A fragment of the Xanthomonas oryzae pv. oryzicola harpin HpaG Xooc reduces disease and increases yield of rice in extensive grower plantings

Harpins of phytopathogenic bacteria stimulate defense and plant growth in many types of plants, conferring disease resistance and enhanced yield. In a previous study, we characterized nine fragments of the harpin protein HpaG(Xooc) from Xanthomonas oryzae pv. oryzicola for plant defense elicitation and plant growth stimulation activity relative to the intact protein. In plants grown under controlled conditions, the fragment HpaG10-42 was more active in both regards than HpaG(Xooc). Here, we demonstrate that the activity of HpaG10-42 in rice under field conditions significantly exceeds that of HpaG(Xooc), stimulating resistance to three important diseases and increasing grain yield. We carried out tests in 672 experimental plots with nine cultivars of rice planted at three locations. Application protocols were optimized by testing variations in application rate, frequency, and timing with respect to rice growth stage. Of the concentrations (24, 24, 12, and 6 microg/ml), and number and timing of applications (at one to four different stages of growth) tested, HpaG10-42 at 6 microg/ml applied to plants once at nursery seedling stage and three times in the field was most effective. Bacterial blight, rice blast, and sheath blight were reduced 61.6 and 56.4, 93.6 and 76.0, and 93.2 and 55.0% in indica and japonica cultivars, respectively, relative to controls. Grain yields were 22 to 27% greater. These results are similar to results obtained with typical local management practices, including use of chemicals, to decrease disease severities and increase yield in rice. Our results demonstrate that the HpaG10-42 protein fragment can be used effectively to control diseases and increase yield of this staple food crop.     

四种柑桔盾蚧的室内人工饲养

在室内利用马铃薯及南瓜为寄主成功地饲养了4种柑桔盾蚧:红圆蚧、黄丘圆蚧、黑褐圆蚧和橙褐圆蚧;但黑点蚧不能正常生长发育。实验还表明,黄丘圆蚧可大批量繁殖多种寄生蜂。~(80)Co定量照射马铃薯能防止其发芽,南瓜可用200倍甲基托布津溶液处理,以延长存放期。以红圆蚧在南瓜上的生长发育较好。     

Increased abundance of patatins,lipoxygenase and miraculins in a thaxtomin A-habituated potato Russet Burbank somaclone with enhanced resistance to common scab

Potato common scab caused by the actinobacterium Streptomyces scabiei is characterized by the formation of corky lesions on tubers that reduce their marketability. Management of common scab is very complex and often ineffective under various environmental conditions. Using potato varieties that are more resistant to common scab remains one of the most efficient strategies to control this disease. However, very little is known about the factors associated with resistance to common scab. Somaclone RB9 regenerated from thaxtomin A-habituated potato Russet Burbank calli produced tubers more resistant to common scab than the original variety. Comparison of the RB9 tuber proteome with that of Russet Burbank using label-free quantitative proteomic analysis revealed changes in the accumulation of defence-related proteins from the patatin and lipoxygenase (LOX) families, which are involved in the metabolism of lipids, and of two miraculins of the Kunitz-type protease inhibitors family. The implication of LOX during common scab infection was studied using synchronized minitubers developed from leaf-bud cuttings. S. scabiei infection stimulated the accumulation of LOX in both Russet Burbank and RB9 minitubers, but this accumulation was intensified in RB9 minitubers. Infection also increased LOX activity in Russet Burbank and RB9 minitubers. However, LOX activity measured in noninfected RB9 minitubers was similar to that of infected Russet Burbank minitubers, indicating endogenous activation of LOX activity in RB9 minitubers. We discuss how increased LOX abundance and activity in the somaclone RB9 may contribute to improving tuber defence against common scab.     

Erwinia amylovora hrpN mutants,blocked in harpin synthesis,express a reduced virulence on host plants and elicit variable hypersensitive reactions on tobacco

Erwinia amylovora is the bacterium responsible for fire blight, a necrotic disease affecting many rosaceous plants and especially pear tree and apple tree. A protein named harpin, secreted through the Hrp secretion pathway and able to elicit an hypersensitive reaction (HR) on tobacco has recently been isolated. Mutants inhrpN, the gene encoding harpin were described as non pathogenic on immature pear fruit and unable to elicit an HR on tobacco [Weiet al., 1992; Wei and Beer, 1993]. In this paper, the phenotype on plant ofhrpN mutants was carefully determined.hrpN mutants expressed a weak but significant virulence on host plants. Furthermore, when infiltrated into tobacco leaf mesophyll, thehrpN mutants elicited varied responses that fluctuated from null reaction to full necrosis of the infiltrated area. These results show that harpin is not absolutely required neither for pathogenicity on host plant nor for elicitation of an hypersensitive reaction on tobacco. Furthermore, in all the tests performed, mutant blocked in harpin secretion remained non pathogenic and unable to elicit an HR on tobacco. This suggests that factor(s), different from harpin, involved both in pathogenicity and HR eliciting ability is (are) secreted through the Hrp secretion pathway.Abbreviations HR hypersensitive reaction - NSI necrosis severity index - CFU colonie forming units     

Uptake and distribution of thiabendazole by seed potatoes

The uptake of thiabendazole into potato seed tubers (cultivar King Edward) was determined by the method of application, more being taken up from solutions and suspensions than from dusts. Penetration and uptake were greatest with solutions at pH 3, and less at higher or lower pHs. The addition of inorganic salts (KCl, CaCl₂) had no effect. Treatment of tubers with weak solutions of mineral acid increased uptake of thiabendazole. There was little movement and no metabolism of the chemical when potatoes were stored for 3 months.     

The effect of foliar application of phosphonate formulations on the susceptibility of potato tubers to late blight

Foliar sprays of potato plants with phosphonic acid (partially neutralised with potassium hydroxide to pH 6.4) substantially reduced infection of the tubers by Phytophthora infestans, the cause of late blight, in glasshouse and field experiments over a 4-year period. Healthy tubers of blight-susceptible cultivars removed from treated plants and artificially inoculated by spraying with sporangial/zoospore suspensions of P infestans did not develop disease symptoms, demonstrating that the phosphonate applications had directly reduced the susceptibility of tubers to infection, probably as a result of translocation into tuber tissue. In contrast, foliar application of fosetyl-aluminium did not significantly reduce tuber blight development following inoculation. Five to six sprays of partially neutralised phosphonic acid (2 kg ha-1) applied at 10-14 day intervals resulted in the least tuber infection, but such a treatment regime may not be economic. In trials where the effect of timing and rate of application of 2-4 kg phosphonic acid ha-1 was examined, a single treatment of 4 kg ha-1 applied mid- or late-season proved the most effective. A spray programme in which one or two applications of phosphonic acid are combined with use of a non-systemic or systemic fungicide to enhance foliar protection offers the possibility of controlling both foliage and tuber blight and could have a major impact in reducing overwinter survival of P infestans in tubers.     

Defense induced by a bis-aryl methanone compound leads to resistance in potato against Phytophthora infestans

Plants recognize certain microbial compounds as elicitors of their active defense mechanisms. In the present study, NUBS-4190, a synthetic bis-aryl-methanone compound elicited NO and ROS generation in potato suspension cultured cells and intact potato leaves. Hypersensitive cell death was found in these cultured cells and in potato leaves without the accumulation of phytoalexins in the tubers. Defense-related genes such as StrbohB, StrbohC, StNR1, StNR5, Sthsr203J and StPR1 were expressed in potato suspension cultured cells treated with NUBS-4190. Resistance against Phytophthora infestans also increased in NUBS-4190-treated potato leaves.     

Expression of harpin(xoo) in transgenic tobacco induces pathogen defense in the absence of hypersensitive cell death

ABSTRACT Harpin(Xoo), encoded by the hpaG(Xoo) gene of Xanthomonas oryzae pv. oryzae, is a member of the harpin group of proteins that induce pathogen resistance and hypersensitive cell death (HCD) in plants. We elaborated whether both processes are correlated in hpaG(Xoo)-expressing tobacco (HARTOB) plants, which produced harpin(Xoo) intracellularly. Resistance to fungal, bacterial, and viral pathogens increased in HARTOB, in correlation with the expression of hpaG(Xoo), the gene NPR1 that regulates several resistance pathways, and defense genes GST1, Chia5, PR-1a, and PR-1b that are mediated by different signals. However, reactive oxygen intermediate burst, the expression of HCD marker genes hsr203 and hin1, and cell death did not occur spontaneously in HARTOB, though they did in untransformed and HARTOB plants treated exogenously with harpin(Xoo). Thus, the transgenic expression of harpin(Xoo) confers nonspecific pathogen defense in the absence of HCD.     

缓控释肥不同施用量和施用方式对旱作区全膜马铃薯生长及产量的影响

为掌握缓控释肥的施用对全膜马铃薯关键生长指标及产量的影响,分别在高(900 kg·hm-2)、中(750 kg·hm-2)、低(600 kg·hm-2)3种施肥条件下进行了2种不同施用方式即集中沟施(F)与表层撒施(B)的田间试验,并测定了马铃薯主要生长指标和产量。结果表明:缓控释肥施用量对旱作全膜马铃薯主要生长指标、产量及商品率的影响均达显著性差异水平,在施用900 kg·hm-2的高肥量时,其沟施处理HF的马铃薯产量、大薯率和商品率均达到最大,分别达43 608.85 kg·hm-2、72.1%和92.7%。施用方式对马铃薯地下主要生长指标、产量和大薯率均表现出显著性差异,且在3种施肥量下,F处理的马铃薯产量、大薯率和商品率均高于B处理,其中在高肥水平下F处理较B处理平均增产达2 834.54 kg·hm-2,增产率为6.4%。施用量×施用方式对马铃薯各生长指标和产量影响均不显著。综合来看,缓控释肥在较高水平的施肥(900 kg·hm-2)条件下,集中沟施处理有利于旱作区全膜马铃薯产量的提高。     

Investigations on potato mop-top furovirus in Sweden1

Potato mop-top furovirus (PMTV) was isolated for the first time in Sweden in 1985. During 1986 and 1987, soil samples from potato fields, mainly in the south-west of Sweden, were tested for PMTV. The virus was detected in 49 out of 305 samples tested. PMTV was also found to be present in soil collected from potato riddles during potato grading in early spring. The soil samples were tested by using Nicotiana clevelandii and N. debneyi as bait plants in a glasshouse. Seven potato cultivars were grown in a PMTV-infested field. PMTV symptoms in the tubers, i.e. external and/or internal brown rings and arcs (spraing), were observed in cvs Evergood, Provita, Saturna and Ukama but not in cvs Bellona, Bintje or King Edward. Very few, or no, PMTV symptoms were found in tubers of susceptible cultivars when examined 2-3 weeks after lifting. In later examinations spraing appeared and increased. The increase was greater in tubers stored at 9°C than in those at 4°C, and in cut than in uncut tubers.     

Biocontrol of powdery scab of potato by seed tuber application of an antagonistic fungus, <Emphasis Type="Italic">Aspergillus versicolor</Emphasis>, isolated from potato roots

An environmentally friendly measure to control potato powdery scab caused by a protozoan pathogen Spongospora subterranea f.sp. subterranea (Sss) was developed by focusing on antagonistic microorganisms that were considered compatible with potato root. Five hundred and eight soil fungi, isolated from potato root cultivated in soil suspensions from four potato fields in Hokkaido, were screened for suppressiveness of root infection by Sss in a hydroponic culture system and for powdery scab severity in greenhouse and field experiments. Antagonistic isolate Im6-50, identified as Aspergillus versicolor, was selected as a potent biological control agent. In a 3-year field test, A. versicolor Im6-50 suppressed powdery scab with a protection value of 54–70 (100?=?complete protection) when applied directly on seed tubers compared with a protection value of 77–93 by the synthetic fungicide fluazinam. A. versicolor Im6-50 was detected from the surface of daughter tubers and from the soil in which the inoculated seed tubers were cultivated by PCR using species-specific primers. The establishment of A. versicolor Im6-50 on the stolon of inoculated potato plants and in the rhizosphere is considered to contribute to the mechanism for disease suppression.     

Effects of seed potato tuber treatment with imidacloprid on some soil and foliar insects in Croatia

The treatment of seed potato tubers by imidacloprid, as well as the standard granular insecticides applied in the furrow, did not, in four years of trials, sufficiently prevent damage from wireworms, noctuid larvae and mole crickets to potato tubers. The main reason for this failure is the spatial and temporal distance between the application of insecticides and the moment when their action is needed. On the other hand, the effects of imidacloprid applied as seed tuber treatment on the larvae of the Colorado potato beetle was very good: depending upon conditions, it lasted from at least 55, to 70 days at the most. It protected the foliage from any substantial damage from the entire first generation, and ensured high yields. Therefore a decision on seed tuber treatment with imidacloprid in Croatia should primarily depend upon the cost/benefit calculation based on damage expected from the CPB (or aphids), and not from soil insect pests.     

Sensitivity of different methods for the detection of Ralstonia solanacearum in potato tuber extracts

The sensitivities of various methods for the detection of Ralstonia solanacearum following dilution in healthy potato tuber tissue macerate were compared. Estimated pathogen populations in undiluted macerates, from samples of 200 heel-end vascular cores each containing a single diseased and 199 healthy tubers, ranged from 1.2 × 10⁶–7.4 × 10⁷ colony-forming units per ml. Following concentration by high-speed centrifugation and resuspension in phosphate buffer, the pathogen was detected by all methods studied, including culture on semi-selective media, enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescent-antibody staining (IFAS) of fixed cells, immunofluorescent colony staining (IFCS), detection of specific DNA sequences following amplification by the polymerase chain reaction (PCR) and bioassay in tomato seedlings. Both ELISA and PCR methods were improved by pre-enrichment of samples in semi-selective broth prior to testing. A nested PCR method was evaluated which could detect fewer than 10 cells per ml in the potato extracts. Of the other methods only dilution plating on semi-selective medium and tomato bioassay could detect fewer than 10⁴ cells per ml. In order to combine ease and speed of use with sensitive detection, it was recommended that a series of methods be used for routine screening of potato tuber stocks for infection by R. solanacearum.     

Effects of seed potato tuber treatment with imidacloprid on some soil and foliar insects in Croatia

The treatment of seed potato tubers by imidacloprid, as well as the standard granular insecticides applied in the furrow, did not, in four years of trials, sufficiently prevent damage from wireworms, noctuid larvae and mole crickets to potato tubers. The main reason for this failure is the spatial and temporal distance between the application of insecticides and the moment when their action is needed. On the other hand, the effects of imidacloprid applied as seed tuber treatment on the larvae of the Colorado potato beetle was very good: depending upon conditions, it lasted from at least 55, to 70 days at the most. It protected the foliage from any substantial damage from the entire first generation, and ensured high yields. Therefore a decision on seed tuber treatment with imidacloprid in Croatia should primarily depend upon the cost/benefit calculation based on damage expected from the CPB (or aphids), and not from soil insect pests.     

受黄色镰刀菌侵染的马铃薯薯块的抗氧化酶、细胞壁防御酶及StLTPa1基因的表达特性

黄色镰刀菌Fusarium culmorum为黑龙江省马铃薯干腐病主要致病菌,干腐病可以导致马铃薯在窖藏过程中发生腐烂,影响薯块的商品价值和食用价值,为进一步研究马铃薯干腐病的发生和防治,本研究采用黄色镰刀菌对不同抗性的马铃薯块茎进行侵染,对病原菌侵染过程中薯块的抗氧化酶及细胞壁降解酶变化及病程相关基因的表特性进行了研究。结果表明,当黄色镰刀菌侵染块茎时,块茎中的可溶性蛋白和丙二醛(malondlaldehyde,MDA)含量,超氧化物歧化酶(superoxide dismutase,SOD)、过氧化物酶(peroxidase,POD)活性、几丁质酶(chitinase)和β-1,3-葡聚糖酶(β-1,3-glucanase)活性都呈现不同程度的上升趋势。非特异性脂质转移蛋白基因StLTPa1表达量随着黄色镰刀菌的侵染时间呈波动性,在植物防御反应中发挥一定的作用。     

Detection of spore balls of Spongospora subterranea on potato tubers by enzyme-linked immunosorbent assay

A rabbit was immunized with a homogenate of spore balls (cystosori) of Spongospora subterranea which also contained some potato tuber debris. The resultant polyclonal antiserum contained antibodies which reacted against extracts of spore balls and healthy tubers. Antibodies to the tuber debris were removed by incubating the blood serum with an extract from a S. subterranea -free tuber. In enzyme-linked immunosorbent assay (ELISA), the γ-globulin fraction of the absorbed antiserum reacted with dilute tuber extracts containing the equivalent of as little as 0·08 spore balls per ml. The reaction with spore balls was inhibited in high concentrations of tuber sap. Extracts of peel from symptomless tubers which had been stored in contact with scabbed tubers also reacted with the γ-globulin, presumably because the symptomless tubers became contaminated with spore balls. However, spore balls in soil were only weakly detected when high numbers were present. The γ-globulin did not react with the plasmodial stage of the pathogen, or with 15 other micro-organisms tested, including resting spores of Plasmodiophora brassicae.     

<Emphasis Type="Italic">N</Emphasis>,<Emphasis Type="Italic">N</Emphasis>-dimethylsphingosine,an inhibitor of sphingosine kinase,induces phytoalexin production and hypersensitive cell death of Solanaceae plants without generation of reactive oxygen species

Plant recognition of elicitors derived from pathogens induces various resistant reactions, including production of reactive oxygen species, hypersensitive cell death and accumulation of phytoalexins. Previously, we isolated a ceramide elicitor from Phytophthora infestans, which activates O₂ ⁻ production of potato suspension-cultured cells. In this study, we employed nine ceramide-related chemicals to test their elicitor activity. Although, none of the tested chemicals induced O₂ ⁻ production, N,N-dimethylsphingosine (DMS) induced accumulation of phytoalexin in potato tubers. In potato, tobacco and Nicotiana benthamiana, DMS also induced rapid cell death. DMS-treated potato cells stained with 4′,6-diamidino-2-phenylindole (DAPI) showed chromatin condensation, and isolated DNA from DMS-treated cells had ladder pattern, confirming that DMS-induced plant cell death is a hypersensitive reaction-like programmed cell death. Involvement of ceramide signaling in induction of plant defense reactions is discussed. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.