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Oat stem rust, caused by Puccinia graminis f. sp. avenae, is one of the most devastating diseases of oat. The most cost-effective and environmentally friendly strategy to control this disease is the use of resistant cultivars. However, P. graminis f. sp. avenae can overcome the resistance of cultivars by rapidly changing its virulence. Thus, information on the virulence of P. graminis f. sp. avenae populations and resistance of cultivars is critical to control the disease. The current study was conducted to monitor the virulence composition and dynamics of the P. graminis f. sp. avenae population in China and to evaluate resistance of oat cultivars. Oat leaves naturally infected by P. graminis f. sp. avenae were collected in 2018 and 2019, and 159 isolates were derived from single uredinia. The isolates were tested on 12 international differential lines, and eight races, TJJ, TBD, TJB, TJD, TJL, TJN, TGD, and TKN, were identified for the first time in China. The predominant race was TJD, virulent against Pg1, Pg2, Pg3, Pg4, Pg8, Pg9, and Pg15, accounting for 35.8 and 37.8% in 2018 and 2019, respectively. The sub-predominant races were TJN (30.2% in 2018, 28.3% in 2019) and TKN (20.8% in 2018, 12.3% in 2019). All isolates were virulent to Pg1, Pg2, Pg3, and Pg4, and avirulent to Pg6 and Pg16. The three predominant races (TJD, TJN, and TKN) were used to evaluate resistance in 30 Chinese oat cultivars at the seedling and adult plant stages. Five cultivars, Bayan 1, Baiyan 2, Baiyan 3, Baiyan 5, and Baiyan 9, were highly resistant to the three races at both seedling and adult plant stages. The results of the virulences and frequencies of P. graminis f. sp. avenae races and the resistant cultivars will be useful in elucidating the pathogen migration and evolution and for breeding oat cultivars with stem rust resistance.

Sheath blight (ShB) significantly threatens rice yield production. However, the underlying mechanism of ShB defence in rice remains largely unknown. Here, we identified a highly ShB-susceptible mutant Ds-m which contained a mutation at the ammonium transporter 1;1 (AMT1;1) D

Sheath blight (ShB) is one of the most common diseases in rice that significantly affects yield production. However, the underlying mechanisms of rice defense remain largely unknown. Our previous transcriptome analysis identified that infection with Rhizoctonia solani significantly induced the expression level of SWEET2a, a member of the SWEET sugar transporter. The sweet2a genome-editing mutants were less susceptible to ShB. Further yeast-one hybrid, ChIP, and transient assays demonstrated that WRKY53 binds to the SWEET2a promoter to activate its expression. WRKY53 is a key brassinosteroid (BR) signaling transcription factor. Similar to the BR receptor gene BRI1 and biosynthetic gene D2 mutants, the WRKY53 mutant and overexpressor were less and more susceptible to ShB compared to wild-type, respectively. Inoculation with R. solani induced expression of BRI1, D2, and WRKY53, but inhibited MPK6 (a BR-signaling regulator) activity. Also, MPK6 is known to phosphorylate WRKY53 to enhance its transcription activation activity. Transient assay results indicated that co-expression of MPK6 and WRKY53 enhanced WRKY53 trans-activation activity to SWEET2a. Furthermore, expression of WRKY53 SD (the active phosphorylated forms of WRKY53) but not WRKY53 SA (the inactive phosphorylated forms of WRKY53), enhanced WRKY53-mediated activation of SWEET2a compared to expression of WRKY53 alone. Taken together, our analyses showed that R. solani infection may activate BR signaling to induce SWEET2a expression via WRKY53 through negative regulation of ShB resistance in rice.

Phytochrome (Phy)-regulated light signaling plays important roles in plant growth, development, and stress responses. However, its function in rice defense against sheath blight disease (ShB) remains unclear. Here, we found that PhyB mutation or shade treatment promoted rice resistance to ShB, while resistance was reduced by PhyB overexpression. Further analysis showed that PhyB interacts with phytochrome-interacting factor-like 15 (PIL15), brassinazole resistant 1 (BZR1), and vascular plant one-zinc-finger 2 (VOZ2). Plants overexpressing PIL15 were more susceptible to ShB in contrast to bzr1-D-overexpressing plants compared with the wild-type, suggesting that PhyB may inhibit BZR1 to negatively regulate rice resistance to ShB. Although BZR1 is known to regulate brassinosteroid (BR) signaling, the observation that BR signaling negatively regulated resistance to ShB indicated an independent role for BZR1 in controlling rice resistance. It was also found that the BZR1 ligand NAC028 positively regulated resistance to ShB. RNA sequencing showed that cinnamyl alcohol dehydrogenase 8B (CAD8B), involved in lignin biosynthesis was up-regulated in both bzr1-D- and NAC028-overexpressing plants compared with the wild-type. Yeast-one hybrid, ChIP, and transactivation assays demonstrated that BZR1 and NAC028 activate CAD8B directly. Taken together, the analyses demonstrated that PhyB-mediated light signaling inhibits the BZR1-NAC028-CAD8B pathway to regulate rice resistance to ShB. This article is protected by copyright. All rights reserved.

Phytochrome B (PhyB), a red-light receptor, plays important roles in diverse biological processes in plants; however, its function in NH

Rhizoctonia solani, the causative agent of the sheath blight disease (ShB), invades rice to obtain nutrients, especially sugars; however, the molecular mechanism via which R. solani hijacks sugars from rice remains unclear.In this study, rice-R. solani interaction model was used to explore whether pathogen effector proteins affect plant sugar absorption during infection.Yeast one-hybrid assay was used to identify Activator of SWEET2a (AOS2) from R. solani. Localization and invertase secretion assays showed that nuclear localization and secreted function of AOS2. Hexose transport assays verified the hexose transporter activity of SWEET2a and SWEET3a. Yeast two-hybrid assays, Bimolecular fluorescence complementation (BiFC) and transactivation assay were conducted to verify the AOS2-WRKY53-Grassy tiller 1 (GT1) transcriptional complex and its activation of SWEET2a and SWEET3a. Genetic analysis is used to detect the response of GT1, WRKY53, SWEET2a, and SWEET3a to ShB infestation. Also, the soluble sugar contents were measured in the mutants and overexpression plants before and after the inoculation of R. solani.The present study found that R. solani protein AOS2 activates rice SWEET2a and localized in the nucleus of tobacco cells and secreted in yeast. AOS2 interacts with rice transcription factor WRKY53 and GT1 to form a complex that activates the hexose transporter gene SWEET2a and SWEET3a and negatively regulate rice resistance to ShB.These data collectively suggest that AOS2 secreted by R. solani interacts with rice WRKY53 and GT1 to form a transcriptional complex that activates SWEETs to efflux sugars to apoplast; R. solani acquires more sugars and subsequently accelerates host invasion.

Rice blast disease caused by infection with Magnaporthe oryzae, a hemibiotrophic fungal pathogen, significantly reduces the yield production. However, the rice defense mechanism against blast disease remains elusive. To identify the genes involved in the regulation of rice defense to blast disease, dissociation (Ds) transposon tagging mutant lines were analyzed in terms of their response to M. oryzae isolate Guy11. Among them, CBL-interacting protein kinase 31 (CIPK31) mutants were more susceptible than wild-type plants to blast. The CIPK31 transcript was found to be insensitive to Guy11 infection, and the CIPK31-GFP was localized to the cytosol and nucleus. Overexpression of CIPK31 promoted rice defense to blast. Further analysis indicated that CIPK31 interacts with Calcineurin B-like 2 (CBL2) and CBL6 at the plasma membrane, and cbl2 mutants are more susceptible to blast compared with wild-type plants, suggesting that calcium signaling might partially through the CBL2-CIPK31 signaling regulate rice defense. Yeast two-hybrid results showed that AKT1-like (AKT1L), a potential potassium (K+) channel protein, interacted with CIPK31, and the K+ level was significantly lower in the cipk31 mutants than in the wild-type control. In addition, exogenous potassium application increased rice resistance to blast, suggesting that CIPK31 might interact with AKT1L to increase K+ uptake, thereby promoting resistance to blast. Taken together, the results presented here demonstrate that CBL2-CIPK31-AKT1L is a new signaling pathway that regulates rice defense to blast disease.

Tillering is one of the most important determinants of biomass and yield in rice (Oryza sativa L.). The capacity of plants to develop tillers from primordial meristems or buds is determined not only by the genotype but also by environmental cues. Here, we characterized the function of rice grassy tiller1 (OsGT1) and its interaction with other genetic and biological factors involved in tiller bud outgrowth in rice by generating OsGT1 RNA interference (RNAi) and overexpression (OX) lines. The tiller number was increased in OsGT1-RNAi mutants but strongly suppressed in OsGT1-OX lines. Expression analysis of OsGT1 in rice phyB mutants and in genotypes carrying various genetic combinations of GT1 RNAi and phyB demonstrated that OsGT1 is not involved in phyB-mediated suppression of tiller development in rice. Expression analysis of fine culm1 (fc1), a rice tb1 homolog, and molecular assays demonstrated that FC1 enhances the expression of OsGT1 by directly binding to its promoter. Comparison of the transcriptomic profiles of fc1 and OsGT1-RNAi mutants revealed differentially expressed genes (DEGs) common to both genotypes. Finally, analysis of tillering phenotypes of OX and RNAi seedlings treated with various phytohormones implied a possible role of OsGT1 in strigolactone-mediated tiller outgrowth. Overall, this study enhances our understanding of the diverse mechanisms of tiller development in grasses.

Rice bacterial blight caused by Xanthomonas oryzae pv. oryzae (Xoo) seriously affects rice yield production. The discovery and application of broad-spectrum resistance genes are of great advance for disease resistance breeding. Previously, we identified that multiple receptor-like kinase (RLK) family gene deletions induced by the Ac/Ds system resulted in a lesion mimic symptom. In this study, the mutant #29 showed that this lesion mimic symptom was isolated. Further analysis identified that four RLK genes (RLK19-22) were deleted in the #29 mutant. The #29 mutant exhibited broad-spectrum resistance to Xoo and subsequent analyses identified that pathogenesis-related genes PR1a, PBZ1, and cellular H2O2 levels were significantly induced in the mutant compared to wild-type plants. A genetic analysis revealed that reconstruction of RLK20, RLK21, or RLK22 rescued the lesion mimic symptom of the #29 mutant, indicating that these three RLKs are responsible for broad-spectrum resistance in rice. Further yeast two hybrid and bimolecular fluorescence complementation assays demonstrated that RLK20 interacts with RBOHB, which is a ROS producer in plants. Compared to wild-type plants, the #29 mutant was more, while #29/RLK20ox was less, susceptible to MV (methyl-viologen), an ROS inducer. Co-expression of RLK20 and RBOHB reduced RBOHB-promoted H2O2 accumulation in the cells. Taken together, our research indicated that the RLKs may inhibit RBOHB activity to negatively regulate rice resistance to Xoo. These results provide the theoretical basis and valuable information about the target genes necessary for the successful breeding of rice cultivars resistant to bacterial blight.

Wheat stem rust caused by Puccinia graminis f. sp. tritici is an important wheat disease with sudden and devastating characteristics. The appearance and spread of new P. graminis f. sp. tritici races (Ug99, TKTTF, and TTTTF) have once again renewed the interest in the prevention and control of wheat stem rust. Fungicides can effectively control the epidemics of this disease in a short period of time. However, the fungal pathogen is prone to developing resistance. Therefore, we collected 89 isolates of P. graminis f. sp. tritici from four provinces in China and used the spore germination method to test the sensitivity of the isolates to fungicide triadimefon. Seven relatively triadimefon-sensitive isolates and six relatively triadimefon-resistant isolates were further tested for sensitivity to fungicides carbendazim, mancozeb, thiophanate-methyl, and kresoxim-methyl. The results showed that the mean concentration for 50% of maximal effect of the isolates to triadimefon was 16.14 mg·liter−1, and the mean resistance factor was 4.48. Only 29 isolates were resistant to triadimefon in which 27 isolates had low levels of resistance and 2 isolates had moderate levels of resistance. However, most of the 89 isolates had no resistance to triadimefon. There was a positive correlation between resistance to triadimefon and carbendazim, but there was no cross-resistance between triadimefon resistance with thiophanate-methyl or kresoxim-methyl resistance. This study provides valuable information for managing fungicide resistant isolates of P. graminis f. sp. tritici.

The AMT1 family comprises major ammonium transporters in rice roots. In this study, we utilized AMT1 RNAi mutants (amt1) to explore how AMT1 affects NH4+- and NO3–-mediated morphological development and NH4+-responsive gene expression in roots. In the presence of NH4+, amt1 showed inhibition of NO3–- dependent lateral root development. The inhibitory action of NH4+ on lateral root growth was independent of the NO3– concentrations supplied to amt1 roots. The results of split root assays indicated that NH4+ exerts systemic action in inhibiting NO3–-dependent lateral root development in amt1. Further study with NAA and NOA, a potent auxin flux inhibitor, suggested that perturbation of membrane dynamics might not be the primary cause of the inhibitory action of NH4+ on NO3–-mediated lateral root growth in amt1 mutants. RNA-seq analysis of NH4+-responsive genes showed that approximately half of DEGs observed in wild-type roots were not detected in the DEGs of amt1 roots. Gene ontology enrichment analysis suggested that the expression of specific functional gene groups were affected by amt1 during the early response to NH4+. Auxin-responsive gene expression and root gravity responses were altered in amt1. This study demonstrated that AMT1 affects the interactions not only between ammonium and nitrate in lateral root growth but also between auxin and NH4+ in rice roots.

Sheath blight disease (ShB) severely affects rice production; however, the details of defense against ShB remain unclear. To understand the rice defense mechanism against ShB, an RNA sequencing analysis was performed using Rhizoctonia solani inoculated rice leaves after 48 h of inoculation. Among them, 3417 genes were upregulated and 2532 were downregulated when compared with the control group (> twofold or

Root-knot nematode (Meloidogyne incognita) is chief plant parasitic nematode of various crops globally. Meanwhile, the negative side effects on human health and environmental concerns associated with haphazard uses of chemical nematicides. Hence, the search for a safe and effective approach is more relevant. The present study was aimed to evaluate the nematicidal potential of Snef1216 (Penicillium chrysogenum) against M. incognita at different concentrations (5%, 10%, 25%, 50%, 75% and 100%) and with the nutritious medium. The egg hatching inhibition and mortality of second stage juveniles of M. incognita were assessed after 6, 12, 24, 48 and 72 h exposure. Results revealed that egg hatching inhibition and percent mortality of M. incognita increased with increasing concentration and exposure time. The highest mortality of juveniles was recorded at 100% conc. i.e., 24.20%, 36%, 66%, 78% and 97.8% at 6, 12, 24, 48 and 72 h, respectively. The highest ovicidal activity was recorded at 100% concentration with 5.20% of eggs hatching. The outcome suggested that Snef1216 (P. chrysogenum) resulted in the lowest LC50 value was recorded as 3718.496 at 6 h exposure period followed by 10479.87, 11186.682, 14838.58 and 24001.430 at 72, 12, 48 and 24 h respectively via ovicidal assay. Whereas, in the larvicidal assay, the lowest LC50 value demonstrated at 72 h being 17.628% exposure period followed by 28.345, 50.490, 215.710 and 482.595% at 48, 24, 12 and 6 h respectively. It is concluded that Snef1216 has potential being used as a biocontrol agent against M. incognita and can serve as a source of a novel nematicidal agent of fungal origin.

Soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is responsible for causing a major soybean disease globally. The fungal strain Penicillium janthinellum Snef1650 was evaluated against H. glycines. However, the effective determinants of the P. janthinellum strain are unknown. By performing pot experiments, a functioning compound was isolated from P. janthinellum Snef1650 through organic solvent extraction, semi-preparative HPLC, Sephadex LH-20 column chromatography, and silica gel column chromatography, and the isolated compound was identified to be scopoletin through 1H NMR, 13C NMR, and HPLC–MS. The pot experiments indicated that the treatment of soybean seeds with scopoletin drastically reduced the SCN population. The field experiments performed in 2017 and 2018 revealed that scopoletin decreased over 43.7% juveniles in the roots and over 61.55% cysts in the soil. Scopoletin treatment also promoted soybean growth and improved its yield, with an increase in plot yield by &gt, 5.33%. Scopoletin obtained from P. janthinellum Snef1650 could be used as an anti-H. glycines biocontrol agent.

Sugars Will Eventually be Exported Transporter (SWEET) is a newly characterized family of sugar transporters, which plays critical roles in plant-pathogen interactions. However, the function of SWEET in tobacco and its interaction with Fusarium oxysporum, a causal agent of root rot, remain unclear. This study aimed to dissect the function of NtSWEETs in tobacco root rot using stem bases from tobacco plants inoculated with F. oxysporum. RNA-sequencing (RNA-Seq) analysis was performed, and the results indicated that Sucrose Transporter 4 (NtSUC4), Sugar Transporter 12 (NtSTP12), Hexose Transporter 6 (NtHEX6), NtSWEET1, NtSWEET3b, and NtSWEET12 were downregulated by infection with F. oxysporum. The expression of NtSWEET1, but not of NtSUC4, NtSTP12, NtHEX6, NtSWEET3b, or NtSWEET12, was suppressed at all the time points tested after inoculation with F. oxysporum. The NtSWEET1-green fluorescent protein was localized on the plasma membrane and possessed the ability to transport glucose, fructose and galactose. Compared with the wild-type plants, NtSWEET1 RNAi plants were more susceptible to root rot, indicating that NtSWEET1 positively regulated the defense of tobacco against root rot. This study identified the role of SWEETs in tobacco and their interaction with F. oxysporum. The results might be useful in protecting tobacco plants from root rot.

Because glucose is an essential energy source for living organisms, glucose transporters (GLUTs) are present in all species worldwide. Encoded by the solute carrier family 2 gene family, the GLUT proteins generally have 12 transmembrane helices (TMHs). In total, 14 GLUT proteins have been identified in humans (hGLUTs), and they are divided into 3 classes on the basis of their transport characteristics and sequence similarities. Herein, we report the use of protein sequence similarity networks (SSNs) to visualize the sequence trends of 4101 GLUT proteins across the Metazoa. The SSNs separated the metazoan proteins into 3 new classes that were different from the traditional classification system. In the new system, 9 of the 14 hGLUTs (hGLUT1-5, 7, 9, 11, and 14) were grouped into class I, 3 (hGLUT10, 12, and 13) were grouped into class II, and 2 (hGLUT6 and 8) were grouped into class III, as also supported by the phylogenetic tree. Multiple sequence alignments further showed that the conserved residues in each class were different. Furthermore, the hGLUTs in each class showed unique evolutionary characteristics, with similar nonsynonymous-to-synonymous divergence ratios and similar regions under conservative selection pressure. Of note, GLUTs with 3, 6, 18, 24, and 36 TMHs were identified among the metazoan genomes, and 1 Chinese hamster protein with 6 TMHs showed GLUT activity. In summary, this large-scale sequence analysis provided new insights into the classification and evolution of GLUTs and further showed that gene duplication and fusion could have been important drivers during the evolution of these transporter molecules.-Jia, B., Yuan, D. P., Lan, W. J., Xuan, Y. H., Jeon, C. O. New insight into the classification and evolution of glucose transporters in the Metazoa.

Although it is known that brassinosteroids (BRs) play pleiotropic roles in plant growth and development, their roles in plant nutrient uptake remain unknown. Here, we hypothesized that BRs directly regulate ammonium uptake by activating the expression of rice AMT1-type genes. Exogenous BR treatment upregulated both AMT1;1 and AMT1;2 expression, while this induction was impaired in the BR-receptor gene BRI1 mutant d61-1. We then focused on brassinazole-resistant 1 (BZR1), a central hub of the BR signaling pathway, demonstrating the important role of this signaling pathway in regulating AMT1 expression and rice roots NH4+ uptake. The results showed that BR-induced expression of AMT1;2 was suppressed in BZR1 RNAi plants but was increased in bzr1-D, a gain-of-function BZR1 mutant. Further EMSA and ChIP analyses showed that BZR1 bound directly to the BRRE motif located in the promoter region of AMT1;2. Moreover, cellular ammonium contents, 15NH4+ uptake, and the regulatory effect of methyl-ammonium on root growth are strongly dependent on the levels of BZR1. Overexpression lines of BRI1 and BZR1 and Genetic combination of them mutants showed that BZR1 activates AMT1;2 expression downstream of BRI1. In conclusion, the findings suggest that BRs regulation of NH4+ uptake in rice involves transcription regulation of ammonium transporters.

Background: Tomato root-knot nematode is a soil-borne disease caused by Meloidogyne incognita. Enhancement of natural plant-defense mechanisms to provide resistance against pathogens may be a promising alternative environmentally friendly nematode management approach. Recently, the biocontrol effect against different pathogens in the presence of Penicillium chrysogenum has been reported in a wide range of plants and pathogens. For understanding the molecular mechanisms of the resistance induced by P. chrysogenum Snef1216 to RKN, transcriptomes of inducer control ‘IRCK’ (induced by Snef1216 only) and pathogen + inducer ‘IRN’ were compared to those of control groups, namely negative control ‘CKCK’ (no inoculum), pathogen control ‘CKN’ (inoculum of RKN only).Results: Numerous high-quality reads were generated by Novogene, means of the RNA-seq method. After being aligned to the reference genome, four comparative transcriptomic profile maps between any pairwise comparisons were obtained to find significantly differentially expressed genes (DEGs) and three databases of induced resistance (IR)-related, nematode pathogenesis (NP)-related and basal resistance (BR)-related genes were gotten. By hierarchal clustering, the phylogenetic relationship between highly and fewer DGEs were obtained and classified the resistance and susceptible responses into two clusters after inoculation with RKN. Finally, the results were verified by RT-qPCR and analysis of important plant defense enzymes.Conclusions: Within an integrated and more sustainable management approach, the use of biocontrol organisms, like P. chrysogenum, seems to be a promising alternative.

Plant architecture and disease resistance are the key factors that control the production of yield. However, the mechanism behind these factors is largely unknown. In this study, we identified that indeterminate domain 3 (IDD3) was obviously induced by inoculation of Rhizoctonia solani AG1-IA. Plants that overexpressed IDD3 (IDD3 OX) were more susceptible, while idd3 mutants showed a similar response to sheath blight disease compared with wild-type plants. Interestingly, IDD3 OX plants developed a wider tiller angle and exhibited altered shoot gravitropism, while idd3 knock-out mutants showed no visible morphological differences compared with the wild-type plants. IDD3 is ubiquitously expressed in different tissues and stages, and the IDD3 transcript was induced by exogenously applied auxin. Expression of the PIN-FORMED (PIN) and Aux/IAA genes was altered in IDD3 OX compared with wild-type plants. Furthermore, IDD3 OX plants are sensitive to auxin and the polar auxin transporter inhibitor N-1-naphthylphalamic acid (NPA). Further yeast-one hybrid, chromatin immunoprecipitation (ChIP) and transient assays revealed that IDD3 directly represses PIN1b via promoter binding. Inoculation with R. solani indicated that PIN1b RNAi plants are more susceptible to sheath blight disease (ShB) compared with the wild-type. Taken together, our analyses suggest that IDD3 controls plant architecture and the resistance of rice to ShB via the regulation of PIN auxin transporter genes.

Wheat powdery mildew, caused by the biotrophic fungus Blumeria graminis f. sp. tritici (Bgt), is a serious disease of wheat worldwide that can cause significant yield losses. Growing resistant cultivars is the most cost-effective and eco-soundly strategy to manage the disease. Therefore, a high breeding priority is to identify genes that can be readily used either singly or in combination for effective resistance to powdery mildew and also in combination with genes for resistance to other diseases. Yunnan Province, with complex and diverse ecological environments and climates, is one of the main wheat growing regions in China. This region provides initial inoculum for starting epidemics of wheat powdery mildew in the region and other regions and thus, plays a key role in the regional and large-scale epidemics of the disease throughout China. The objectives of this study were to evaluate seedling resistance of 69 main wheat cultivars to powdery mildew and to determine the presence of resistance genes Pm3, Pm8, Pm13, Pm16, and Pm21in these cultivars using gene specific DNA markers. Evaluation of 69 wheat cultivars with six Bgt isolates showed that only four cultivars were resistant to all tested isolates, indicating that the overall level of powdery mildew resistance of Yunnan wheat cultivars is inadequate. The molecular marker results showed that 27 cultivars likely have at least one of these genes. Six cultivars were found likely to have Pm3,18 likely to have Pm8,5 likely to have Pm16,and 3 likely to have Pm21. No cultivar was found to carry Pm13. The information on the presence of the Pmresistance genes in Yunnan wheat cultivars can be used in future wheat disease breeding programs. In particular, cultivars carrying Pm21, which is effective against all Bgtraces in China, should be pyramided with other effective genes to developing new cultivars with durable resistance to powdery mildew.

Plant architecture and disease resistance are the key factors that control the production of yield. However, the mechanism behind these factors is largely unknown. In this study, we identified that indeterminate domain 3 (IDD3) was obviously induced by inoculation of Rhizoctonia solani AG1-IA. Plants that overexpressed IDD3 (IDD3 OX) were more susceptible, while idd3 mutants showed a similar response to sheath blight disease compared with wild-type plants. Interestingly, IDD3 OX plants developed a wider tiller angle and exhibited altered shoot gravitropism, while idd3 knock-out mutants showed no visible morphological differences compared with the wild-type plants. IDD3 is ubiquitously expressed in different tissues and stages, and the IDD3 transcript was induced by exogenously applied auxin. Expression of the PIN-FORMED (PIN) and Aux/IAA genes was altered in IDD3 OX compared with wild-type plants. Furthermore, IDD3 OX plants are sensitive to auxin and the polar auxin transporter inhibitor N-1-naphthylphalamic acid (NPA). Further yeast-one hybrid, chromatin immunoprecipitation (ChIP) and transient assays revealed that IDD3 directly represses PIN1b via promoter binding. Inoculation with R. solani indicated that PIN1b RNAi plants are more susceptible to sheath blight disease (ShB) compared with the wild-type. Taken together, our analyses suggest that IDD3 controls plant architecture and the resistance of rice to ShB via the regulation of PIN auxin transporter genes.

Background Loose Plant Architecture 1 (LPA1), an indeterminate domain (IDD) protein, exhibits almost no expression in the leaves, but the overexpression of LPA1 significantly increases the resistance of rice to sheath blight disease (ShB) via the activation of PIN-FORMED 1a (PIN1a). Results In this study, we determined that Rhizoctonia solani infection significantly induced LPA1 expression in the leaves, and lpa1 was more susceptible to R. solani compared with the wild-type and revertant plants. In addition, infection with R. solani altered the expression of IDD3, IDD5, IDD10, and IDD13, and yeast two-hybrid, split-GFP, and coimmunoprecipitation assays showed that LPA1 interacts with IDD3 and IDD13. IDD13 RNAi plants were more susceptible, while IDD13 overexpressors were less susceptible to ShB compared with the wild-type. In parallel, idd3 exhibited no significant differences, while IDD3 overexpressors were more susceptible compared to the wild-type response to ShB. Additional chromatin-immunoprecipitation and electrophoretic mobility shift assay experiments indicated that IDD13 and IDD3 bound to the PIN1a promoter, and the transient assay indicated that IDD13 and IDD3 positively and negatively regulate PIN1a expression, respectively. Moreover, IDD13, IDD3, and LPA1 form a transcription factor complex that regulates PIN1a. A genetic study showed that the LPA1 repressor lines were similar to lpa1/IDD13 RNAi and were more susceptible than the lpa1 and IDD13 RNAi plants in response to ShB. The overexpression of IDD13 increased resistance to ShB in the lpa1 background. Conclusions Taken together, our analyses established that IDD3, IDD13, and LPA1 form a transcription factor complex to regulate the defense of rice against ShB possibly via the regulation of PIN1a.

Root-knot nematodes (RKNs) severely affect plants growth and productivity, and several commercial biocontrol bacteria can improve plants resistance to RKNs. Pseudomonas putida Sneb821 isolate was found to induce tomatoes resistance against Meloidogyne incognita. However, the molecular functions behind induced resistance remains unclear. Long non-coding RNA (lncRNA) is considered to be a new component that regulates the molecular functions of plant immunity. We found lncRNA was involved in Sneb821-induced tomato resistance to M. incognita. Compared with tomato inoculated with M. incognita, high-throughput sequencing showed that 43 lncRNAs were upregulated, while 35 lncRNAs were downregulated in tomatoes previously inoculated with Sneb821. A regulation network of lncRNAs was constructed, and the results indicated that 12 lncRNAs were found to act as sponges of their corresponding miRNAs. By using qRT-PCR and the overexpression vector pBI121, we found the expression of lncRNA44664 correlated with miR396/GRFs (growth-regulating factors) and lncRNA48734 was correlated with miR156/SPL (squamosal promoter-binding protein-like) transcription factors. These observations provided a novel molecular model in biocontrol bacteria-induced tomato resistance to M. incognita.

Meloidogyne incognita and Meloidogyne graminicola are root-knot nematodes (RKNs) infecting rice (Oryza sativa L.) roots and severely decreasing yield, whose mechanisms of action remain unclear. We investigated RKN invasion and development in rice roots through RNA-seq transcriptome analysis. The results showed that 952 and 647 genes were differently expressed after 6 (invasion stage) and 18 (development stage) days post inoculation, respectively. Gene annotation showed that the differentially expressed genes were classified into diverse metabolic and stress response categories. Furthermore, phytohormone, transcription factor, redox signaling, and defense response pathways were enriched upon RKN infection. RNA-seq validation using qRT-PCR confirmed that CBL-interacting protein kinase (CIPK) genes (CIPK5, 8, 9, 11, 14, 23, 24, and 31) as well as brassinosteroid (BR)-related genes (OsBAK1, OsBRI1, D2, and D11) were altered by RKN infection. Analysis of the CIPK9 mutant and overexpressor indicated that the RKN populations were smaller in cipk9 and larger in CIPK9 OX, while more galls were produced in CIPK9 OX plant roots than the in wild-type roots. Significantly fewer numbers of second-stage infective juveniles (J2s) were observed in the plants expressing the BR biosynthesis gene D2 mutant and the BR receptor BRI1 activation-tagged mutant (bri1-D), and fewer galls were observed in bri1-D roots than in wild-type roots. The roots of plants expressing the regulator of ethylene signaling ERS1 (ethylene response sensor 1) mutant contained higher numbers of J2s and developed more galls compared with wild-type roots, suggesting that these signals function in RKN invasion or development. Our findings broaden our understanding of rice responses to RKN invasion and provide useful information for further research on RKN defense mechanisms.

Rhizoctonia solani causes sheath blight disease in rice; however, the defense mechanism of rice plants against R. solani remains elusive. To analyze the roles of brassinosteroid (BR) and ethylene signaling on rice defense to R. solani, wild-type (WT) rice and several mutants and overexpressing (OX) lines were inoculated with R. solani. Mutants d61-1 and d2 were less susceptible to sheath blight disease, bri1-D was more susceptible, and ravl1 and d61-1/EIL1 Ri5 were similarly susceptible compared with WT. The double mutant ravl1/d61-1 was phenotypically similar to the ravl1 mutant. Transcriptome analysis, chromatin immunoprecipitation assay, electrophoretic mobility shift assay, and transient assays indicted that RAVL1 might directly activate Ethylene insensitive 3-like 1 (EIL1), a master regulator of ethylene signaling. Mutants ers1 and d61-1/RAVL1 OX were resistant to sheath blight disease, whereas EIL1 RNAi mutants and RAVL1 OX were more susceptible than WT. BRI1 and D2 expression in EIL1 Ri5/RAVL1 OX and EIL1 expression in d61-1/RAVL1 OX indicated that RAVL1 activates BRI1/D2 and EIL1, respectively, independent of BR and ethylene signaling. Our analyses provide information on how BR and ethylene signaling regulate sheath blight disease and on the regulatory function of RAVL1 in rice sheath blight disease.

Pathogen–host interaction is a complicated process; pathogens mainly infect host plants to acquire nutrients, especially sugars. Rhizoctonia solani, the causative agent of sheath blight disease, is a major pathogen of rice. However, it is not known how this pathogen obtains sugar from rice plants. In this study, we found that the rice sugar transporter OsSWEET11 is involved in the pathogenesis of sheath blight disease. Quantitative real‐time polymerase chain reaction (qRT‐PCR) and β‐d‐glucuronidase expression analyses showed that R. solani infection significantly enhanced OsSWEET11 expression in leaves amongst the clade III SWEET members. The analyses of transgenic plants revealed that Ossweet11 mutants were less susceptible, whereas plants overexpressing OsSWEET11 were more susceptible, to sheath blight compared with wild‐type controls, but the yield of OsSWEET11 mutants and overexpressors was reduced. SWEETs become active on oligomerization. Split‐ubiquitin yeast two‐hybrid, bimolecular fluorescence complementation and co‐immunoprecipitation assays showed that mutated OsSWEET11 interacted with normal OsSWEET11. In addition, expression of conserved residue mutated AtSWEET1 inhibited normal AtSWEET1 activity. To analyse whether inhibition of OsSWEET11 function in mesophyll cells is related to defence against this disease, mutated OsSWEET11 was expressed under the control of the Rubisco promoter, which is specific for green tissues. The resistance of transgenic plants to sheath blight disease, but not other disease, was improved, whereas yield production was not obviously affected. Overall, these results suggest that R. solani might acquire sugar from rice leaves by the activation of OsSWEET11 expression. The plants can be protected from infection by manipulation of the expression of OsSWEET11 without affecting the crop yield.

The root-knot nematode (RKN) severely affects plant growth and productivity. Thus, protection of plants from RKNs has become an important task, especially for increasing crop yields. The objective of this study was to screen plant growth-promoting rhizobacteria (PGPR) for the ability to control RKNs. In this study, 860 strains of bacteria were collected from rhizosphere soil and five bacteria isolates were identified with high efficacy against RKNs: Bacillus cereus (Sneb 560), B. subtilis (Sneb 815), Pseudomonas putida (Sneb 821), P. fluorescens (Sneb 825) and Serratia proteamaculans (Sneb 851). The five bacteria isolates had high larvicidal and ovicidal activity in vitro. In a novel result, Sneb 851 showed especially high potential as a biocontrol agent against Meloidogyne incognita, causing 99.17% juvenile mortality and 61.11% egg mortality. In the pot experiment, tomato seeds treated with Sneb 825 displayed significantly higher levels of growth in root and shoot compared to control plants. Meanwhile, treatment with Sneb 815, Sneb 821 and Sneb 825 exhibited higher efficacies in reducing the number of galls and juveniles in the soil. In the field experiment, application of the five bacteria isolates increased the plant biomass and showed high biocontrol efficacy against M. incognita. The lowest gall index was observed in the treatment with Sneb 815 and Sneb 825 both in 2014 and 2015. Taken together, the five PGPR isolates can be regarded as potential biocontrol agents against RKNs in the future.

NH4+ is an important nitrogen resource for rice plants in paddy soil. Therefore, it is likely that NH4+-triggered plant growth interacts with phytohormone-mediated developmental mechanisms. Our previous transcriptomic analysis revealed that many genes involved in auxin signaling and efflux are sensitive to NH4+. In the current study, we found that NH4+ treatment causes a delayed gravity response in rice roots. To further elucidate the interlocking relationship between NH4+ and auxin signaling during root development, we utilized mutants and overexpressors of a key NH4+ signaling transcription factor INDETERMINATE DOMAIN 10 (IDD10), encoding a transcription factor that regulates the expression of NH4+ uptake and N-assimilation genes. We obtained several lines of evidence that auxin affects NH4+-mediated gene expression and root development in rice plants via IDD10. First, the gravity response was delayed in idd10 roots and accelerated in IDD10 overexpressor (IDD10 OX) roots in the absence and (especially) presence of NH4+. Second, idd10 plants showed strong root coiling only in the presence of NH4+. However, treatment of 1-N-naphthylphthalamic acid (NPA), a polar auxin transport inhibitor suppressed the NH4+-specific root phenotype of idd10. Third, the expression of NH4+-responsive auxin-related genes was affected in idd10 and IDD10 overexpressors. Finally, IDD10 expression was induced by IAA and suppressed by NPA. These findings suggest that the gene expression patterns and phenotypes triggered by NH4+ are influenced by the actions of auxin during root development, pointing to a regulatory circuit between NH4+ and auxin signaling that functions in root development in rice.

Glycine max (soybean) is an extremely important crop, representing a major source of oil and protein for human beings. Heterodera glycines (soybean cyst nematode, SCN) infection severely reduces soybean production; therefore, protecting soybean from SCN has become an issue for breeders. Black soybean has exhibited a different grade of resistance to SCN. However, the underlying mechanism of Huipizhi Heidou resistance against SCN remains elusive. The Huipizhi Heidou (ZDD2315) and race 3 of Heterodera glycines were chosen to study the mechanism of resistance via examination of transcriptomic changes. After 5, 10, and 15days of SCN infection, whole roots were sampled for RNA extraction, and uninfected samples were simultaneously collected as a control. 740, 1413, and 4925 genes were isolated by padj (p-value adjusted)

NH 4 + is important for the growth and the yield production of paddy-soil grown rice. However, the mechanism of NH 4 + signaling is largely unknown. Previously, we performed a microarray analysis to examine genomic responses and signaling pathways in rice roots exposed to NH 4 + , and interestingly found that transcription of a large number of genes including kinase, phosphatase, and transcription factors were sensitive to NH 4 + . In this study, we further analyzed transcription factors respond to NH 4 + , and identified an NH 4 + -repressed bHLH transcription factor involved in NH 4 + acquisition process. bHLH is a nuclear protein and has transcription-activation activity in yeast and plants. RNAi-mediated suppression of bHLH and bHLH overexpression ( bHLH OX ) resulted in the accumulation of lower and higher NH 4 + contents in transgenic rice roots, respectively. NH 4 + -mediated induction of AMT1 ; 1 and AMT1 ; 2 is lower in bHLH RNAi , but higher in bHLH OX as compared to in wild-type plant roots. However, bHLH does not directly activate AMT1 ; 1 and AMT1 ; 2 transcriptions. IDD10 expression is not altered in bHLH RNAi plant roots, and bHLH maintained similar expression level in idd10 mutant roots compared to in wild-type plant roots. Furthermore, NH 4 + contents in idd10 ; bHLH RNAi double mutant roots were much lower than in each single mutant and wild-type plant roots, indicating that IDD10 and bHLH independently regulates NH 4 + uptake. Taken together, these results provide a new regulatory path by which bHLH regulates NH 4 + uptake in rice.

The Fusarium graminearum species complex (FGSC) is an important group of pathogens distributed in maize-producing areas worldwide. This study investigated the genetic diversity and pathogenicity of 40 FGSC isolates obtained from stalk rot and ear rot samples collected from 42 locations in northeastern China during 2013 and 2014. A phylogenetic tree of translation elongation factor (EF-la) sequences designated the 40 isolates as F. graminearum sensu stricto (67.5%) and F. boothii (32.5%). By using inter-simple sequence repeat analysis (ISSR), it was shown that the isolates were divided into two clades, which corresponded to the species identity of the isolates. However, the isolates from the two different diseases could not be distinguished in pathogenicity. The disease severity index of seedlings inoculated with stalk isolates was slightly higher than that of seedlings inoculated with isolates from infected ears, whereas the pathogenicity of the stalk and ear isolates were identical.