Combining physical, chemical and biological methods for synergistic control of postharvest diseases: A case study of Black Root Rot of carrot

Combining different control methods can improve control efficacy, increase the spectrum of controlled pathogens and reduce the possibility of resistance development. To be successful, however, the different methods need to be compatible: the first treatment should not have any deleterious effect on the succeeding one; preferably, it should contribute to its efficacy. In the last few years, carrot growers in Israel have begun to brush carrots before storage to remove the outer peel of the root. In the present study we show that this practice enhances the appearance of Black Root Rot during storage, a postharvest disease caused by the fungus Thielaviopsis basicola. The chemical fungicide iprodione is usually applied before storage to reduce the development of postharvest diseases. We evaluated the efficacy of combining physical, low-residue chemical and biological control agents as an alternative to the conventional chemical control approach. A technology for the precise application of steam and combined application with stabilized hydrogen peroxide (Tsunami^® 100) or a yeast commercial product (Shemer™) were tested. Used alone, both the steam and Tsunami were highly effective at reducing disease decay but were phytotoxic to the roots. Application of combined treatments of sublethal steam followed by a sublethal dosage of Tsunami or Shemer improved efficacy and disease control by 80 and 86%, respectively. These combinations showed a synergistic effect as compared to each of the treatments alone. The same pattern, effecting up to 54% disease control, was observed with the non-compatible combination of applying Tsunami first, washing it off with water and then applying Shemer. Thus disease-control agents can potentially be used for a short period, then washed off, if needed, and efficiently followed by application of a biological control agent. The biological pathway and mode of action are still under investigation but to the best of our knowledge, this is the first study to mathematically demonstrate synergistic effects of sublethal treatments applied sequentially to control postharvest disease as a potential method to reduce the use of chemicals in fruit and vegetables.     

椰子果腐病菌的鉴定及生物学特性

[目的]确定椰子果实腐烂的病原菌及其生物学特性,为椰子贮运及防病提供理论依据.[方法]用稀释分离法从病果中分离的纯培养物进行致病性测定确定病原菌,通过形态学和分子生物学方法对病原菌进行鉴定,并研究其生物学特性.[结果]椰子果腐病菌为Thielaviopsis paradoxa,可侵染菠萝蜜、甘蔗、木薯、香蕉、豆薯、龙血...     

Prevalence of Berkeleyomyces basicola infections in black rot-affected carrot determined using the MCM7 gene region

Thielaviopsis and related taxa are responsible for losses in several agricultural crops. In carrot these pathogens can cause black rot, mainly after harvesting. Molecular studies using species-specific DNA sequences are the principal tools for accurate identification of these pathogens. The objective of this work was to investigate which taxa are associated with black rot in carrot in Brazil using gene markers and Bayesian phylogenetic analysis. Eighteen isolates were obtained from carrots with symptoms of black rot. Through morphological characteristics all isolates were classified as Thielaviopsis-like. Bayesian inference using only MCM7 sequences and a concatenated data set (LSU, MCM7 and 60S rRNA) allowed identification of all isolates as belonging to Berkeleyomyces basicola. All isolates induced black rot in carrot. Although our studies include a small number of isolates, the results indicate the likely predominance of B. basicola in causing black rot in carrot in Brazil, and that MCM7 sequences are enough to distinguish the Berkeleyomyces species by Bayesian inference.     

土壤中烟草根黑腐病菌的实时定量PCR检测技术研究

 Thielaviopsis basicola is a soil-borne plant pathogen which causes root rot disease in tobacco plants. Detection and monitoring of T. basicolain soil is of great significance to control this disease. Based on the differences in internal transcribed spacer (ITS) sequences of T. basicola and other fungal pathogens, a specific primer pair Tb1/Tb2 for T. basicolawas developed. The results showed that the primer pair gave a single amplicon of 330 bp from T. basicola and revealed no undesirable cross-reaction with other seven soil-borne pathogen isolates and three tobacco rhizosphere dominant fungi isolates. With a series of 10-fold genomic DNA dilutions of T. basicola, the detection limit of 1 pg/μL in conventional PCRand100 fg/μL in real-time quantitative PCR was achieved. With DNA from the soil inoculated with different numbers of T. basicola conidia, the detection limit was 10 conidia per reaction in conventional PCR and 0.4 conidia per reaction in real-time quantitative PCR.     

Morphological and chemical characteristics of doubled haploids of flue-cured tobacco combining resistance to Thielaviopsis basicola and TSWV

Thielaviopsis basicola and Tomato spotted wilt virus (TSWV) are the most important problems in a moderate climate zone. Previously obtained doubled haploids (DH) of F₁ hybrids of the flue-cured line WGL3 resistant to Th. basicola and the dark-cured line PW-834 carrying RTSW-al gene provided the research material. Biological tests and SCAR markers linked with TSWV were applied to confirm resistance of DH. Lines combining resistance to TSWV and Th. basicola were evaluated for morphological and chemical characteristics. Most of DH were significantly shorter than parents but two lines, 31/A/2 and 31/B/3, were close to the flue-cured WGL3. Usually DH possessed fewer leaves while one of them 31/B/3, exceeded parental forms. The doubled haploids flowered later than their parents. The most negative effect was reduced area of mid-position leaves of DH. It might be explained by a recombination during microsporogenesis in F₁, however the influence of ‘Polalta’-derived RTSW-al gene cannot be excluded. Extensive line to line variation for nicotine and sugars content was not associated with the genes for TSWV and Th. basicola resistance. Biological tests and field performance of DH revealed potential to overcome the negative effect of coupling between the RSTV-al gene and genes responsible for the morphological deformations.     

香龙血树茎腐病的调查与鉴定

对从哥斯达黎加进境的香龙血树上的茎腐病进行了调查和鉴定，确定其病原菌为根串珠霉Ｔｈｉｅｌａｖｉｏｐｓｉｓｐａｒａｄｏｘａ。迄今为止，该病尚未见报道     

Pathogenic fungi involved in root rot of peas in the Netherlands and their physiological specialization

Research on root rot pathogens of peas in the Netherlands has confirmed the prevalence ofFusarium solani, F. oxysporum, Pythium spp.,Mycosphaerella pinodes andPhoma medicaginis var.pinodella. Aphanomyces euteiches andThielaviopsis basicola were identified for the first time as pea pathogens in the Netherlands. Other pathogens such asRhizoctonia solani andCylindrocarpon destructans were also found on diseased parts of roots. F. solani existed in different degrees of pathogenicity, and was sometimes highly specific to pea, dwarf bean of field bean, depending on the cropping history of the field.A. euteiches was specific to peas, whereasT. basicola showed some degree of physiological specialization.     

加拿利海枣和丝葵萎缩病的病原鉴定

加拿利海枣和丝葵萎缩病发生于厦门的行道树和苗圃 ,病树基部的茎干内变黑腐烂 ;从发病部位分离纯化获得根串珠霉。将此菌回接于苗圃和盆栽的加拿利海枣 ,产生相同症状并重新分离到该病原真菌。因此 ,通过柯赫法则证实该病的病原物为根串珠霉 ,本文对病害症状和病原形态特征进行了描述。     

芪合酶基因的遗传转化及其转基因烟草抗根黑腐病的研究

【目的】利用根癌农杆菌(Agrobacterium tumefaciens,GV3101)介导法获得转芪合酶基因烟草,改良烟草对根黑腐病的抗性。【方法】以普通烟草品种“97204”叶片为受体,采用根癌农杆菌介导法,将芪合酶基因导入烟草基因组并进行分子检测,利用灌根法对4株转芪合酶基因烟草植株接种烟草根黑腐病菌,观察烟草地上部分和根部的变化,并测定苯丙氨酸解氨酶（PAL）、多酚氧化酶（PPO）活性。【结果】通过根癌农杆菌介导、抗性选择和分子检测,获得32株转芪合酶基因烟草。从中选取长势一致的4株转基因烟草用于抗病性鉴定,结果表明,2株在灌根接种后生长正常,另外2株在接种第7天时出现轻微黄化,3株根部无发病症状,另外1株在接种16 d后根部出现发黑症状,而对照植株接种后第3天叶片就出现黄化及萎蔫症状,随着接种时间的延长,叶片黄化加剧,到第7天时已出现明显的发病症状,15 d植株几乎死亡;转基因烟草PAL及PPO活性本底均显著高于对照,PAL活性在接种后4～10 d保持在一个较高的水平,10 d后又迅速下降,PPO活性在接种2 d后迅速升高,在接种6 d达到峰值,之后开始缓慢下降,而非转基因烟草PAL、PPO活性与转基因烟草相似,但变化幅度较小。【结论】用于抗性鉴定的4株转芪合酶基因烟草中,有3株对烟草根黑腐病的抗性较好。