Neo-tetraploid rice with high fertility is a useful germplasm for polyploid rice breeding, which was developed from the crossing of different autotetraploid rice lines. However, little information is available on the molecular mechanism underlying the fertility of neo-tetraploid rice. Here, two contrasting populations of tetraploid rice, including one with high fertility (hereafter referred to as JG) and another with low fertility (hereafter referred to as JD), were generated by crossing Huaduo 3 (H3), a high fertility neo-tetraploid rice that was developed by crossing Jackson-4x with 96025-4x, and Huajingxian74-4x (T452), a low fertility autotetraploid rice parent. Cytological, global genome sequencing-based bulked-segregant (BSA-seq) and CRISPR/Cas9 technology were employed to study the genes associated with pollen fertility in neo-tetraploid rice. The embryo sacs of JG and JD lines were normal; however, pollen fertility was low in JD, which led to scarce fertilization and low seed setting. Cytological observations displayed low pollen fertility (25.1%) and approximately 31.3 and 27.2% chromosome lagging at metaphase I and II, and 28.8 and 24.8% chromosome straggling at anaphase I and II in JD, respectively. BSA-seq of F_2–3 generations and RNA-seq of F₄ generation detected a common fragment, i.e., 18,915,234–19,500,000, at chromosome 7, which was comprised of 78 genes associated with fertility. Among 78 genes, 9 genes had been known to be involved in meiosis and pollen development. Two mutants ny1 (LOC_Os07g32406) and ny2 (LOC_Os07g32040) were generated by CRISPR/Cas9 knockout in neo-tetraploid rice, and which exhibited low pollen fertility and abnormal chromosome behavior. Our study revealed that two unknown genes, LOC_Os07g32406 (NY1) and LOC_Os07g32040 (NY2) play an important role in pollen development of neo-tetraploid rice and provides a new perspective about the genetic mechanisms of fertility in polyploid rice.

The management of fertilizers in a context of climate change and the preservation of the environment is strongly related to the regulation and accumulation of 2-acetyl-1-pyrroline (2AP) in fragrant rice. However, the feasibility of such management strategies in terms of enhancing the accumulation of 2AP has not yet been explored in aromatic cultivars. Here, we investigated the impact of the application of two fertilizers at three depth (surface, 5 cm and 10 cm) levels of placements to improve the aromatic rice quality, including such aspects as the 2AP content-, protein-, amylose- and yield-related traits. For this purpose, two known rice cultivars, Basmati 385 (B-385) and Yunjingyou (YJY), were grown in pots during 2019 and 2020 under fluctuating climates. The deep application of fertilizer at 10 cm significantly affected the 2AP content with such values as 127.53 μg kg⁻¹ and 111.91 μg kg⁻¹ obtained for Fragrant Fertilizer (FF) and Urea in B-385 cultivar, and 126.5 μg kg⁻¹ and 114.24 μg kg⁻¹ being observed for FF and Urea in YJY, respectively, during 2019. In addition, values of 108.41 μg kg⁻¹ and 117.35 μg kg⁻¹ were recorded for FF and Urea in B-385, while 125.91-μg kg⁻¹ and 90.71-μg kg⁻¹ were measured for FF and Urea in YJY, respectively, during 2020. Similarly, B-385 had better 2AP content and yield-related traits, as well as amylose content and cooked rice elongation, as compared to the YJY rice cultivar. The 2AP accumulation and its related biochemical parameters, and their relationships in different plant tissues at different growth stages under FF and Urea treatments, were also improved. Further, the 2AP content and the P5C activity demonstrated strong correlations during the grain filling periods in both fragrant rice cultivars. In conclusion, our findings have the potential to provide useful information to farmers and agriculture extension workers in terms of the saving of fertilizers and the improvement of rice grain quality under fluctuating climate conditions.

Autotetraploid rice is a useful germplasm for the breeding of polyploid rice; however, low fertility is a major hindrance for its utilization. Neo-tetraploid rice with high fertility was developed from the crossing of different autotetraploid rice lines. Our previous research showed that the mutant (ny1) of LOC_Os07g32406 (NY1), which was generated by CRISPR/Cas9 knock-out in neo-tetraploid rice, showed low pollen fertility, low seed set, and defective chromosome behavior during meiosis. However, the molecular genetic mechanism underlying the fertility remains largely unknown. Here, cytological observations of the NY1 mutant (ny1) indicated that ny1 exhibited abnormal tapetum and middle layer development. RNA-seq analysis displayed a total of 5606 differentially expressed genes (DEGs) in ny1 compared to wild type (H1) during meiosis, of which 2977 were up-regulated and 2629 were down-regulated. Among the down-regulated genes, 16 important genes associated with tapetal development were detected, including EAT1, CYP703A3, CYP704B2, DPW, PTC1, OsABCG26, OsAGO2, SAW1, OsPKS1, OsPKS2, and OsTKPR1. The mutant of EAT1 was generated by CRISPR/Cas9 that showed abnormal tapetum and pollen wall formation, which was similar to ny1. Moreover, 478 meiosis-related genes displayed down-regulation at same stage, including 9 important meiosis-related genes, such as OsREC8, OsSHOC1, SMC1, SMC6a and DCM1, and their expression levels were validated by qRT-PCR. Taken together, these results will aid in identifying the key genes associated with pollen fertility, which offered insights into the molecular mechanism underlying pollen development in tetraploid rice.

The number one goal of most rice improvement programmes around the world is breeding for high potential to take care of the increasing consumption of rice. The objective of this experiment was to study the inheritance of yield and yield components and to estimate the heritabilities of important quantitative traits in rice (Oryza sativa L.). Six generations viz., P1, P2, F1, F2, BCP1 and BCP2 of a cross between IET6279 and IR70445-146-3-3 were used for the study. Generation mean analysis suggested that additive effects had a major role for the expression of plant height, number of panicles, number of spikelets per panicle, percentage spikelets fertility per plant and grain yield per plant, which further suggested that phenotypic selection was possible at an early stage. Both additive and non-additive gene actions were important for the expression of number of fertile spikelets per panicle and 100-grain weight. Therefore, selection for these characters would be fruitful, if delayed till epistasis effects are reduced to a minimum. High broad sense heritability estimates were observed for characters viz. plant height and 100-grain weight, suggesting that the characters under study are less influenced by environment in their expression. Thus, selection based on phenotypic value of these characters would be reliable and effective. Low estimates of broad sense heritability were observed for number of panicles per plant, number of spikelets per panicle, number of fertile spikelets per panicle, percentage spikelets fertility per plant and grain yield per plant, indicating that influence of the environment was high for these characters. Therefore, superior genotypes selection based on phenotypic performance for these characters may not effective. Key Words: Additive, gene action, heritability, inheritance, non- additive, rice

Discrete element model (DEM) parameters are crucial for accurately predicting soil properties and disturbance levels. This study aimed to provide an efficient method for accurately determining DEM parameters for paddy soil. The Hertz-Mindlin and JKR contact models were used to simulate the paddy soil, and the Plackett-Burman, the Steepest Ascent, and the Box-Behnken tests were used to determine the DEM parameters. The accuracy of the established discrete element model was evaluated using actual slump test. The Plackett-Burman test results showed that the soil surface energy, soil-soil rolling friction coefficient, and soil-steel static friction coefficient had a significant impact on the total relative error between the simulation results and the test results. Box-Behnken test optimization results show that the soil surface energy, soil-soil rolling friction coefficient, and soil-steel static friction coefficient are 0.869 J.m², 0.109, and 0.651, respectively. Comparison with actual soil slump test results shows that the calibrated DEM model has an overall relative error of 5.59% and a coefficient of variation of 3.49%. This research can provide a theoretical basis and technical support for subsequent research on soil-machine interaction mechanisms.