A nonspecific lipid transfer protein,StLTP10, mediates resistance to Phytophthora infestans in potato

Nonspecific lipidtransfer proteins (nsLTPs), which are small, cysteine-rich proteins, belong to the pathogenesis-related protein family, and several of them act as positive regulators during plant disease resistance. However, the underlying molecular mechanisms of these proteins in plant immune responses are unclear. In this study, a typical nsLTP gene, StLTP10, was identified and functionally analysed in potato. StLTP10 expression was significantly induced by Phytophthora infestans, which causes late blight in potato, and defence-related phytohormones, including abscisic acid (ABA), salicylic acid, and jasmonic acid. Characterization of StLTP10-overexpressing and knockdown lines indicated that StLTP10 positively regulates plant resistance to P. infestans. This resistance was coupled with enhanced expression of reactive oxygen species scavenging- and defence-related genes. Furthermore, we identified that StLTP10 physically interacts with ABA receptor PYL4 and affects its subcellular localization. These two proteins work together to regulate stomatal closure during pathogen infection. Interestingly, we also found that wound-induced protein kinase interacts with StLTP10 and positively regulates its protein abundance. Taken together, our results provide insight into the role of StLTP10 in resistance to P. infestans and suggest candidates to enhance broad-spectrum resistance to pathogens in potato.     

Evaluating between-plant interference in field trials for assessing potato genotypes for resistance to late blight

Field trials were done with four cultivars over 3 years to assess the extent to which the amount of late blight on the foliage of a potato plant could be influenced by that on a neighbouring plant of the same or a different cultivar. Drills containing the test plants were interspersed with those of spreader plants (cv. King Edward) which were artificially inoculated with Phytophthora infestans. The intensity of blight on the test plants was recorded on several occasions.
Resistant cultivars tended to be scored as less resistant in mixtures with other cultivars than in pure stands, and susceptible cultivars tended likewise to be scored as more resistant in mixed stands. However, standard analysis of variance indicated no systematic evidence of a significant effect due to neighbour cultivars, nor of interaction between cultivars and neighbour cultivars. In contrast, Kempton's (1982) neighbour model indicated a significant and positive interference coefficient (β) in each trial, which generally decreased over time. Predicted pure stand scores for each cultivar indicated that the adjustment was greatest for the most resistant and most susceptible cultivars. There was no advantage in using two-plant rather than one-plant plots in withstanding neighbour effects.     

Management of the late blight (Phytophthora infestans) disease of potato in the southern hills of India

Late blight of potato is considered to be the most devastating problem causing severe yield losses in potato worldwide. Among the different management strategies, the use of resistant cultivars is the most viable option, but the non‐availability of enough quantity of quality seed materials of resistant cultivars forces the farmers to grow susceptible cultivars with proper fungicide scheduling. Therefore, in the present study, chemical control using fungicide has been attempted with newer molecules in the susceptible cultivar along with a resistant cultivar as a positive control. All the tested fungicides were found safe, and no phytotoxicity was observed with any chemical at the applied rate. In resistant cultivar, no late blight was appeared in both the years, whereas maximum AUDPC was observed in the untreated control (276.3) and minimum (41.7) in mancozeb‐cymoxanil + mancozeb based scheduling which was found on par with chlorothalonil‐famoxadone + cymoxanil (51.3) and chlorothalonil‐ametoctradin + dimethomorph (53.5) based scheduling. Among the treatments, resistant cultivar, Kufri Girdhari followed by chlorothalonil‐ametoctradin + dimethomorph and mancozeb‐cymoxanil + mancozeb based fungicidal scheduling were proven as the best treatments for both the crop seasons resulting in the highest yield parameters. The disease severity showed a strong negative correlation with the tuber yield of potatoes in both the years. Based on overall observations including BC ratio, it can be concluded that, wherever seed material of resistant cultivar is available farmers should grow the same or else with susceptible cultivars the fungicidal scheduling based on mancozeb‐cymoxanil + mancozeb or chlorothalonil‐ametoctradin + dimethomorph can be followed to obtain the maximum returns with effective management of late blight at the southern hills of India.     

Genome shuffling enhances biocontrol abilities of Streptomyces strains against two potato pathogens

Aims: To employ the genome shuffling technique for improving the phenotype of a biocontrol control agent of the genus Streptomyces. Methods and Results: Two rounds of genome shuffling (GS) were carried out with Streptomyces melanosporofaciens EF‐76, a geldanamycin producer. Six fusants that showed optimized in vitro antagonistic activity against Streptomyces scabies or Phytophthora infestans, two important pathogens of potato crops, were selected. All selected fusants retained the capacity to produce geldanamycin, but none overproduced this antibiotic. The higher antagonism ability appeared to result from a diversification of secreted metabolites. Seven or eight metabolites were detected in the HPLC profiles of parental strains, whereas 12–15 were detected in fusant strains. Biocontrol assays revealed that four of six fusants protected tubers more efficiently than parental strains. Conclusions: GS emerged as an elegant and rapid tool to optimize the antagonistic ability of Streptomyces strains. Optimization of the in vitro antagonistic activity against plant pathogens appears to be an effective approach to select for improved biocontrol agents. The enhanced phenotype did not depend on an overproduction of a specific antibiotic but rather on the secretion of a wider variety of secondary metabolites. Significance and Impact of the Study: Improved capacities of a biocontrol agent compensate for the lack of efficient chemical control of potato scab.     

植物提取物对大豆疫霉菌抗菌活性的初步研究

采用Paper Disk法测定了云南省200种植物的提取物对大豆疫霉(Phytophthora sojae)的抑菌活性。结果表明,光叶合欢(Albizia lucidior)、白钩藤(Uncaria sessilifructus)及黄豆树(Albizia procera)的提取物在浓度为10μg/μL时有较好的抗菌活性,而在2.5μg/μL时只有黄豆树提取物显示出抗菌活性。并从黄豆树中分离出其抗菌的主要活性成分3-O-[α-L-arabinopyranosyl-(1→2)-β-D-fucopyranosyl-(1→6)-2-acetamidO-2-deoxy-β-D-glu-copyranosyl]echinocystic acid。     

Mitochondrial DNA polymorphisms in Phytophthora infestans: New haplotypes are identified and re-defined by PCR

Polymorphisms of mitochondrial DNA (mt-DNA) are particularly useful for monitoring specific pathogen populations like Phytophthora infestans. Basically type I and II of P. infestans mt-DNA were categorized by means of polymorphism lengths caused by an ~ 2 kb insertion, which can be detected via restriction enzyme digestion. In addition genome sequencing of haplotype Ib has been used as a simple Polymerase Chain Reaction–Restriction Fragment Length Polymorphism (PCR–RFLP) method to indirectly identify type I and II alterations through EcoR I restriction enzyme DNA fragment patterns of the genomic P4 area. However, with the common method, wrong mt-DNA typing occurs due to an EcoR I recognition site mutation in the P4 genomic area. Genome sequencing of the four haplotypes (Ia, Ib, IIa, and IIb) allowed us to thoroughly examine mt-DNA polymorphisms and we indentified two hypervariable regions (HVRs) named HVRi and HVRii. The HVRi length polymorphism caused by a 2 kb insertion/deletion was utilized to identify mt-DNA types I and II, while another length polymorphism in the HVRii region is caused by a variable number of tandem repeats (n = 1, 2, or 3) of a 36 bp sized DNA stretch and was further used to determine mt-DNA sub-types, which were described as R_{n=1, 2, or 3}. Finally, the P. infestans mt-DNA haplotypes were re-defined as IR₁ or IIR₂ according to PCR derived HVRi and HVRii length polymorphisms. Twenty-three isolates were chosen to verify the feasibility of our new approach for identifying mt-DNA haplotypes and a total of five haplotypes (IR₁, IR₂, IR₃, IIR₂ and IIR₃) were identified. Additionally, we found that six isolates determined as type I by our method were mistakenly identified as type II by the PCR–RFLP technique. In conclusion, we propose a simple and rapid PCR method for identification of mt-DNA haplotypes based on sequence analyses of the mitochondrial P. infestans genome.     

Analysis of internal transcribed spacer (ITS) region of Phytophthora tropicalis causing quick wilt disease of black pepper in Vietnam

Phytophthora quick wilt is a devastating disease of black peppers in Vietnam. The internal transcribed spacer (ITS) region of the ribosomal DNA of four Phytophthora samples isolated from the diseased vines in Daknong province of the central highland part of Vietnam was Polymerase chain reaction-amplified, cloned, sequenced and characterised. Database search have showed that they are most closely related to an isolate of Phytophthora tropicalis from Taiwan. Sequence comparisons and phylogenetic analyses based on the ITS region of the four Vietnamese and other GenBank isolates of P. tropicalis and a closely related species, P. capsici, provide strong evidences that the Vietnamese isolates are all different isolates of P. tropicalis.     

Temperature linked degenerative tissue rots in Capsicum annuum (L.) varieties diseased with Phytophthora capsici

Five-week-old seedlings of Capsicum annuum variety SAMPEP 4, Californian Wonder and Ex Dandamasa drenched with 15,000 infectious units per ml of Phytophthora capsici were incubated at 5°C, 20°C, 30°C and 35°C in alternating light–dark cool cycle Gallenkamp incubators and monitored for root rot development. Each host–pathogen system was replicated five times. Successful disease development was contingent on been incubated at ambient temperature for not less than 3.5 ± 0.5 h. Depending on variety, degenerate tissue rots were aggravated ≤2–3 days after a preconditioning temperature treatment for 24 h possibly linked to cell wall constitution, composition and permeability. Lesion development on stem heightened (27.8%) when incubated at temperatures above 20°C. Ten days after treatment, plant mortality and disease severity were not affected significantly by post-inoculation temperature.     

Greenhouse and field evaluations of potassium phosphonate: the control of Phytophthora foot rot of black pepper in Vietnam

Phytophthora foot rot of black pepper caused by Phytophthora capsici is a major disease of black pepper throughout production areas in Vietnam. The disease causes collar, foot and tap root rots and eventual death of the infected vine. Potassium phosphonate was evaluated for the control of this disease in greenhouse and field trials. In greenhouse trials three-month-old vines treated with phosphonate by soil drenching (10–20 g a.i./l) and then inoculated with P. capsici mycelium (2% v/v soil) had significantly less foot rot compared to vines grown in non-treated soil. In field trials mature vines were treated with phosphonate at 50–100 g a.i/pole soil drenching or 10 g a.i./l by root infusion. After 10 days root, stem and leaf specimens were removed for bioassay by inoculation with 5 ml of P. capsici zoospores suspension (10⁶–10⁸ spores/ml). Soil drenching with phosphonate inhibited the colonisation of pathogen on excised leaf, stem and root tissues, significantly more than phosphonate root infusion. Our study provides further evidence supporting the efficacy of potassium phosphonate in the management of black pepper foot rot caused by P. capsici. The excised leaf and stem bioassay used in this study is a rapid and useful technique for testing the efficacy of systemic fungicides in controlling this disease.     

Effect of tissue wounding and time of soil inoculation on pepper root rot development

Five-week-old pepper plants with wounds created on stems and roots were transplanted to soils having inoculum of Phytophthora capsici incorporated for different lengths of time. Disease severity (39.99%) on root trimmed seedlings was not significantly different (P ≤ 0.05) from the severity (36.24%) obtained on stem lacerated seedlings. The wound treatments did not result in significantly different rates of lesion extension per day; stem lacerated seedling had the fastest, 1.99 mm/day lesion extension rate, followed by 1.90 and 1.89 mm/day extension rates obtained on root trimmed and unwounded treatments, respectively. However, time of soil inoculation had significant effect on severity; root trimmed and stem lacerated treatments had 46.3% and 39.8% severities, respectively. Tissue wounding × time of soil inoculation interaction did not have significant effect on disease severity; stem lacerated seedlings transplanted to 1-day and 3-day inoculated soils gave highest severity (49.9%), followed by seedlings inoculated at the time of transplantation. Root trimmed seedlings inoculated at the time of transplantation had highest severity (61.1%), while the lowest severity was obtained on seedlings transplanted to 5-day inoculated soil.