Genomic characterization of a novel torradovirus infecting Arctium lappa L. in China.

Burdock (Arctium lappa L.) is not only a popular vegetable crop but also an important medicinal plant. In burdock plants with symptoms of leaf mosaic, a novel torradovirus tentatively named "burdock mosaic virus" (BdMV) was identified by high-throughput sequencing. The complete genomic sequence of BdMV was further determined using RT-PCR and the rapid amplification of cDNA ends (RACE) method. The genome is composed of two positive-sense single-stranded RNAs. RNA1 (6991 nt) encodes a polyprotein of 2186 aa, and RNA2 (4700 nt) encodes a protein of 201 aa and a polyprotein of 1212 aa that is predicted to be processed into one movement protein (MP) and three coat proteins (CPs). The Pro-Pol region of RNA1 and the CP region of RNA2 shared the highest amino acid sequence identity of 74.0% and 70.6%, respectively, with the corresponding sequences of lettuce necrotic leaf curl virus (LNLCV) isolate JG3. Phylogenetic analysis based on the amino acid sequences of the Pro-Pol and CP regions showed that BdMV clustered with other non-tomato-infecting torradoviruses. Taken together, these results suggest that BdMV is a new member of the genus Torradovirus.

First Report of Powdery Mildew Caused by Podosphaera xanthii on Hulless Cucurbita pepo of Inner Mongolia and Xinjiang Autonomous Region in China.

Hulless Cucurbita pepo is an annual herb in the Cucurbitaceae family and is one of the main economic vegetable crops in China, and is a raw material for cosmetics and health care products. It is also called hulless pumpkin because its seeds have no seed coat, which is a rare variation of the Cucurbita. In July 2010, powdery mildew was observed on hulless Cucurbita pepo 'Tianran' in fields of Wuwei District, China(Liang et al.2010). Disease incidence when first observed was 65.67%, but increased to 100% in July 2019. Early disease symptoms appeared as circular or irregular white powdery areas on both leaf surfaces. At later infection stages, entire leaves，petioles, and stems were covered with white fungal mycelia that resulted in leaf yellowing and senescence，but not defoliation . Fungal hyphae were septate, branched and flexuous to straigh. Conidiophores were unbranched, straight and grew vertically to the mycelium. Conidiophore foot cells of the were cylindrical with slight constriction at basal septa and followed by one to four short cells that eventually became conidia. Conidia were barrel-shaped with ends darker than the middle and measured 20 to 32 × 12 to 19µm.T Cleistothecium formed at 20 ℃, 70% relative humidity and light intensity of 4,400 lx. They were scattered, spherical, dark brown, with parietal cells irregularly rectangular or polygonal, and 70 to 75×90 to 95µm in diameter. Cleistothecia had four to eight appendages that were colorless to partially brown, each with three to five septa and lengths 0.5 to 3.0 times the diameter of cleistothecia. The powdery mildew fungus was tentatively identified as Podosphaera xanthii and showed characteristics similar to those reported by others (Cui et al. 2018; Choi et al. 2020). rDNA was extracted from pools of fungus conidia and the ITS region amplified using primers ITS 1:5'-TCCGTAGGT GAACCTGCGG-3'/ITS 4:5'-TCCTCCGCTTATTGATATGC-3' and then sequenced. BLASTn analysis of the 540-bp (MT250855) amplicon revealed 100% sequence identity with respective rDNA sequences of Podosphaera xanthii isolates from Momordica charantia(AB774158.1). Based on the morphological characteristics and ITS sequences, the fungal species was identified as P. xanthii .Pathogenicity of the powdery mildew fungus was tested by dusting conidia from infected hulless pumpkin leaves onto three asymptomatic plants. Three noninoculated plants was used as a controls. The Infection process of P.xanthii on pumpkins observed that the conidia began to germinate at 12h after inoculation.Twenty four to 72 h post inoculation, powdery mildew mycelia appeared on inoculated leaves. After 73 to 96 h, chains of conidia formed and these germinated to form secondary infection sites. In later stages of the disease cycle, dark brown cleistothecia formed on the yellowing plant foliage. Fungus morphology from inoculated leaves was identical to that observed on original naturally infected plants. Uninoculated plants remained healthy. Powdery mildew caused by P. xanthii is a major foliage disease that affects members of the Cucurbitaceae family worldwide. The fungus has previously been reported from China on Cucurbita moschata (DQ490752), Cucurbita maxima (DQ490759), Cucurbita pepo(DQ490750), Cucumis sativus(DQ490755)(Park et al.2010; Liang et al.2007) and Cucurbita maxima, Abelmoschus esculentus (okra), Sechium edule (mirliton, vegetable pear), and Lagenaria siceraria (bottle gourd) (Choi et al. 2020; Fan et al. 2019; Xu et al. 2020; Cui et al. 2018).To our knowledge, this is the first report on the occurrence of P. xanthii on hulless Cucurbita pepo in China.

Induction of resistance of Podosphaera xanthii (hull-less pumpkin powdery mildew) to triazole fungicides and its resistance mechanism.

The aim of this study was to uncover the molecular mechanism through which fungicide resistance develops in Podosphaera xanthii, a fungi that causes powdery mildew in hull-less pumpkin. Treatments of inoculated P. xanthii were carried out on leaves of hull-less pumpkin and subsequently treated with kinds of triazole fungicide for seven generations. Resistant strains of P. xanthii thus obtained were evaluated for their resistance levels. The resistance levels of the fungi to four fungicides of were high except that of the propiconazole-resistant strain, which showed moderate resistance. The F7 generations of five resistant strains thus obtained were cultured continuously for five generations without fungicide induction, and their resistance level were found to be relatively stable. The DNA of the sensitive strain and the five kinds of resistant strains were extracted by the sodium dodecyl sulfate (SDS) method and its internal transcribed spacer (ITS) region was amplified by using ITS1/ITS4 primer and specific primer F/R and they were sequenced respectively. The DNA sequence comparison of resistant and sensitive strains showed that the base pairs of tebuconazole-resistant strains and flusilazole-resistant strains were mutated, with mutation rates of 4.8% and 1.6%, respectively. The base pairs of the other three resistant strains did not change.

[Efficiency of Trichoderma longibrachiatum T6 in the control of Meloidogyne incognita and its rhizosphere colonization in cucumber].

Efficiency of different concentrations of Trichoderma longibrachiatum T6 against Meloidogyne incognita and its rhizosphere colonization in cucumber were determined in greenhouse experiments. The results of rhizosphere colonization experiments showed that the number of colonies in cucumber soil and root increased significantly ten weeks after inoculation with the second stage juveniles of M. incognita and different concentrations of T. longibrachiatum T6, and there was significant difference in different concentrations of T. longibrachiatum T6, e.g., the maximum numbers of colonies in soil and root were 7.8 x 107 and 6.3 x 105 CFU · mL⁻¹ respectively after treated with the spore suspension of 1.5 x 107 CFU · mL⁻¹. Greenhouse experiments results showed that different concentrations of T. longibrachiatum T6 had significant control effect on different life stages of M. incognita, and the control effect increased with the concentration of T. longibrachiatum T6. T. longibrachiatum T6 significantly increased plant height, root length, above-ground and root fresh mass o cucumber inoculated by M. incognita. T. longibrachiatum T6 could colonize in cucumber rhizosphere, had control effect on M. incognita, and significantly improved the growth of cucumber.