Breeding of Brassica napus Cultivar Zhongshuang9 with High-Resistance to Sclerotinia sclerotiorum and Dynamics of Its Important Defense Enzyme Activity

Zhongshuang9, a new semi-winter Brassica napus variety with high resistance to Sclerotinia sclerotiorum and lodging, high-yield, double-low quality and extensive adaptability, was bred by multiple crossing and microspore culture technique. It was registered and released in China in 2002. In regional trial of Hubei Province in China, Zhongshuang9 yielded 2 482. 2 kg ha-1 averagely in 2000 - 2002, 15. 33% higher than the control variety Zhongyou821. Erucic acid, glucosinolates and oil contents of Zhongshuang9 were 0.23%, 22.69 μmol g-1(in meal)and 42%, respectively. In field assessment of resistance to S. Sclerotiorum , the disease incidence and disease index of Zhongshuang9 averaged 13.31 % and 6.47, respectively, which were lower than those of Zhongyou821 by 28% and 36%, respectively. After inoculation of detached leaves with mycelia, the lesion size of Zhongshuang9 was 4. 709 cm2, which was significantly smaller than that of the mid-resistant variety Zhongyou821(5. 933 cm2). The stem lesion length of Zhongshuang9 after match-stick inoculation was 1.275 cm, which was significantly lower than that of Zhongyou821(1.943 cm). The possible mechanism of resistance to S. sclerotiorum was studied through comparing the activities of phenylalanine ammonia lyase(PAL), exo-chitinase, β-1, 3-glucanase, peroxidase(POD)and polyphenoloxidase(PPO)in Zhongshuang9 with those in other resistant, mid-resistant and susceptible cultivars.

Defensive Responses of Rice Genotypes for Resistance Against Rice Leaffolder Cnaphalocrocis medinalis

The experiment was carried out to assess the reaction of different categories of rice genotypes viz., resistant, susceptible, hybrid, scented, popular and wild in response to the infestation by rice leaffolder （RLF）, Cnaphalocrocis medinalis （Guenee） and to explore the possible use of these genotypes in developing RLF-resistant rice varieties. The changes of various biochemical constituents such as leaf soluble protein, phenol, ortho-dihydroxy phenol, tannin and enzymes viz., peroxidase, phenyl alanine ammonia lyase （PAL） were assessed spectrophotometrically in all the rice genotypes before and after RLF infestation. The protein profile was analyzed using sodium dodecyl sulphate-poly acrylamide gel electrophoresis （SDS-PAGE） method. A significant constituent of biochemical content such as tannin, phenol and ortho-dihydroxy phenol has been increased along with enzyme activities of peroxidase and PAL in the infested resistant （Ptb 33, TKM6 and LFR831311） and wild rice genotypes （Oryza minuta and O. rhizomatis）. A decrease in leaf protein content was evident invariably in all the infested rice genotypes. It is also evident that the contents of biochemicals such as phenol, ortho- dihydroxy phenol and tannin were negatively correlated with leaffolder damage. However, leaf protein content was positively correlated with the damage by rice leaffolder. SDS-PAGE analysis for total protein profiling of healthy and C. medinalis-infested genotypes revealed the enhanced expression of a high molecular weight （〉 97 kDa） protein in all the genotypes. Besides, there was also an increased induction of a 38 kDa protein in C. medinalis infested resistant genotypes, which was absent in uninfested plants. The present investigation proved that the elevated levels of biochemicals and enzymes may play a vital role in rice plants resistance to RLF.

Novel 18β-glycyrrhetinic acid amide derivatives show dual-acting capabilities for controlling plant bacterial diseases through ROS-mediated antibacterial efficiency and activating plant defense responses

Natural products have long been a crucial source of,or provided inspiration for new agrochemical discovery.Naturally occurring 18β-glycyrrhetinic acid shows broad-spectrum bioactivities and is a potential skeleton for novel drug discovery.To extend the utility of 18β-glycyrrhetinic acid for agricultural uses,a series of novel 18β-glycyrrhetinic acid amide derivatives were prepared and evaluated for their antibacterial potency.Notably,compound 5k showed good antibacterial activity in vitro against Xanthomonas oryzae pv.oryzae(Xoo,EC50=3.64 mg L–1),and excellent protective activity(54.68%)against Xoo in vivo.Compound 5k induced excessive production and accumulation of reactive oxygen species in the tested pathogens,resulting in damaging the bacterial cell envelope.More interestingly,compound 5k could increase the activities of plant defense enzymes including catalase,superoxide dismutase,peroxidase,and phenylalanine ammonia lyase.Taken together,these enjoyable results suggested that designed compounds derived from 18β-glycyrrhetinic acid showed potential for controlling intractable plant bacterial diseases by disturbing the balance of the phytopathogen’s redox system and activating the plant defense system.

Exploring the nano-fungicidal efficacy of green synthesized magnesium oxide nanoparticles(MgO NPs)on the development,physiology,and infection of carrot(Daucus carota L.)with Alternaria leaf blight(ALB):Molecular docking

In this research,green synthesized magnesium oxide nanoparticles(MgO NPs)from lemon fruit extracts and their fungicidal potential was evaluated against Alternaria dauci infection on carrot(Daucus carota L.)under greenhouse conditions.The scanning and transmission electron microscopy(SEM and TEM)and ultra-violet(UV)visible spectroscopy were used to validate and characterize MgO NPs.The crystalline nature of MgONPs was determined using selected area electron diffraction(SAED).MgO NPs triggered substantial antifungal activity against A.dauci when exposed to 50 and 100 mg L^(–1)concentrations but the higher antifungal potential was noticed in 100 mg L^(–1)under invitro conditions.In fungal inoculated plants,a marked decrease in growth,photosynthetic pigments,and an increase in phenol,proline contents,and defense-related enzymes of carrot were seen over control(distilled water).However,foliar application of MgO NPs at 50 and 100 mg L^(–1)resulted in significant improvement of plant growth,photosynthetic pigments,phenol and proline contents,and defense enzymes activity of carrots with and without A.dauci infection.Spraying of MgO NPs at 100 mg L^(–1)had more plant length(17.11%),shoot dry weight(34.38%),plant fresh weight(20.46%),and root dry weight(49.09%)in carrots when challenged with A.dauci over inoculated control.The leaf blight indices and percent disease severity were also reduced in A.dauci inoculated plants when sprayed with MgO NPs.The non-bonding interactions of Alternaria genus protein with nanoparticles were studied using molecular docking.

Identification of Resistance to Pathogenesis Related Protein GmPR1L in Tobacco Botrytis cinerea Infection

Soybean(Glycine max(Linn.)Merr.)annual leguminous crop is cultivated all over the world.The occurrence of diseases has a great impact on the yield and quality of soybean.In this study,based on the RNA-seq of soybean variety M18,a complete CDS(Coding sequence)GmPR1L of the pathogenesis-related protein 1 family was obtained,which has the ability to resist fungal diseases.The overexpression vector and interference expression vector were transferred into tobacco NC89,and the resistance of transgenic tobacco(Nicotiana tabacum L.)to Botrytis cinerea infection was identified.The results show that:Compared with the control,the activities of related defense enzymes SOD(Superoxide dismutase),POD(Peroxidase),PAL(L-phenylalanine ammonia-lyase)and PPO(Polyphenol oxidase)in the over-expressed transgenic tobacco OEA1 and OEA2 increased to different degrees,and increased significantly at different infection time points.The activities of defense enzymes in the interfering strains IEA1 and IEA2 were significantly lower than those in the control strains.The results of resistance level identification showed that the disease spot rate of OEA1 was significantly lower than that of the control line,and the disease spot rate of OEA2 was significantly lower than that of the control line.The plaque rate of the interfering expression line IEA1-IEA2 was significantly higher than that of the control line.It is preliminarily believed that the process related protein GmPR1L can improve the resistance of tobacco to B.cinerea.

A study on JA-and BTH-induced resistance of Rosa rugosa‘Plena’to powdery mildew(Sphaerotheca pannosa)

Different concentrations of jasmonic acid(JA)and benzothiadiazole(BTH) were sprayed on 2-year-old Rosa rugosa‘Plena’ seedlings. The induced resistance of JA and BTH to Sphaerotheca pannosa(Wallr.) and the changes of their related physiological indices were investigated. Results showed that JA and BTH treatments had inhibitory impacts on S. pannosa infection. The optimal concentration of JA and BTH was 0.5 mmol/L for the disease-resistance induction of the leaves, its inductive effect was up to 66.36% for BTH and 54.49% for JA. Our results confirmed that exogenous JA and BTH significantly improved R. rugose ‘Plena’ resistance to S. pannosa. When treated with JA and BTH, activities of the three defense enzymes(POD, PPO, and PAL) increased significantly.Contents of total phenolics, flavonoids, and lignin also increased significantly. It is inferred from these results that exogenous JA and BTH could improve the resistance of R.rugose ‘Plena’ to S. pannosa through enhancing activities of the defensive enzymes and accumulation of secondary metabolites in the leaves.

Field Efficacy of Agricultural Antibiotic 702 against Rice Leaf Blast and Panicle Blast

The control effects of agricultural antibiotic 702 （ ag-antibiotic 702） on rice leaf blast and panicle blast were evaluated in field trials. With ag-antibiotic 702 and the two-line early hybrid rice 287 as test materials, different concentrations of ag-antibiotic 702 liquids were sprayed at tiUering stage and full heading stage of rice. In leaf blast test, the disease indexes of leaf blast at tillering stage were surveyed at 1 d prior to the first spraying and at 15 d post spraying, respectively. The disease indexes （DI） and the rice yield in panicle blast test was investigated at milky stage. Meanwhile, enzyme activities, including peroxidase （ POD）, catalase （CAT）, superoxide dismutase （SOD）, phenylalanine ammonia-lyase （PAL）, and polyphanol oxidase （PPO）, were measured. Kasugamycin （45 μg/mL） and deionized water were designed as positive and negative controls. When the concentrations of ag-antibiotie 702 were 15, 30 and 45 μg/mL, the control effects against leaf blast were 72.14%, 80.32% and 85.22%, respectively; the control effects on panicle blast were 74.08%, 84.87% and 86.58%, respectively; and the yield increase rates were 17.28%, 21.40% and 22.57%, respectively. So, ag-antibiotic 702, as a new agricultural antibiotic fungicide, could effectively cantrel rice blast by inducing defensive enzymes, and increase rice yield. The results will contribute to further development of ag-antibiotic 702 and its application in rice blast control.

Physiological Response of Oncidium hybridum‘Gower Ramsey’Infected by Erwinia carotovora

[Objective]The paper was to study the physiological mechanism of Oncidium hybridum‘Gower Ramsey’infected by Erwinia carotovora pv.carotovora.[Method]O.hybridum‘Gower Ramsey’was inoculated with E.carotovora by needle-punching method.The leaves inoculated with sterile water were used as control,and samples were collected at 12,24,36 and 48 h post inoculations to determine the physiological indexes,respectively.[Result]The contents of MDA,flavonoid,lignin,soluble protein and the activities of POD,SOD,CAT,PPO and PAL in O.hybridum leaves first increased then decreased.At 12 h post inoculation,the activities of SOD and CAT increased significantly by 40.49%and 37.77%(P<0.01,the same below),respectively;the contents of flavonoid increased significantly by 21.56%(P<0.05,the same below);and the activities of PAL,POD and PPO increased significantly by 42.29%,81.8%and 37.98%,respectively.The contents of MDA and lignin increased significantly by 57.98%and 58.72%at 24 h,respectively;the content of soluble protein increased by 87.71%at 24 h,and 61.28%at 36 h,while the content of Pro increased at 24 h(P>0.05,the same below),and increased significantly by 23.66%at 36 h;the content of soluble sugar content had no significant difference compared with the control.Additionally,the chlorophyll a and carotenoid contents tended to decrease dramatically,while chlorophyll b increased significantly by 95.33%at 24 h,then decreased.The initial fluorescence parameter of Fm/Fv and Y(II)decreased significantly by 47.14%and 54.61%,respectively,while the Y(NO)increased significantly by 33.61%,compared with the control.[Conclusion]The contents of Pro,MDA,lignin,soluble protein and the activities of POD,SOD,CAT,PPO and PAL in O.hybridum leaves are closely related to the infection of E.carotovora,which provide new clues for revealing the infection mechanism of E.carotovora in O.hybridum.

Foliar Application of Benzothiadiazole and Salicylic Acid to Combat Sheath Blight Disease of Rice

A field study was undertaken to ascertain the effects of elicitors viz. benzothiadiazole （BTH） and salicylic acid （SA） on defense related enzymes viz. peroxidase, phenylalanine ammonia lyase, superoxide dismutase, chitinase and 13-1,3-glucanase, and phenols in rice （Pusa Basmati I） plants. First foliar spray of BTH （50 mg/kg） and SA （50 mg/kg） was done at the maximum tillering stage and inoculation with Rhizoctonia solani was carried 24 h after elicitor treatment. Elicitors were further sprayed at every growth stage. Time course analysis showed peak accumulation of defense related enzymes and phenols in the rice leaves treated with BTH and SA, and accumulation was the highest at the flowering stage. Higher enzymatic activity was observed in elicitor treated plants inoculated with R. solani. Compared to the untreated control plants, application of elicitors before R. solani inoculation significantly elicited the defense related enzymes and phenols. Moreover, application of elicitors had a positive effect on yield and disease reduction. It is suggested that pretreatment of rice leaves with BTH and SA could be used to enhance the level of protection against sheath blight and to improve rice yield in the fields.

Quantitative Gene Expression of Peroxidase, Polyphenoloxidase and Catalase as Molecular Markers for Resistance against Ralstonia solanacearum

Brown rot or bacterial wilt of potato caused by Ralstonia solanacearum, is an economically important disease. Potato, cv. Nicola, was found to be relatively highly resistant to the infection with R. solanacearum and showed 15.12% wilt disease index, meantime, cv. Kara showed intermediate resistance with 37.40% disease index while, cv. Spunta was susceptible and showed 80.33% disease index. The role of defense-related enzymes in imparting resistance in potato against R. solanacearum was investigated by quantifying enzymes activity and gene expression of three defense- related enzymes, peroxidase, polyphenol oxidase and catalase. Peroxidase showed maximum activity 0.488 min-1·g-1 early at 12 h after pathogen inoculation in the cv. Nicola, whereas in susceptible cultivar Spunta showed lower activity of maximum 0.226 min-1·g-1 later at 48 h after inoculation. While, the moderately resistant cultivar Kara showed intermediate activity for the peak and its time. Meanwhile, polyphenol oxidase showed similar trends to that of peroxidase. On the contrary, catalase showed the highest activity values in the susceptible, cv. Spunta, while, in relatively highly resistant (cv. Nicola) and the moderately resistant (cv. Kara) showed lower values of activity and up to 96 h after inoculation. Meanwhile, gene expression of related enzymes the RT-PCR was used. At zero time, the relatively highly resistant potato cultivar, Nicola, showed the highest values of gene expression for both Peroxidase (POD) and Poly Phenol Oxidase (PPO). While, the susceptible potato cultivar, Spunta showed the lowest values. On the contrary, Catalase (CAT) gene expression was the highest in the susceptible, cv. Spunta, and was the lowest in the relatively highly resistant, cv. Nicola, while, was of intermediate values in the intermediate resistance, cv. Kara. Results show that peroxidase and polyphenol oxidase activities can be used as biochemical markers to reveal the resistance and susceptibility nature of potato cultivars against bacterial wilt disease of potato caused by R. solanacaerum.

Influence of rice black streaked dwarf virus on the ecological fitness of non-vector planthopper Nilaparvata lugens （Hemiptera： Delphacidae）

Rice black streak dwarf virus （RBSDV） is transmitted by the small brown planthopper （SBPH）, Laodelphax striatellus （Fallen）. Non-vector rice brown planthopper （BPH）, Nilaparvata lugens （Stal）, shares the same host rice plants with SBPH in paddy fields. The changes in nutritional composition of rice plants infected by RBSDV and the ecological fitness of BPH feeding on the infected plants were studied under both artificial climate chamber and field conditions. Contents of 16 detected amino acids and soluble sugar in RBSDV infected rice plants were higher than those in the healthy ones. On the diseased plants BPH had significantly higher nymphal survival rates, nymphal duration of the males, weight of the female adults, as well as egg hatchability compared to BPH being fed on healthy plants. However, there was no obvious difference in female nymph duration, longevity and fecundity. Defense enzymes （superoxidase dismutase, SOD and catalase, CAT） and detoxifying enzymes （carboxylesterase, CAE and glutathione S-transferase, GST） in BPH adults fed on diseased plants had markedly higher activities. The results indicate rice plants infected by RBSDV improved the ecological fitness of the brown planthopper, a serious pest but not a transmitter of the RBSDV virus.