Development of Powdery Mildew Resistance in Cucumber Using CRISPR/Cas9-Mediated Mutagenesis of <i>CsaMLO8</i>

Shnaider, Yulia; Elad, Y.; David, Dalia Rav; Pashkovsky, Ekaterina; Leibman, Diana; Kravchik, Michael; Shtarkman-Cohen, Meital; Gal‐On, Amit; Spiegelman, Ziv

doi:10.1094/phyto-06-22-0193-fi

Cited by 9 publications

(3 citation statements)

References 36 publications

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“…The symptoms are characterized by the presence of powdery-looking patches or a diffusive layer of white, yellow, brown, or grayish mycelia and conidiophores on the plant surfaces of the plants. This disease is a serious threat to the production of various crops, including those belonging to the Cucurbitaceae family [30,31]. Currently, some biofungicides have been registered and recommended for the control of cucumber powdery mildew disease in many countries [9,32].…”

Section: Discussionmentioning

confidence: 99%

Evaluating the Efficacy of the Fermentation Formula of Bacillus velezensis Strain Tcb43 in Controlling Cucumber Powdery Mildew

2023

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Powdery mildew is a prevalent disease that affects cucumber crops, leading to yield reductions and declines in quality. Bacillus sp. strain Tcb43 culture medium was shown to be capable of inhibiting fungal spore germination in previous reports. In this study, the identification of the Tcb43 strain was updated to Bacillus velezensis by using whole-genome sequencing. The strain exhibited tolerance to a wide range of temperatures (12–40 °C), salinities (7–10%), and pH levels (ranging from 5 to 11). Additionally, Tcb43 demonstrated insensitivity to most tested fungicides. A new fermentation formula, Tcb43FBSO, was developed by adding 0.25% soybean oil to the fermented formula (Tcb43FB). This new formula exhibited a shelf life of up to 12 months with the decrease in bacterial count from 5.35 × 108 to 1.97 × 108 cfu/mL. Greenhouse assays showed that the treatment of potted cucumber plants with a 100-fold dilution (100×) of Tcb43FBSO for four weeks resulted in a significant reduction (64.64%) of cucumber powdery mildew compared to the mock group. In large-scale greenhouse trials, the treatment of cucumber plants with 200× of Tcb43FBSO for 5 weeks effectively suppressed powdery mildew disease, with a control rate that reached 76.6% compared to the mock group. These findings highlight the potential of Tcb43 as a biocontrol agent for managing cucumber powdery mildew and suggest its promising application in agriculture.

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Section: Discussionmentioning

confidence: 99%

Evaluating the Efficacy of the Fermentation Formula of Bacillus velezensis Strain Tcb43 in Controlling Cucumber Powdery Mildew

2023

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“…Powdery mildew is one of the devastating diseases of Cucurbitaceae family. Two transgene-free Csamlo8 CRISPR mutant lines (Csamlo-cr-1 and Csamlo-cr-2) showing high resistance to PM were isolated from susceptible cucumber cultivar 'Ilan' using CRISPR/Cas9-mediated mutagenesis (Shnaider et al 2022). Genome editing especially by CRISPR-Cas9 is a cost-effective, highly efficient method due to its targeted approach.…”

Section: Progress and Prospects Of Genome Editingmentioning

confidence: 99%

Cucumber: Breeding and Genomics

Dey,

Sagar,

Kujur

et al. 2023

isvs

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Cucumber (Cucumis sativus L.) is one of the most important vegetable crops grown worldwide for immature fruits. Several improved varieties with a number of economically important traits were developed through conventional breeding. The availability of draft genome sequences has facilitated the wide application of genomics tools in cucumber improvement in last two decades. C. sativus var. hardwickii widely distributed in the northern foothills of the Himalayas is the progenitor of the present-day cultivated cucumbers. A large number of commercially cultivated varieties and hybrids were developed through conventional breeding approaches. In hybrid breeding, gynoecious sex expression has been used widely for the development of more productive hybrids economically. Genetic inheritance and molecular characterization were done for a large number of traits including agro-morphological, quality, biotic and abiotic stress tolerance. Besides, candidate genes were also identified for the number of traits and later on validated through transformation and knock-out. Sex expression has been studied widely in this crop and cucumber serves as the model crop to study sex expression. A number of genes for sex expression and other economically important traits were characterized and cloned. Development of cucumber inbreds with multiple disease resistance, better quality and tolerance to abiotic stress tolerance are the major focus in future cucumber improvement programme.

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“…Powdery Mildew affects the correct development of the plant, severely affecting the quality of the fruit and causing yield losses. Nonetheless, several studies have reported genetic resistance against this disease in several species of cucurbits (He et al, 2013;Wang et al, 2016;Shnaider et al, 2022;López-Martín et al, 2022).…”

Section: Pestsmentioning

confidence: 99%

First report of Cucurbit Chlorotic Yellows Virus (CCYV) and Tomato Leaf Curl New Delhi Virus (ToLCNDV) in Algeria and lack of evidence for seed transmission of ToLCNDV in melon and pumpkin.

Kheireddine

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Emerging plant viruses in the Mediterranean Basin have arisen in the last decades becoming an important threat to crop production in this region. The spread of new viral diseases has become one of the main factors limiting agronomic and economic yields of cultivated species of Cucurbitaceae family, the second in importance after Solanaceae. A wide cluster of elements modulates and favors the emergence of these pathogens, including biological, ecological, social, and cultural factors. Global warming favors vector movement to new geographies, while global commercial networks lead to germplasm trade among countries, allowing the long-distance dissemination of viruses. Moreover, the intensive agricultural model, in which prevail genetically homogeneous cultivars, prompts pathogens outbreaks within new geographic areas at a fast rate.One of the key components to develop control strategies, and secure horticultural production, is to identify the type and distribution of viruses present in crops. However, such information in Algeria is scarce and very limited. The objective of this Doctoral Thesis was to identify and determine the prevalence and distribution of the most common viruses on cultivated cucurbit plants (zucchini, cucumber, and melon), and get information about their genetic variability and mode of dispersion.In this work, cucurbit chlorotic yellows virus (CCYV) was first detected infecting cucumber and zucchini in Algeria. CCYV (genus Crinivirus, family Closteroviridae) is part of a complex of whitefly-transmitted viruses that cause yellowing disease in cucurbits. Determination of the complete CP, and partial RdRp and Hsp70 sequences of an Algerian CCYV isolate was conducted to unveil the evolutionary relationships with the published isolates in databases. The phylogenetic analysis showed that the Algerian isolate clustered into group I together with the majority of the reported CCYV isolates. The genetic variation within this group was very low (nucleotide identity higher than 99.5%) despite their extensive and discontinuous geographical distribution, plant species (cultivated or weeds) and year of collection. This low genetic divergence could be due to the recentness of this virus that did not face major changes for the present time. In our case, the presence of CCYV was only detected in mixed infection with tomato leaf curl New Delhi virus (ToLCNDV). In future works, it will be interesting to check the likely synergistic or antagonistic interaction between these two viruses that may affect the symptoms development and spread between crops. SUMMARY viiiToLCNDV (genus Begomovirus, family Geminiviridae) was also first detected infecting cucurbit plants in Algeria. ToLCNDV is a bipartite begomovirus that causes destructive epidemics on economically valuable cucurbit and solanaceous crops. The complete DNA-A and DNA-B genomic sequences of a ToLCNDV isolate from cucumber (ToLCNDV-Biskra) were obtained.Alignments of the nucleotide sequences of DNA-A and DNA-B revealed identities of 98.7 and 97.6%, respectively, ...

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Development of Powdery Mildew Resistance in Cucumber Using CRISPR/Cas9-Mediated Mutagenesis of CsaMLO8

Cited by 9 publications

References 36 publications

Evaluating the Efficacy of the Fermentation Formula of Bacillus velezensis Strain Tcb43 in Controlling Cucumber Powdery Mildew

Evaluating the Efficacy of the Fermentation Formula of Bacillus velezensis Strain Tcb43 in Controlling Cucumber Powdery Mildew

Cucumber: Breeding and Genomics

First report of Cucurbit Chlorotic Yellows Virus (CCYV) and Tomato Leaf Curl New Delhi Virus (ToLCNDV) in Algeria and lack of evidence for seed transmission of ToLCNDV in melon and pumpkin.

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