Increased panicle nitrogen application improves rice yield by alleviating high-temperature damage during panicle initiation to anther development.

The grain yield is closely associated with spikelet fertility in rice (Oryza sativa L.) under high temperatures, and nitrogen (N) plays a crucial role in yield formation. To investigate the effect of panicle N application on yield formation under high temperatures at the panicle initiation stage, two rice varieties [Liangyoupeijiu (LYPJ, heat susceptible) and Shanyou63 (SY63, heat tolerant)] were grown and exposed to high daytime temperature (HT) and control temperature (Control) during the panicle initiation stage. Low (LPN) and high (HPN) panicle N applications were conducted. HT markedly decreased the yields by 87% at LPN and 48% at HPN in LYPJ and 31% at LPN and 36% at HPN in SY63. The decrease in grain yield under HT was primarily attributed to the decline in spikelet fertility, HPN increased spikelet fertility. HT resulted in the abnormal development of anthers, which included disordered, enlarged, and broken anther wall layers, degraded and irregularly shaped microspores, delayed tapetum degradation, less vacuolated microspores per locule, abnormal and aborted pollen grains; however, HPN improved the development of anthers under HT, particularly in LYPJ. A high rate of evapotranspiration resulted in an approximately 1°C decrease in panicle temperatures at HPN compared with that at LPN in both varieties under HT. Overall, these results demonstrate that the increased panicle N application favors normal anther development in LYPJ by decreasing the panicle temperature, which results in high pollen viability and spikelet fertility, and consequently less yield loss under HT.

Phloem Unloading in Developing Rice Caryopses and its Contribution to Non-Structural Carbohydrate Translocation from Stems and Grain Yield Formation.

Phloem unloading plays an important role in photoassimilate partitioning and grain yield improvements in cereal crops. The phloem unloading strategy and its effects on photoassimilate translocation and yield formation remain unclear in rice. In this study, plasmodesmata were observed at the interface between the sieve elements (SEs) and companion cells (CCs), and between the SE-CC complex and surrounding parenchyma cells (PCs) in phloem of the dorsal vascular bundle in developing caryopses. Carboxyfluorescein (CF) signal was detected in the phloem of caryopses, which showed that CF was unloaded into caryopses. These results indicated that the SE-CC complex was symplasmically connected with adjacent PCs by plasmodesmata. Gene expression for sucrose transporter (SUT) and cell wall invertase (CWI), and OsSUT1 and OsCIN1 proteins were detected in developing caryopses, indicating that rice plants might actively unload sucrose into caryopses by the apoplasmic pathway. Among three rice recombinant inbred lines, R201 exhibited lower plasmodesmal densities at the boundaries between cell types (SE-CC, SE-PC and CC-PC) in developing caryopses than R91 and R156. R201 also had lower expression of SUT and CWI genes and lower protein levels of OsSUT1 and OsCIN1, as well as CWI activity, than R91 and R156. These data agreed with stem non-structural carbohydrate (NSC) translocation and grain yields for the three lines. The nitrogen application rate had no significant effect on plasmodesmal densities at the interfaces between different cells types, and did not affect CF unloading in the phloem of developing caryopses. Low nitrogen treatment enhanced expression levels of OsSUT and OsCIN genes in the three lines. These results suggested that nitrogen application had no substantial effect on symplasmic unloading but affected apoplasmic unloading. Therefore, we concluded that poor symplasmic and apoplasmic unloading in developing caryopses might result in low stem NSC translocation and poor grain yield formation of R201.

Abnormal anther development leads to lower spikelet fertility in rice (Oryza sativa L.) under high temperature during the panicle initiation stage.

BACKGROUND: 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. RESULTS: 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. CONCLUSIONS: 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.

Estimating the yield stability of heat-tolerant rice genotypes under various heat conditions across reproductive stages: a 5-year case study.

Heat events during the reproductive stages of rice plants induce great yield losses. Cultivating heat-tolerant varieties is a promising strategy for guaranteeing grain security under global warming scenarios. Most heat-tolerant rice genotypes were identified under heat during the flowering stage, but it is unclear whether these currently screened heat-tolerant rice genotypes maintain stable high grain yields when heat stress occurs during the other reproductive stages. In the present study, two notable heat-tolerant rice cultivars, Nagina22 and Shanyou63, and one typical heat-sensitive cultivar, Liangyoupeijiu, were evaluated for their yield response and yield stability under heat treatments during the panicle initiation, flowering, and grain filling stages during 2010-2014. Our results revealed that rice cultivars respond differently to heat stress during different reproductive stages. Nagina22 was the most tolerant to heat stress during the flowering and grain filling stages but was susceptible during panicle initiation; Shanyou63 was the most tolerant to heat stress during panicle initiation and grain filling and was moderately tolerant to heat stress during the flowering stages. Genotype and genotype-by-environment interaction biplot yield analysis revealed that Shanyou63 exhibited the highest stability in high grain yield, followed by Nagina22, and Liangyoupeijiu exhibited stable low grain yield when experiencing heat stress across the three reproductive stages. Our results indicate that the heat tolerance of different rice cultivars depends on the reproductive stage during which heat stress occurs, and the effects manifest as reductions in grain yields and seed setting rates. Future efforts to develop heat-tolerant varieties should strive to breed varieties that are comprehensively tolerant to heat stress during any reproductive stage to cope with the unpredictable occurrence of future heat events.

Low Nitrogen Application Enhances Starch-Metabolizing Enzyme Activity and Improves Accumulation and Translocation of Non-structural Carbohydrates in Rice Stems.

More than 4 billion inhabitants in Asia depend on rice for 35-60% of the calories consumed in their diets, but new rice cultivars frequently do not reach expected yields because of poor rice grain filling. Here, we quantified the activities of enzymes involved in starch metabolization in rice to investigate the mechanisms regulating the accumulation and translocation of stem non-structural carbohydrates (NSC) under different levels of nitrogen fertilizer application. A pot experiment was conducted using two rice cultivars, Liangyoupeijiu (LYPJ) and Shanyou63 (SY63), under high and low nitrogen applications. Compared with high nitrogen application (HN), low nitrogen application (LN) increased stem NSC concentration before the heading stage and NSC translocation during the grain filling stage; concomitantly, LN significantly shortened the active grain filling period and increased the grain filling rate in superior spikelets. Compared with the LYPJ cultivar, SY63 exhibited a higher grain weight, higher grain filling percentage, and higher stem NSC concentration before heading and greater NSC translocation after heading. During the period between panicle initiation and heading, the activities of adenosine diphosphate-glucose pyrophosphorylase (AGP), starch synthase (StS), and starch branching enzyme (SBE), all enzymes involved in starch synthesis, increased under the LN treatment and positively correlated with increases in stem NSC. During grain filling, the activities of enzymes involved in starch-to-sucrose conversion [α-amylase, ß-amylase, and sucrose phosphate synthase (SPS)] increased under the LN treatment and positively correlated with stem NSC remobilization. Overall, the investigated enzymes exhibited higher activities in SY63 than in LYPJ. Our results suggest that low nitrogen increases the activities of AGP, StS, SBE, α-amylase, ß-amylase, and SPS, leading to increased accumulation and remobilization of stem starch and NSC in SY63. We conclude that calculated reductions in nitrogen application and the choice of an appropriate cultivar may improve rice grain yields via enhanced stem NSC accumulation and translocation, thereby reducing the costs and increasing the sustainability of rice production.

Limitation of Unloading in the Developing Grains Is a Possible Cause Responsible for Low Stem Non-structural Carbohydrate Translocation and Poor Grain Yield Formation in Rice through Verification of Recombinant Inbred Lines.

Remobilisation of non-structural carbohydrates (NSC) from leaves and stems and unloading into developing grains are essential for yield formation of rice. In present study, three recombinant inbred lines of rice, R91, R156 and R201 have been tested for source-flow-sink related attributes determining the nature of NSC accumulation and translocation at two nitrogen levels in the field. Compared to R91 and R156, R201 had lower grain filling percentage, harvest index, and grain yield. Meanwhile, R201 had significantly lower stem NSC translocation during grain filling stage. Grain filling percentage, harvest index, and grain yield showed the consistent trend with stem NSC translocation among the three lines. In comparison with R91 and R156, R201 had similarity in leaf area index, specific leaf weight, stem NSC concentration at heading, biomass, panicles m-2, spikelets per panicle, remobilization capability of assimilation in stems, sink capacity, sink activity, number and cross sectional area of small vascular bundles, greater number and cross sectional area of large vascular bundles, and higher SPAD, suggesting that source, flow, and sink were not the limiting factors for low stem NSC translocation and grain filling percentage of R201. However, R201 had significant higher stem and rachis NSC concentrations at maturity, which implied that unloading in the developing grains might result in low NSC translocation in R201. The results indicate that stem NSC translocation could be beneficial for enhancement of grain yield potential, and poor unloading into caryopsis may be the possible cause of low stem NSC translocation, poor grain filling and yield formation in R201.

Heat-Induced Cytokinin Transportation and Degradation Are Associated with Reduced Panicle Cytokinin Expression and Fewer Spikelets per Panicle in Rice.

Cytokinins (CTKs) regulate panicle size and mediate heat tolerance in crops. To investigate the effect of high temperature on panicle CTK expression and the role of such expression in panicle differentiation in rice, four rice varieties (Nagina22, N22; Huanghuazhan, HHZ; Liangyoupeijiu, LYPJ; and Shanyou63, SY63) were grown under normal conditions and subjected to three high temperature treatments and one control treatment in temperature-controlled greenhouses for 15 days during the early reproductive stage. The high temperature treatments significantly reduced panicle CTK abundance in heat-susceptible LYPJ, HHZ, and N22 varieties, which showed fewer spikelets per panicle in comparison with control plants. Exogenous 6-benzylaminopurine application mitigated the effect of heat injury on the number of spikelets per panicle. The high temperature treatments significantly decreased the xylem sap flow rate and CTK transportation rate, but enhanced cytokinin oxidase/dehydrogenase (CKX) activity in heat-susceptible varieties. In comparison with the heat-susceptible varieties, heat-tolerant variety SY63 showed less reduction in panicle CTK abundance, an enhanced xylem sap flow rate, an improved CTK transport rate, and stable CKX activity under the high temperature treatments. Enzymes involved in CTK synthesis (isopentenyltransferase, LONELY GUY, and cytochrome P450 monooxygenase) were inhibited by the high temperature treatments. Heat-induced changes in CTK transportation from root to shoot through xylem sap flow and panicle CTK degradation via CKX were closely associated with the effects of heat on panicle CTK abundance and panicle size. Heat-tolerant variety SY63 showed stable panicle size under the high temperature treatments because of enhanced transport of root-derived CTKs and stable panicle CKX activity. Our results provide insight into rice heat tolerance that will facilitate the development of rice varieties with tolerance to high temperature.

Heat-induced phytohormone changes are associated with disrupted early reproductive development and reduced yield in rice.

Heat stress causes morphological and physiological changes and reduces crop yield in rice (Oryza sativa). To investigate changes in phytohormones and their relationships with yield and other attributes under heat stress, four rice varieties (Nagina22, Huanghuazhan, Liangyoupeijiu, and Shanyou 63) were grown in pots and subjected to three high temperature treatments plus control in temperature-controlled greenhouses for 15 d during the early reproductive phase. Yield reductions in Nagina22, Huanghuazhan, and Liangyoupeijiu were attributed to reductions in spikelet fertility, spikelets per panicle, and grain weight. The adverse effects of high temperature were alleviated by application of exogenous 6-benzylaminopurine (6-BA) in the heat-susceptible Liangyoupeijiu. High temperature stress reduced active cytokinins, gibberellin A1 (GA1), and indole-3-acetic acid (IAA), but increased abscisic acid (ABA) and bound cytokinins in young panicles. Correlation analyses and application of exogenous 6-BA revealed that high temperature-induced cytokinin changes may regulate yield components by modulating the differentiation and degradation of branches and spikelets, panicle exsertion, pollen vigor, anther dehiscence, and grain size. Heat-tolerant Shanyou 63 displayed minor changes in phytohormones, panicle formation, and grain yield under high temperature compared with those of the other three varieties. These results suggest that phytohormone changes are closely associated with yield formation, and a small reduction or stability in phytohormone content is required to avoid large yield losses under heat stress.