Characterization of Endophytic Fungi, Acremonium sp., from Lilium davidii and Analysis of Its Antifungal and Plant Growth-Promoting Effects.

The present study was aimed at isolating endophytic fungi from the Asian culinary and medicinal plant Lilium davidii and analyzing its antifungal and plant growth-promoting effects. In this study, the fungal endophyte Acremonium sp. Ld-03 was isolated from the bulbs of L. davidii and identified through morphological and molecular analysis. The molecular and morphological analysis confirmed the endophytic fungal strain as Acremonium sp. Ld-03. Antifungal effects of Ld-03 were observed against Fusarium oxysporum, Botrytis cinerea, Botryosphaeria dothidea, and Fusarium fujikuroi. The highest growth inhibition, i.e., 78.39 ± 4.21%, was observed for B. dothidea followed by 56.68 ± 4.38%, 43.62 ± 3.81%, and 20.12 ± 2.45% for B. cinerea, F. fujikuroi, and F. oxysporum, respectively. Analysis of the ethyl acetate fraction through UHPLC-LTQ-IT-MS/MS revealed putative secondary metabolites which included xanthurenic acid, valyl aspartic acid, gancidin W, peptides, and cyclic dipeptides such as valylarginine, cyclo-[L-(4-hydroxy-Pro)-L-leu], cyclo(Pro-Phe), and (3S,6S)-3-benzyl-6-(4-hydroxybenzyl)piperazine-2,5-dione. Other metabolites included (S)-3-(4-hydroxyphenyl)-2-((S)-pyrrolidine-2-carboxamido)propanoic acid, dibutyl phthalate (DBP), 9-octadecenamide, D-erythro-C18-Sphingosine, N-palmitoyl sphinganine, and hydroxypalmitoyl sphinganine. The strain Ld-03 showed indole acetic acid (IAA) production with or without the application of exogenous tryptophan. The IAA ranged from 53.12 ± 3.20 µg ml-1 to 167.71 ± 7.12 µg ml-1 under different tryptophan concentrations. The strain was able to produce siderophore, and its production was significantly decreased with increasing Fe(III) citrate concentrations in the medium. The endophytic fungal strain also showed production of organic acids and phosphate solubilization activity. Plant growth-promoting effects of the strain were evaluated on in vitro seedling growth of Allium tuberosum. Application of 40% culture dilution resulted in a significant increase in root and shoot length, i.e., 24.03 ± 2.71 mm and 37.27 ± 1.86 mm, respectively, compared to nontreated control plants. The fungal endophyte Ld-03 demonstrated the potential of conferring disease resistance and plant growth promotion. Therefore, we conclude that the isolated Acremonium sp. Ld-03 should be further investigated before utilization as a biocontrol agent and plant growth stimulator.

Control of Panama disease of banana by intercropping with Chinese chive (Allium tuberosum Rottler): cultivar differences.

Panama disease (Fusarium wilt disease) caused by Fusarium oxysporum f. sp. cubense race 4 (FOC) severely threatens banana (Musa spp.) production worldwide. Intercropping of banana with Allium plants has shown a potential to reduce Panama disease. In this study, six cultivars of Chinese chive (Allium tuberosum Rottler) were selected to compare their differences in antifungal activity and active compounds. Three cultivars Duokang Fujiu 11, Fujiuhuang 2, and Duokang Sijiqing with higher levels of antifungal compounds were further used for intercropping with banana in the pots and field to compare their effects on growth and disease incidence of banana.The six cultivars showed significant differences in antifungal activity against FOC mycelia growth in both leaf volatiles and aqueous leachates. The aqueous leachates displayed stronger antifungal activity than the volatiles. FJH cultivar showed the best inhibitory effect among all six cultivars. Contents of three main antifungal compounds dipropyl trisulfide (DPT), dimethyl trisulfide (DMT), and 2-methyl-2-pentenal (MP) in volatiles and aqueous leachates varied considerably among cultivars. Pot and field experiments showed that intercropping with three selected Chinese chive cultivars significantly improved banana vegetative growth, increased photosynthetic characteristics and yield but decreased disease incidence of Panama disease.Our results indicate that intercropping with Chinese chive shows potential to reduce banana Panama disease and selection of appropriate cultivars is vital for effective disease control.

Chinese chives and garlic in intercropping in strawberry high tunnels for Neopamera bilobata Say (Hemiptera: Rhyparochromidae) control.

Strawberry is affected by several pests and diseases. Neopamera bilobata is an emerging pest that has been reported by several strawberry growers, usually associated with catfacing symptoms in fruits. We evaluated intercropping garlic or Chinese chives on N. bilobata populations on strawberry crops grown in high tunnels in two experiments. In the first experiment, we evaluated N. bilobata populations on strawberry intercropping with garlic plants (three densities: 8, 16, 24 GP - garlic plant per plot) on the bags by taking 12 samples from December 2015 to April 2017. N. bilobata populations on strawberry were also assessed when Chinese chives were grown under the suspended wooden structures in which strawberry plants are grown ('undercropping') (14 samples), in two high tunnels, from November 2016 to March 2017. The number of nymphs and adults on 14 randomly selected fruits per plot were assessed. During the garlic intercropping experiment, the treatments of three densities of garlic reduced N. bilobata populations; however, the 24 GP treatment caused a greater reduction than the 8 GP treatment. Garlic densities reduced N. bilobata populations by 35, 50, and 64% for the 8, 16, and 24 GP treatments, respectively. Chinese chives cultivated under the structures reduced N. bilobata populations by 47%. The results suggest that intercropping garlic or undercropping Chinese chives are suitable tools to be tested in integrated pest management in strawberry crops.

Identification of a novel cytochrome P450 CYP3356A1 linked with insecticide detoxification in Bradysia odoriphaga.

BACKGROUND: Cytochrome P450 monooxygenases play an important role in the metabolic detoxification of insecticides in insect pests. However, little is known about the role of a specific P450 gene and its responses to insecticide exposure in Bradysia odoriphaga, a major pest in Chinese chive production. RESULTS: In this study, a novel P450 gene, CYP3356A1, was cloned from Bradysia odoriphaga. The full-length cDNA sequence of CYP3356A1 is 2153 bp and its open reading frame (ORF) encodes 508 amino acids. Quantitative real time PCR(qRT-PCR) analyses in different tissues showed that CYP3356A1 expression was the highest in the Malpighian tubule. Moreover, among the different developmental stages of the insect, the highest expression of CYP3356A1 was found in fourth-instar larvae. Expression of CYP3356A1 was upregulated by treatment with imidacloprid, thiamethoxam, and ß-cypermethrin at median lethal concentrations (LC50 ). RNA interference (RNAi)-mediated silencing of CYP3356A1 significantly increased mortality by 36.90%, 25.17%, and 36.73 when fourth-instar B. odoriphaga larvae were exposed to imidacloprid, thiamethoxam, and ß-cypermethrin, respectively, at the LC50 dose. CONCLUSION: These results demonstrate that CYP3356A1 is related to the detoxification of imidacloprid, thiamethoxam, and ß-cypermethrin in B. odoriphaga. Moreover, the study also increased our understanding of the molecular mechanisms of insecticide detoxification in this pest insect. © 2018 Society of Chemical Industry.

Highly sensitive optical interferometric technique reveals stress-dependent instantaneous nanometric growth fluctuations of Chinese chive leaf under heavy metal stress.

Plant growth apart from being a complex and highly dynamic is dependent on its immediate environment. Leaf expansion measurements using Statistical Interferometry Technique, a sensitive interferometric technique at nanometric accuracy and at sub-second levels revealed the presence of characteristic nanometric intrinsic fluctuations [Plant Biotechnology 31, 195 (2014)]. In this paper, we demonstrate that the nanometric intrinsic fluctuations are sensitive enough that they change under exposure of heavy metals, essential micronutrient zinc and non-essential element cadmium, at relatively low concentrations in the leaves of Chinese chive (Allium tuberosum). The nanometric intrinsic fluctuations of leaves were observed for 4h under three cadmium concentrations or two zinc concentrations. Results showed significant reduction of nanometric intrinsic fluctuations for all cadmium concentrations, and in contrast significant increase of nanometric intrinsic fluctuations for all zinc concentrations. There was significant reduction of nanometric intrinsic fluctuations for cadmium exposure of concentrations of 0.001mM for even an hour, and significant increment of nanometric intrinsic fluctuations under 0.75mM zinc from 1hr exposure. For comparison, antioxidative enzymes and metal uptake were also measured under 4hr exposure of cadmium or zinc. However, no significant changes could be seen in antioxidative enzymes within 4h under the smaller concentration of 0.001mM cadmium as seen for nanometric intrinsic fluctuations. The results imply that nanometric intrinsic fluctuations can be not only used as a measure for heavy metal stress but also it can be more sensitive to detect the toxic as well as positive effects of smaller amounts of heavy metal on plants at an early stage.

Shoot Injury Increases the Level of Persistence of Salmonella enterica Serovar Sofia and Listeria innocua on Cos Lettuce and of Salmonella enterica Serovar Sofia on Chive.

Minor shoot injury significantly (P < 0.05) increased the level at which Salmonella enterica serovar Sofia persisted on cos lettuce in the greenhouse. Initial mean counts of the Salmonella on the injured and uninjured cos lettuce were on the order of 6 log CFU/g. After 3 days, the mean count decreased to 4.8 log CFU/g on the injured plants compared with the significantly (P < 0.05) smaller count of 3.4 log CFU/g on the uninjured plants. By the end of the 3-week experiment, the count from the injured plants was 2.9 log CFU/g compared with a count of below the level of detection from the uninjured plants. A similar pattern of bacterial persistence was observed on injured versus uninjured plants by using Listeria innocua on cos lettuce and S. enterica serovar Sofia on chive. The findings reaffirm earlier results with Escherichia coli and increase the impetus to avoid shoot injury during the production of cos lettuce and chive, if bacteria of food safety concern are present.

[Cytogenetic effects in Allium schoenoprasum growing on the anthropogenically contaminated soil].

Cytogenetic effects in Allium schoenoprasum meristematic root tip cells grown for a year on the territory contaminated with 235U, 238U and 232Th decay series radionuclides, heavy metals and As were studied. The area is characterized with different concentrations of chemical compounds in soil affecting a toxic element migration in biocoenosis. Analysis of the chromosome aberration spectrum showed an ambiguous cell response to soil contamination. Within the weighted absorbed dose range up to 1.2 Gy the higher the dose the aberrant cell frequency increase was shown. But further increase in the dose resulted in a genotoxic effect decrease due to high toxic effects of heavy metals and radionuclides in soil. This was registered as a mitotic index decrease that can provoke a chromosome aberration frequency underestimation and result in erroneous conclusions about genotoxic effects in A. schoenoprasum used as a bioindicator.

Cadmium accumulation in Allium schoenoprasum L. grown in an aqueous medium.

The ability of Allium schoenoprasum L. (chives) to accumulate and tolerate cadmium in aqueous Hoagland medium at 50microM and 250microM was tested under continuous growth or several successive harvests of shoots. After 28 days of continuous growth, chives accumulated the metal up to 0.2% and 0.5% of its dry weight, when grown in 50microM and 250microM, respectively. In experiments that the leaves were successively harvested every 16 days, there were no obvious stress symptoms after six harvests during a period of 96 days at 50microM Cd. At 250microM, after 64 days and four harvests, inhibition of growth occurred. In each treatment, a total of 1.2g kg(-1) DW and 2.4g kg(-1) DW was accumulated in the leaves, respectively. Total SH compounds concentration in leaf was found significantly higher by 3 and 7.4 times in plants treated with Cd at 50microM and 250microM in comparison to the control, respectively, while no difference in the concentration of glutathione (GSH+GSSG) was found. Thus, it is assumed that sulphur-containing compounds, yet unknown, are involved in defensive mechanisms against heavy metals in chives. The results presented, point to chives phytoremediation potential, but also on the potential risk in accumulation of heavy metals in a commonly edible plant.