马铃薯细菌性软腐病研究进展

细菌性软腐病是一种可引起果蔬严重危害的世界性植物病害,其频发已成为制约马铃薯种植业发展的瓶颈。本文基于国内外细菌性软腐病最新的研究报道,对马铃薯细菌性软腐病的病原鉴定、发病症状、致病机理、致病因子及防治措施等进行综述,并提出了进一步的研究方向,以期为马铃薯细菌性软腐病的相关研究提供科学参考。     

茉莉酸甲酯调控防御酶活性诱导猕猴桃果实抗采后软腐病

以‘金魁’猕猴桃果实为试验材料,研究茉莉酸甲酯(methyl jasmonate,MeJA)调控防御酶活性抗猕猴桃采后软腐病的效应。测定了MeJA对猕猴桃软腐病病斑直径、软腐病菌Botryosphaeria dothidea抑菌作用及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)和多酚氧化酶(PPO)等防御酶活性的影响。结果表明:在0.001～10 mmol/L浓度范围内,MeJA对猕猴桃软腐病菌B.dothidea的抑制作用随浓度升高而增强;MeJA对猕猴桃果实最佳诱导浓度和熏蒸时间分别为0.1 mmol/L和24 h,其诱导效果分别为26.01%和26.85%;猕猴桃果实经0.1 mmol/L MeJA熏蒸处理24 h后,SOD、POD、CAT、APX和PPO活性提高,其中SOD和POD活性分别较对照增加33.85%和61.61%,差异达显著水平(P<0.05)。以上结果暗示MeJA诱导猕猴桃果实抗采后软腐病可能与其提高防御酶活性有关。     

11种杀菌剂对马铃薯软腐病的防治效果

由果胶杆菌属细菌Pectobacterium spp.引起的软腐病是世界范围内马铃薯生产上重要的细菌性病害之一。本研究分别采用碟片法和生长速率法探究了11种常用的杀菌剂对胡萝卜果胶杆菌胡萝卜亚种菌株Pcc20181的离体抑菌效果; 采用马铃薯半薯接种法评价了供试杀菌剂对马铃薯软腐病的防控效果。碟片法和生长速率法测定结果均显示:11种供试杀菌剂中72%农用硫酸链霉素SP、0.3%四霉素AS和3%噻霉酮WP离体抑菌效果最好, 抑菌圈直径介于0.13~1.47 cm, EC50介于1.695~44.363 mg/L; 新鲜半薯接种法测定结果表明, 11种供试杀菌剂中有8种对软腐病均有控病效果, 防效为33.33~92.50%。其中, 3%噻霉酮WP对马铃薯软腐病的防效最高为92.50%, 随后依次为72%农用硫酸链霉素SP （89.76%）、20%叶枯唑WP （58.81%）、0.3%四霉素AS （51.60%）。综合评价, 3%噻霉酮WP、20%叶枯唑WP和0.3%四霉素AS对马铃薯软腐病具有较好的防控效果。     

Induction of defence responses against Erwinia soft rot by an endogenous pectate lyase in potatoes

Pectate lyase (PL) enzymes are major virulence factors of Erwinia carotovora (Ec) bacteria. They degrade plant cell wall pectin into unsaturated oligogalacturonates (OG) known to elicit plant defence responses. Therefore, a gene encoding the isoenzyme PL3 of Ec ssp. atroseptica was transformed by means of Agrobacterium into potatoes of cv. Désirée. Four PL-transgenic potato plant lines selected on the basis of greenhouse experiments were grown over a period of 4 years (1997–2000) in the field. It is shown that the heterologous PL enzyme mediated an enhanced resistance to Erwinia soft rot in field grown tubers. Thus compared to the non-transgenic counterpart extension of rotting was significantly reduced (P < 0.001) on the wound surface of PL-expressing tubers. Moreover, the threshold density ofEc -bacteria causing a progressive soft rot was up to 19-fold higher on tuber tissue containing the PL enzyme. An induction of plant defence responses in PL-transgenic potatoes may be indicated by an enhanced resistance of tuber tissue cell walls to Erwinia -derived enzymes, an increased PPO- and PAL-activity in tuber tissue as well as by a strengthened formation of necrosis on the wound surface of tubers after infection with Ec -bacteria.     

Protection against bacterial soft rot by olive extracts is related to general defence induction in potato tubers

Controlling bacterial plant diseases remains a challenge, as direct chemical control is usually not possible. Obtaining new biocontrol methods, in particular efficient biomolecules able to boost defence reactions and limit infection or symptom development, is therefore of major importance. This study screened extracts from leaves of two Mediterranean plants rich in bioactive phenolic compounds, olive and carob, for their ability to reduce soft rot severity and to trigger phenylalanine ammonia‐lyase (PAL) activity in potato tuber slices. Extracts from olive leaves significantly reduced disease severity caused by Pectobacterium atrosepticum or Pectobacterium carotovorum, whereas carob leaf extracts significantly increased it. Olive extracts and its main phenolic components, oleuropein and hydroxytyrosol, also significantly increased PAL activity 7.5 h after application. None of the extracts or purified molecules reduced bacterial growth in vitro. Furthermore, the effect of these extracts varied according to potato cultivars. These data therefore open new ways for the biological control of soft rot bacteria, but stress the importance of understanding the causes of response difference in different potato varieties in order to obtain optimal efficacy.     

Etiology of potato soft rot and blackleg diseases complex in Egypt

Journal of Plant Diseases and Protection - Recent observations of increasing incidences of potato plants showing blackleg symptoms have caused serious concern for potato growers and traders in...     

The biochemical basis of resistance of potato to soft rot bacteria

Evidence is presented that a number of mechanisms are involved in the resistance of commercial cultivars of potato to soft rottingErwinia spp. These include phytoalexins, phenolics, cell wall modification, and possibly proteins. The role of elicitor-active oligogalacturonides, which are released from plant cell walls by pectic enzymes, is discussed.     

Identification and differentiation of soft rot and blackleg bacteria from potato using nested and multiplex PCR

Journal of Plant Diseases and Protection - The bacterial genera Pectobacterium, Dickeya and Enterobacter are plant pathogens that cause soft rot and blackleg of potato. In the present study, 25...     

广东省马铃薯块茎软腐病病原菌鉴定

为明确广东省惠州市马铃薯块茎软腐病的病原菌,采用常规病组织分离法获得4株菌株,通过生理生化特征和分子生物学特性对其进行鉴定,并测定了该病原菌对20个马铃薯品种的致病力。结果显示,该病原菌为革兰氏阳性菌,在LB平板上菌落有透明和不透明2种形态;除葡萄糖发酵阳性、对链霉素和青霉素具有抗性等特征不同外,BP-hd-1、BP-hd-2、BP-hd-3和BP-hd-4菌株与短小芽胞杆菌Bacillus pumilus菌株的其它生理生化特征均相同。系统进化分析结果表明,4株菌株的16S rDNA序列分别与短小芽胞杆菌AI-Khrj18(KY123871)、ML270(KC692158)、NCTC10337(LT906438)和ARD21(KX023236)的相似性为100.0%;4株菌株的gyrB基因序列均与短小芽胞杆菌AUEC29菌株(HM585095)的gyrB基因相似性最高,为99.7%～99.8%。生理生化特征及分子生物学鉴定结果表明,引起广东省马铃薯块茎软腐病的病原菌为短小芽胞杆菌。致病力测定结果显示,接种菌株BP-1后20个马铃薯品种的发病率均为100.0%,表明该病原菌对马铃薯有强致病力。     

Detection,quantification and classification of soft rot erwinias associated with potato tubers

An improved method for detection, quantification and classification of soft rot bacteria associated with potato seed tubers, plant material, soil and water has been developed. The method is based on the use of a modified version of the crystal violet pectate selective medium (CVP), enrichment cultures under anaerobic conditions using pectate as the sole carbon source for recovery improvement, and the quantitative estimation ofErwinia spp. by employing a new solid medium - most probable number (MPN) method. The use of this method enabled an improvement in the recovery and identification of specificErwinia spp. in mixed populations. This was done by incubating CVP plates — used for the MPN counting — at three different temperatures (15, 28 and 39°C). These combined techniques were used for estimating low level populations at less than one cell per gram or ml tested ofErwinia carotovora subsp.carotovora, E. carotovora subsp.atroseptica, andE. chrysanthemi.     

Pectinolytic bacteria associated with potato soft rot and blackleg in South Africa and Zimbabwe

Soft rot and blackleg can cause severe economic losses in potato production in South Africa and Zimbabwe depending on climatic conditions. The aim of the study was to identify the predominant bacteria causing potato soft rot and blackleg in these countries. Samples, comprising of stems and tubers from potato plants with blackleg and soft rot symptoms were collected from 2006?C2009 from potato production areas where disease outbreaks occurred. The isolates from these plants and tubers yielded Gram negative, pectinolytic bacteria on crystal violet pectate and inoculated tubers. Identification was based on biochemical and phenotypic characteristics, rep-PCR, Amplified Fragment Length Polymorphisms and sequences of gyrB and recA genes. Isolates from Zimbabwe were identified as Pectobacterium carotovorum subsp. brasiliensis (Pcb) (21 isolates), Dickeya dadantii subsp. dadantii (Dd) (20 isolates), P. c. subsp. carotovorum (Pcc) (16 isolates) and P. atrosepticum (Pa) (4 isolates). Pcb, Pcc and Dd subsp. dadantii were isolated from samples collected from all the regions, while Pa was isolated from Nyanga the coolest region in Zimbabwe. In South Africa, however, Pcb was the most common causal agent of soft rot and blackleg. P. atrosepticum was the only pathogen isolated from samples collected in Nyanga, Zimbabwe, and was not isolated from any South African samples. AFLP analysis separated the Pcb strains into 12 clusters, reflecting subdivision in terms of geographic origin, and Pcc isolates were clearly differentiated from Pcb isolates. A large degree of DNA polymorphism was evident among these 12 clusters. The study identified all the pathogens associated with the blackleg/soft rot disease complex.     

A rapid method to identify and quantify soft rot erwinias on seed potato tubers1

Potato tubers are usually contaminated by more than one species or pathovar of soft rot erwinia and, because blackleg incidence is related to the contamination level of seed tubers, the disease potential of seed stocks may be assessed by determining seed-tuber contamination level. A method is described for identifying and quantifying directly from tubers the three soft rot erwinias commonly associated with potatoes. Replicate lots of 10–15 tubers are peeled by dry abrasion in a commercial potato peeler and the peel weight determined by weighing the tubers before and after peeling. Sap is expressed from the peel, an antioxidant (0.075% dithiothreitol) added, and the sap is dilution-plated on a diagnostic selective medium (crystal violet pectate [CVP]). After incubating for 24 h at 20°C, the plates are velvet-replicated onto fresh plates of CVP with or without 35 μg ml^-1 erythromycin and incubated for 48 h at 27°C and 24 h at 33.5 or 37°C. Soft-rot erwinias typically form deep cup-like cavities on CVP and they can be identified and enumerated according to the pattern of cavity formation. Cavities are formed by Erwinia carotovora pv. atroseptica only at 27°C, by E. carotovora pv. carotovora at 27 and 33.5°C but not at 37°C, whereas E. chrysanthemi forms cavities at all temperatures but fails to grow in the presence of erythromycin. Contamination levels can be expressed as the number of different erwinias per tuber or per g peel.     

一株马铃薯干腐病生防菌的鉴定及其抑菌活性研究

镰孢菌引起的马铃薯干腐病是影响马铃薯贮存期间产量与品质的重要病害之一。寻找高效低毒的防治马铃薯干腐病的生防药剂对提高马铃薯的产量与质量具有重要意义。本研究以腐皮镰孢Fusarium solani为病原菌, 测定分离自马铃薯块茎的菌株2-1-9-CJK-2的抑菌活性。该菌株发酵液、无菌发酵液、菌悬液、胞内组分以及挥发性有机物对腐皮镰孢均具有抑菌活性, 抑制率分别为93.37%、61.92%、80.82%、44.01%和49.16%。结合形态学、生理生化特征和多基因系统发育树构建鉴定其为贝莱斯芽胞杆菌Bacillus velezensis。马铃薯切片试验和马铃薯块茎试验表明, 当菌株发酵液浓度为1×109 cfu/mL时, 对病原菌的抑制效果最好。除此之外, 菌株可以分泌蛋白酶、淀粉酶、纤维素酶, 具有溶磷能力和生物膜形成能力, 但不具有溶血性, 并且对供试的其他6种病原真菌有较强的抑菌活性。研究表明贝莱斯芽胞杆菌2-1-9-CJK-2抑菌谱广, 具有生物安全性, 对腐皮镰孢有较强的抑菌活性, 在植物病害防治中具有一定的应用潜力, 可为马铃薯干腐病生防制剂的开发奠定基础。     

马铃薯内生细菌的分离及环腐病拮抗菌的筛选鉴定

 本研究从大同、太原和内蒙古等地采集马铃薯块茎,分离到240株内生细菌,通过离体抑菌作用测定,共得到55株对环腐病菌有拮抗作用的菌株,占菌株总数的22.9%,抑菌圈半径最大的可达13 mm。按抑菌圈半径大小将拮抗菌分为强、中、弱三类。从中筛选出9个对环腐病等病菌具有较强拮抗作用的内生菌株进行了细菌学鉴定,结果表明:118为荧光假单胞生物型V (Pseudomonas fluorescens biovar V);110为短小芽孢杆菌(Bacillus pumilus);085为嗜热脂肪芽孢杆菌(Bacillus stearothermophilus);069为草生欧文氏菌(Erwinia herbicola);043为草莓黄单胞菌(Xanthomonas fragariae);A-10'、T3为芽孢杆菌属(Bacillus);H1-6为荧光假单胞菌(Pseudomonas fluorescens);116为短小杆菌属(Curtobacterium)。     

Contamination and subsequent multiplication of soft rot erwinias on healthy potato leaves and debris after haulm destruction

Small plots of potatoes were inoculated with a mixture of Erwinia carotovora (E. c.) subsp. carotovora and E. carotovora subsp, atroseptica strains resistant to rifampicin. Subsequently the population off, c. subsp, carotovora and E. c. subsp, atroseptica (rifampicin-resistant and wild types) present as epiphytes on the surface of potato leaves was assessed using three methods, qualitative, semi-quantitative and quantitative, during 1986 and 1987. The population was generally low (< 10² colony forming units (> 10⁴cfu/g leaves) but reached higher levels (> 10⁴ cfu/g) on occasions later in the growing season, Rifampicin-resistant erwinias were reisolated only infrequently throughout this study. Different methods of haulm destruction (herbicide, pulverization, sulphuric acid treatment and natural senescence) greatly influenced the number of erwinias present in the resulting plant debris. Pulverization resulted in the highest population (10⁶-10⁷ wild-type cfu/g) in both seasons. In 1987. the wettest of the two seasons of this study, herbicide treatment resulted in similarly high populations. The results suggest that the high numbers of erwinias found in the haulm debris were probably derived from the generally low populations of epiphytic bacteria previously present on healthy leaves, E. c. subsp, carotovora was the most frequent subspecies in the rotting plant debris; E. c. subsp, atroseptica was more commonly found on healthy leaves. The implications of the results are discussed in relation to the production of seed potatoes with a low risk of blackleg.     

Control of blackleg and tuber soft rot of potato caused by Pectobacterium and Dickeya species: a review

This paper briefly reviews research on the causative agents of blackleg and soft rot diseases of potato, namely Pectobacterium and Dickeya species, and the disease syndrome, including epidemiological and aetiological aspects. It critically evaluates control methods used in practice based on the avoidance of the contamination of plants, in particular the use of seed testing programmes and the application of hygienic procedures during crop production. It considers the perspective of breeding and genetic modification to introduce resistance. It also evaluates the application of physical and chemical tuber treatments to reduce inoculum load and examines the possibility of biocontrol using antagonistic bacteria and bacteriophages.     

Methods of testing potato cultivars for resistance to soft rot of tubers caused by Erwinia carotovora subsp. atroseptica

The resistance of eight potato cultivars to tuber soft rot caused by E. carotovora subsp. atroseptica was assessed in 2 years using three different test methods. Similar cultivar resistance rankings were obtained for any one method within a year and between years for two methods (single site, infectivity titration), but not for the third (vacuum infiltration). However, the ranking of cultivars differed for the three methods. Ranking was not affected by inoculating the cortex or the more susceptible medullary tissue, or by assessing rotting in terms of infection frequency or lesion size, but it was affected by oxygen concentration during incubation. Differences among cultivars were greater when inoculated tubers were incubated anaerobically than when incubated with 5% oxygen. There was no relationship between the relative susceptibilities of cultivars to tuber soft rot in storage in January/February and those of mother tubers after planting.     

Evidence that DL-3-aminobutyric acid and acibenzolar-S-methyl induce resistance against bacterial head rot disease of broccoli

Head rot of broccoli caused by Pseudomonas marginalis (Brown) Stevens and P. fluorescens Migula is a major disease in Brittany (France). To date, no accession with a satisfactory field resistance has been identified, and available pesticides are not effective in controlling the disease. The aim of this study was to test whether acibenzolar-S-methyl (ASM), DL-3-aminobutyric acid (BABA) and potassium phosphonate (K(2)HPO(3)), known to induce resistance against various diseases, can help protect broccoli against head rot. The susceptible broccoli F1 hybrids Marathon and Shogun were grown in a greenhouse until head formation. They were then sprayed with ASM (0.23 mM AI), BABA (20 mM AI) or potassium phosphonate (37.41 mM AI) until runoff. In one experiment, heads from treated plants were excised, inoculated (10(4) cfu ml(-1)) and incubated in Magenta GA7 vessels. In another experiment, heads were inoculated on treated living plants. Disease ratings were made 5 days after inoculation. Antibiotic- and water-treated plants served as controls. Results obtained showed that, on excised treated heads, potassium phosphonate was not protective and disease scores were comparable with those of the water control. BABA- and ASM-treated excised heads were poorly, but significantly, protected. On whole plants with heads attached, the latter two compounds were much more effective. ASM-induced resistance increased in effectiveness over 8 days after inoculation, whilst that induced by BABA decreased. This result suggests that testing disease resistance inducers on excised broccoli heads is not accurate. ASM and BABA may offer alternative methods for controlling head rot of broccoli.     

甲基硫菌灵和百菌清浸苗防治甘薯黑斑病的影响因素分析

为分析杀菌剂浸苗时各因素对甘薯黑斑病发生的影响,探讨根据药剂类型优化浸苗技术的必要性,选择内吸性的甲基硫菌灵和非内吸性的百菌清开展浸苗试验,通过单因素试验确定浸苗时间和药液质量浓度对防治效果的影响,通过正交试验比较浸苗时间、药液浓度、孢子浓度对甘薯黑斑病发病的影响,利用浸苗时间与药液浓度的组合试验,探讨浸苗条件与防治效...