Effect of immunomodulators on potato resistance and susceptibility to Phytophthora infestans

The mechanisms of induced resistance and susceptibility of potato (Solanum tuberosum L.) tubers to late blight agent (Phytophthora infestans Mont de Bary) were studied using an elicitor chitosan and an immunosuppressor laminarin. It was elucidated that treatment of disks from potato tubers with chitosan resulted in salicyclic acid (SA) accumulation due to activation of benzoate-2-hydroxylase and hydrolysis of SA conjugates. Such SA accumulation in potato tissues inhibited one of the antioxidant enzymes, catalase, inducing an oxidative burst and resistance development. The mechanisms of induced susceptibility to the late blight causal agent were studied using an unspecific immunosuppressor, laminarin, an analogue of natural specific suppressor of potato immune responses, β-1,3,β-1,6-glucan. It was established that the development of immunosuppression in tissues treated with laminarin did not affect the SA level in tissues. However, catalase sensitivity to SA reduced in laminarin-treated tissues, and the enzyme activity increased. In its turn, this might result in the reduced level of hydrogen peroxide in the cells and, as a sequence, in the increased potato susceptibility to late blight.     

The R1 gene for potato resistance to late blight (Phytophthora infestans) belongs to the leucine zipper/NBS/LRR class of plant resistance genes

Late blight caused by the oomycete Phytophthora infestans is the most destructive disease in potato cultivation worldwide. New, more virulent P. infestans strains have evolved which overcome the genetic resistance that has been introgressed by conventional breeding from wild potato species into commercial varieties. R genes (for single-gene resistance) and genes for quantitative resistance to late blight are present in the germplasm of wild and cultivated potato. The molecular basis of single-gene and quantitative resistance to late blight is unknown. We have cloned R1, the first gene for resistance to late blight, by combining positional cloning with a candidate gene approach. The R1 gene is member of a gene family. It encodes a protein of 1293 amino acids with a molecular mass of 149.4 kDa. The R1 gene belongs to the class of plant genes for pathogen resistance that have a leucine zipper motif, a putative nucleotide binding domain and a leucine-rich repeat domain. The most closely related plant resistance gene (36% identity) is the Prf gene for resistance to Pseudomonas syringae of tomato. R1 is located within a hot spot for pathogen resistance on potato chromosome V. In comparison to the susceptibility allele, the resistance allele at the R1 locus represents a large insertion of a functional R gene.     

Effects of ethylene and auxin on polyamine levels in suspension-cultured tobacco cells

The effects of ethylene and auxin on polyamine levels were studied in suspension-cultured cells of tobacco ( Nicotiana tabacum . L). Treatment of 4-day-cultured cells with ethylene increased the levels of spermidine and spermine. The activities of arginine decarboxylase (ADC; EC 4.1.1.19), ornithine decarboxylase (ODC: EC 4.1.1.17), and S -adenosylmethionine decarboxylase (SAMDC: EC 4.1.1.50) rapidly increased between 3 and 12 h. An auxin, indole-3-acetic acid (IAA), increased polyamine levels and activities of ADC, ODC and SAMDC. The spermine level continued to increase significantly during a 24-h incubation with IAA. The increases in polyamine accumulation induced by ethylene were partially offset by an inhibitor of ethylene action, 2,5-norbornadiene. It is suggested that the auxin-induced polyamine accumulation occurred directly, without metabolic competition between ethylene and polyamine biosynthesis, and indirectly, through auxin-induced ethylene formation.     

Alteration of pathogenicity-linked life-history traits by resistance of its host Solanum tuberosum impacts sexual reproduction of the plant pathogenic oomycete Phytophthora infestans

Although sexual reproduction implies a cost, it represents an evolutionary advantage for the adaptation and survival of facultative sexual pathogens. Understanding the maintenance of sex in pathogens requires to analyse how host resistance will impact their sexual reproduction through the alteration of their life-history traits. We explored this experimentally using potato (Solanum tuberosum) and one of its pathogens, the heterothallic oomycete Phytophthora infestans. Sexual reproduction was highest on hosts favouring asexual multiplication of the pathogen, suggesting similar nutritional requirements for both sexual and asexual sporulation. Sexual reproduction was also highest on hosts decreasing the latent period, probably because of a trade-off between growth and reproduction. Distinguishing host effects on each pathogenic trait remains however uneasy, as most life-history traits linked to pathogenicity were not independent of each other. We argue that sexual reproduction of P. infestans is an adaptation to survive when the host is susceptible and rapidly destroyed.     

Ribonuclease and proteinase activities in potato leaves immunized against Phytophthora infestans

Local treatment of potato plants with arachidonic acid caused a repeated increase of ribonuclease and proteinase activities in leaves that were directly sprayed with this inducer. Immunization caused no significant systemic changes in the enzyme activity of other potato leaves. However, leaves above the induction zone in the first days after the challenge were found to have a pronounced ribonuclease activity increase in comparison to that gradually rising with disease development in inoculated leaves from plants that were not induced. The proteolytic activity in the leaves of immunized plants after the challenge was decisively on a lower level than that in the leaves of noninduced plants subjected to inoculation.     

An elicitor-and pathogen-induced cdna from potato encodes a stress-responsive cyclophilin

Differential mRNA display was used to identify pathogen-responsive, stress-related genes in potato cell suspensions treated with salicylic acid and a cell wall-derived elicitor from Phytophthora infestans. Among the positive clones identified, one was found to be expressed at a significantly higher level in elicited cells than in control cells. DNA sequencing of this amplicon revealed high homology and identified it as a potato cyclophilin cDNA. The maximum amount of the cyclophilin mRNA was found 9 to 12 h after elicitation. Cyclophilin (CyP) mRNA synthesis was also up-regulated from 12 to 24 h in potato leaves locally infected with zoospores from Phytophthora infestans. However, untreated leaves responding systemically to the pathogen showed only a weak, delayed response at 24 h post infection. The observed accumulation of potato CyP mRNA in response to salicylic acid, P. infestans elicitor and P. infestans infection, suggest that CyPs play an important role in plant stress responses.     

福建省部分地区马铃薯致病疫霉群体遗传多样性分析

致病疫霉(Phytophthora infestans)引起的晚疫病是马铃薯的一种毁灭性病害。有效控制马铃薯晚疫病需要明确致病疫霉的群体遗传结构特征。采用8对SSR引物对采自福建省福州、长乐、漳州2010年分离的95株马铃薯致病疫霉进行遗传多样性分析。结果共检测出21个等位基因和26个基因型。三个地点致病疫霉菌群体间的平均遗传分化系数FST为0.22,在8个位点中有5个位点的等位基因频率分布差异显著。三个群体的观测纯合度小于期望纯合度,观测杂合度大于期望杂合度,以无性生殖为主。结果表明福建群体的遗传多样性高,群体间的存在较高的遗传分化度。     

Efficacy of essential oils and biopesticides on Phytophthora infestans suppression in laboratory and growth chamber studies

Control of late blight by alternative products is important for reduction of fungicide inputs and potato production costs. The efficacy of essential oils and biopesticides for inhibition of growth of Phytophthora infestans on media and suppression of late blight on potato plants in growth chambers was examined. Growth of pathogen isolates of diverse genotypes was evaluated on Rye B media amended with essential oils (lavender, thyme, thyme borneal, and oregano) and the biopesticide Serenade (Bacillus subtilis strain QST 713). Over 90% inhibition of pathogen growth was achieved with oregano and Serenade amendments. The protective foliar application of Serenade, an aerated compost tea (ACT), Effective Microorganism mix (EM), and oregano, resulted in disease suppression of 5-40% relative to the untreated control. ACT had no significant suppressive effects (0-15% reduction), EM resulted in mild suppression (15-30% reduction), and oregano and Serenade consistently resulted in moderate disease reduction (20-40%). No oil or biological treatment produced disease control comparable to the chemical control chlorothalonil, which resulted in disease reductions of 80-98%. Both oregano and Serenade resulted in some phytotoxicity at high doses. These results suggest that the natural products and biological amendments tested are not sufficient for effective late blight control by themselves; however, when used in combination with other established disease control practices, these approaches may contribute to improved, integrated, and more sustainable management options for late blight.     

马铃薯POTHR-1 cDNA的克隆及晚疫病菌和其他非生物因子诱导表达分析

利用RACE和重叠延伸相结合的方法,从经晚疫病菌接种诱导的马铃薯水平抗性材料叶片中克隆了一个POTHR-I基因(potato Phytophthora infestans-induced hypersensitive response related protein gene)的全长cDNA。序列分析表明,该基因编码225个氨基酸,与烟草harpin诱导蛋白基因hinI有很高的同源性(编码区核苷酸和氨基酸序列分别为83％和81％)。Southern杂交结果显示在马铃薯基因组中有2、3个拷贝。对其诱导表达模式研究表明：晚疫病病原菌接种36h后,该基因表达迅速增加;机械伤害及茉莉酸(JA)处理能够诱导表达;渗透胁迫(NaCI浸泡)能够诱导其微弱表达;但水杨酸(SA)不能诱导表达。该基因可能和病原与寄主互作时寄主产生过敏反应及细胞生理性死亡有关。     

Paraquat toxicity is reduced by polyamines in rice leaves

The protective effect of polyamines against paraquat (PQ) toxicity of rice (Oryza sativa) leaves was investigated. PQ treatment resulted in a higher putrescine (PUT) and lower spermidine (SPD) and spermine (SPM) levels in rice leaves. Pretreatment with SPD and SPM, which resulted in a 10- and 20-fold increase in endogenous level of SPD and SPM, respectively, reduced PQ toxicity (30%). Limited reduction of PQ toxicity by exogenous SPD and SPM is most likely due to the fact that they are not readily transported in rice leaf cells and localized to those areas along the cut edges of detached rice leaves [4]. PUT pretreatment did not increase endogenous SPD and SPM levels and had no effect on reducing PQ toxicity. It was found that 1,10-phenanthroline, an iron chelator, treatment reduced the toxicity of PQ (35%) and increased the levels of SPD (27%). The results indicate that reduction of PQ toxicity by SPD and SPM is due to increased activities of catalase (18%) and peroxidase (40%).     

Tuber surface microorganisms influence the susceptibility of potato tubers to late blight

Potato tubers (cvs Cara and Bintje) were grown in compost in a glasshouse and immature tubers harvested 57, 68 and 78 days after planting. Two moisture levels were imposed after the first harvest by disconnecting the water supply to one of the treatments and allowing the soil in that treatment to dry naturally. Tubers from wetter compost (59.4% moisture holding capacity) were more resistant to Phytophthora infestans than those from drier compost (14.7% moisture holding capacity) 78 days after planting. The potential causes of this difference were investigated. Aqueous extracts of wet compost did not inhibit the growth of P. infestans. The susceptibility of the internal tuber tissue, from which the periderm had been removed, was different to whole tuber susceptibility. The internal tissue of tubers from wet compost was more susceptible (cv. Cara), or as susceptible (cv. Bintje) as that of tubers from dry compost 78 days after planting. Fungi were isolated from the surface of whole tubers and there were no differences between the populations of potentially antagonistic fungal genera on tubers from wet and dry compost. As the experiment progressed, the number of bacteria per gram fresh weight on tubers grown in wet compost increased, whereas that on tubers from drier compost decreased (cv. Bintje) or remained similar (cv. Cara). There were significantly (P= 0.008) more bacteria on the surface of tubers from wet compost 78 days after planting. When P. infestans was co-cultured in Petri dishes with randomly selected tuber surface bacteria, some isolates (≤ 16.7%) inhibited the growth of the fungus. The percentage of the total bacterial population that was antagonistic to P. infestans was not significantly affected by soil moisture level (P= 0.368). The greater numbers of bacteria, of which a proportion are antagonistic to P. infestans, on the surface of tubers grown in wet compost may account for the greater resistance to tuber blight in that instance.     

Somatic hybrids between Solanum bulbocastanum and potato: a new source of resistance to late blight

 Solanum bulbocastanum, a wild, diploid (2n=2x=24) Mexican species, is highly resistant to Phytophthora infestans, the fungus that causes late blight of potato. However this 1 EBN species is virtually impossible to cross directly with potato. PEG-mediated fusion of leaf cells of S. bulbocastanum PI 245310 and the tetraploid potato line S. tuberosum PI 203900 (2n=4x=48) yielded hexaploid (2n= 6x=72) somatic hybrids that retained the high resistance of the S. bulbocastanum parent. RFLP and RAPD analyses confirmed the hybridity of the materials. Four of the somatic hybrids were crossed with potato cultivars Katahdin or Atlantic. The BC₁ progeny segregated for resistance to the US8 genotype (A-2 mating type) of P. Infestans. Resistant BC₁ lines crossed with susceptible cultivars again yielded populations that segregated for resistance to the fungus. In a 1996 field-plot in Wisconsin, to which no fungicide was applied, two of the BC₁ lines, from two different somatic hybrids, yielded 1.36 and 1.32 kg/plant under a severe late-blight epidemic. In contrast, under these same conditions the cultivar Russet Burbank yielded only 0.86 kg/plant. These results indicate that effective resistance to the late-blight fungus in a sexually incompatible Solanum species can be transferred into potato breeding lines by somatic hybridization and that this resistance can then be further transmitted into potato breeding lines by sexual crossing. Received: 27 October 1997 / Accepted: 11 November 1997     

Polyamines and carnation flower senescence: Endogenous levels and the effect of applied polyamines on senescence

The polyamines putrescine and spermine were effective in delaying the senescence of carnation buds, but were ineffective when applied to flowers of which the petals had already opened, and were 90° with respect to the stem. Higher levels of endogenous putrescine were detected in the open flowers than in the buds, and this may explain the negative effect obtained when polyamines were applied to open flowers that were picked for commercial distribution.Abbreviations AOA amino-oxyacetic acid - AVG amino-ethoxyvinyl-glycine - SAM S-adenosyl methionine - STS silver thiosulphate     

Genetic Diversity of Phytophthora infestans (Mont.) de Bary in the Eastern and Western Highlands of Uganda

Eight isolates of Phytophthora infestans were recovered from late blight infected samples collected from the districts of Mbale and Mbarara in the Eastern and Western highlands of Uganda in 2001 and analysed using mitochondrial deoxyribonucleic acid (DNA) haplotype and Amplified Fragment Length Polymorphism (AFLP) markers. Polymerase chain reaction amplification with the P2 primer followed by digestion with MspI yielded a three‐fragment pattern characteristic of isolates belonging to the US‐1 clonal lineage; the polymorphism was confirmed by DNA sequencing. AFLP analysis yielded 60 markers, analysis of which clustered the Ugandan isolates with reference to US‐1 isolates (US930258 and US940501). These results suggest that the examined Ugandan isolates belong to the US‐1 clonage lineage.     

Polyamine Uptake and Translocation in Plants

Recently, evidence has increased for both long- and short-distance transport of polyamines (PAs) in living organisms, but the mechanisms involved and physiological significance of PAs translocation are still not well understood. This review deals with various aspects of polyamine uptake and transport in higher plant tissues.