2022 Vol. 37, No. 10
Display Method:
2022, 37(10): 1245-1249.
doi: 10.19303/j.issn.1008-0384.2022.010.001
Abstract:
Objective Role of traf6 like gene in innate immunity regulation of Procambarus clarkia(Pc-traf6 like) was investigated. Methods RNAi technology was employed to interfere the expression of Pc-traf6 like gene. Bacterial simulation was performed to determine the changes of apoptosis-related gene expression by real-time quantitative PCR (qRT-PCR). Result The interference on the gene was effective after 48 h for the subsequent experiments. The interference-induced pro-apoptotic gene Bax, anti-apoptotic gene Bcl 2, and apoptotic protease Caspase 3 were upregulated in varying degrees with the stimulation of Staphylococcus aureus or Edwardsiella ictaluri. Conclusion After the Pc-traf6 like gene was effectively interfered, the relative expressions of Bax, Bcl 2, and
2022, 37(10): 1250-1255.
doi: 10.19303/j.issn.1008-0384.2022.010.002
Abstract:
Objective Prokaryotic expression and acetylation modification of the LysR family transcription factor AphB protein were studied. Method Primers of AphB of Vibrio alginolyticus HY9901 were designed based on the GenBank database No:WP_005380599.1. Heterologous induction was carried out by using E. coli expression system with optimized time, temperature, and IPTG concentration gradient. AphB protein was then purified to determine the degree of acetylation modification using anti-acetyl lysine specific antibody. Result The full length of aphB gene was approximately 876 bp. After the bacterial solution was induced with 0.1 mmol·L−1 of IPTG at 37 ℃ for 6 h, the expression of the fusion protein peaked. The purified fusion protein was 37.3 kD with an acetylation modification site. However, in vitro, the degree of acetylation was not regulated by deacetylase CobB. Conclusion This study preliminarily determined AphB protein to be an acetylated protein that could not be enzymatically deacetylated by deacetylase CobB in vitro. The results reinforced the existing theory of acetylation modification in prokaryotic Vibrio and provided a valuable reference for studying the post-translation regulation mechanism of virulence gene aphB of V. alginolyticus.
2022, 37(10): 1256-1265.
doi: 10.19303/j.issn.1008-0384.2022.010.003
Abstract:
Objective Effects of vitamin C addition in water to improve cold-tolerance of turbot ( Scophthalmus maximus ) on the physiology, biochemistry, and muscles of the fish in chilling preservation were studied. Method Live turbots were kept in water containing 0, 25, 50, or 100 mg·L−1 of vitamin C for 12 h prior to chilling to 3 ℃ by circulating the water through a refrigeration system and holding for 0, 12, 24, 36, 48, 60, or 72 h out of water to observe the survival rate of the fish and determine the optimal vitamin C concentration for the pretreatment. In water with the optimized vitamin C concentration, turbots were kept for 12 h followed by cooling the water to 18, 13, 8, and 3 ℃ to measure the physiological and biochemical indexes, such as white blood cells, epinephrine, cortisol, and muscle texture, of the turbots under cold stress. Result The turbots kept in water containing 50 mg·L−1 vitamin C then chilled to 3 ℃ for 72 h had the highest survival rate at 40.0%. The white blood cells, red blood cells, hemoglobin, and platelets of the vitamin C-treated fish were significantly higher than those of control (P＜0.05). During cooling from 18 ℃ to 3 ℃, the contents of serum epinephrine, cortisol, and total protein in fish were on an increasing trend, while serum glucose, liver glycogen, and muscle pH decreasing; the physiological and biochemical indexes changed significantly less on the pretreated turbots than control (P＜0.05); and the muscle firmness, elasticity, and chewiness of the treatment fish were significantly higher than those of control (P＜0.05). Conclusion With a 50 mg·L−1 vitamin C pretreatment, the negative impacts of cold stress on liver and kidney of the chilled turbots were significantly reduced and the energy metabolism and meat quality of the fish relatively unaffected.
2022, 37(10): 1275-1280.
doi: 10.19303/j.issn.1008-0384.2022.010.005
Abstract:
Objective Optimal number of canopy leaves on exit-nursery Hevea brasiliensis buddings cultivated using large polytubes was investigated. Method Rubber plant buddings raised in large polytubes with 6 to 8 leaves on the canopy were grown by hanging root control method in separate batches denoted as Y6, Y7, and Y8, respectively. In addition, plants with retained one canopy leaf, P1, two leaves, P2, 3 leaves, P3, and 4 leaves, P4 were also included. The growth vigor, exit-nursery performance, and size of stem girth after 0.5 years of large polytube cultivation of the plants were evaluated by a fuzzy membership function method. Result With significant differences between treatments, the plant height, canopy leaf count, and total leaf number of the buddings ranked Y6＜Y7＜Y8. In contrast, the average distance between two canopy leaves was Y6＞Y7＞Y8. The sprouting rates of the buddings with varied exit-nursery canopy leaf counts ranging from 93.34% to 94.25% were not significantly different among treatments. The rates of successful exiting nursery of the buddings were Y6＞Y7＞Y8, while the planting survival rates Y7＞Y6＞Y8. Upon leaving the nursery, the leaf retention proportions, which ranked P3＞P4＞P2＞P1, and the planting survival rates, which ranked P4＞P3＞P1＞P2, were both statistically significant. The significantly different stem girths of the exited plants were Y6＜Y7＜Y8 with Y6 being significantly smaller than the others after cultivating for half a years, but not significantly different between Y7 and Y8. The stem diameters of P1 and P2 were significantly smaller than those of P3 and P4 at time of exiting the nursery, whereas that of P2 significantly smaller than those of others after the 0.5-year cultivation. Conclusion Considering the rates of successful nursery exiting, leaf retention, and survival after planting as well as the stem diameter and exit-nursery canopy leaf count, the large polytube-raised H. brasiliensis buddings with 7 canopy leaves (Y7) had a higher survival rate and stronger field growth than the others. After exiting the nursery, the 3-canopy-leaf P3 buddings appeared to be most successful among all regarding the exit-nursery proportion and large after 0.5-year-cultivation stem girth. These parameters could be used as an indicator for determining numbers of kept canopy leaves to be retained on the large polytube-raised H. brasiliensis buddings.
2022, 37(10): 1281-1287.
doi: 10.19303/j.issn.1008-0384.2022.010.006
Abstract:
Objective Physiological mechanism and exogenous melatonin (MT) application to improve cold tolerance of green pepper seedlings were investigated. Method Seedlings of green pepper cultivar, Britney, were planted in matrix pots and sprayed with a MT solution of 0 (CK), 25, 50, 75, or 100 µmol·L−1. Effects on the growth, antioxidant enzyme activity, osmotic regulatory substances, and photosynthetic characteristics of the seedlings under chilling at 4 ℃ were determined. Result Comparing to CK, spraying MT increased the contents of catalase (CAT), superoxide dismutase (SOD), and proline (Pro), as well as the net photosynthetic rate (Pn), transpiration rate (Tr), stomatal conductance (Gs), maximum quantum yield (Fv/Fm), maximum photosynthetic potential (Fv/Fo), electron transport activity (Fm/Fo), leaf SPAD, while decreased malondialdehyde (MDA), relative electrical conductivity, and intercellular CO2 concentration in leaf. At 50 µmol·L−1 level, the MT spray showed the greatest effect in alleviating cold injury on the seedlings. The treatment also allowed the underground dry weight, CAT, SOD, MDA, relative conductivity, Pn, Ci, leaf SPAD, Fv/Fm, Fv/Fo, and Fm/Fo to reach significant heights and Pro, Tr, and Gs at extremely significant levels. Conclusion The exogenous MT spray mitigated injury by chilling temperature and improved cold tolerance of the green pepper seedlings through mechanisms such as a heightened antioxidant enzyme activity and photosynthetic rate as well as suppressed membrane lipid peroxidation and regulated osmotic substances. The optimum MT application on the green peeper plants was determined to be at 50 µmol·L−1.
2022, 37(10): 1288-1297.
doi: 10.19303/j.issn.1008-0384.2022.010.007
Abstract:
Objective Based on SSR markers, the genetic diversity of passion fruit varieties in Fujian province were analyzed, and SSR fingerprints of passion fruit varieties were constructed. Method The SSR markers of passion fruit were developed using transcriptome sequencing. 15 SSR markers with good repeatability and high polymorphism were screened for the identification and genetic polymorphism analysis of 17 Fujian passion fruit varieties, and the SSR molecular fingerprint was constructed. Result Using the MISA software, 24319 unigenes above 1 kilo base pairs of passion fruit were analyzed, and a total of 11385 SSRs were identified from 8742 unigenes, with an appearing frequency of 46.82 % and an average distribution distance of 7.15 kb. Mononucleotide, dinucleotide and trinucleotide accounted for 63.72 %, 20.40 % and 14.28 % of the total SSR, respectively. Furthermore, the A/T、 AG/CT and AAG/CTT were the predominant dinucleotide repeat types for mononucleotide, dinucleotide and trinucleotide repeat types, respectively. There were 28257 SSR primers designed using Primer 3.0. 15 pairs of primers with good repeatability were selected from 26 pairs of effective primers to verify the polymorphism of 17 passion fruit cultivars. A total of 235 polymorphic bands were generated by 15 pairs of SSR primers, and the average PIC was 0.95. The 17 cultivars were divided into 7 groups at the genetic distance of 0.765 by UPGMA cluster analysis. According to the 15 pairs of polymorphic primer amplification electrophoresis map, PFSSR20 and PFSSR29 primer combinations were selected to construct DNA fingerprinting, and 17 passion fruit varieties could be identified. Conclusion The SSR molecular markers constructed using passion fruit transcriptome data have high polymorphism potential and rich types. 17 Passiflora varieties (lines) were successfully identified based on the DNA fingerprints constructed in this study.
2022, 37(10): 1298-1304.
doi: 10.19303/j.issn.1008-0384.2022.010.008
Abstract:
Objective Molecular identification and genetic diversity of Camellia impressinervis were studied to improve the preservation and utilization of the endangered species. Method A highly valuable ornamental and medicinal, but extremely endangered as classified by the International Union for the Conservation of Nature, C. impressinervis was compared with the closely related C. nitidissima, C. euphlebia, and C. tunghinensis. Using 19 specimens of the 4 species, validity of applying trnL-F barcode to confidently identify them was evaluated, and their genetic diversity examined based on the inter-simple sequence repeat (ISSR) primers. Result TrnL-F could accurately differentiate C. impressinervis from those closely related camellia cultivars and clustered all in a same group. The ISSR analysis grouped the 4 C. impressinervis specimens together and showed a similarity index of 0.57 with the other 3 camellia species. Conclusion The genetic fingerprint in terms of trnL-F barcode and ISSR primers successfully identified the close relationship of C. impressinervis with, but distinguishable from, C. nitidissima, C. euphlebia, and C. tunghinensis.
2022, 37(10): 1305-1311.
doi: 10.19303/j.issn.1008-0384.2022.010.009
Abstract:
Objective a new bougainvillea cultivar was bred by seedling selection to add to the germplasm collection in China for landscaping. Method In 2009, the seeds of natural hybrid Bougainvillea glabra Mrs. Eva Mauve Variegata were sown, and seedlings planted a year later. In the field, the selected seedlings were propagated with cut tissues from 2010 to 2013 and named Minhong 1. Subsequently, they were compared to the parent in 2013–2014, and subjected to multi-point tests in 2017–2019. Genetic relationship among cultivars was examined by GBS in 2022. Result The Minhong 1 planted in Xiamen, Zhangzhou, and other localities bloomed from September to following April with an average internode distance of 1.39 cm and thorn length of 4.01 mm. The leaves were reddish orange when young and green at mature stage with an average length of 7.42 cm and width of 3.73 cm. The inflorescences borne on top and middle of the plant with purplish red bracts of an average length of 3.97 cm and width of 3.19 cm. It had 232 flowering days in a year. Minhong 1 and its parent had a close genetic relationship according to the molecular sequences. Conclusion Minhong 1 was a newly bred bougainvillea cultivar with prolonged florescence, abundant flowers, and high ornamental value for landscaping. It has officially been certified as a new bougainvillea variety issued by the National Forestry and Grassland Administration with the Variety Right of No. 20210523 in October 2021.
2022, 37(10): 1312-1317.
doi: 10.19303/j.issn.1008-0384.2022.010.010
Abstract:
Objective Biological characteristics of the pathogen causing sour rot disease on Pseudostellaria heterophylla were studied. Method Diameters of Geotrichum candidum FJAT-32652 colonies on culture medium under varied temperature, pH, light, and carbon and nitrogen sources were measured to determine the optimal growth conditions for the pathogen. Lethal temperature and time of the spores were determined, and spore germination observed under a microscope. Results The optimum growth conditions for the pathogen included 30 ℃, pH 6, maltose, lactose, and glucose for carbon source, and ammonium sulfate for nitrogen source. Light exposure had no significant effect on the mycelial growth. Lethality occurred by submerging the spores in 65 ℃ hot water for 10 min. Spore germination started 3 h after incubation, and mycelia became visible in 9 h. Schizogamy was the reproduction mode of the pathogen. Conclusion The biological characteristics of G. candidum were determined for subsequent studies on the sour rot disease of P. heterophylla.
2022, 37(10): 1318-1325.
doi: 10.19303/j.issn.1008-0384.2022.010.011
Abstract:
Objective The pathogen and its biological characteristics of camellia petal blight disease were investigated for improving the quality and protecting the ornamental value of the floral plant. Method A pathogen was isolated from of diseased Camellia japonica at the Botanical Garden in Kunming City by tissue separation method and confirmed by pathogenicity assay as well as morphological and molecular biological identifications. Biological characteristics of the isolate were studied. Result A suspected microbe was isolated from the diseased tissue and inoculated onto healthy camellia flower petals. With the appearance of yellowing and browning petals with dry or wet rots and fungal mycelial growth, the similar symptoms shown on the infected plants in the field, the isolate was studied further for a confirmed identification. On a PDA medium, the fungal colonies displayed yellow mycelia and produced black spores and lemon-like ascospores at late stage. The phylogenetic analysis of multiple genes of ITS, LSU, and EF1 clustered the isolate with Chaetomium pseudocochliodes clade. Subsequently, the optimal conditions for the growth of the isolated fungus were found to be at 25 ℃ on LB and OMA culture medium,without special requirement of light,beef paste as a nitrogen source and fructose as a carbon source have the highest utilization rates, and the lethality at 55 ℃ in 10 min. Conclusion C. pseudocochliodes was identified as the pathogen that caused floral petal blight disease on C. japonica. The pathogen grew optimally at 25 ℃ on LB and OMA culture medium ,without a specific light requirement, beef paste as a nitrogen source and fructose as a carbon source have the highest utilization rates, and it was killed by exposure to 55 ℃ for 10 min.
2022, 37(10): 1326-1334.
doi: 10.19303/j.issn.1008-0384.2022.010.012
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Objective Allelopathic effect of Ambrosia trifida on vegetables was studied by an in vitro experimentation. Method Genotoxicity and effects of the aqueous extracts of A. trifida stems and leaves on the seed germination and seedling growth of cabbage (Brassica chinensis L.) and rape (Brassica napus L.) were determined. Seed germination and seedling growth were conducted using the box filter paper method under the mass concentrations of A. trifida extracts at 0.025, 0.050, 0.075, and 0.100 g·mL−1, along with distilled water as control. Genotoxicity of the extracts was compared according to the root tip micronucleus assay on broad beans. Result The germination of the cabbage and rape seeds were inhibited by the presence of the extract which was mainly displayed by the alternation in allelopathy. The length of main cabbage and rape seedling roots was promoted by the extract at low concentrations and inhibited at high concentrations. The strongest enhancing effect was shown at 0.025 g·mL−1, while the greatest inhibition occurred at 0.100 g·mL−1. On the other hand, the plant height and fresh weight of the cabbage and rape seedlings increased with the increasing extract concentration. The photosynthetic pigment in the cabbage and rape seedling leaves increased with increasing extract as well. The micronucleus experiments on broad beans showed a variety of chromosomal aberrations, such as micronuclei, chromosomal bridges, and chromosomal lagging, on the bean root tip cells after treatment. Conclusion The aqueous extract of A. trifida tissues exhibited allelopathic effects on the growth and development of cabbage and rape seedlings with the cabbage responded more strongly than the rape. The micronucleus of broad bean root tips under the extract treatments showed a significant damaging effect on the genetic materials.
2022, 37(10): 1335-1343.
doi: 10.19303/j.issn.1008-0384.2022.010.013
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Objective Fermentation of Bacillus velezensis FJ17-4 known with a high inhibitory activity against several pathogens was optimized for potential application as a biocontrol agent. Methods Spectrometric measurement at OD600 of the fermentation broth was used as the index for evaluation. Culture medium and conditions were optimized by single factor and orthogonal experiments. Bacterial quantity, pathogenic inhibition, and indoor potted control effect of the optimized fermentation product were determined. Results The optimized FJ17-4 fermentation used a medium that consisted of 12.5 g·L−1 soybean meal, 5.0 g·L−1 corn flour, and 12.5 g·L−1 potassium dihydrogen phosphate at an initial pH of 7.0 to ferment a liquid loading (medium volume) of 50 mL with an inoculum size of 12.5% for 40 h at 30 ℃ in a 250 mL flask that rotated at a constant speed of 180 r·min−1. The OD600 of the optimized fermentation broth was 1.52 representing a cell load of 1.03×1010 cfu·mL−1, which were 25.62% and 21.95%, respectively, higher than those prior to the optimization. The inhibition rate on the mycelial growth of Fusarium oxysporum f. sp. cucumerinum and the indoor potted control on cucumber fusarium wilt of the 50× optimized fermentation broth were 42.35% and 72.14%, representing 56.38% and 13.46% increases, respectively, over the original. Conclusion The FJ17-4 fermentation was significantly improved by the optimization. An operational cost reduction was also achieved.
2022, 37(10): 1344-1353.
doi: 10.19303/j.issn.1008-0384.2022.010.014
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Objective Potential of applying native Xinjiang plants in combination with microbes as a bioagent to treat low concentration oil sludge pollution was explored. Method Three treatments of microbes, plants, and plant-microbe combination that used plant including Cynodon dactylon, Festuca elata, Lolium Perenne, or Sorghum sudanense and the microbes including Candida lipolytica (Microbe A), Bacillus subtilis PL-2 (Bacterium B), and/or B. subtilis XJ-16 (Bacterium C) were conducted along with control in a pot experiment for 120 d. Effects of the treatments on petroleum hydrocarbon residues, and microbial population, as well as biomass and chlorophyll content of the plants, were determined. Result Under the single-factor treatments, C. dactylon, S. sudanense, Bacterium B, or Bacterium C provided relatively similar effect in degrading petroleum hydrocarbons with 31.39%, 34.19%, 33.71%, and 33.39% reductions, respectively, which were significantly higher than control (P＜0.05). The plant-microbe combinations, on the other hand, showed significantly greater pollution remedying effect, such as achieved by incorporating C. dactylon with Bacterium A, B, and C that delivered the hydrocarbon removal rates of 43.02%, 40.20%, and 42.54%, respectively. Meanwhile, the culturable bacteria count increased significantly from 1.50×105 cfu·g−1 to 2.59×105 cfu·g−1 and fungi count from 4.32×104 cfu·g−1 to 5.53×104 cfu·g−1. In addition, the dry weight and total chlorophyll content of C. dactylon rose significantly in the presence of the microbes (P＜0.05). Conclusion In view of petroleum hydrocarbon residue, microbial population, plant biomass, and leaf chlorophyll content, it appeared that the combined applications of Xinjiang native plant C. dactylon with yeast C. lipolytica and bacterium B. subtilis PL-2 or XJ-16 could be feasible for cleaning the environmental pollution caused by low concentration oil sludge in the area.
2022, 37(10): 1354-1361.
doi: 10.19303/j.issn.1008-0384.2022.010.015
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Objective Effects of different treatments of soybean plants on the microbial community in tea plantation soil were studied. Method MiSeq high-throughput sequencing method was employed to analyze the differentiations on the microbial structure and diversity in rhizosphere soils at tea plantations with or without soybeans plants grown on the land and the use of cut plant parts. The treatments applied for the study were USB (removing aboveground parts of planted soybean plants), ASB (ground-mulching with cut plant parts from USB treatment), and WSB (covering soybean growing ground with cut aboveground plant parts). Result The rhizosphere soil of the treatment fields had significantly higher OTU, ACE index, and Chao1 index than that of no-treatment CK (P＜0.05). ASB or WSB significantly increased the fungal OTU, while all 3 treatments significantly raised the fungal ACE index. The Bray clustering and ternary phase plots on the 3 treatments showed a similarity on soil bacterial and fungal structures that had Sphingomonas being the dominant bacteria at 7.53% and positively correlated with Bryobacter. The relative abundance of the fungal communities, on the other hand, significantly differed from that of CK as the dominant species Arnium at 7.21% found only in the treatment soils which was negatively correlated with Penicillium. In addition, the abundance of Condenascus increased significantly. Conclusion The various treatments on the soybean plants at the tea plantations altered the soil microbial community. They increased the diversity and abundance of microbes, especially, WSB significantly enriched the microbial population in in the rhizosphere soil.
2022, 37(10): 1362-1370.
doi: 10.19303/j.issn.1008-0384.2022.010.016
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Objective Contents of major anions and cations in the Jiuquxi River at the Wuyishan National Park were analyzed to unveil the spatial and temporal distributions, action mechanism, and lithological control of the surface water. Method Thirty water samples from different river sections were collected at different hydrological periods for chemical determination. Gibbs, triangle, and endmember diagrams were applied to analyze the hydrochemical mechanism and lithological control. Principal component analysis and model calculation were applied to quantify the hydrochemical contributions. Result The river water was weakly acidic with an average total dissolved solids (TDS) of 25.30 mg·L−1, which was well below the world average of 100.00 mg·L−1. The contents of major cations in the water ranked in the order of Ca2+＞Na+＞K+＞Mg2+, while that of anions HCO3−＞Cl−＞NO3−＞SO42−. In different hydrological periods, TDS and most of the ions were dry period＞normal period＞wet period and increased from upstream to downstream of the river. Conclusion The hydrochemistry of Jiuquxi water in the national park was HCO3−-Ca2++Na+ type and, basically, of a rock weathering action mechanism that affected by precipitation, and to a certain extent, human activities. The rock weathering altered the hydrochemistry more on the upper reach than the middle or lower reach of the river. Precipitation affected the hydrochemistry most greatly in wet season. The lithological control on the area waters was largely weathering of carbonate and silicate rocks that contributed 58.82% of the controlling factors of the hydrochemistry.
