Species of Botryosphaeriaceae fungi are important plant pathogens causing cankers, blight, and fruit rot in an extremely wide range of host. In recent years, kiwifruit rot has been a serious problem in Sichuan Province, one of the important kiwifruit production areas of China. Botryosphaeria dothidea has previously been associated with kiwifruit rot but little is known regarding whether other Botryosphaeriaceae genera also constitute kiwifruit rot pathogens in China. Accordingly, diseased fruit were collected from six different areas of Sichuan Province. Based on morphological characteristics, pathogenicity testing, and comparisons of DNA sequences of the internal transcribed spacer, transcription elongation factor 1-α, and β-tubulin genes, 135 isolates of Botryosphaeriaceae were identified as B. dothidea, Lasiodiplodia theobromae, and Neofusicoccum parvum. All of these species were found to cause kiwifruit rot. To understand the infection cycle of kiwifruit rot pathogens, these three species were used to inoculate leaves and shoots of kiwifruit. The results showed that these species could cause spots on leaves and lesions on shoots, producing abundant pycnidia on leaves and shoots surfaces. Moreover, B. dothidea conidia and ascospores from overwintered pycnidia and pseudothecia in kiwifruit orchards in April and August could cause fruit rot and spots on leaves of kiwifruit. Therefore, we concluded that overwintered pycnidia and pseudothecia of B. dothidea in kiwifruit orchards are the primary inoculum for kiwifruit rot, with new pycnidia that develop during the growing season serving as a secondary inoculum. This is the first report of N. parvum and L. theobromae causing kiwifruit rot in China and is also the first report that B. dothidea is able to overwinter as pycnidia and pseudothecia in kiwifruit orchards and serve as the primary inoculum for kiwifruit rot.
Loss of functional diversity has been demonstrated to have a variety of impacts on ecosystem functioning. However, most studies have been implemented in artificially assembled communities by removing the original vegetation and seeding with desired species or functional group compositions. Such approaches could significantly disturb belowground biomass, especially roots, making it difficult to examine belowground responses to diversity manipulations. To circumvent this issue, plant diversity gradients were established by in situ removal of aboveground biomass of different plant functional groups (PFGs) in a typical steppe, and belowground processes related to roots and soil were examined. Root nutrient pools exhibited contrasting patterns, with the phosphorus (P) pool decreasing linearly upon increased PFG removal while the nitrogen (N) pool had a humpshaped response. Soil NO 3− increased while net N mineralization decreased with PFG removal. In contrast, soil P showed little response to PFG removal. Furthermore, both the identity and number of PFG removed had a significant influence on root and soil properties. The results of this study showed that loss of a combination of PFGs was important in natural grassland, and an approach with minimal influence on belowground processes is promising in studying diversity loss effects in natural ecosystems.
In this study, a simple, fast, and reliable method to fabricate a micro free-flow electrophoresis (μFFE) device on glass is presented. The two-dimensional depth channel in the chip was easily achieved by using a photocurable monomer (NOA 81) that served as the bonding material. In such a geometrical structure (two-dimensional depth channel), the effect of fluid behavior on the separation efficiency of micro free-flow zone electrophoresis (μFFZE) was simulated. The results of numerical simulation indicate that the pressure at the inlets may play an important role in the separation performance. Under the optimum separation conditions, four FITC-labeled amino acids were well separated, indicating the validity of the performance of the chip. Since the chip was fabricated by organic polymer bonding, it was easily recyclable through a simple re-fabrication process. The reproducibility of results from these recycling re-fabrication chips was investigated. The RSD of the resolution between FITC-L-glycine and FITC-L-phenylalanine was 5.3%. Furthermore, three FITC-labeled proteins were successfully separated with the resolution of 2.2 and 5.46, respectively, by using the coating of neutral liposome.
Botrytis cinerea (anamorph of Botryotinia fuckeliana) causes gray mold on numerous plants, including kiwifruit. The primary aim of this study was to investigate the phenotypic and genetic characteristics of the Botrytis cinerea population from kiwifruit in Sichuan Province, China. In all, 176 isolates were collected from kiwifruit orchards from eight geographic regions in Sichuan. All isolates were identified as B. cinerea sensu stricto based on the combined datasets, including morphological criteria, determination of the Bc-hch allele, and phylogenetic analysis of the genes RPB2, G3PDH, and HSP60. Three colony types (i.e., sclerotial, mycelial, and conidial) were observed on potato dextrose agar after 2 weeks, with sclerotial isolates, the predominant category, accounting for 40.91%. No obvious differences in microscopic characteristics were observed among the three types. Three genotypes of transposable elements were identified in the B. cinerea population: boty, flipper, and transposa types. The most prevalent genotype from different geographic populations of B. cinerea was transposa; in contrast, the flipper genotype accounted for only 3.98% of the total population, whereas the vacuma genotype was absent. According to MAT locus amplification, 87 and 89 isolates are MAT1-1 and MAT1-2 type, respectively, and the two mating types were found to be balanced overall in the population. Forty-eight representative isolates were all able to cause gray mold to some extent, and disease severities were significantly different between the cultivars Hongyang and Hort16A (P < 0.01). Disease severity was significantly greater on young leaves than on mature leaves (P < 0.01). No significant relationship was found between pathogenicity and geographical region, colony type, or transposon distribution. The results obtained in the present study suggest a relatively uniform species diversity of Botrytis but rich phenotypic and genetic differentiation within the B. cinerea population on kiwifruit in China. Utilizing resistant cultivars and rain-shelter cultivation instead of fungicides may be an effective approach to delaying pathogen variability.
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