Discovery and structure-activity relationship studies of novel tetrahydro-β-carboline derivatives as apoptosis initiators for treating bacterial infections

Developing and excavating new agrochemicals with highly active and safe is an important tactic for protecting crop health and food safety.In this paper, to discover the new bactericide candidates, we designed, prepared a new type of1,2,3,4-tetrahydro-β-carboline(THC)derivatives and evaluated the in vitro and in vivo bioactivities against the Xanthomonas oryzae pv.oryzae(Xoo),Xanthomonas axonopodis pv.citri(Xac),and Pseudomonas syringae pv.actinidiae (Psa).The in vitro bioassay results exhibited ...     

Functional identification of phenazine biosynthesis genes in plant pathogenic bacteria Pseudomonas syringae pv. tomato and Xanthomonas oryzae pv. oryzae

Phenazines are secondary metabolites with broad spectrum antibiotic activity and thus show high potential in biological control of pathogens.In this study,we identified phenazine biosynthesis（phz）genes in two genome-completed plant pathogenic bacteria Pseudomonas syringae pv.tomato（Pst）DC3000 and Xanthomonas oryzae pv.oryzae（Xoo）PXO99^A.Unlike the phz genes in typical phenazine-producing pseudomonads,phz homologs in Pst DC3000 and Xoo PXO99^A consisted of phzC/D/E/F/G and phzC/E1/E2/F/G,respectively,and the both were not organized into an operon.Detection experiments demonstrated that phenazine-1-carboxylic acid（PCA）of Pst DC3000 accumulated to 13.4μg L^-1,while that of Xoo PXO99^A was almost undetectable.Moreover,Pst DC3000 was resistant to 1 mg mL^-1PCA,while Xoo PXO99^A was sensitive to50μg mL^-1PCA.Furthermore,mutation of phzF blocked the PCA production and significantly reduced the pathogenicity of Pst DC3000 in tomato,while the complementary strains restored these phenotypes.These results revealed that Pst DC3000 produces low level of and is resistant to phenazines and thus is unable to be biologically controlled by phenazines.Additionally,phz-mediated PCA production is required for full pathogenicity of Pst DC3000.To our knowledge,this is the first report of PCA production and its function in pathogenicity of a plant pathogenic P.syringae strain.     

Evaluation of the Pathotypes of Xanthomonas oryzae pv. oryzae and Preliminary Analysis of the Resistant Reactions of Main Japonica Rice in the Yunnan Plateau, China

The pathogenicity of 36 isolates of Xanthomonas oryzae pv. oryzae （Xoo）, which were collected from japonica rice varieties in the Yunnan Plateau, China, was evaluated. It was evaluated on 29 rice varieties including a set of seven varieties to identify pathogenicity, i.e., Haonuoyang, TN1, Kogyoku, Zhenzhu＇ai, IR26, Nanjing 33, and Kinmaze, which may be considered as a set of differential varieties for Xoo races from Yunnan japonica rice. The efficiency of the seven varieties was further confirmed. The results showed reversible and specific interactions between isolates and varieties. The isolates were classified into nine pathotypes from pathotyp Ⅰ to Ⅸ according to their pathogenic reactions on the seven rice varieties. The pathotype V was the epidemic, whereas pathogen Ⅶ was the most pathogenic. Most japonica varieties grown in the Yunnan Plateau were susceptible to Xoo. The rice lines IRBB21 （Xa-21）, Zhachanglong （Xa-22,, Xa- 24,）, and IR1545-339 （xa-5）, which were resistant to all the isolates tested, can be used as donors of resistant genes for bacterial blight in japonica rice breeding in the Yunnan Plateau.     

Genetical and Physiological Basis of Plant Type Model of Erect and Large Panicle Japonica Super Rice in Northern China

The historical changes in rice yields across China were explored. The physiological mechanisms and genetic basis of the erect and large panicle super-high-yield plant type model for breeding japonica super rice were analyzed mainly on the panicle type, number of large vascular bundles （LVB） in the panicle neck, and the panicle type index （PTI）. In the production point of view, we suggested that, for the breeding of super-high-yield japonica rice, the erect panicle types with more LVB numbers in the panicle neck and superior upper grains in the secondary branches would be the key factors. The information has potential significance in the rice breeding and productivity not only in China but also throughout the rice production areas of the world.     

Intercropping of rice varieties increases the eff iciency of blast control through reduced disease occurrence and variability

Creating a crop-heterogeneous system by intraspecific mixtures of different rice varieties can substantially reduce blast diseases.Such variety mixtures provide an ecological approach for effective disease control,maintaining high yields with the minimum fungicide applications.Whether such an approach is universally applicable for random rice variety combinations and what is the variation pattern of the diseases under intercropping still remains unclear.We conducted two-year large-scale field experiments involving 47 rice varieties/lines and 98 variety-combinations to compare the occurrence of rice blast in monoculture and intercropping plots at multiple sites.In the experiments,the plant height of the selected traditional varieties was about 30 cm taller,and their life cycle was 10 days longer,than that of the improved rice varieties.The monoculture included either traditional or modern rice varieties grown in separate plots.The intercropping included both traditional and modern rice varieties planted together in the same plots.Results from the field experiments under natural disease conditions demonstrated significant reduction for rice blast disease in intercropping plots,compared with that in monoculture plots.For traditional varieties,the average blast incidence reduced from~26%in monoculture to~10%in intercropping,and the disease severity reduced from~17 in monoculture to~5 in intercropping.For modern varieties,the average blast incidence reduced from~19%in monoculture to~10%in intercropping,and the severity from~10 in monoculture to~4 in intercropping.Traditional rice varieties(~72%)had a much greater increase in the efficiency of disease control than modern varieties(~60%).In addition,substantially lower values of variance in the blast incidence and severity was detected among the variety combinations in intercropping plots than in monoculture plots.Based on these results,we conclude that the intercropping or mixture of rice varieties greatly reduces the occurrence and variation of rice blast disease in particular variety combinations,which makes the intercropping system more stable and consistent for disease suppression on a large scale of rice cultivation.     

Functional characterization of the catalytic and bromodomain of FgGCN5 in development,DON production and virulence of Fusarium graminearum

FgGCN5, a GCN5 homolog in Fusarium graminearum, plays a critical role in hyphal vegetative growth, asexual and sexual reproduction, deoxynivalenol(DON) biosynthesis and plant infection. For nuclear localized GCN5, four conserved sequence motifs(I–IV) are presented in the catalytic domain and a bromodomain in the carboxy-terminus. As a lysine acetyltransferase, conserved negatively charged residues are present to neutralize the protons from lysine substrates. However, the role of conserved motifs/domains and residues in FgGCN5 are unclear. Here, we generated deletion mutant strains for each the conserved motifs/domains and a glutamate residue 130(E130) replacement mutant. Deletion of each conserved motif in the catalytic domain and replacement of E130 site resulted in manifold defects in hyphae growth, asexual and sexual development, DON biosynthesis, and plant infection. Phenotypic defects in the mutant strains were similar to deletion mutants. The deletion of the bromodomain led a significant reduction in DON production and virulence, with no effects on hyphae growth, asexual or sexual reproduction. FgGCN5 was further found to localize to the nucleus in conidia and hyphae cells. In conclusion, FgGCN5 encodes a nuclear localized acetyltransferase. The conserved motifs in the catalytic domain and E130 are essential for correct functions of the gene. The conserved bromodomain is important for DON production and pathogen virulence. This was the first report to identify the functions of conserved motifs/domains in FgGCN5, which will contribute to our understanding of the mechanism(s) by which FgGCN5 regulates F. graminearum.     

Antimicrobial activity from Piper sarmentosum Roxb.against rice pathogenic bacteria and fungi

In vitro antimicrobial activity of the dichloromethane and methanol extracts of Piper sarmentosum leaves,fruits,stems,and the essential oil obtained from the fresh leaves together with their constituents were investigated against two rice pathogenic fungi:Rhizoctonia solani(sheath blight causal agent)and Bipolaris oryzae(brown spot causal agent),and two rice pathogenic bacteria:Xanthomonas oryzae pv.oryzae(Xoo)(bacterial leaf blight causal agent)and pv.oryzicola(Xoc)(bacterial leaf streak causal agent).Among them,the dichloromethane extracts of the leaves and fruits,and the essential oil showed significantly high potential anti-rice microbial activity.Based on bioassay-guided fractionation of the dichloromethane leave and fruit extracts,myristicin,sarmentine,brachystamide B,brachyamide B,and piperonal were isolated.Moreover,the major constituent of its oil was also myristicin.Myristicin and brachyamide B revealed the highest potent inhibition against R.solani and B.oryzae(half maximal inhibitory concentration(IC_(50))of 0.69 and 0.12 mmol L~(–1)),respectively.Moreover,brachyamide B and piperonal displayed most antibacterial activity against Xoo(MIC/MBC 7.62/1.90 mmol L~(–1))and Xoc(MIC/MBC 2.59/20.75 mmol L~(–1)),respectively.Additionally,the essential oil also exhibited the antimicrobial activity against all tested rice pathogenic bacteria and fungi.These compounds and the oil were first evaluated for anti-rice pathogenic microbial activity.     

Production of Transgenic Anliucheng Sweet Orange （Citrus sinensis Osbeck） with Xa21 Gene for Potential Canker Resistance

Citrus canker, an epidemic quarantine disease caused by Xanthomonas axonopodis pv. citri, has brought a great damage in citrus production worldwide. Herein, a rice PRR （pattern recognition receptor） gene Xa21 together with GUS reporter gene and hygromycin phosphotransferase gene （HPT） was introduced into Anliucheng sweet orange （Citrus sinensis Osbeck） via Agrobacterium-mediated transformation of embryogenic callus. The transgenic calluses were screened on MT basal medium containing hygromycin （HYG） and detected by histochemical GUS staining. The transgenic plantlets were recovered through somatic embryogenesis pathway. The regenerated plantlets were accustomed to and maintained in the greenhouse. The transgene integration of recovered plantlets was identiifed by PCR and Southern blot hybridization. It showed that all the transgenic plantlets tested had undergone single copy integration, the expression of Xa21 in eight different transgenic lines detected by qRT-PCR can be divided into three grades, high for T5 and T6, middle for T4 and low for the rest. The tolerance to citrus canker disease of the three recovered transgenic lines T2, T4 and T6 was assessed by in vitro pin-puncture inoculation. The results showed that all the three transgenic lines conferred improved resistance to citrus canker bacterium infection and the T4 transgenic line displayed the highest resistance. The mechanism and feasibility of rice Xa21 in triggering innate immunity in citrus was brielfy discussed.     

Identification and Genetic Analysis of a Novel Rice Spotted-Leaf Mutant with Broad-Spectrum Resistance to Xanthomonas oryzae pv. oryzae

A spotted-leaf mutant of rice HM143 was isolated from an EMS-induced IR64 mutant bank. Brown lesions randomly distributed on leaf blades were observed about 3 wk after sowing. The symptom lasted for the whole plant growth duration. Histochemical analysis indicated that cell death occurred in and around the site of necrotic lesions accompanied with accumulation of hydrogen hyperoxide. Agronomic traits were largely similar to the wild type IR64 except seed setting rate and 1 000-grain weight which were significantly decreased in the mutant. Disease resistance of the mutant to multiple races of Xanthomonas oryzae pv. oryzae was significantly enhanced. Genetic analysis showed that the mutation was controlled by a single recessive gene, tentatively termed splHM143. In addition, using molecular markers and 1023 mutant type individuals from an F2 segregating population derived from the cross HM143/R9308, the spotted-leaf gene was finally delimited to an interval of 149 kb between markers XX25 and ID40 on the long arm of chromosome 4. splHM143 is likely a novel rice spotted-leaf gene since no other similar genes have been identified near the chromosomal region.     

NBS-LRR Proteins and Their Partners:Molecular Switches of Plant Defense

Specificity of the plant innate immune system is often conferred by resistance（R）proteins.Most plant disease resistance （R）proteins contain a series of leucine-rich repeats（LRRs）,a nucleotide-binding site（NBS）,and a putative amino-terminal signaling domain.They are termed NBS-LRR proteins.The LRRs are mainly involved in recognition,and the amino-terminal domain determines signaling specificity,whereas the NBS domain presumably functions as a molecular switch.During the past years,the most important discoveries are the role of partners in NBS-LRR gene mediated defenses,mounting support for the so-called＂guard hypothesis＂of R gene function,and providing evidence for intramolecular interactions and intermolecular interactions within NBS- LRR proteins as a mode of signaling regulation.The outcome of these interactions determines whether a plant activates its defense responses.     

Effects of silicon amendment on the occurrence of rice insect pests and diseases in a field test

Rice is one of the most important staple foods for the world population,but it is attacked by a number of destructive pests.While evidence from greenhouse and laboratory tests has shown that silicon(Si)amendment can confer enhanced resistance to pests in rice,few studies have directly demonstrated the Si-mediated protection from pests in a field situation.In this study,field plots with silicon amendments at 0,75,150 and 300 kg SiO_2 ha~(–1) in early-and late-season rice were employed to evaluate the effects of silicon amendment on the occurrence of major insect pests and diseases and rice yield.Compared with the control plots without silicon amendment,plant damage by stem borer and leaf folder and population size of planthopper were significantly lower in three to five of the seven monitoring observations in each season in the plots amended with 300 kg SiO_2 ha~(–1).The disease index of rice blast in the early-season rice was lower in the plots amended with Si at 300 kg SiO_2 ha~(–1) than in the control plots,while Si protection from rice blast in the late-season rice and from rice sheath blight in the early-season rice were not apparent.An insignificant increase of rice yield by 16.4%(604 kg ha~(–1))was observed in the plots amended with 300 kg SiO_2 ha~(–1) over the control plots.Our results indicate that Si amendment at 300kg SiO_(2 )ha~(–1) can provide substantial protection from some of the rice pests under field conditions.These findings support the recommendation of silicon amendment as a key component of integrated management of rice pests.     

Characteristics of Growth and Yield Formation of Rice in Rice-Fish Farming System

By using single rice cultivation as a control, the effect of rice-fish culture on growth dynamic, plant type and yield formation of rice was studied. The results showed as follows： rice-fish culture improved the physical-chemical properties of arable layer soil of paddy field, extended growth period of rice, increased dry matter and LAI of different growth stages, improved three top leaves area, deterred the degeneration of leaves function, increased the diameter of stem, promoted the growth of roots and the formation of roots in the extended stem. At the same time, rice-fish culture extended the length of basal internodes, increased the number of internodes, uplifted the gravity of plant, and depressed the root vigor. For the grain yield and yield structure office, rice-fish culture decreased ear/tillering ratio, spikelet/panicle and seed set percentage, increased grain weight. If variety choice and cultivation technology were controlled appropriately, rice-fish culture could increase the effective panicles and improve grain yield of rice.     

Pathotype Structure of Magnaporthe grisea in the Fields of Chongyang and Yuan’an in Hubei Province,China

[Objective] This study aimed to explore the pathotype structure of Magnaporthe grisea in Chongyang and Yuan’an in Hubei Province,China.[Method] From the rice-growing fields of Chongyang and Yuan’an in Hubei Province where rice blast occurs frequently,60 isolates which were pathotyped against two sets of host differentials:Chinese host differentials and CO39 NILs,were obtained.Then,20 pathotypes with the six indica host differentials(CO39 NILs) were observed,while 13 pathotypes in four race groups were observed out of the same single spore isolates with Chinese host differentials which consists of three indica cultivars and four japonica cultivars.The diversity of the pathotypes of M.grisea populations tested by CO39 NILs was 2.54 and the pathotype 137.1 occurred at predominantly high frequency(21.67%).The diversity of physiological races of M.grisea populations tested by Chinese host differentials was 1.22 and the race group ZA occurred at predominantly high frequency(73.33%).The diversity of physiological races of M.grisea in Chongyang and Yuan’an were also calculated.Overall,the diversity of pathotypes of M.grisea in Yuan’an was higher than that in Chongyang with the two sets of the host differentials.[Conclusion] This study provided current information on the pathotype spectrum of M.grisea populations in the rice fields of Hubei Province to allow the formulation of viable strategies for blast resistance breeding programs in Hubei Province.     

The Empidemiology of Wheat Rosette Stunt in Hulunbeir District of Inner—Mongolia

The virus disease threatening wheat production in Hulunbeir District of Inner-Mongolia was identified as NCMV(Northern Cereal Mosaic Virus)or WRSV(Wheat Rosette Stunt Virus)by test of host range,transmission,virus particle and serology.The main vector for spreading the virus is planthopper,Laodelphax striatellus(Fallen),Which overwinters in the stubbles of wheat underground.The overwintered viruliferous nymphs emerged in late Pril is responsible for the early infection of the disease.Agropyron repens,an important perennial weed in cultivated regions,is also an important wild host of the virus and its vector.Severe loss is induced when wheat is infected before 3-leaf stage.The disease incidence could be predicted by a model with the population and viruliferous rate of overwintered vectors as independent variables.     

Cloning and Expression Analysis of a Brassinosteroid Biosynthetic Enzyme Gene, GhDWF1, from Cotton （Gossypium hirsuturm L.）

Brassinosteroids （BRs） are an important class of plant steroidal hormones that are essential in a wide variety of physiological processes. To determine the effects of BRs on the development of cotton fibers, through screening cotton fiber EST database and contigging the candidate ESTs, a key gene （GhDWF1） involved in the upstream biosynthetic pathway of BRs was cloned from developing fibers of upland cotton （Gossypium hirsutum L.） cv. Xuzhou 142. The full length of the cloned cDNA is 1 849 bp, including a 37 bp 5＇-untranslated region, an ORF of 1 692 bp, and a 120 bp 3＇-untranslated region. The cDNA encodes a polypeptide of 563 amino acid residues with a predicted molecular mass of 65 kD. The deduced amino acid sequence has high homology with the BR biosynthetic enzyme, DWARF1/DIMINUTO, from rice, maize, pea, tomato, and Arabidopsis. Furthermore, the typical conserved structures, such as the transmembrane domain, the FAD- dependent oxidase domain, and the FAD-binding site, are present in the GhDWF1 protein. The Southern blot indicated that the GhDWF1 gene is a single copy in upland cotton genome. RT-PCR analysis revealed that the highest level of GhDWF1 expression was detected in 0 DPA （day post anthesis） ovule （with fibers） while the lowest level was observed in cotyledon. The GhDWF1 gene presents high expression levels in root, young stem, and fiber, especially, at the fiber developmental stage of secondary cell wall accumulation. Moreover, the expression level was higher in ovules （with fibers） of wildtype （Xuzhou 142） than in ovules of fuzzless-lintless mutant at the same developmental stages （0 and 4 DPA）. The results suggest that the GhDWF1 gene plays a crucial role in fiber development.     

Problems,challenges and future of plant disease management: from an ecological point of view

Plant disease management faces ever-growing challenges due to:(i)increasing demands for total,safe and diverse foods to support the booming global population and its improving living standards;(ii)reducing production potential in agriculture due to competition for land in fertile areas and exhaustion of marginal arable lands;(iii)deteriorating ecology of agro-ecosystems and depletion of natural resources;and(iv)increased risk of disease epidemics resulting from agricultural intensification and monocultures.Future plant disease management should aim to strengthen food security for a stable society while simultaneously safeguarding the health of associated ecosystems and reducing dependency on natural resources.To achieve these multiple functionalities,sustainable plant disease management should place emphases on rational adaptation of resistance,avoidance,elimination and remediation strategies individually and collectively,guided by traits of specific host-pathogen associations using evolutionary ecology principles to create environmental(biotic and abiotic)conditions favorable for host growth and development while adverse to pathogen reproduction and evolution.