TabHLH27 orchestrates root growth and drought tolerance to enhance water use efficiency in wheat.

Cultivating high-yield wheat under limited water resources is crucial for sustainable agriculture in semiarid regions. Amid water scarcity, plants activate drought response signaling, yet the delicate balance between drought tolerance and development remains unclear. Through genome-wide association studies and transcriptome profiling, we identified a wheat atypical basic helix-loop-helix (bHLH) transcription factor (TF), TabHLH27-A1, as a promising quantitative trait locus candidate for both relative root dry weight and spikelet number per spike in wheat. TabHLH27-A1/B1/D1 knock-out reduced wheat drought tolerance, yield, and water use efficiency (WUE). TabHLH27-A1 exhibited rapid induction with polyethylene glycol (PEG) treatment, gradually declining over days. It activated stress response genes such as TaCBL8-B1 and TaCPI2-A1 while inhibiting root growth genes like TaSH15-B1 and TaWRKY70-B1 under short-term PEG stimulus. The distinct transcriptional regulation of TabHLH27-A1 involved diverse interacting factors such as TaABI3-D1 and TabZIP62-D1. Natural variations of TabHLH27-A1 influence its transcriptional responses to drought stress, with TabHLH27-A1Hap-II associated with stronger drought tolerance, larger root system, more spikelets, and higher WUE in wheat. Significantly, the excellent TabHLH27-A1Hap-II was selected during the breeding process in China, and introgression of TabHLH27-A1Hap-II allele improved drought tolerance and grain yield, especially under water-limited conditions. Our study highlights TabHLH27-A1's role in balancing root growth and drought tolerance, providing a genetic manipulation locus for enhancing WUE in wheat.

Low light stress promotes new tiller regeneration by changing source-sink relationship and activating expression of expansin genes in wheat.

Low light stress seriously decreased wheat grain number through the formation of aborted spike during the reproductive period and induced new tiller regeneration to offset the loss of grain number. However, the mechanism by which plants coordinate spike aborted growth and the regeneration of new tillers remains unknown. To better understand this coordinated process, morphological, physiological and transcriptomic analyses were performed under low light stress at the young microspore stage. Our findings indicated that leaves exhausted most stored carbohydrates in 1 day of darkness. However, spike and uppermost internode (UI) were converted from sink to source, due to increased abscisic acid (ABA) content and decreased cytokinin content. During this process, genes encoding amylases, Sugars Will Eventually be Exported Transporters (SWEET) and sucrose transporters or sucrose carriers (SUT/SUC) were upregulated in spike and UI, which degraded starch into soluble sugars and loaded them into the phloem. Subsequently, soluble sugars were transported to tiller node (TN) where cytokinin and auxin content increased and ABA content decreased, followed by unloading into TN cells by upregulated cell wall invertase (CWINV) genes and highly expressed H+ /hexose symporter genes. Finally, expansin genes integrated the sugar pathway and hormone pathway, and regulate the formation of new tillers directly.

Development and application of a relative soil water content - transpiration efficiency curve for screening high water use efficiency wheat cultivars.

Improving water use efficiency (WUE) has been proven to be a prosperous way to produce more grain in drought-prone areas. Transpiration efficiency (TE) has been proposed as a criterion for screening cultivars with high WUE. This study quantifies the relations of TE to relative soil water content (RSWC) gradients using pot experiments and evaluates the capability of the relations of TE-RSWC on assessing the cultivar performance in field yield and WUE. Twelve winter wheat cultivars were grown at 6 RSWC, 12.1, 24.2, 36.3, 48.4, 60.5, and 72.6% of field capacity (FC = 24.8 g/g) for 33 days in tightly sealed pots preventing soil evaporation. The results showed that TE decreased power functionally following the increase in RSWC for all cultivars. The relationship could be described as TE = TE FC × (RSWC) b , named TE-RSWC curve. This curve could be divided into an orderly area where the rank of cultivars was stable when RSWC ≤ 12.1% or RSWC ≥ 72.6% and a disorderly area where the rank was unstable when 12.1% < RSWC < 72.6%. To assess the consistency of pot TE to field yield and WUE, the same 12 varieties were grown under rainfed and two irrigations (75 mm at the jointing and flowering stages, respectively). TE FC was found to be positively related to field yield and WUE independent of irrigation. TE measured near the wilting point was negatively related to field yield and WUE. These results indicated that TE FC could be used as a surrogate for screening high-yield and high-WUE cultivars. The consistency and inconsistency can be attributed to the orderly area and disorderly area of the TE-RSWC curves.

Wheat Escapes Low Light Stress by Altering Pollination Types.

Although low light stress seriously affects florets fertility and grain number during the reproductive period, crops can be fertilized by heterologous pollen to alleviate the reduction of grain number. However, wheat is strongly autogamous, how to change to outcross after low light remains unclear. To understand the mechanisms of this change process, an approach combined morphological, physiological, and transcriptomic analyses was performed under low light stress imposed at the young microspore stage the booting stage from tetrad to uni-nucleate microspores stage. The results showed that low light stress caused pollen abortion, and the unfertilized ovary is fertilized by heterologous pollen after floret opening. Compared to control, the opening angle of lemma and glume were increased by 11.6-48.6 and 48.4-78.5%, respectively. The outcross of stressed wheat compensated for the 2.1-18.0% of grain number loss. During this process, phytohormones played an important role. Jasmonic acid (JA) and methyl jasmonate (MeJA) levels in spikelets were increased. Meanwhile, lignin and cellulose content decreased, and genes associated with cell wall related GO terms were enriched. Among the differentially expressed genes (DEGs), were identified 88-710 transcription factors genes, of which some homologs in Arabidopsis are proposed to function in lignin and cellulose, influencing the glume and lemma opening. Our finding can provide new insight into a survival mechanism to set seeds through pollination way alteration in the absence of self-fertilization after the stress of adversity.

Wheat FRIZZY PANICLE activates VERNALIZATION1-A and HOMEOBOX4-A to regulate spike development in wheat.

Kernel number per spike determined by the spike or inflorescence development is one important agricultural trait for wheat yield that is critical for global food security. While a few important genes for wheat spike development were identified, the genetic regulatory mechanism underlying supernumerary spikelets (SSs) is still unclear. Here, we cloned the wheat FRIZZY PANICLE (WFZP) gene from one local wheat cultivar. WFZP is specifically expressed at the sites where the spikelet meristem and floral meristem are initiated, which differs from the expression patterns of its homologs FZP/BD1 in rice and maize, indicative of its functional divergence during species differentiation. Moreover, WFZP directly activates VERNALIZATION1 (VRN1) and wheat HOMEOBOX4 (TaHOX4) to regulate the initiation and development of spikelet. The haplotypes analysis showed that the favourable alleles of WFZP associated with spikelet number per spike (SNS) were preferentially selected during breeding. Our findings provide insights into the molecular and genetic mechanisms underlying wheat spike development and characterize the WFZP as elite resource for wheat molecular breeding with enhanced crop yield.

Photosynthetic base of reduced grain yield by shading stress during the early reproductive stage of two wheat cultivars.

The young microspore (YM) stage is the most sensitive period for wheat grain formation to abiotic stress. Shading stress during YM stage reduces grain yield mainly due to grain number decrease. However, the photosynthetic base for grain number decrease is still unclear. In this study, 100% (control), 40% (S1), and 10% (S2) of natural light were applied for 1, 3, 5, and 7 days (D1, D3, D5 and D7) during YM stage of two wheat cultivars (Henong825, Kenong9204). The results showed that grain number in Henong825 and Kenong9204 was reduced by - 3.6 to 33.3% and 14.2-72.7%, respectively. The leaf photosynthetic rate (Pn) in Henong825 and Kenong9204 was deducted by 4.5-93.9% and 26.4-99.0%. Under S1-D1, the leaf Pn of Henong825 reducing was mainly due to the reduction of light intensity. With shading intensity and duration increasing, the reasons for leaf Pn decrease were the low light intensity, the low Gs (stomatal conductance) and chlorophyll content, the damage of ultrastructure of chloroplast and photosynthetic system. Under S2-D7, the chlorophyll content, Fv/Fm (maximal photochemical efficiency of photosystem II) and Jmax (maximum electron transport) were reduced by 19.6%, 5.2% and 28.8% in Henong825, and by 29.9%, 7.8% and 33.1% in Kenong9204. After shading removal, the leaf Pn of Kenong9204 under D5 and D7 could not reach to the level of CK. This study indicated that the reduction of leaf Pn was mainly due to the low light intensity under short shading duration (shorter than 3 days), and due to low light intensity and damage of the leaf photosynthetic system under longer shading duration (longer than 5 days), especially for Kenong9204 (shade-sensitive cultivar).

Effects of Drought Stress on Pollen Sterility, Grain Yield, Abscisic Acid and Protective Enzymes in Two Winter Wheat Cultivars.

Drought stress induced pollen sterility is a detrimental factor reducing grain number in wheat. Exploring the mechanisms underlying pollen fertility under drought conditions could assist breeding high-yielding wheat cultivars with stress tolerance. Here, by using two Chinese wheat cultivars subjected to different levels of polyethylene glycol (PEG)-induced drought stress, possible links between pollen fertility and stress tolerance were analyzed under different levels of drought stress at the young microspore stage. In both cultivars, higher grain number reduction was observed under condition of lower water availability. Overall, the drought tolerant cultivar (Jinmai47) exhibited less grain number reduction than the drought sensitive cultivar (Shiluan02-1) under all stress conditions. Compared with Shiluan02-1, Jinmai47 exhibited superior physiological performance in terms of leaf photosynthetic rate, ear carbohydrate accumulation, pollen sink strength, pollen development and fertility under stress. Moreover, Jinmai47 showed a lower increase in endogenous abscisic acid in ears than Shiluan02-1. Furthermore, higher levels of superoxide dismutase (SOD) and peroxidase (POD) activities were also found in the drought tolerant cultivar Jinmai47 under PEG stress, compared with the drought sensitive cultivar Shiluan02-1. Changes in these physiological traits could contribute to better pollen development and male fertility, ultimately leading to the maintenance of grain number under drought stress.

The Wheat GT Factor TaGT2L1D Negatively Regulates Drought Tolerance and Plant Development.

GT factors are trihelix transcription factors that specifically regulate plant development and stress responses. Recently, several GT factors have been characterized in different plant species; however, little is known about the role of GT factors in wheat. Here, we show that TaGT2L1A, TaGT2L1B, and TaGT2L1D are highly homologous in hexaploid wheat, and are localized to wheat chromosomes 2A, 2B, and 2D, respectively. These TaGT2L1 genes encode proteins containing two SANT domains and one central helix. All three homologs were ubiquitously expressed during wheat development and were responsive to osmotic stress. Functional analyses demonstrated that TaGT2L1D acts as a transcriptional repressor; it was able to suppress the expression of AtSDD1 in Arabidopsis by binding directly to the GT3 box in its promoter that negatively regulates drought tolerance. TaGT2L1D overexpression markedly increased the number of stomata and reduced drought tolerance in gtl1-3 plants. Notably, ectopic expression of TaGT2L1D also affected floral organ development and overall plant growth. These results demonstrate that TaGT2L1 is an ortholog of AtGTL1, and that it plays an evolutionarily conserved role in drought resistance by fine tuning stomatal density in wheat. Our data also highlight the role of TaGT2L1 in plant growth and development.

Comparison of the effects of symmetric and asymmetric temperature elevation and CO2 enrichment on yield and evapotranspiration of winter wheat (Triticum aestivum L.).

Under the changing climate, asymmetric warming pattern would be more likely during day and night time, instead of symmetric one. Concurrently, the growth responses and water use of plants may be different compared with those estimated based on symmetric warming. In this work, it was compared with the effects of symmetric (ETs) and asymmetric (ETa) elevation of temperature alone, and in interaction with elevated carbon dioxide concentration (EC), on the grain yield (GY) and evapotranspiration in winter wheat (Triticum aestivum L.) based on pot experiment in the North China Plain (NCP). The experiment was carried out in six enclosed-top chambers with following climate treatments: (1) ambient temperature and ambient CO2 (CON), (2) ambient temperature and elevated CO2 (EC), (3) elevated temperature and ambient CO2 (ETs; ETa), and (4) elevated temperature and elevated CO2 (ECETs, ECETa). In symmetric warming, temperature was increased by 3°C and in asymmetric one by 3.5°C during night and 2.5°C during daytime, respectively. As a result, GY was in ETa and ETs 15.6 (P < 0.05) and 10.3% (P < 0.05) lower than that in CON. In ECETs and ECETa treatments, GY was 14.9 (P < 0.05) and 9.1% (P < 0.05) higher than that in CON. Opposite to GY, evapotranspiration was 7.8 (P < 0.05) and 17.9% (P < 0.05) higher in ETa and ETs treatments and 7.2 (P < 0.05) and 2.1% (P > 0.05) lower in ECETs and ECETa treatments compared with CON. Thus, GY of wheat could be expected to increase under the changing climate with concurrent elevation of CO2 and temperature as a result of increased WUE under the elevated CO2. However, the gain would be lower under ETa than that estimated based on ETs due to higher evapotranspiration.

[Effects of drought stress, high temperature and elevated CO2 concentration on the growth of winter wheat].

The impacts of climate change on the grain yield, photosynthesis, and water conditions of winter wheat were assessed based on an experiment, in which wheat plants were subjected to ambient and elevated CO2 concentrations, ambient and elevated temperatures, and low and high water conditions independently and in combination. The CO2 enrichment alone had no effect on the photosynthesis of winter wheat, whereas higher temperature and drought significantly decreased the photosynthetic rate. Water conditions in flag leaves were not significantly changed at the elevated CO2 concentration or elevated temperature. However, drought stress decreased the relative water content in flag leaves, and the combination of elevated temperature and drought reduced the water potential in flag leaves. The combination of elevated CO2 concentration, elevated temperature, and drought significantly reduced the photosynthetic rate and water conditions, and led to a 41.4% decrease in grain yield. The elevated CO2 concentration alone increased the grain yield by 21.2%, whereas the elevated temperature decreased the grain yield by 12.3%. The grain yield was not affected by the combination of elevated CO2 concentration and temperature, but the grain yield was significantly decreased by the drought stress if combined with any of the climate scenarios applied in this study. These findings suggested that maintaining high soil water content might be a vital means of reducing the potential harm caused by the climate change.

Control of abscisic acid catabolism and abscisic acid homeostasis is important for reproductive stage stress tolerance in cereals.

Drought stress at the reproductive stage causes pollen sterility and grain loss in wheat (Triticum aestivum). Drought stress induces abscisic acid (ABA) biosynthesis genes in anthers and ABA accumulation in spikes of drought-sensitive wheat varieties. In contrast, drought-tolerant wheat accumulates lower ABA levels, which correlates with lower ABA biosynthesis and higher ABA catabolic gene expression (ABA 8'-hydroxylase). Wheat TaABA8'OH1 deletion lines accumulate higher spike ABA levels and are more drought sensitive. ABA treatment of the spike mimics the effect of drought, causing high levels of sterility. ABA treatment represses the anther cell wall invertase gene TaIVR1, and drought-tolerant lines appeared to be more sensitive to the effect of ABA. Drought-induced sterility shows similarity to cold-induced sterility in rice (Oryza sativa). In cold-stressed rice, the rate of ABA accumulation was similar in cold-sensitive and cold-tolerant lines during the first 8 h of cold treatment, but in the tolerant line, ABA catabolism reduced ABA levels between 8 and 16 h of cold treatment. The ABA biosynthesis gene encoding 9-cis-epoxycarotenoid dioxygenase in anthers is mainly expressed in parenchyma cells surrounding the vascular bundle of the anther. Transgenic rice lines expressing the wheat TaABA8'OH1 gene under the control of the OsG6B tapetum-specific promoter resulted in reduced anther ABA levels under cold conditions. The transgenic lines showed that anther sink strength (OsINV4) was maintained under cold conditions and that this correlated with improved cold stress tolerance. Our data indicate that ABA and ABA 8'-hydroxylase play an important role in controlling anther ABA homeostasis and reproductive stage abiotic stress tolerance in cereals.