Interaction Between Ustilaginoidea virens and Rice and Its Sustainable Control

Ustilaginoidea virens is a common rice pathogen that can easily lead to a decline in rice quality and the production of toxins pose potential risks to human health.In this review,we present a comprehensive literature review of research since the discovery of rice false smut.We provide a comprehensive and,at times,critical overview of the main results and findings from related research,and propose future research directions.Firstly,we delve into the interaction between U.virens and rice,including the regulation of transcription factors,the process of U.virens infecting rice panicles,and the plant immune response caused by rice infection.Following that,we discuss the identification and characterization of mycotoxins produced by the pathogenic fungus,as well as strategies for disease management.We emphasize the importance of comprehensive agricultural prevention and control methods for the sustainable management of U.virens.This knowledge will update our understanding of the interaction between U.virens and rice plants,offering a valuable perspective for those interested in U.virens.

Rapid detection of the rice false smut fungus Ustilaginoidea virens by lateral flow strip-based recombinase polymerase amplification assay

Rice false smut,caused by Ustilaginoidea virens,is a devastating disease that greatly reduces rice yield and quality.However,controlling rice false smut disease is challenging due to the unique infection mode of U.virens.Therefore,there is a need for early diagnosis and monitoring techniques to prevent the spread of this disease.Lateral flow strip-based recombinase polymerase amplification(LF-RPA)overcomes the limitations of current U.virens detection technologies,which are time-consuming,require delicate equipment,and have a high false-positive rate.In this study,we used a comparative genomics approach to identify Uv_3611,a specific gene of U.virens,as the target for the LF-RPA assay.The designed primers and probe efffectively detected the genomic DNA(gDNA)of U.virens and demonstrated no cross-reactivity with related pathogens.Under optimal conditions,the LF-RPA assay demonstrated a sensitivity of 10 pg of U.virens gDNA.Additionally,by incorporating a simplified PEG-NaOH method for plant DNA extraction,the LF-RPA assay enabled the detection of U.virens in rice spikelets within 30 min,without the need for specialized equipment.Furthermore,the LF-RPA assay successfully detected U.virens in naturally infected rice and seed samples in the field.Therefore,the LF-RPA assay is sensitive,efficient,and convenient,and could be developed as a kit for monitoring rice false smut disease in the field.

Carbon Catabolite Repressor UvCreA is Required for Development and Pathogenicity in Ustilaginoidea virens

The rice false smut disease, caused by Ustilaginoidea virens, has emerged as a significantglobal threat to rice production. The mechanism of carbon catabolite repression plays a crucial role in theefficient utilization of carbon nutrients and enzyme regulation in the presence of complex nutritionalconditions. Although significant progress has been made in understanding carbon catabolite repression infungi such as Aspergillus nidulans and Magnaporthe oryzae, its role in U. virens remains unclear. Toaddress this knowledge gap, we identified UvCreA, a pivotal component of carbon catabolite repression,in U. virens. Our investigation revealed that UvCreA localized to the nucleus. Deletion of UvCreA resultedin decreased growth and pathogenicity in U. virens. Through RNA-seq analysis, it was found that theknockout of UvCreA led to the up-regulation of 514 genes and down-regulation of 640 genes. Moreover,UvCreA was found to be involved in the transcriptional regulation of pathogenic genes and genesassociated with carbon metabolism in U. virens. In summary, our findings indicated that UvCreA isimportant in fungal development, virulence, and the utilization of carbon sources through transcriptionalregulation, thus making it a critical element of carbon catabolite repression.

Uses of Quenchers Methodology in the Management of Loose Smut Disease(Ustilago nuda)of Barley(Hordeum vulgare)through Seed Dressing and SRAP Markers Associated with Resistance in Some Egyptian Barley Genotypes

Barley production has been constrained by various factors,of which loose smut(Ustilago nuda)is the main biotic factor.Molecular and agronomical screening analyses were evaluated to study the similarity levels and marker assisted selection associated with resistance to loose smut among eight barley genotypes using sequence-related amplified polymorphism(SRAP).Agronomical parameters during two consecutive 2021 and 2022 seasons were studied to identify their reaction to loose smut.The results exhibited significant differences among all studied genotypes for all studied characters,and the highest mean values for all studied traits were detected in Giza 136 and Giza 137.Six SRAP selected primer combinations were amplified and gave 56 total fragments,where primer combination me1+em3 gave the highest polymorphism(100%)and the highest polymorphic information content of PIC was 0.96.The dendrogram of SRAP markers had clustered all studied genotypes into two main clusters.Cluster I includes all the resistance genotypes Giza 136,Giza 137,Giza 123,Giza 132,Giza 138 and Line 2.However,cluster II includes only Line 1 and Line 3 as susceptible genotypes.Thus,SRAP marker could be efficiently used to assess genetic variation among barley genotypes and useful for barley germplasm management in terms of biodiversity protection and design of new crosses for loose smut breeding programs,and seed dressings are commonly used to prevent infected seed from developing smutted heads.Some effective seed dressings include Triticonazole,Black seed oil and gamma rays 150,200,250 Gy.The tested fungicide was highly effective in controlling the disease and gave more than 99%disease control with high grain yield while nature product as black seed oil and utilizing radiation gamma ray 250 GY was the most effective.Utilizing radiation and natural ingredients to manage loose barley smut is economical.

Analysis on Quantitative Trait Loci for Resistance to Rice False Smut under Different Environmental Conditions

A recombinant inbred line （RILs） population with 157 lines derived from an inter-subspecies cross of Daguandao （japonica）/IR28 （indica） by the single seed de-scent method was used to detect quantitative trait loci （QTLs） conferring resistance to rice false smut caused by Ustilaginoidea virens（Cooke） Takahashi in Nanjing and Yangzhou. The disease rate index of the two parents and 157 RILs caused by rice false smut were scored and the QTLs for rice false smut resistance were detected accordingly by QTL Cartographer software. Eight QTLs control ing false smut resis-tance were detected on chromosomes 1, 2, 4, 8, 10, 11 and 12, respectively, with the phenotypic variance of 8.6%-22.5%. There were five QTLs detected in Nanjing and Yangzhou, respectively, and only two QTLs were found in both two years, the phenotypic variation was explained by individual QTL ranged from 18.0% to 18.9% for these two QTLs, and the additive effects of these two QTLs contributed to the 8.0%-14.6% decrease of disease index and therefore the disease resistance increased. The direction of the additive effects at six loci qFsr1, qFsr2, qFsr8, qFsr10a, qFsr11 and qFsr12 coincided with that predicted by phenotypes of the parents, and the IR28 al eles at these loci had positive effect against rice false smut while the negative effects were found in Daguandao al eles at qFsr4 and qFsr10b. Both qFsr10a and qFsr11 should be useful in rice breeding for resistance to rice false smut in marker-assisted selection （MAS） program.

Biological Activity of Several Fungicides against Ustilaginoidea virens

[Objective] This study aims to screen for the high effective fungicides which could significantly decrease the disease incidence and disease index of rice false smut. [Method] The inhibitory activities of the fungicide against mycelial growth of Ustilaginoidea virens were measured to in vitro evaluate the ECho values. And 17 fungicides were sprayed to evaluate the efficacy and effect of the fungicides tested in the field trials on the rice characters, [Result] The results showed that epoxicona- zole, difenoconazole, propiconazole and procloraz exhibited high inhibitory activity against mycelial growth of Ustilaginoidea virens with the ECso values 0.04, 0.07, 0.12 and 0.11 pg/ml, respectively. The results of field trials showed that the efficacy of Wen- quning, and fungicides such as difenoconazole, prochloraz, propiconazole, epoxi- conazole and their mixtures in controlling rice false smut were all 70% or more. [Conclusion] The 17 tested fungicides behaved efficacy in controlling rice false smut and did not cause drug injury on leaves and grains of rice plants, sprayed when flag leaves of rice fully expanded.

Establishment of Near Isogenic Lines of Huangzaosi Maize with Resistance against Maize Head Smut

[ Objective ] The paper was to establish the near isogenic lines of Huangzaosi maize with resistance against maize head smut, and to provide basis for its wide application in breeding and production. [Method] Combing with hybrid, backcress and self-cross method, as well as molecular markers, Huangzaosi maize was successfully introduced with head smut resistance, and its near isogenic lines were studied. The characteristics of resistant and susceptible variation and the changes of combining ability of near isogenic lines after backcrossing breeding were observed. [ Result ] The incidence rate of Huangzansi maize in control was 46% ; the incidence rates of the selected 24 near isogenic lines of resistant Huangzaosi were all lower than 10%, and the selected rate was 92.6% ; the combining ability of most near isogenic lines of resistant Huangzaosi maize had no significant difference with that of H^i maize in control; the yields of hybrids prepared by several inbred lines such as M135 and M140 were outstanding, which exceeded the control Zhengdan 958, showing a higher combining ability. [ Conclusion] The phenotype and combining ability of near isngenic lines of Huangzaosi maize with resistance against maize head smut obtained in the test were very close to Huan- gzaosi, and its resistance against maize head smut was greatly increased, thus solving the problems of Huangzaesi without resistance against maize head smut.

Analysis of Quantitative Trait Loci for Pathogenic Strain Pi-1 of Resistance to Rice False Smut in Rice(Oryza sativa)

In order to identify the resistant gene of rice false smut in rice, a recombi- nant inbred line （RILs） population with 157 lines derived from an inter-subspecies cross of Daguandao/IR28 by the single seed descent method was used to detect quantitative trait loci （QTLs） conferring resistance to strain Pi-1 of rice false smut caused by Usti/aginoiclea virens （Cooke） Takahashi in 2012 and 2013. The disease rate indexes of the two parents and 157 RILs caused by the strain Pi-1 of rice false smut were scored and the QTLs for rice false smut resistance were detected accordingly by QTL Cartographer software. Seven QTLs controlling false smut re- sistance were detected on chromosomes 2, 7, 8, 11 and 12, respectively, with the phenotypic variance of 8.5%-17.2%. There were four QTLs detected in 2012 and 2013, respectively, and only one QTL was found in both two years, the phenotypic variation explained by this individual QTL was 13.5% and 17.2%, and the additive effects of this QTL contributed to the 9.9% and 14.3% decrease of disease index and therefore the disease resistance increased. The direction of the additive effects at five loci qFsr2a, qFsr8a, qFsr8b, qFsr11 and qFsr12 coincided with that predicted by phenotypes of the parents, and the IR28 alleles at these loci had positive effect against rice false smut while the negative effects were found in Daguandao alleles at qFsr2b and qFsr7. The qFsr11 should be useful in rice breeding for resistance to rice false smut in marker-assisted selection （MAS） program.

Control of Maize Head Smut with 6% Tebuconazole FS

In order to test the control effect of 6% tebuconazole FS on maize head smut and to select its optimal dose for mixing seed, field tests were performed in 2012 and 2013. The results showed that 6% tebuconazole FS was effective to control maize head smut, and the control effect ranged from 73.4% to 91.1%. In addition, it also improved the maize yield by 36.7%-44.1%, compared with the blank control. The optimal dose of 6% tebuconazole FS for seed dressing was 166.5 g/100 kg to control head smut on maize.

稻曲病侵染规律研究进展与展望

水稻稻曲病(Rice false smut,RFS)发生情况日趋严重,已经上升为水稻主要病害之一。目前对稻曲病的研究已经取得一定的进展,但对其病菌侵染方式和侵染机理的了解还有局限性。通过激光共聚焦和透射电镜等方法已经基本明确了稻曲病菌在颖花内的侵染过程,然而稻曲病是否具有系统侵染、种子带菌以及幼苗根部侵染对病害发生的影响程度仍有待明确。对稻曲病菌研究中存在的亟待解决的问题进行了探讨分析,同时也对稻曲病主要的防控手段和病原监测方法进行了总结。

Ustilaginoidea virens: A Fungus Infects Rice Flower and Threats World Rice Production

Rice false smut disease, which is caused by the fungus Ustilaginoidea virens, is currently one of the most devastating rice fungal diseases in the world. Rice false smut disease not only causes severe yield loss and grain quality reduction, but also threatens food safety due to its production of mycotoxins. In this review, the most recent progresses regarding the life cycle, infection processes, genome and genetic diversity, pathogenic gene and disease resistance in rice were summarized in order to provide theoretical basis for the control of U. virens. We also proposed some future directions and key questions that need to be addressed for a better understanding of the molecular mechanism that leads to rice false smut disease and the prospects for sustainable control of rice false smut.

Effect of temperature on the development of sclerotia in Villosiclava virens

The sclerotia of Villosiclava virens were found commonly in high altitude and the temperate regions, where the temperatures are relatively low in rice filling stage. To make sure if low temperature induce the sclerotial formation in V. virens, the inoculated rice panicles in laboratory and the diseased rice panicles cut from paddy fields were treated under different temperatures. The results showed that 3 days of night temperature at 15°C were enough to induce the sclerotial formation. The low temperature was much more effective for young balls with intact membranes. After appearance of chlamydospores on the ball surfaces, the sclerotium could not differentiate anymore. The sclerotia began to differentiate below the chlamydospore layer and gradually grew onto the ball surfaces. This suggests that low temperature in the early development stage of false smut balls is an important factor to induce the sclerotial differentiation, and rice cultivars with long growth periods are able to produce more sclerotium-bearing balls, which will produce mass of spores in paddy field in the coming year.

Progress in the Study of False Smut Disease in Rice

Rice false smut disease is an increasing concern for research and production, not only because of the increasing epidemic occurrence in rice production, but also the intriguing specific pathogenesis of the disease to be a unique pathological system to enrich the molecular mechanism of plant-microbe interaction. Progresses have been achieved in the pathogen phylogenetic placement, the alternative hosts, the pathogen morphology and diversity, the toxins generated by false smut balls, the artificial inoculation method, and the pathogen transformation as well as rice resistance to the disease. However, it is still controversy on the infection process. It is not clear how the life cycle of this pathogen is coupled with the disease cycle. This review summarized our current understanding on the pathogen, the pathogenesis, and the rice resistance to the disease. Future work should pay attention to developing a more rapid and effective system to evaluate rice resistance and susceptibility to the disease, screening of rice germplasm for disease-resistance breeding, studying the resistance inheritance, and investigating the molecular mechanism of rice-false smut fungus interaction.

Rice False Smut Fungus, <i>Ustilaginoidea virens</i>, Inhibits Pollen Germination and Degrades the Integuments of Rice Ovule

Ustilaginoidea virens is a flower-infecting fungus that forms false smut balls in rice panicle. Rice false smut has long been considered a minor disease, but recently it occurred frequently and emerged as a major disease in rice production. In vitro co-cultivation of U. virens strain with young rice panicles showed that U. virens enters inside of spikelets from the apex and then grows downward to infect floral organs. In response to U. virens infection, rice host exhibits elevated ROS accumulation and enhanced callose deposition. The secreted compounds of U. virens can suppress rice pollen germination. Examination of sectioning slides of freshly collected smut balls demonstrated that both pistil and stamens of rice flower are infected by U. virens, hyphae degraded the contents of the pollen cells, and also invaded the filaments. In addition, U. virens entered rice ovary through the thin-walled papillary cells of the stigma, then decomposed the integuments and infected the ovary. The invaded pathogen could not penetrate the epidermis and other layers of the ovary. Transverse section of the pedicel just below the smut balls showed that there were no fungal hyphae observed in the vascular bundles of the pedicel, implicating that U. virens is not a systemic flower-infecting fungus.

Diversity in metagenomic sequences reveals new pathogenic fungus associated with smut in Job's tears

Smut is a serious disease in Job's tears,also known as adlay,and contributes to the reduction of crop yield in agricultural fields.In this study,the key pathogenic fungi in adlay smut disease were first identified by internal transcribed spacer region(ITS)high-throughput sequencing and then used to elucidate the composition and diversity of the fungal community in adlay smut.Results indicate that an abundance of operational taxonomic units(OTUs)were detected in the infected involucres of flowering plants and the OTUs were classified to nine phyla,20 classes,45 orders,90 families and 119 genera.A total of 4986 OTUs clustered together,sharing six core OTUs in all samples,while 145 OTUs were shared among four geographical groups.The Shannon and Simpson indices ranged from 0.137–1.629 and 0.357–0.970,respectively.Community diversity ranked as Anshun(AS)>Qinglong(QL)>Xingren(XR)>Xingyi(XY)among the four geographical groups by Shannon and Simpson indices,exhibiting complex community diversities among accessions and geographical groups.The richness and diversity data imply a weak relationship between the accession community richness and geographical origins of samples.Two closely related fungal genera,Sporisorium and Ustilago,were implicated as causes of smut disease.The genus Sporisorium appears to be more commonly found among accessions and thus is more likely to be the fungal pathogen causing smut in adlay.This work can facilitate strategies to control and prevent smut infection to improve adlay yield.

Putative Phosphatase UvPsr1 Is Required for Mycelial Growth, Conidiation, Stress Response and Pathogenicity in Ustilaginonidea virens

Ustilaginoidea virens is the causal agent of rice false smut,which can be a highly destructive disease of rice.The plasma membrane phosphatase Psr1 proteins,which act as a regulator of the salinity stress response in yeast,are widely distributed across fungi,but their functional characterization is sketchy.In this study,we characterized the functions of Psr1 protein,UvPsr1,in U.virens.Analyses of the AUvpsr1 and its complementation strain showed that UvPsr1 is required for normal mycelial growth,conidiation and tolerance to oxidative,osmotic and cell wall stresses.When rice panicles were inoculated with the AUvpsr1 strains,no symptoms of false smut disease developed,showing that UvPSR1 also contributes to the pathogenicity of the fungus.The deletion mutant of UvPSR1 also appeared to produce a smaller titer of toxic compounds able to inhibit elongation of the germinated seeds.In conclusion,our results indicated that UvPsr1 is a new pathogenic factor of U.virens.

Inhibition of the spread of endophytic Sporisorium reilianum renders maize resistance to head smut

Head smut, caused by the fungal pathogen Sporisorium reilianum, poses a grave threat to maize(Zea mays) production worldwide. Here we report cytological and molecular evidence for maize resistance to head smut. During early stages of root infection, S. reilianum mycelium was capable of penetrating the root epidermis of both resistant(Ji1037) and susceptible(HZ4) inbred lines. S. reilianum hyphae were observed in the root–stem junction at 6 days after inoculation. In an attempt to monitor hyphal spread within the maize plant,a highly specific and sensitive real-time PCR method was established to estimate the hyphal content in infected maize tissues. During the upward growth of endophytic S.reilianum, the extent of hyphal spread was markedly different between Ji1037 and HZ4. Very little or no pathogen was detected in aerial parts of Ji1037, whereas large amounts of pathogen accumulated in aboveground tissues, particularly inflorescences, of HZ4. Thus,maize resistance to S. reilianum was achieved mainly by inhibition of endophytic hyphal growth rather than by prevention of early-root penetration by the pathogen.

UvWhi2 Is Required for Stress Response and Pathogenicity in Ustilaginoidea virens

Some stress response-related genes have been identified in Ustilaginoidea virens,but it is not clear whether and how defects of stress responses affect the pathogenesis processes of U.virens.In this study,we identified a general stress response factor UvWHI2 as a homolog of Saccharomyces cerevisiae Whi2 in U.virens.The relative expression level of Uv Whi2 was significantly up-regulated during infection,suggesting that UvWHI2 may be involved in pathogenesis.Furthermore,knockout of Uv Whi2 showed decreased mycelial growth,increased conidiation in the potato sucrose medium and a defect in pathogenicity.In addition,the RNA-Seq and phenotypic analysis showed that UvWHI2 was involved in response to oxidative,hyperosmotic,cell wall stress and nutrient limitation.Further studies revealed that the defects of stress responses of the?Uvwhi2 mutant affected the formation of secondary spores on the nutrient limited surface and the rice surface,resulting in a significant reduction of pathogenicity of U.virens.Our results suggest that UvWHI2 is necessary for fungal growth,stress responses and the formation of secondary spores in U.virens.In addition,the defects of stress responses can affect the formation of secondary spores on the rice surface,and then compromise the pathogenicity of U.virens.

Inheritance Pattern of Two-line Hybrid Rice Resistance to False Smut

[Objective] The aim of this study was to explore the inheritance pattern of the resistance of two-line hybrid rice to false smut. [Method] A moderate susceptible TGMS line 33S as the female parent, was crossed with 30 susceptible restorer lines and 23 resistant restorer lines. Then false smut resistance of the hybrid F1 and parents were identified. [Result] All F1 of 33S crossed with 30 susceptible re- storer lines were susceptible to rice false smut. The F1 of 33S crossed with 23 re- sistant restorer lines showed dominant, incompletely dominant or recessive inheri- tance. Dominant and incompletely dominant inheritance accounted for 87.0%. The diseased panicle rate, disease index, average diseased grains per panicle, and the highest diseased grains per panicle of F1 shared highly significant positive correla- tions with corresponding indicators of restorer lines. [Conclusion] This study provides references for genetic improvement of two-line hybrid rice.

Study on Sugarcane Smut

The pathogen and characteristics, infection cycle, occurrence regularity and damage symptoms of sugarcane smut were introduced in the paper. Moreover, the comprehensive prevention measures were proposed on the basis of planting disease-resistant species, strengthening cultivation managements to improve dis- ease-resistant ability of sugarcane, and adopting fungicide control as assistant measures. The study provided certain basis for reducing damages of sugarcane smut and imorovin yield and quality of sugarcane.