Rice Shoots Research Articles

Antixenosis and antibiosis mechanisms of resistance to Asian rice gall midge, Orseolia oryzae (Wood‐Mason) in rice land races

AbstractAsian rice gall midge is one of the important pests of rice, which attacks the crop from nursery to the end of the tillering stage. Managing this pest through plant resistance is the most viable and economical. Two hundred‐two rice genotypes were phenotyped against Asian rice gall midge, Orseolia oryzae (Wood‐Mason) and the mechanism of resistance in terms of antixenosis and antibiosis was studied. Antixenosis mechanism for adult settlement and egg laying indicated that the susceptible genotypes were preferred more than the resistant genotypes in a free choice test. First instar maggots were found and did not continue their growth on resistant genotypes further. However, in susceptible genotypes, they molted successfully and emerged as adults. No choice tests revealed that the emergence of adults in susceptible genotypes took less time than in resistant genotypes. Higher adult sex ratio was found in susceptible genotypes. Estimation of biochemical components in rice shoot apices of selected genotypes revealed that higher levels of total phenols, wax content, total flavonoids and total free amino acids were present in the resistant genotypes. Still, the number of total sugars, reducing sugars and total protein contents were significantly higher in the susceptible genotypes. Hence, the resistance mechanism in rice gall midge was displayed as a combination of antixenosis and antibiosis mechanism. The output of the study would be helpful in breeding for rice varieties resistant to Asian rice gall midge.

Kinetin-mediated reduction of cadmium accumulation in rice (Oryza sativa L.) via modulation of cell wall binding capacity in a NO-dependent manner

Cadmium (Cd) is a heavy metal that is toxic to both plant growth and human health. Kinetin, a precursor of cytokinin, plays crucial roles in the response of plants to abiotic stress. However, the mechanism underlying kinetin detoxification in Cd-induced stress remains poorly understood. In this study, we observed a significant increase in endogenous kinetin levels in both rice shoots and roots under Cd stress. Furthermore, the application of exogenous kinetin resulted in a reduction in hemicellulose content within cell walls, thereby reducing the cell wall's capacity to bind Cd and limit its entry into cells. In addition, exogenous kinetin downregulated the transcript levels of genes associated with Cd uptake and root-to-shoot translocation while simultaneously upregulating the transcript levels of genes involved in Cd efflux from cells and sequestration into vacuoles, consequently leading to reduced Cd accumulation in rice plants. Moreover, kinetin significantly decreased nitric oxide (NO) accumulation in response to Cd stress, potentially facilitating detoxification during Cd-induced stress through a NO-dependent mechanism. Overall, our findings suggest that kinetin enhances rice's tolerance to Cd by increasing the cell wall's ability to bind Cd and regulating gene expression related to Cd accumulation. These mechanisms may be NO-dependent and could potentially inform the development of new strategies for mitigating Cd toxicity in agricultural systems.

Dynamics of Humic Acid, Silicon, and Biochar under Heavy Metal, Drought, and Salinity with Special Reference to Phytohormones, Antioxidants, and Melatonin Synthesis in Rice.

This study aimed to develop a biostimulant formulation using humic acid (HA), silicon, and biochar alone or in combination to alleviate the lethality induced by combined heavy metals (HM-C; As, Cd, and Pb), drought stress (DS; 30-40% soil moisture), and salt stress (SS; 150 mM NaCl) in rice. The results showed that HA, Si, and biochar application alone or in combination improved plant growth under normal, DS, and SS conditions significantly. However, HA increased the lethality of rice by increasing the As, Cd, and Pb uptake significantly, thereby elevating lipid peroxidation. Co-application reduced abscisic acid, elevated salicylic acid, and optimized the Ca2+ and Si uptake. This subsequently elevated the K+/Na+ influx and efflux by regulating the metal ion regulators (Si: Lsi1 and Lsi2; K+/Na+: OsNHX1) and increased the expressions of the stress-response genes OsMTP1 and OsNramp in the rice shoots. Melatonin synthesis was significantly elevated by HM-C (130%), which was reduced by 50% with the HA + Si + biochar treatment. However, in the SS- and DS-induced crops, the melatonin content showed only minor differences. These findings suggest that the biostimulant formulation could be used to mitigate SS and DS, and precautions should be taken when using HA for heavy metal detoxification.

Co-inoculation effects of B. licheniformis and P. aeruginosa on soil Cd and As availability and rice accumulation

There have been studies reporting the effects of multiple bacterial strains on the Cd/As immobilization and transformation in culture media. However, there is limited research to validate the effects of microbial strain combination on plant Cd/As accumulation and antioxidant system in the soil-plant system. By planting the rice (Zhefu 7) with the co-inoculation of bacterial strains (i.e. Bacillus licheniformis and Pseudomonas aeruginosa) after two months with the contaminations of Cd (2 mg/kg), As (80 mg/kg) and Cd + As (2 + 80 mg/kg), we found that the bacterial co-inoculation decreased Cd concentrations in the rhizosphere soil porewater, but had limited effects on mitigating plant Cd accumulation. By contrast, the co-inoculation did not affect the As(III) and As(V) concentrations in the rhizosphere soil porewater, but decreased As(III) and As(V) concentrations by 17% and 17% in the root respectively and by 17% and 37% in rice shoot respectively. Using DNA sequencing, we found the increased abundance in both exogenous Bacillus licheniformis and native microorganisms, indicating that the added strains had synergetic interactions with soil native microorganisms. Regarding on plant antioxidant enzyme system, the bacterial co-inoculation decreased the concentrations of superoxide dismutase (SOD), hydrogen peroxide (H2O2) and malondialdehyde (MDA) by 75%, 74% and 22%, mitigating the As damage to rice root and promote plant growth. However, under Cd and As co-stress, the effects of co-inoculation on mitigating plant As accumulation and enhancing plant stress resistance appear to be diminished. Our findings underscore the importance of microbial co-inoculation in reducing plant As accumulation and preserving plant health under heavy metal stress.

GenX uptake by wheat and rice in flooded and non-flooded soils: a greenhouse experiment

GenX (hexafluoropropylene oxide dimer acid) belongs to the group of per- and poly-fluoroalkyl substance (PFAS) compounds introduced to replace perfluorooctanoic acid (PFOA), which has been phased out in industrial and consumer product formulations. While GenX has been investigated in lab animals, there is limited information available regarding its uptake and translocation in wheat and rice. This study reports on a greenhouse experiment in which wheat and rice grown under flooded and non-flooded conditions were exposed to two GenX concentrations in the soil (0.4 mg kg−1 and 2 mg kg−1). GenX was analysed in the soil, porewater and shoots using targeted liquid chromatography-tandem mass spectroscopy (LC-MS/MS) analysis. Extractable organic fluorine (EOF) was determined using high-resolution continuum source graphite furnace molecular absorption spectrometry (HR-GFMAS) instrument. Results showed that different species took up different amounts of GenX. The GenX concentration in rice shoots was found to be 2.34 (± 0.45) and 4.11 (± 0.87) μg g−1 under flooded and non-flooded conditions, respectively, at a low exposure level. At high exposure, the GenX concentrations in flooded and non-flooded rice shoots increased threefold to 10.4 (± 0.41) and 13.4 (± 0.72) μg g−1, respectively. Wheat shoots showed similar concentrations and increases between low- and high-level exposure. The translocation factor was significantly higher (P = 0.013) in non-flooded rice compared to flooded rice. The GenX bioaccumulation behaviours under the same culture conditions (e.g. temperature, humidity, light, same GenX concentration in the soil) were significantly different in non-flooded and flooded rice (P < 0.001). Non-flooded rice plants displayed a higher level of GenX bioaccumulation than flooded ones. Following exposure to GenX, flooded rice plants showed a reduction in biomass (25%) compared to the control plants (P < 0.014). Our findings indicate that GenX is a bioaccumulative compound, the presence of which likely inhibits the growth of plants.

More effective than direct contact: Nano hydroxyapatite pre-treatment regulates the growth and Cd uptake of rice (Oryza sativa L.) seedlings

Cd contamination in rice urgently needs to be addressed. Nano hydroxyapatite (n-HAP) is an eco-friendly material with excellent Cd fixation ability. However, due to its own high reactivity, innovative application of n-HAP in the treatment of Cd contamination in rice is needed. In this study, we proposed a new application, namely n-HAP pre-treatment, which can effectively reduce Cd accumulation in rice and alleviate Cd stress. The results showed that 80 mg/L n-HAP pre-treatment significantly reduced Cd content in rice shoot by 35.1%. Biochemical and combined transcriptomic-proteomic analysis revealed the possible molecular mechanisms by which n-HAP pre-treatment promoted rice growth and reduced Cd accumulation. (1) n-HAP pre-treatment regulated gibberellin and jasmonic acid synthesis-related pathways, increased gibberellin content and decreased jasmonic acid content in rice root, which promoted rice growth; (2) n-HAP pre-treatment up-regulated gene CATA1 expression and down-regulated gene OsGpx1 expression, which increased rice CAT activity and GSH content; (3) n-HAP pre-treatment up-regulated gene OsZIP1 expression and down-regulated gene OsNramp1 expression, which reduced Cd uptake, increased Cd efflux from rice root cells.

Low phosphorus promotes NSP1–NSP2 heterodimerization to enhance strigolactone biosynthesis and regulate shoot and root architecture in rice

Phosphorus is an essential macronutrient for plant development and metabolism, and plants have evolved ingenious mechanisms to overcome phosphate (Pi) starvation. However, the molecular mechanisms underlying the regulation of shoot and root architecture by low phosphorus conditions and the coordinated utilization of Pi and nitrogen remain largely unclear. Here, we show that Nodulation Signaling Pathway 1 (NSP1) and NSP2 regulate rice tiller number by promoting the biosynthesis of strigolactones (SLs), a class of phytohormones with fundamental effects on plant architecture and environmental responses. We found that NSP1 and NSP2 are induced by Oryza sativa PHOSPHATE STARVATION RESPONSE2 (OsPHR2) in response to low-Pi stress and form a complex to directly bind the promoters of SL biosynthesis genes, thus markedly increasing SL biosynthesis in rice. Interestingly, the NSP1/2–SL signaling module represses the expression of CROWN ROOTLESS 1 (CRL1), a newly identified early SL-responsive gene in roots, to restrain lateral root density under Pi deficiency. We also demonstrated that GR244DO treatment under normal conditions inhibits the expression of OsNRTs and OsAMTs to suppress nitrogen absorption but enhances the expression of OsPTs to promote Pi absorption, thus facilitating the balance between nitrogen and phosphorus uptake in rice. Importantly, we found that NSP1p:NSP1 and NSP2p:NSP2 transgenic plants show improved agronomic traits and grain yield under low- and medium-phosphorus conditions. Taken together, these results revealed a novel regulatory mechanism of SL biosynthesis and signaling in response to Pi starvation, providing genetic resources for improving plant architecture and nutrient-use efficiency in low-Pi environments.

A citrate efflux transporter important for manganese distribution and phosphorus uptake in rice.

The plant citrate transporters, functional in mineral nutrient uptake and homeostasis, usually belong to the multidrug and toxic compound extrusion transporter family. We identified and functionally characterized a rice (Oryza sativa) citrate transporter, OsCT1, which differs from known plant citrate transporters and is structurally close to rice silicon transporters. Domain analysis depicted that OsCT1 carries a bacterial citrate-metal transporter domain, CitMHS. OsCT1 showed citrate efflux activity when expressed in Xenopus laevis oocytes and is localized to the cell plasma membrane. It is highly expressed in the shoot and reproductive tissues of rice, and its promoter activity was visible in cells surrounding the vasculature. The OsCT1 knockout (KO) lines showed a reduced citrate content in the shoots and the root exudates, whereas overexpression (OE) line showed higher citrate exudation from their roots. Further, the KO and OE lines showed variations in the manganese (Mn) distribution leading to changes in their agronomical traits. Under deficient conditions (Mn-sufficient conditions followed by 8 days of 0 μm MnCl2 · 4H2 O treatment), the supply of manganese towards the newer leaf was found to be obstructed in the KO line. There were no significant differences in phosphorus (P) distribution; however, P uptake was reduced in the KO and increased in OE lines at the vegetative stage. Further, experiments in Xenopus oocytes revealed that OsCT1 could efflux citrate with Mn. In this way, we provide insights into a mechanism of citrate-metal transport in plants and its role in mineral homeostasis, which remains conserved with their bacterial counterparts.

Dissecting the promotional effect of zinc on cadmium translocation from roots to shoots in rice.

It is often expected that Zn decreases Cd accumulation in plants due to competition for the same transporters. Here, we found that increasing Zn supply markedly increased the root-to-shoot translocation of Cd in rice. RNA sequencing showed that high Zn up-regulated expression of genes involved in glutathione biosynthesis and metabolism and the Zn/Cd transporter gene OsHMA2, but down-regulated expression of genes related to Zn uptake. Knockout of the iron or Zn transporter genes OsIRT1, OsIRT2, or OsZIP9 did not affect the Zn promotional effect on Cd translocation. Knockout of the manganese/Cd transporter gene OsNRAMP5 greatly reduced Cd uptake but did not affect the Zn promotional effect. Variation in the tonoplast transporter gene OsHMA3 affected Cd translocation but did not change the Zn promotional effect. Knockout of the Zn/Cd transporter gene OsHMA2 not only decreased Cd and Zn translocation, but also abolished the Zn promotional effect. Increased expression of OsHMA2 under high Zn conditions supports the hypothesis that this transporter participates in the promotional effect of Zn on Cd translocation. The results also show that OsIRT1, OsIRT2, and OsZIP9 made only small contributions to Cd uptake under low Zn conditions but not under high Zn conditions, whereas the dominant role of OsNRAMP5 in Cd uptake diminished under low Zn conditions.

Hormones and the antioxidant transduction pathway and gene expression, mediated by Serratia marcescens DB1, lessen the lethality of heavy metals (As, Ni, and Cr) in Oryza sativa L.

Microorganisms have recently gained recognition as efficient biological tool for reducing heavy metal toxicity in crops. In this experiment, we isolated a potent heavy metal (As, Ni, and Cr) resistant rhizobacterium Serratia marcescens DB1 and detected its plant growth promoting traits such as phosphate solubilization, gibberellin synthesis, organic acid production and amino acid regulation. Based on these findings, DB1 was further investigated for application in a rice var. Hwayeongbyeo subjected to 1 mM As, 4 mM Ni, and 4 mM Cr stress. The rice plants treated with Cr and Ni appeared healthy but were lethal, indicating unfitness for consumption due to toxic metal deposition, whereas the plants treated with > 1 mM As instantaneously died. Our results showed that DB1 inoculation significantly decreased metal accumulation in the rice shoots. Particularly, Cr uptake dropped by 16.55% and 22.12% in (Cr + DB1) and (Cr + As + Ni + DB1), respectively, As dropped by 48.90% and 35.82% in (As + DB1) and (Cr + As + Ni + DB1), respectively, and Ni dropped by 7.95% and 19.56% in (Ni + DB1) and (Cr + As + Ni + DB1), respectively. These findings were further validated by gene expression analysis results, which showed that DB1 inoculation significantly decreased the expression of OsPCS1 (a phytochelatin synthase gene), OsMTP1 (a metal transporting gene), and OsMTP5 (a gene for the expulsion of excess metal). Moreover, DB1 inoculation considerably enhanced the morphological growth of rice through modulation of endogenous phytohormones (abscisic acid, salicylic acid, and jasmonic acid) and uptake of essential elements such as K and P. These findings indicate that DB1 is an effective biofertilizer that can mitigate heavy metal toxicity in rice crops.

As(III)-Oxidizing Bacteria Alleviate Arsenite Toxicity via Reducing As Accumulation, Elevating Antioxidative Activities and Modulating Ionome in Rice (Oryza sativa L.).

Paddy rice trends to accumulate more arsenic (As) from soils than other terrestrial crops. The toxicity and mobility of As mainly depend on its chemical species. Transformation of arsenite [As(III)] into arsenate [As(V)] would be a promising method to mitigate As toxicity. In the current study, As(III)-oxidizing strain SMS11 isolated from As-contaminated soils was employed for As remediation. Co-cultured with SMS11 alleviated As(III) stress to the rice plants by increasing the length and biomass of rice shoots up to 10% and 15%, respectively. Evaluation of oxidative stress indices showed that the activity of catalase in the rice shoots was weakened when exposed to As(III), increasing the risk of hydroxyl radical (·OH) formation. When co-cultivated with the bacteria, ·OH formation was significantly inhibited in the rice shoots. The ionomes of the rice plants were impacted by the external conditions. As(III) stress significantly disturbed ionome homeostasis in the rice plants. Uptake of As simultaneously elevated the levels of macro and nutrient elements such as Mg, P, K, Ca, and Zn in the rice shoots. The ionomic variation in the rice plants under As(III) stress was mitigated by inoculated with SMS11. The results represented that the As(III)-oxidizing bacteria alleviated external As(III) stress to the rice plants through elevating antioxidative activities and modulating ionome homeostasis.

Towards increased shading capacity: A combined phenotypic and genetic analysis of rice shoot architecture

Societal Impact StatementRice farming is transitioning from transplanting rice seedlings towards the less labour‐intensive and less water‐demanding method of directly seeding rice. This, however, is accompanied by increased weed proliferation. To tackle this issue, this study seeks to identify how the crop itself can better suppress weeds, with a focus on light competition via shading. Using a rice diversity panel, traits were identified that contribute to enhanced shading capacity, and these traits were encapsulated into a single shading capacity metric. This was followed by the identification of the genetic loci underpinning variation in the core traits. The identified haplotypes can be used in breeding programmes to improve weed suppression by rice, thus contributing to sustainable agriculture.Summary In modern rice farming, one of the major constraints is weed proliferation and the entailed ecological impact of herbicide application. This requires increased weed competitiveness in current rice varieties, achieved via enhanced shade casting to limit the growth of shade‐sensitive weeds. To identify traits that increase rice shading capacity, we exhaustively phenotyped a rice diversity panel of 344 varieties at an early vegetative stage. A genome‐wide association study (GWAS) revealed genetic loci underlying variation in canopy architecture traits linked with shading capacity. The screen shows considerable natural variation in shoot architecture for 13 examined traits, of which shading potential is mostly determined by projected shoot area, number of leaves, culm height and canopy solidity. The shading rank, a metric based on these core traits, identifies varieties with the highest shading potential. Five genetic loci were found to be associated with canopy architecture, shading potential and early vigour. Identification of traits contributing to shading capacity and underlying allelic variation will serve future genomic‐assisted breeding programmes. Implementing the presented genetic resources for increased shading and weed competitiveness in rice breeding will make its farming less dependent on herbicides and contribute towards more environmentally sustainable agriculture.

Pelapisan Benih dengan Aktinobakteri untuk Meningkatkan Pertumbuhan Tanaman Padi

The need for rice continues to increase along with the increase in population. Efforts to increase rice production is generally carried out through proper and balanced fertilization. Other than that, plant growth-promoting actinobacterial (PGPB) inoculants can be used as an alternative solution. This study aimed to examine the effect of the application of actinobacterial seed dressing and NPK fertilizers on the growth of rice plants grown in a glass house. A randomized block design with three factors was conducted, consisting of actinobacterial seed dressing (added and not added), type of carrier (zeolite, peat, and combinations) and doses of NPK fertilizer (0 g/pot, 0.375 g/pot, and 0.75 g/pot). The addition of actinobacteria consortium, peat-zeolite combination 1:3, and NPK fertilizer at a dose of 0.75 g/pot (A2C3P3) gave consistent results in increasing the average yield of rice vegetative and reproductive parameters observed in the glass house. The A2C3P3 treatment had a significant effect on the number of tillers, width of flag leaf, dry weight of roots and shoots of rice observed at 10 WAP compared to other treatment combinations. The actinobacterial seed coating plays a pivotal role in supporting rice plant growth.

Physiological and transcriptome analysis reveals that prohexadione-calcium promotes rice seedling's development under salt stress by regulating antioxidant processes and photosynthesis.

Prohexadione-calcium (Pro-Ca) has been proved to play an important role in releasing abiotic stress in plants. However, there is still a lack of research on the mechanism of Pro-Ca alleviating salt stress in rice. To explore the protective effects of Pro-Ca on rice seedlings under salt stress, we investigated the effect of exogenous Pro-Ca on rice seedling under salt stress by conducting the following three treatment experiments: CK (control), S (50 mmol·L-1 NaCl saline solution) and S + Pro-Ca (50 mmol·L-1 NaCl saline solution + 100 mg·L-1 Pro-Ca). The results indicated that Pro-Ca modulated the expression of antioxidant enzyme-related genes (such as SOD2, PXMP2, MPV17, E1.11.1.7). Spraying Pro-Ca under salt stress significantly increased in ascorbate peroxidase, superoxide dismutase, and peroxidase activity by 84.2%, 75.2%, and 3.5% as compared to the salt treatment, as demonstrated by an example of a 24-hour treatment. Malondialdehyde level in Pro-Ca was also dramatically decreased by 5.8%. Moreover, spraying Pro-Ca under salt stress regulated the expression of photosynthesis genes (such as PsbS, PsbD) and chlorophyll metabolism genes (heml, PPD). Compared to salt stress treatment, spraying Pro-Ca under salt stress significantly increased in net photosynthetic rate by 167.2%. In addition, when rice shoots were sprayed with Pro-Ca under salt stress, the Na+ concentration was considerably reduced by 17.1% compared to salt treatment. In conclusion, Pro-Ca regulates antioxidant mechanisms and photosynthesis to aid in the growth of rice seedlings under salt stress.

Growth rate and competitive ability of susceptible and multiple-resistant late watergrass (Echinochloa phyllopogon) biotypes to rice

Abstract Late watergrass [Echinochloa phyllopogon (Stapf) Koss.] has evolved multiple-resistance (MR) to herbicides in many areas worldwide and scientists wonder if this resistance has imposed fitness cost or advantage on the resistant (MR) biotypes. As multiple-resistance of this weed has also been recorded in Greece, one susceptible (S), one with multiple-resistance to ALS (penoxsulam, bispyribac, imazamox and nicosulfuron + rimsulfuron) and ACCase-cyhalofop (MRAC) and two with multiple-resistance to ALS and synthetic auxin-quinclorac (MRAQ1, MRAQ2) late watergrass biotypes collected from rice fields (northern Greece) were studied for their growth rate and competitive ability (regarding stem number and fresh weight) in the absence and presence of rice. Also, the competition indices relative yield (RY), competitive ratio (CR) and aggressiveness (AI) were evaluated. The S and the three MR biotypes grown in absence of competition (one plant/pot) showed different shoot number but similar fresh weight growth rates. In contrast, grown under intra-specific competition (one, two, four and six plants/pot), the S late watergrass biotype produced similar fresh weight with that of MRAQ1 but lower than that of the MRAC and MRAQ2 biotypes. However, grown under inter-specific competition (one, two, four and six plants of late watergrass/pot in competition with six rice plants/pot), the MRAQ2 late watergrass biotype produced more shoot number and fresh weight than the other biotypes at all weed densities, but the rice shoot number and fresh weight reduction were not followed the opposite trend of the late watergrass biotypes. Also, the competition indices of the MR biotypes (based on both intra- and inter-specific completion data) were found similar in most cases to that of the S biotype. These findings strongly suggest that there is no clear association between the growth rate and competitive ability of the late watergrass biotypes and their herbicide resistance.

Hydrogen peroxide reduces root cadmium uptake but facilitates root-to-shoot cadmium translocation in rice through modulating cadmium transporters

Cadmium (Cd) contamination in agricultural soils has become a serious worldwide environmental problem threatening crop production and human health. Hydrogen peroxide (H2O2) is a critical second messenger in plant response to Cd exposure. However, its role in Cd accumulation in various organs of plants and the mechanistic basis of this regulation remains to be elucidated. In this study, we used electrophysiological and molecular approaches to understand how H2O2 regulates Cd uptake and translocation in rice plants. Our results showed that the pretreatment of H2O2 significantly reduced Cd uptake by rice roots, which was associated with the downregulation of OsNRAMP1 and OsNRAMP5. On the other hand, H2O2 promoted the root-to-shoot translocation of Cd, which might be attributed to the upregulation of OsHMA2 critical for Cd2+ phloem loading and the downregulation of OsHMA3 involved in the vacuolar compartmentalization of Cd2+, leading to the increased Cd accumulation in rice shoots. Furthermore, such regulatory effects of H2O2 on Cd uptake and translocation were notably amplified by the elevated level of exogenous calcium (Ca). Collectively, our results suggest that H2O2 can inhibit Cd uptake but increase root to shoot translocation through modulating the transcriptional levels of genes encoding Cd transporters, furthermore, application of Ca can amplify this effect. These findings will broaden our understanding of the regulatory mechanisms of Cd transport in rice plants and provide theoretical foundation for breeding rice for low Cd accumulation.

The SMC5/6 complex subunit MMS21 regulates stem cell proliferation in rice.

SMC5/6 complex subunit OsMMS21 is involved in cell cycle and hormone signaling and required for stem cell proliferation during shoot and root development in rice. The structural maintenance of chromosome (SMC)5/6 complex is required for nucleolar integrity and DNA metabolism. Moreover, METHYL METHANESULFONATE SENSITIVITY GENE 21 (MMS21), a SUMO E3 ligase that is part of the SMC5/6 complex, is essential for the root stem cell niche and cell cycle transition in Arabidopsis. However, its specific role in rice remains unclear. Here, OsSMC5 and OsSMC6 single heterozygous mutants were generated using CRISPR/Cas9 technology to elucidate the function of SMC5/6 subunits, including OsSMC5, OsSMC6, and OsMMS21, in cell proliferation in rice. ossmc5/ + and ossmc6/ + heterozygous single mutants did not yield homozygous mutants in their progeny, indicating that OsSMC5 and OsSMC6 both play necessary roles during embryo formation. Loss of OsMMS21 caused severe defects in both the shoot and roots in rice. Transcriptome analysis showed a significant decrease in the expression of genes involved in auxin signaling in the roots of osmms21 mutants. Moreover, the expression levels of the cycB2-1 and MCM genes, which are involved the cell cycle, were significantly lower in the shoots of the mutants, indicating that OsMMS21 was involved in both hormone signaling pathways and the cell cycle. Overall, these findings indicate that the SUMO E3 ligase OsMMS21 is required for both shoot and root stem cell niches, improving the understanding of the function of the SMC5/6 complex in rice.

Transcription elongation factors OsSPT4 and OsSPT5 are essential for rice growth and development and act with APO2.

The transcription elongation factor SPT4/SPT5 complex is essential for rice vegetative and reproductive growth and that OsSPT5-1, with its interactor APO2, is involved in multiple phytohormone pathways. The SPT4/SPT5 complex is a transcription elongation factor that regulates the processivity of transcription elongation. However, our understanding of the role of SPT4/SPT5 complex in developmental regulation remains limited. Here, we identified three SPT4/SPT5 genes (OsSPT4, OsSPT5-1, and OsSPT5-2) in rice, and investigated their roles in vegetative and reproductive growth. These genes are highly conserved with their orthologs in other species. OsSPT4 and OsSPT5-1 are widely expressed in various tissues. By contrast, OsSPT5-2 is expressed at a relatively low level, which could cause osspt5-2 null mutants have no phenotypes. Loss-of-function mutants of OsSPT4 and OsSPT5-1 could not be obtained; their heterozygotes showed severe reproductive growth defects. An incomplete mutant line (osspt5-1#12) displayed gibberellin-related dwarfed defects and a weak root system at an early vegetative phase, and a short life cycle in different planting environments. Furthermore, OsSPT5-1 interacts with the transcription factor ABERRANT PANICLE ORGANIZATION 2 (APO2) and plays a similar role in regulating the growth of rice shoots. RNA sequencing analysis verified that OsSPT5-1 is involved in multiple phytohormone pathways, including gibberellin, auxin, and cytokinin. Therefore, the SPT4/SPT5 complex is essential for both vegetative and reproductive growth in rice.

Effects of endophytic Bacillus spp. on accumulation and distribution of iron in the shoots of lowland rice grown under iron toxic conditions

AbstractBackgroundThe tolerance of plants against abiotic stresses can be greatly influenced by their interaction with microbes. In lowland rice (Oryza sativa) production, the iron toxicity of the soils constitutes a major constraint. Although there are tolerant cultivars, the mechanisms underlying the tolerance against excess iron are not fully understood. Even less is known about the role of microbes in the response to iron toxicity.AimIn the study presented here, the effects of different Bacillus isolates on the accumulation and distribution of iron within the shoots of different rice cultivars grown under iron toxicity were analyzed.MethodsThree lowland rice cultivars with contrasting tolerance to iron toxicity (IR31785‐58‐1‐2‐3‐3, Sahel 108, Suakoko 8) were inoculated with three Bacillus isolates (two B. pumilus and one B. megaterium) and, after 1 week, exposed to excess iron (1,000 ppm) for 8 days. Tolerance was evaluated by leaf symptom scoring.ResultsBacterial inoculation mitigated leaf symptoms in the sensitive cultivar IR31785‐58‐1‐2‐3‐3 despite no significant differences in shoot iron concentration between inoculated and noninoculated plants. In the tolerant excluder cultivar, Suakoko 8, leaf symptoms were exacerbated when inoculated with B. pumilus Ni9MO12. While the total shoot Fe concentration was not affected in this bacteria × cultivar combination, the distribution of iron within the shoot was clearly disturbed. Tolerance to iron toxicity of the tolerant includer cultivar, Sahel 108, was not affected by Bacillus inoculation.ConclusionIn conclusion, our results show that Bacillus inoculation can affect the tolerance of lowland rice to iron toxicity and that the effects strongly depend on the bacteria × cultivar combination.

OsNPF3.1, a nitrate, abscisic acid and gibberellin transporter gene, is essential for rice tillering and nitrogen utilization efficiency

Low-affinity nitrate transporter genes have been identified in subfamilies 4–8 of the rice nitrate transporter 1 (NRT1)/peptide transporter family (NPF), but the OsNPF3 subfamily responsible for nitrate and phytohormone transport and rice growth and development remains unknown. In this study, we described OsNPF3.1 as an essential nitrate and phytohormone transporter gene for rice tillering and nitrogen utilization efficiency (NUtE). OsNPF3.1 possesses four major haplotypes of its promoter sequence in 517 cultivars, and its expression is positively associated with tiller number. Its expression was higher in the basal part, culm, and leaf blade than in other parts of the plant, and was strongly induced by nitrate, abscisic acid (ABA) and gibberellin 3 (GA3) in the root and shoot of rice. Electrophysiological experiments demonstrated that OsNPF3.1 is a pH-dependent low-affinity nitrate transporter, with rice protoplast uptake assays showing it to be an ABA and GA3 transporter. OsNPF3.1 overexpression significantly promoted ABA accumulation in the roots and GA accumulation in the basal part of the plant which inhibited axillary bud outgrowth and rice tillering, especially at high nitrate concentrations. The NUtE of OsNPF3.1-overexpressing plants was enhanced under low and medium nitrate concentrations, whereas the NUtE of OsNPF3.1 clustered regularly interspaced short palindromic repeats (CRISPR) plants was increased under high nitrate concentrations. The results indicate that OsNPF3.1 transports nitrate and phytohormones in different rice tissues under different nitrate concentrations. The altered OsNPF3.1 expression improves NUtE in the OsNPF3.1-overexpressing and CRISPR lines at low and high nitrate concentrations, respectively.