericarp was removed with a scalpel. Whole grain and pericarp fresh and dry weight were weighed with a precision balance. At harvest, 20 grains from ten spikes were weighed and grain dimensions were measured. KEY RESULTS: Fresh weight, dry matter and water content of pericarp dynamics showed a maximum between 110 and 235 °Cd. Maximum dry matter of the pericarp ranged between 4.3 and 5.7 mg, while water content achieved values of up to 12.5 mg. Maximum values and their timings were affected by the genotype, environmental condition and grain position. Final grain weight was closely associated with maximum dry matter and water content of the pericarp. CONCLUSIONS: Maximum pericarp weight is a determinant of grain weight and size in wheat, which is earlier than other traits considered as key determinants of grain weight during grain filling. Better growing conditions increased maximum pericarp weight, while higher temperature negatively affected this trait.

Expansin genes expression in growing ovaries and grains of sunflower are tissue-specific and associate with final grain weight.

BACKGROUND: Grain weight (GW) is a key component of sunflower yield and quality, but may be limited by maternal tissues. Cell growth is influenced by expansin proteins that loosen the plant cell wall. This study aimed to identify spatio-temporal expression of EXPN genes in sunflower reproductive organ tissues (ovary, pericarp, and embryo) and evaluate correlations between reproductive organ growth and expansin genes expression. Evaluations involved eight different developmental stages, two genotypes, two source-sink treatments and two experiments. The genotypes evaluated are contrasting in GW (Alybro and confection variety RHA280) under two source-sink treatments (control and shaded) to study the interactions between grain growth and expansin genes expression. RESULTS: Ovaries and grains were sampled at pre- and post-anthesis, respectively. Final GW differed between genotypes and shading treatments. Shading treatment decreased final GW by 16.4 and 19.5% in RHA280 and Alybro, respectively. Relative expression of eight expansin genes were evaluated in grain tissues. EXPN4 was the most abundant expansin in the ovary tissue, while EXPN10 and EXPN7 act predominantly in ovary and pericarp tissues, and EXPN1 and EXPN15 in the embryo tissues. CONCLUSIONS: Specific expansin genes were expressed in ovary, pericarp and embryo in a tissue-specific manner. Differential expression among grain tissues was consistent between genotypes, source-sink treatments and experiments. The correlation analysis suggests that EXPN genes could be specifically involved in grain tissue extension, and their expression could be linked to grain size in sunflower.

The trade-off between grain weight and grain number in wheat is explained by the overlapping of the key phases determining these major yield components.

Enhancing grain yield is a primary goal in the cultivation of major staple crops, including wheat. Recent research has focused on identifying the physiological and molecular factors that influence grain weight, a critical determinant of crop yield. However, a bottleneck has arisen due to the trade-off between grain weight and grain number, whose underlying causes remain elusive. In a novel approach, a wheat expansin gene, TaExpA6, known for its expression in root tissues, was engineered to express in the grains of the spring wheat cultivar Fielder. This modification led to increases in both grain weight and yield without adversely affecting grain number. Conversely, a triple mutant line targeting the gene TaGW2, a known negative regulator of grain weight, resulted in increased grain weight but decreased grain number, potentially offsetting yield gains. This study aimed to evaluate the two aforementioned modified wheat genotypes (TaExpA6 and TaGW2) alongside their respective wild-type counterparts. Conducted in southern Chile, the study employed a Complete Randomized Block Design with four replications, under well-managed field conditions. The primary metrics assessed were grain yield, grain number, and average grain weight per spike, along with detailed measurements of grain weight and dimensions across the spike, ovary weight at pollination (Waddington's scale 10), and post-anthesis expression levels of TaExpA6 and TaGW2. Results indicated that both the TaExpA6 and the triple mutant lines achieved significantly higher average grain weights compared to their respective wild types. Notably, the TaExpA6 line did not exhibit a reduction in grain number, thereby enhancing grain yield per spike. By contrast, the triple mutant line showed a reduced grain number per spike, with no significant change in overall yield. TaExpA6 expression peaked at 10 days after anthesis (DAA), and its effect on grain weight over the WT became apparent after 15 DAA. In contrast, TaGW2 gene disruption in the triple mutant line increased ovary size at anthesis, leading to improved grain weight above the WT from the onset of grain filling. These findings suggest that the trade-off between grain weight and number could be attributed to the overlapping of the critical periods for the determination of these traits.

Integrated Transcriptome Analysis Identified Key Expansin Genes Associated with Wheat Cell Wall, Grain Weight and Yield.

This research elucidates the dynamic expression of expansin genes during the wheat grain (Triticum aestivum L.) development process using comprehensive meta-analysis and experimental validation. We leveraged RNA-seq data from multiple public databases, applying stringent criteria for selection, and identified 60,852 differentially expressed genes across developmental stages. From this pool, 28,558 DEGs were found to exhibit significant temporal regulation in at least two different datasets and were enriched for processes integral to grain development such as carbohydrate metabolism and cell wall organization. Notably, 30% of the 241 known expansin genes showed differential expression during grain growth. Hierarchical clustering and expression level analysis revealed temporal regulation and distinct contributions of expansin subfamilies during the early stages of grain development. Further analysis using co-expression networks underscored the significance of expansin genes, revealing their substantial co-expression with genes involved in cell wall modification. Finally, qPCR validation and grain morphological analysis under field conditions indicated a significant negative correlation between the expression of select expansin genes, and grain size and weight. This study illuminates the potential role of expansin genes in wheat grain development and provides new avenues for targeted genetic improvements in wheat.

Raising yield potential of wheat. III. Optimizing partitioning to grain while maintaining lodging resistance.

A substantial increase in grain yield potential is required, along with better use of water and fertilizer, to ensure food security and environmental protection in future decades. For improvements in photosynthetic capacity to result in additional wheat yield, extra assimilates must be partitioned to developing spikes and grains and/or potential grain weight increased to accommodate the extra assimilates. At the same time, improvement in dry matter partitioning to spikes should ensure that it does not increase stem or root lodging. It is therefore crucial that improvements in structural and reproductive aspects of growth accompany increases in photosynthesis to enhance the net agronomic benefits of genetic modifications. In this article, six complementary approaches are proposed, namely: (i) optimizing developmental pattern to maximize spike fertility and grain number, (ii) optimizing spike growth to maximize grain number and dry matter harvest index, (iii) improving spike fertility through desensitizing floret abortion to environmental cues, (iv) improving potential grain size and grain filling, and (v) improving lodging resistance. Since many of the traits tackled in these approaches interact strongly, an integrative modelling approach is also proposed, to (vi) identify any trade-offs between key traits, hence to define target ideotypes in quantitative terms. The potential for genetic dissection of key traits via quantitative trait loci analysis is discussed for the efficient deployment of existing variation in breeding programmes. These proposals should maximize returns in food production from investments in increased crop biomass by increasing spike fertility, grain number per unit area and harvest index whilst optimizing the trade-offs with potential grain weight and lodging resistance.

Transcriptomic and Physiological Response of Durum Wheat Grain to Short-Term Heat Stress during Early Grain Filling.

In a changing climate, extreme weather events such as heatwaves will be more frequent and could affect grain weight and the quality of crops such as wheat, one of the most significant crops in terms of global food security. In this work, we characterized the response of Triticum turgidum L. spp. durum wheat to short-term heat stress (HS) treatment at transcriptomic and physiological levels during early grain filling in glasshouse experiments. We found a significant reduction in grain weight (23.9%) and grain dimensions from HS treatment. Grain quality was also affected, showing a decrease in starch content (20.8%), in addition to increments in grain protein levels (14.6%), with respect to the control condition. Moreover, RNA-seq analysis of durum wheat grains allowed us to identify 1590 differentially expressed genes related to photosynthesis, response to heat, and carbohydrate metabolic process. A gene regulatory network analysis of HS-responsive genes uncovered novel transcription factors (TFs) controlling the expression of genes involved in abiotic stress response and grain quality, such as a member of the DOF family predicted to regulate glycogen and starch biosynthetic processes in response to HS in grains. In summary, our results provide new insights into the extensive transcriptome reprogramming that occurs during short-term HS in durum wheat grains.

Expansins expression is associated with grain size dynamics in wheat (Triticum aestivum L.).

Grain weight is one of the most important components of cereal yield and quality. A clearer understanding of the physiological and molecular determinants of this complex trait would provide an insight into the potential benefits for plant breeding. In the present study, the dynamics of dry matter accumulation, water uptake, and grain size in parallel with the expression of expansins during grain growth in wheat were analysed. The stabilized water content of grains showed a strong association with final grain weight (r(2)=0.88, P <0.01). Grain length was found to be the trait that best correlated with final grain weight (r(2)=0.98, P <0.01) and volume (r(2)=0.94, P <0.01). The main events that defined final grain weight occurred during the first third of grain-filling when maternal tissues (the pericarp of grains) undergo considerable expansion. Eight expansin coding sequences were isolated from pericarp RNA and the temporal profiles of accumulation of these transcripts were monitored. Sequences showing high homology with TaExpA6 were notably abundant during early grain expansion and declined as maturity was reached. RNA in situ hybridization studies revealed that the transcript for TaExpA6 was principally found in the pericarp during early growth in grain development and, subsequently, in both the endosperm and pericarp. The signal in these images is likely to be the sum of the transcript levels of all three sequences with high similarity to the TaExpA6 gene. The early part of the expression profile of this putative expansin gene correlates well with the critical periods of early grain expansion, suggesting it as a possible factor in the final determination of grain size.

Increased Night Temperature Negatively Affects Grain Yield, Biomass and Grain Number in Chilean Quinoa.

Quinoa high nutritive value increases interest worldwide, especially as a crop that could potentially feature in different cropping systems, however, climate change, particularly rising temperatures, challenges this and other crop species. Currently, only limited knowledge exists regarding the grain yield and other key traits response to higher temperatures of this crop, especially to increased night temperatures. In this context, the main objective of this study was to evaluate the effect of increased night temperature on quinoa yield, grain number, individual grain weight and processes involved in crop growth under the environmental conditions (control treatment) and night thermal increase at two phases: flowering (T1) and grain filling (T2) in southern Chile. A commercial genotype, Regalona, and a quinoa accession (Cod. BO5, N°191, grain bank from Semillas Baer, hereby referred to as Accession) were used, due to their adaptability to Southern Chilean conditions and contrasting grain yield potential, grain weight and size of plants. Temperature was increased ≈4°C above the ambient from 8 pm until 9 am the next morning. Control treatments reached a high grain yield (600 and 397 g m-2, i.e., Regalona and Accession). Temperature increase reduced grain yield by 31% under T1 treatment and 12% when under T2 in Regalona and 23 and 26% in Accession, respectively. Aboveground biomass was negatively affected by the thermal treatments and a positive linear association was found between grain yield and aboveground biomass across treatments. By contrast, the harvest index was unaffected either by genotype, or by thermal treatments. Grain number was significantly affected between treatments and this key trait was linearly associated with grain yield. On the other hand, grain weight showed a narrow range of variation across treatments. Additionally, leaf area index was not affected, but significant differences were found in SPAD values at the end of T1 treatment, compared to control. Little change was found in the harvest index, individual grain weight, grain protein content or water soluble carbohydrates in response to the increased night temperature in this crop.

Genetic dissection of grain size and grain number trade-offs in CIMMYT wheat germplasm.

Grain weight (GW) and number per unit area of land (GN) are the primary components of grain yield in wheat. In segregating populations both yield components often show a negative correlation among themselves. Here we use a recombinant doubled haploid population of 105 individuals developed from the CIMMYT varieties Weebill and Bacanora to understand the relative contribution of these components to grain yield and their interaction with each other. Weebill was chosen for its high GW and Bacanora for high GN. The population was phenotyped in Mexico, Argentina, Chile and the UK. Two loci influencing grain yield were indicated on 1B and 7B after QTL analysis. Weebill contributed the increasing alleles. The 1B effect, which is probably caused by to the 1BL.1RS rye introgression in Bacanora, was a result of increased GN, whereas, the 7B QTL controls GW. We concluded that increased in GW from Weebill 7B allele is not accompanied by a significant reduction in grain number. The extent of the GW and GN trade-off is reduced. This makes this locus an attractive target for marker assisted selection to develop high yielding bold grain varieties like Weebill. AMMI analysis was used to show that the 7B Weebill allele appears to contribute to yield stability.