Enhancing root physiology for increased yield in water-saving and drought-resistance rice with optimal irrigation and nitrogen.

Objectives: Water-saving and drought-resistance rice (WDR) plays a vital role in the sustainable development of agriculture. Nevertheless, the impacts and processes of water and nitrogen on grain yield in WDR remain unclear. Methods: In this study, Hanyou 73 (WDR) and Hyou 518 (rice) were used as materials. Three kinds of nitrogen fertilizer application rate (NFAR) were set in the pot experiment, including no NFAR (nitrogen as urea applied at 0 g/pot), medium NFAR (nitrogen as urea applied at 15.6 g/pot), and high NFAR (nitrogen as urea applied at 31.2 g/pot). Two irrigation regimes, continuous flooding cultivation and water stress, were set under each NFAR. The relationships between root and shoot morphophysiology and grain yield in WDR were explored. Results: The results demonstrated the following: 1) under the same irrigation regime, the grain yield of two varieties increased with the increase of NFAR. Under the same NFAR, the reduction of irrigation amount significantly reduced the grain yield in Hyou 518 (7.1%-15.1%) but had no substantial influence on the grain yield in Hanyou 73. 2) Under the same irrigation regime, increasing the NFAR could improve the root morphophysiology (root dry weight, root oxidation activity, root bleeding rate, root total absorbing surface area, root active absorbing surface area, and zeatin + zeatin riboside contents in roots) and aboveground physiological indexes (leaf photosynthetic rate, non-structural carbohydrate accumulation in stems, and nitrate reductase activity in leaves) in two varieties. Under the same NFAR, increasing the irrigation amount could significantly increase the above indexes in Hyou 518 (except root dry weight) but has little effect on Hanyou 73. 3) Analysis of correlations revealed that the grain yield of Hyou 518 and Hanyou 73 was basically positively correlated with aboveground physiology and root morphophysiology, respectively. Conclusion: The grain yield could be maintained by water stress under medium NFAR in WDR. The improvement of root morphophysiology is a major factor for high yield under the irrigation regime and NFAR treatments in WDR.

Factors Influencing Seed Dormancy and Germination and Advances in Seed Priming Technology.

Seed dormancy and germination play pivotal roles in the agronomic traits of plants, and the degree of dormancy intuitively affects the yield and quality of crops in agricultural production. Seed priming is a pre-sowing seed treatment that enhances and accelerates germination, leading to improved seedling establishment. Seed priming technologies, which are designed to partially activate germination, while preventing full seed germination, have exerted a profound impact on agricultural production. Conventional seed priming relies on external priming agents, which often yield unstable results. What works for one variety might not be effective for another. Therefore, it is necessary to explore the internal factors within the metabolic pathways that influence seed physiology and germination. This review unveils the underlying mechanisms of seed metabolism and germination, the factors affecting seed dormancy and germination, as well as the current seed priming technologies that can result in stable and better germination.

Biochar Coating as a Cost-Effective Delivery Approach to Promoting Seed Quality, Rice Germination, and Seedling Establishment.

The application of high-quality seeds ensures successful crop establishment, healthy growth, and improved production in both quantity and quality. Recently, biochar-based seed coating has been recognized as a new, effective, and environmentally friendly method to enhance seed quality, seedling uniformity, and nutrient availability. To study the impact of biochar coating on the surface mechanical properties of coated seeds, rice emergence and growth, and related physical and physiological metabolic events, laboratory experiments were performed on two water-saving and drought-resistance rice (WDR) varieties (Huhan1512 and Hanyou73) using biochar formulations with varying contents (20%-60%). The results showed that the appropriate concentration of biochar significantly improved emergence traits and seedling performance of the two rice varieties, compared to the uncoated treatment, and that the optimal percentage of biochar coating was 30% (BC30). On average, across both varieties, BC30 enhanced emergence rate (9.5%), emergence index (42.9%), shoot length (19.5%), root length (23.7%), shoot dry weight (25.1%), and root dry weight (49.8%). The improved germination characteristics and vigorous seedling growth induced by biochar coating were strongly associated with higher water uptake by seeds, increased α-amylase activity and respiration rate, and enhanced accumulation of soluble sugar and soluble protein. Moreover, the evaluation results of mechanical properties related to seed coating quality found that increasing the proportion of biochar in the coating blend decreased the integrity and compressive strength of the coated seeds and reduced the time required for coating disintegration. In conclusion, biochar coating is a cost-effective strategy for enhancing crop seed quality and seedling establishment.

Mesophyll conductance and N allocation co-explained the variation in photosynthesis in two canola genotypes under contrasting nitrogen supply.

The application of nitrogen fertilizer within a normal range has been found to increase the leaf nitrogen content and photosynthetic rate of canola plants (Brassica napus L.). Despite numerous studies on the separate effects of CO2 diffusion limitation and nitrogen allocation trade-off on photosynthetic rate, few have examined both these factors in relation to the photosynthetic rate of canola. In this study, two genotypes of canola with varying leaf nitrogen content were analyzed to determine the impact of nitrogen supply on leaf photosynthesis, mesophyll conductance, and nitrogen partitioning. The results showed that the CO2 assimilation rate (A), mesophyll conductance (g m), and photosynthetic nitrogen content (N psn) increased with an increase in nitrogen supply in both genotypes. The relationship between nitrogen content and A followed a linear-plateau regression, while A had linear relationships with both photosynthetic nitrogen content and g m, indicating that the key to enhancing A is increasing the distribution of leaf nitrogen into the photosynthetic apparatus and g m, rather than just increasing nitrogen content. Under high nitrogen treatment, the genotype (QZ) with high nitrogen content had 50.7% more nitrogen than the other genotype (ZY21), but had similar A, which was primarily due to ZY21's higher photosynthetic nitrogen distribution ratio and stomatal conductance (g sw). On the other hand, QZ showed a higher A than ZY21 under low nitrogen treatment as QZ had higher N psn and g m compared to ZY21. Our results indicate that, in selecting high PNUE rapeseed varieties, it is important to consider the higher photosynthetic nitrogen distribution ratio and CO2 diffusion conductance.

Alleviating effects of zinc and 24-epibrassionlide on cadmium accumulation in rice plants under nitrogen application.

Heavy metals such as cadmium (Cd) in farmland soil not only affect crop production, but also endanger human health through the food chain. Rice is the main food crop with the strongest ability to absorb Cd, remediation techniques to reduce soil uptake and grain accumulation of Cd are urgently required, for which the application of foliar spraying seems to be a convenient and auspicious method. This study clarified the effects of nitrogen (N), zinc (Zn), 24-epibrassionlide (EBL) and their combined application on the growth performance and physiological characteristics of Cd and Zn in rice plants under Cd stress. Experimental results showed that N and its combination with Zn, EBL treatments promoted rice growth and yield, especially raised the yield level by 81.12% under N + EBL treatment. Additionally, three EBL treatments (EBL, N + EBL, Zn + EBL) significantly reduced the TF values of Cd in TF stems-grains, TF leaves-grains and TF glumes-grains by 42.70%, 43.67% and 50.33%, while the EF soil-roots under Zn and N + Zn treatments was the lowest, which decreased by 55.39% and 57.71%, respectively. Further, the application of N, Zn, EBL and their combined treatments significantly increased glutathione (GSH) and phytochelatins (PCs) content as well as enhanced Cd distribute into cell walls of rice shoots and roots by 15.18% and 13.20%, respectively. In addition, N, Zn, EBL and their combined application increased Zn concentration, free amino acid and glutelin content, and decreased the Cd accumulation in albumin, glutelin and globulin, thus lowered Cd concentration in grains by 27.55%, 58.29% and 51.56%, respectively. These results comprehensive suggest that the possibility of N management combined with Zn or EBL application for maintaining high yield and alleviating Cd stress by regulating the absorption and remobilization process under mild stress.

Biochar Coating Is a Sustainable and Economical Approach to Promote Seed Coating Technology, Seed Germination, Plant Performance, and Soil Health.

Seed germination and stand establishment are the first steps of crop growth and development. However, low seed vigor, improper seedbed preparation, unfavorable climate, and the occurrence of pests and diseases reduces the germination rate and seedling quality, resulting in insufficient crop populations and undesirable plant growth. Seed coating is an effective method that is being developed and applied in modern agriculture. It has many functions, such as improving seed vigor, promoting seedling growth, and reducing the occurrence of pests and diseases. Yet, during seed coating procedures, several factors, such as difficulty in biodegradation of coating materials and hindrance in the application of chemical ingredients to seeds, force us to explore reliable and efficient coating formulations. Biochar, as a novel material, may be expected to enhance seed germination and seedling establishment, simultaneously ensuring agricultural sustainability, environment, and food safety. Recently, biochar-based seed coating has gained much interest due to biochar possessing high porosity and water holding capacity, as well as wealthy nutrients, and has been proven to be a beneficial agent in seed coating formulations. This review presents an extensive overview on the history, methods, and coating agents of seed coating. Additionally, biochar, as a promising seed coating agent, is also synthesized on its physico-chemical properties. Combining seed coating with biochar, we discussed in detail the agricultural applications of biochar-based seed coating, such as the promotion of seed germination and stand establishment, the improvement of plant growth and nutrition, suitable carriers for microbial inoculants, and increase in herbicide selectivity. Therefore, this paper could be a good source of information on the current advance and future perspectives of biochar-based seed coating for modern agriculture.

Molecular Breeding of Water-Saving and Drought-Resistant Rice for Blast and Bacterial Blight Resistance.

Rice production is often affected by biotic and abiotic stressors. The breeding of resistant cultivars is a cost-cutting and environmentally friendly strategy to maintain a sustainable high production level. An elite water-saving and drought-resistant rice (WDR), Hanhui3, is susceptible to blast and bacterial blight (BB). This study was conducted to introgress three resistance genes (Pi2, xa5, and Xa23) for blast and BB into Hanhui3, using marker-assisted selection (MAS) for the foreground selection and a whole-genome single-nucleotide polymorphism (SNP) array for the background selection. As revealed by the whole-genome SNP array, the recurrent parent genome (RPG) recovery of the improved NIL was 94.2%. The resistance levels to blast and BB of the improved NIL and its derived hybrids were higher than that of the controls. In addition, the improved NIL and its derived hybrids retained the desired agronomic traits from Hanhui3, such as yield. The improved NIL could be useful to enhance resistance against biotic stressors and produce stable grain yields in Oryza sativa subspecies indica rice breeding programs.

How Biochar Affects Nitrogen Assimilation and Dynamics by Interacting Soil and Plant Enzymatic Activities: Quantitative Assessment of 2 Years Potted Study in a Rapeseed-Soil System.

The amendment of biochar has been proposed to improve soil fertility and crop yield. However, consolidated information lacks explaining the role of biochar on soil and plant enzymatic activities involved in nutrients cycling in soil and accumulation in plants improving utilization of applied inorganic fertilizer and crop growth. In the current study, we evaluated the integral effects of biochar levels (B0:0, B15:15, B3:30, and B60:60 t ha-1) and nitrogen fertilizer levels (N0:0, N75:75, N225:225, and N450:450 kg ha-1) on soil physicochemical properties, enzymatic activities, nitrogen use efficiency (NUE) and grain yield of rapeseed for 2 years in the pots during 2020 and 2021. The findings revealed that compared to control (B0 + N0), a combination of B30 + N450 increased soil urease activity by 73 and 75%, and B60 + N450 increased activities of soil catalase by 17 and 16%, and B60 + N225 increased alkaline phosphatase by 17 and 19%, respectively, in the first and second year. Moreover, a single application of high nitrogen at 450 kg ha-1 reduced the activities of plant nitrogen metabolism-related enzymes, however; the integration of biochar at 30 t ha-1 compensated the high nitrogen toxicity and improved the activities of nitrate reductase (NR), nitrite reductase NIR, glutamate synthase (GS) and glutamine synthetase (GOGAT) at seedling stage (SS) and flowering stage (FS) in both years. The integration of biochar at 30 t ha-1 with nitrogen at 450 kg ha-1 induced synergetic effects on rapeseed growth through sorption of excessive nitrogen in soil and significantly improved the plant height up to 11 and 18%, pods plant-1 39 and 32% and grain yield plant-1 54 and 64%, respectively, during the first and second year. Moreover, biochar at 15 t ha-1 along with nitrogen at 225 kg ha-1 resulted in the highest NUE of 29% in both years suggesting that biochar can also offset the deficiency of lower nitrogen. This study highlighted the ameliorative effect of biochar suppressing high nitrogen toxicity and decreasing lower nitrogen deficiency effects on rapeseed growth by improving nitrogen use efficiency via enhancing soil conditions, enzymatic activities and soil nitrogen utilization potential and thus improving rapeseed growth and yield.

Breeding an early maturing, blast resistance water-saving and drought-resistance rice (WDR) cultivar using marker-assisted selection coupled with rapid generation advance.

The japonica water-saving and drought-resistance rice (Oryza sativa L.) (WDR) cultivar Huhan 9 harbors genes for resistance to rice blast (Magnaporthe oryzae), including Pi-ta and Pi-b. The early maturing japonica rice cultivar Suhuxiangjing and the high-yield WDR cultivars Huhan 3 and Huhan 11 were used as the parents to conduct single cross breeding and composite hybridization breeding. Strict drought resistance screening was conducted in the segregating generations, the genotypes of which were determined using functional markers of Pi-ta and Pi-b genes. By combining the rapid generation advance of the industrialized breeding system and multi-site shuttle identification, the new WDR cultivar Huhan 106 with early maturity, blast resistance, high yield, and high quality was bred, and it was certified by the Agricultural Crop Variety Certification Commission of Shanghai in 2020. Molecular marker-assisted selection coupled with rapid generation advance and multi-site shuttle identification is a rapid and efficient breeding method for the value-added improvement of crop varieties. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-022-01319-3.

Molecular Breeding of a Novel PTGMS Line of WDR for Broad-Spectrum Resistance to Blast Using Pi9, Pi5, and Pi54 Genes.

BACKGROUND: The two-line method based on the photoperiod and thermo-sensitive genic male sterile (PTGMS) lines is more cost-effective, simple, and efficient than the three-line system based on cytoplasmic male-sterility. Blast and drought are the most prevalent biotic and abiotic stress factors hampering rice production. Molecular techniques demonstrate higher efficacy in the pyramiding of disease resistance genes, providing green performance under the background of water-saving and drought-resistance rice. RESULTS: This study employed molecular marker-assisted selection, conventional hybridization, and high-intensity stress screening to integrate three broad-spectrum blast resistance genes Pi9, Pi5, and Pi54 into Huhan 1S. Subsequently, a novel water-saving and drought-resistance rice (WDR) PTGMS line Huhan 74S was developed. The drought resistance of the new PTGMS line Huhan 74S was comparable to that of Huhan 1S. Pathogenicity assays involving the inoculation of 14 blast prevalent isolates in the glasshouse showed that the blast resistance frequency of Huhan 74S was 85.7%. Further evaluation under natural blast epidemic field conditions showed that Huhan 74S and its hybrids were resistant to leaf and neck blast. The critical temperature point of fertility-sterility alteration of Huhan 74S was 23 °C daily mean temperature. The complete male sterility under natural growth conditions in 2017 at Shanghai lasted for 67 days. Also, both the agronomic and grain quality traits met the requirement for two-line hybrid rice production. CONCLUSION: These results indicate that the newly bred PTGMS line Huhan 74S can be used to breed high-yielding, good-quality, disease-resistant two-line hybrid water-saving and drought-resistance rice (WDR), hence promoting sustainable rice production in China.

Fine mapping a quantitative trait locus, qSER-7, that controls stigma exsertion rate in rice (Oryza sativa L.).

BACKGROUND: Stigma exsertion rate (SER) is a key determinant of outcrossing in hybrid rice seed production. A quantitative trait locus (QTL) for stigma exsertion rate in rice, qSER-7, has previously been detected on chromosome 7 by using a F2 population derived from two indica cytoplasmic male sterility (CMS) maintainers, Huhan 1B and II-32B. RESULTS: The chromosomal location of qSER-7 was precisely delimited by fine-scale mapping. Near-isogenic lines (NILs) were established, one of which isolated the locus in the qSER-7II-32B line, which contains an introgressed segment of II-32B in the Huhan 1B genetic background, and exhibits a significantly higher stigma exsertion rate than that of the recurrent parent. Using 3192 individuals from the BC4F2 segregation population, the QTL qSER-7 was narrowed down to a 28.4-kb region between the markers RM3859 and Indel4373 on chromosome 7. According to the rice genome annotation database, three genes were predicted within the target region. Real-time PCR analysis showed significantly higher expression levels of LOC_Os07g15370 and LOC_Os07g15390 in II-32B than in Huhan 1B. LOC_Os07g15370(OsNRAMP5) was a previously reported gene for Mn and Cd transporter. The stigma exertion rates of OsNRAMP5-overexpressing plants were significantly higher than that of wild type plants, in contrast, a T-DNA insertion mutant osnramp5 showed a lower stigma exertion rate. CONCLUSIONS: In the present study, the QTL qSER-7 was isolated to a region between the markers RM3859 and Indel4373. Two candidate genes were selected based on the expression difference between the two parents, which can facilitate the further cloning of the gene underlying the quantitative trait associated with qSER-7 as well as the marker-assisted transfer of desirable genes for stigma exsertion rate improvement in rice.

Development and validation of a PCR-based functional marker system for identifying the low amylose content-associated gene Wx hp in rice.

Low amylose content (AC) is a desirable trait for rice (Oryza sativa L.) cooking quality and is selected in soft rice breeding. The Wx hp allele was derived from a Yunnan rice landrace in China, Haopi, with low AC. To efficiently and rapidly utilize the low amylose content-associated gene Wx hp in rice molecular breeding programs, we developed a tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) method, according to the single-nucleotide variation of the Wx hp . Four Wx hp -specific primers were used to perform PCR assays using genomic DNA extracted from several rice varieties. Based on the band pattern of the amplified products after electrophoresis, this method can accurately distinguish three Wx hp -related genotypes (i.e., Wx hp homozygotes, Wx hp heterozygotes, and wild-type), and the genotypes completely correspond to the appearance of mature endosperm. This method represents a novel approach that is both inexpensive and highly efficient and can be widely used for genotyping Wx hp alleles in rice germplasm collections and may aid breeding programs with marker-assisted selection (MAS).

Enhanced rice salinity tolerance via CRISPR/Cas9-targeted mutagenesis of the OsRR22 gene.

Salinity is one of the most important abiotic stress affecting the world rice production. The cultivation of salinity-tolerant cultivars is the most cost-effective and environmentally friendly approach for salinity control. In recent years, CRISPR/Cas9 systems have been widely used for target-site genome editing; however, their application for the improvement of elite rice cultivars has rarely been reported. Here, we report the improvement of the rice salinity tolerance by engineering a Cas9-OsRR22-gRNA expressing vector, targeting the OsRR22 gene in rice. Nine mutant plants were identified from 14 T0 transgenic plants. Sequencing showed that these plants had six mutation types at the target site, all of which were successfully transmitted to the next generations. Mutant plants without transferred DNA (T-DNA) were obtained via segregation in the T1 generations. Two T2 homozygous mutant lines were further examined for their salinity tolerance and agronomic traits. The results showed that, at the seedling stage, the salinity tolerance of T2 homozygous mutant lines was significantly enhanced compared to wild-type plants. Furthermore, no significantly different agronomic traits were found between T2 homozygous mutant lines and wild-type plants. Our results indicate CRISPR/ Cas9 as a useful approach to enhance the salinity tolerance of rice.