2021 Vol. 36, No. 9
Display Method:
2021, 36(9): 989-998.
doi: 10.19303/j.issn.1008-0384.2021.09.001
Abstract:
Objective The reverse transcriptase (RT) gene sequences of Ty3-gypsy-like retrotransposons were isolated from the sugarcane genome, and their characteristics, differences, and phylogenetic relationships analyzed to study the transcriptional activities and regulatory functions associated with the retrotransposons. Method Degenerate primers were designed according to the conserved region of the RT gene sequences of Ty3-gypsy-like retrotransposons. The genomic DNA of sugarcane variety, ROC22, was amplified by PCR. The recovered, cloned, and sequenced target band was analyzed by bioinformatics. Result Thirty-six RT gene sequences were successfully isolated. Only one sequence of the target bands was 430 bp in length, while the remainders were 432 bp. The proportions of AT in the sequence ranged from 56.71% to 64.81%, and the ratio of AT to GC from 1.31 to 1.84. A high heterogeneity existed among the nucleotide sequences, as the similarity was shown to be 46.2% to 99.3%. The cluster analysis divided the 36 RT gene sequences into 5 families. Family I and Family IV contained most of the sequences. After translated into amino acids, 6 sequences displayed nonsense mutations. The similarity between amino acid sequences ranged from 10.1% to 100% exhibiting a high degree of heterogeneity. The conserved motifs of 34 of the sequences were identical, indicating a highly conservative property. The overall configuration of the tertiary structure of the representative proteins in each family was basically similar, however, there were significant differences in the number of hydrogen bonds and turns, suggesting a certain degree of heterogeneity and polymorphism. The phylogenetic tree showed 7 distinct groups among the RT gene sequences. Of which, 18 in Group I were highly homology with BAB40828.1 of Arabidopsis thaliana, and SoRT3-26 in Group VII was genetically closest to the BAB40834.1 of soybean, BAB40833.1 of spinach, and BAB40824.1 of japonica rice. It indicated that a horizontal transfer of Ty3-gypsy-like retrotransposons might have occurred in evolution between sugarcane and those plant species. By comparing the sugarcane EST database, 10 Ty3-gypsy-like retrotransposons were found with transcriptional activity in ROC22. Conclusion Thirty-six RT gene sequences of Ty3-gypsy-like retrotransposons in sugarcane were obtained with 10 of them showing a transcriptional activity. The results provided the basic information for the isolation of full-length Ty3-gypsy-like retrotransposons, further understanding of the transcriptional, transposon activity and functions of the retrotransposons, and the development of sugarcane molecular markers based on LTR retrotransposons.
2021, 36(9): 999-1006.
doi: 10.19303/j.issn.1008-0384.2021.09.002
Abstract:
Objective Functions of ZmCOL3 involving the flowering regulation and other mechanisms in maize were investigated by analyzing the structure and functions of the protein encoded by the gene. Method Based on bioinformatics, the physicochemical properties, conserved domain, secondary and tertiary structures, signal peptide, transmembrane domain, subcellular localization, and cis acting elements of ZmCOL3 protein were predicted. Expression of the gene in various organs of a maize plant were analyzed. Result The ZmCOL3-encoded protein contained 335 amino acids with a molecular weight of 35.39 KD and a theoretical isoelectric point of 5.04. It was an unstable acidic, hydrophilic protein located primarily in the nucleus and consisted of a CCT domain and a B-box domain but no signal peptide or transmembrane domain. Its secondary structure was mostly irregular coils with some α helixes and a small number of extended chains and β turns. The homology modeling similarity was 64.15%. The promoter of ZmCOL3 gene not only contained the basic cis acting elements, such as TATA and CAAT boxes, but also light response elements, such as SP1, as well as hormone response elements, such as abscisic acid and methyl jasmonate. In various tissues, the leaves, followed by the male ears, had the highest and significantly higher expression of ZmCOL3 than the primary roots, internodes, or tassels, while and the seeds the lowest. Conclusion The unstable acidic, hydrophilic ZmCOL3 protein contained CCT and B-box domains consistent with the structural characteristics of COL subfamily in the CCT gene family of which it belonged. It was postulated to participate in the regulation of biological clock that affects maize florescence. The promoter of ZmCOL3 gene contained the elements responding to light and various hormones that suggested a possible involvement on the multiple response network regulating the photoperiod and abiotic stress of a maize plant.
2021, 36(9): 1007-1016.
doi: 10.19303/j.issn.1008-0384.2021.09.003
Abstract:
Objective Myelocytomatosis proteins (MYCs), the key transcription factor in the jasmonic acid signal transduction pathway, in Camellia sinensis were identified and analyzed to help understand the underlying molecular mechanism associated with resistance of tea plants to abiotic stress. Method A genome-wide analysis on the MYC family in tea plant (CsMYC) was conducted using bioinformatics methods. Result Nine CsMYC members were identified in the tea plant genome which distributed unevenly in 5 chromosomes. The phylogenetic analysis suggested that the CsMYCs might originate from terrestrial plants and have undergone lineage specific differentiation. Structurally, the family had only 0 to 3 introns indicating a deletion might have occurred in evolution. Eight of the 9 CsMYCs had homologous genes of dicotyledonous plants like grape, but not of monocotyledonous plants like rice. The transcriptome data showed that, except for CsMYC 2 and CsMYC 9, all members were highly expressed in the buds and leaves. On the fluorescence quantitative tests, all members responded to MeJA stress and some also to drought, low temperature, and/or GA treatments. It indicated a likely close association of tea plants to abiotic stress played by CsMYCs. The KEGG enrichment of the CsMYCs suggested its wide involvement in the secondary metabolism pathways in tea plants. Conclusion Nine CsMYCs in C. sinensis were identified in this study. The structures and potential molecular functions of the members were analyzed and predicted. Combining the results obtained by the real-time fluorescence quantification and functional enrichment analyses, it appeared that the CsMYC family might be closely associated with the response of tea plants to abiotic stress.
2021, 36(9): 1017-1024.
doi: 10.19303/j.issn.1008-0384.2021.09.004
Abstract:
Objective Applicability of a seed-coating Bacillus subtilis suspension for promoting chili pepper seedling growth was investigated. Method The chili pepper, Yangjiao No. 2, was used in the study by applying a B. subtilis agent on the seeds at the pharmacopoeia ratios of suspension to seeds at 1∶2, 1∶4, 1∶6, 1∶8, 1∶10, or 1∶12 (ml∶g). Subsequently, the seed germination as well as the seedling growth and physiological characteristics were monitored to determine the effects brought about by the coating treatments. Result Compared with control, the coating increased the seed germination rate and the plant height, stem diameter, leaf area, plant dry weight, and growth index of the seedlings. The effects peaked at the suspension to seeds ratio at 1∶8. The coating treatments enhanced the net photosynthetic rate (Pn), transpiration rate (Tr), and stomatal conductance (Gs) but reduced the intercellular CO2 concentration (Ci) on the leaves resulting in an uplifted photosynthetic capacity of the plants. Moreover, the treatments decreased the MDA content and O2−production rate, while increased the SOD, POD and CAT activities, of the leaves that were conducive to sustaining high contents of chlorophyll and soluble protein for the plants. The seedling growth index was found positively correlated with the net photosynthetic rate, transpiration rate, and stomatal conductance but inversely correlated with the intercellular CO2 concentration. Conclusion The seed-coating B. subtilis suspension alleviated the peroxidation of membrane lipids and increased antioxidant enzyme activities with a sustained high photosynthetic capacity in the leaves benefiting a vigorous subsequent growth of the seedlings. The pharmacopoeia ratio at 1ml of suspension to 8g of seeds provided the most desirable results.
2021, 36(9): 1025-1032.
doi: 10.19303/j.issn.1008-0384.2021.09.005
Abstract:
Objective Correlations among the color, pH, and epidermal cell morphology of petals of ornamental crabapple flowers were preliminarily analyzed. Method Colors of the petals of 14 varieties of white, reddish purple, pink, and dark-purple crabapples at blooming, big-bud, full-bloom, and end florescence stages were measured by RHSCC and a chromatic meter. Morphology of petal epidermal cells was examined under a scanning electron microscope. Correlations among these properties were statistically analyzed. Result The white and pink lines of crabapples had a petal pH ranging from 5.24 to 6.37, while the dark-purple and reddish purple lines, from 4.98 to 5.53. In general, the pH declined as the color turned from light or white to dark or reddish purple and the colorimeter red-green a* and saturation C* measurements increased. A similar trend was also observed during the flowering process. Aside from the dark-purple variety, the other 3 crabapple series had petal epidermal cells of varied types and, in florescence, the cells became more noticeably flattened as the petal color faded. Conclusion There appeared to be correlations among the pH of petals and the formation and fading of flower color on the 14 ornamental crabapple varieties. The petal epidermal cell morphology did not significantly correlate to the flower color phenotype but possibly to the color fading in florescence of the plants.
2021, 36(9): 1033-1041.
doi: 10.19303/j.issn.1008-0384.2021.09.006
Abstract:
Objective Pathogens causing broiler death by unknown etiology at a chicken farm in Nanping, Fujian were investigated. Methods To establish standard references for this investigation, pathogens on chicken, such as, infectious bronchitis virus (IBV), fowl adenovirus(FadV), chicken parvovirus (ChPV), newcastle disease virus(NDV), chicken infectious anemia virus (CIAV), chicken infectious bursal virus(IBDV), laryngotracheitis virus(LTV), and avian nephritis virus (ANV), were detected by PCR and the amplified fragments cloned, sequenced, and analyzed using bioinformatics software. Results The clinic specimens were found positive on ChPV, IBV, and CIAV but negative on the other viral pathogens. The homology of the isolates was 92.17%–100% with ChPV, 76.9%–84.3% with IBV, and 91.19%–92.14% with CIAV. The isolated IBV and CIAV strains were in their own unique branch, while ChPV closely related to a Guangxi strain as shown by the phylogenetic analysis. Conclusion A case of simultaneous co-infection by ChPV, IBV, and CIAV at a chicken farm was confirmed. The S1 gene of the isolated IBV and the VP1 in the CIAV strain appeared to have mutated prior to the infection incidence.
2021, 36(9): 1042-1047.
doi: 10.19303/j.issn.1008-0384.2021.09.007
Abstract:
Objective A rapid detection and quantification method for bovine viral diarrhea virus (BVDV) was established. Method For the methodology development, specific primers and probes were designed based on the target regions of the BVDV 5′UTR gene published by GenBank. The RNA of in vitro transcription viruses was used as the absolute quantitative standard. Reaction conditions of the fluorescent quantitative RT-PCR were optimized. Result The newly developed assay had a minimum detection limit of 5.0267copies/μL and an intragroup variation coefficient of less than 1% with high repeatability and specificity. Other than BVDV, it amplified no viral nucleic acids of viruses such as swine fever and foot-and-mouth diseases. Conclusion The established fluorescence quantitative RT-PCR method was highly sensitive, specific, and repeatable in detecting and quantifying BVDV. It appeared appropriate for early diagnosis of bovine viral diarrhea.
2021, 36(9): 1048-1053.
doi: 10.19303/j.issn.1008-0384.2021.09.008
Abstract:
Objective Functions of crustacean gene of Procambarus clarkia were studied. Method Pc-crustin6 was targeted for this study with specifically designed primers for PCR. Amino acids alignment and phylogenetic tree analysis were performed with bioinformatics software to decipher the molecular properties of Pc-crustin 6. Expressions and distributions of Pc-crustin 6 in different tissues were analyzed by qRT-PCR. The expression after an Edwardsiella ictaluri challenge was used to determine the disease resistance associated with the gene. Result The full length Pc-crustin 6 Conclusion The Pc-crustin 6 expression in organs of P. clarkia was significantly altered after being stimulated by the induction of E. ictaluri. It indicated that the gene was involved in the pathogenic resistance of P. clarkia.
2021, 36(9): 1054-1062.
doi: 10.19303/j.issn.1008-0384.2021.09.009
Abstract:
Objective Acute toxicities of commonly applied drugs on juvenile Pseudogobio vaillanti were determined for safe disease control in aquaculture ponds. Method Using a static bioassay method, toxicities of 8 antimicrobial agents, i.e., ncluded potassium permanganate, copper sulfate, dipterex, chlorine dioxide, 20% concentrated glutaraldehyde solution, 45% benzalkonium bromide solution, 10% povidone iodine solution, and salt, on juvenile P. vaillanti were studied. The tank water temperature was maintained within 21.3 to 23.5 ℃. Toxicity accumulation coefficient (MAC) was also employed as index for the evaluation. Result The agents displayed toxicities on the fish ranking from the strongest to the weakest as: copper sulfate＞chlorine dioxide＞potassium permanganate＞45% benzalkonium bromide solution＞dipterex＞20% concentrated glutaraldehyde solution＞10% povidone iodine solution＞salt, with the safety mass concentrations (SC) of 0.019, 0.061, 0.346, 0.701, 1.377, 2.674, 6.906, and 910.6 mg·L−1, respectively. However, no consistent pattern on MAC of the agents could be obtained as it varied with the duration of treatment. Conclusion It appeared that most of the commonly used drugs could be safely applied on juvenile P. vaillanti for the aquaculture, but special cautions would be required on the use of salt, potassium permanganate, and copper sulfate.
2021, 36(9): 1063-1068.
doi: 10.19303/j.issn.1008-0384.2021.09.010
Abstract:
Objective Resistance of local sorghum varieties in Guizhou to anthracnose was determined by artificially infecting the plants in the field. Method Using both crushed straws and diseased leaves to induce the anthracnose infection on 283 cultivars of sorghum in Guizhou, a field experimentation was conducted in two separate years to examine and classify the disease resistance of the crop plants. Result Among the cultivars, B39-2 and F41 showed a high, Grade 2 resistance to anthracnose. Seven cultivars were moderately resistant to the disease in varying degrees depending upon the environmental conditions. E36 and F4 exhibited resistance that varied greatly. A few were of heterozygous genotypes displaying multiple grades of resistance. And the remaining 266 specimens were susceptible to the disease. Conclusion Two outstanding local sorghum cultivars in Guizhou were highly and consistently resistant to anthracnose that were to be further studied for breeding programs. This study showed the combined use of crushed straws and diseased leaves to artificially infect sorghum plants to be more effective than applying crushed straws alone in determining the degree of the disease resistance.
2021, 36(9): 1069-1074.
doi: 10.19303/j.issn.1008-0384.2021.09.011
Abstract:
Objective The pathogen that caused the bacterial pith necrosis on greenhouse tomatoes grown in Fujian and Zhejiang provinces during 2017-2019 was identified. Method Biolog analysis, 16S rDNA gene sequencing, and MALDI-TOF-MS were performed to identify the pathogen. The Koch’s rule was employed to determine the pathogenicity of the isolate. Result Two bacteria strains were isolated from the diseased tomatoes. The symptoms of discolored, dark brown vascular tissue and pith of main stem observed on tomatoes in the laboratory were identical to those found in the field. The bacteria were re-isolated from the infected plants and identified as Enterobacter asburiae. Conclusion The pathogen that caused bacterial pith necrosis in tomatoes in Fujian and Zhejiang was identified and reported as E. asburiae for the first time.
2021, 36(9): 1075-1080.
doi: 10.19303/j.issn.1008-0384.2021.09.012
Abstract:
Objective Biological characteristics of the pathogen that caused the plum fruit spot disease, Fusarium fujikuroi, were studied. Various agents for effective control of the disease were tested in the laboratory. Method Effects of culture medium, light exposure, temperature, pH, and carbon/nitrogen sources on the growth of F. fujikuroi were determined. Toxicities of 7 fungicides to the pathogen were compared with the mycelium growth rate method in the laboratory. Result F. fujikuroi grew well on Czapek or PDA medium at pH of 7-8 and 25 ℃. The preferred carbon sources were starch and glucosaccharide, and those for nitrogen, peptone and sodium nitrate for the culture. Different light exposures did not significant affect the fungal growth. The 7 fungicides displayed significant but varying inhibitory effects on the mycelial development. Among them, 45% imidazolide WDG, 20% pyrathiazolide SC, and 30% prothiazolium OD showed high toxicities with EC50 of 0.891 4, 0.9436, and 1.022 0 μg·mL−1, respectively. Conclusion The effects of temperature, pH, light exposure, and nutrients on the growth of F. fujikuroi were clearly shown by this study. The relative efficacies of 7 commonly available fungicides for controlling the plum fruit spot disease were compared.
2021, 36(9): 1081-1086.
doi: 10.19303/j.issn.1008-0384.2021.09.013
Abstract:
Objective Inhibitory effect of a strain of Brevibacillus brevis against Lasiodiplodia theobromae (Pat.) Griffon & Maubl on longan fruits was evaluated. Method In vitro antagonistic activity of B. brevis FJAT-0809-GLX was observed by the inhibition zone and rate on the mycelial growth and morphology of L. theobromae. Efficacy of the strain for biocontrol on the pathogen was determined by in vivo experiments, as well as the activities of defense enzymes in longan peel. Result Supernatant of the fermentation broth showed a pathogenic inhibition zone diameter of 15.77 mm and an inhibition rate that increased with increased fermentation time or treatment dosage. The inhibition rates reached 74.17% when 50 mL of the supernatant was applied. The supernatant inoculation on longan fruits significantly reduced the disease spreading rendering a healthy fruit rate of 68.33%. Meanwhile, the activities of superoxide dismutase and hydrogen peroxidase in the treated fruits became significantly higher than those of control. Conclusion B. brevis FJAT-0809-GLX significantly inhibited the growth of L. theobromae as shown by in vitro plating as well as in the field challenge test. It demonstrated a potential venue for bio-controlling the disease on longan.
2021, 36(9): 1087-1091.
doi: 10.19303/j.issn.1008-0384.2021.09.014
Abstract:
Objective Capability of Oryza sativa L. in accumulating and transforming methylmercury (MeHg) at different growth stages was studied. Method Rice plants without MeHg exposure (as control) and 4 treatments groups with a MeHg exposure at seedling, tillering, jointing, or heading stages were cultivated in a hydroponic setting. Total Hg and MeHg concentrations in tissues of the plants were determined, and percentages of MeHg to total Hg in straws and grains calculated. Result Compared with control, the MeHg exposure increased the MeHg content in rice tissues at all growth stages, but the accumulation significantly varied according to both the growth stage of the exposure and the tissue tested. The total Hg and MeHg contents in all parts of rice at the seedlings or tillering stage were lower than those at the jointing or heading stage. It indicated greater accumulation tended to occur approaching the end of plant growth. And at the seedling and tillering stage, more than 60% of the Hg in the grains was in the form of MeHg, as opposed to more than 80% at the jointing and heading stages. Thus, MeHg was more highly accumulated, or less likely be transformed, in the grains at the heading stage than in the early stage of rice growth. Conclusion As a rice plant maturing, the MeHg accumulation increased and the transformation lessened. Hence, exposure of rice to MeHg would present less severe a safety risk at early than late stage of growth.
2021, 36(9): 1092-1099.
doi: 10.19303/j.issn.1008-0384.2021.09.015
Abstract:
Objective Effects of the application of pig manure combined with chemical fertilizer on the soil properties and aggregate organic carbon distribution on eroded forest lands were studied for soil improvement and waste utilization. Method In a short-term, pot experiment, 5 fertilizations including control and applications of chemical fertilizer at a rate of 500kg·hm−2 without or with an addition of pig manure at low (7.5 t·hm−2), medium (15 t·hm−2), or high (30 t·hm−2) level were incorporated into the soil. Distribution, average weight diameter, geometric average diameter, destruction rate, and organic carbon content of the aggregates in the resulting soil were determined. Results (1) The addition of pig manure promoted the water-stable soil microaggregates transforming into macroaggregates. When pig manure was applied at a rate of 15 t·hm−2, the number of aggregates sized smaller than 0.25 mm decreased by 20.56% and that of aggregates larger than 2 mm increased by 24.92% over control. (2) Pig manure significantly increased the average weight diameter and geometric average diameter of the aggregates (P＜0.05) with a upward trend as the addition rate increased. (3) Pig manure effectively reduced 19.10% on the aggregate destruction rate (PAD) with the 15 t·hm−2 application rate that delivered the most significant effect (P＜0.05). And (4) pig manure significantly raised the organic carbon content in the soil and aggregates (P＜0.05). Conclusion When combined with chemical fertilizer, pig manure could effectively increase the organic carbon content and improved the aggregate stability of the soil on an eroded forest land. At a rate of 15 t·hm−2, the pig manure addition significantly increased the average weight diameter and average diameter of the soil aggregates with the highest stability among all treatments, while effectively reduced PAD. At a higher rate of 30 t·hm−2, the pig manure application significantly increased the organic carbon contents in both non-water-stable and water-stable macroaggregates.
2021, 36(9): 1100-1109.
doi: 10.19303/j.issn.1008-0384.2021.09.016
Abstract:
Purpose Efficiency of biochar made of spent corn straws in adsorbing sulfonamide residues in water solutions of various conditions was studied for aquacultural application. Methods The biochar was made from stover after corn harvest. The physicochemical properties of the biochar were determined by FTIR, SEM, XRD, and elemental analyzer. In water solutions of varied conditions, including biochar addition, sulfonamide concentration, pH, and treatment time and temperature, the kinetics, thermodynamics, and efficiencies of the biochar adsorptions on sulfadiazine (SDZ) and sulfachlorpyridazine (SCP) were studied. Results The sulfonamides removal rate significantly affected by the amount of biochar used, initial antibiotic concentration in solution, adsorption treatment time, and solution pH. At 25oC and pH 5, 94.57% of 10 g·L−1 initial concentration of SDZ in solution was removed by 8 g·L−1 addition of the biochar in 4h, while 98.12% of SCP at 10 g·L−1 initial concentration was adsorbed by 5 g·L−1 biochar. The sulfonamide adsorption kinetics of the biochar was a pseudo-second-order function, and the isotherms fitted a Freundlich equation. The adsorption process is mainly controlled by rapid reaction. Since a greater than 40% removal rate on SDZ or SCP could still be achieved by a same biochar application after 3 cycles, repeated usage of the adsorption material seemed durable. Conclusion The corn straws biochar exhibited a capacity in effectively adsorbing residues of sulfonamide antibiotics in water solutions. It suggested a potential utilization of the low cost, highly efficient, and easily available material in aquaculture ponds for mitigating the antibiotic contamination.
2021, 36(9): 1110-1118.
doi: 10.19303/j.issn.1008-0384.2021.09.017
Abstract:
Objective Changes incurred to the fungal community during the natural fermentation of tomatoes were studied to pave the way for product and process development. Methods Composition and diversity of fungi presented at various stages of the naturally occurring fermentation on tomatoes were analyzed by high-throughput sequencing technique. Physical and chemical parameters for indexing the process were determined. Results A total of 705 309 effective sequences, including 11 phyla and 31 genera, were found in the fermentation broth during the entire process. The richly diverse fungal community was dominated by the families of Ascomycota and Basidiomycota. At the genus level, Penicillium predominated in the early stage of the fermentation which was followed by Verticillium in the middle stage and Hanseniaspora in the final stage. It indicated continual differentiations on fungal growth happened as the fermentation progressed. Along with the changing microbial community, the pH and soluble solids declined with fluctuations on the contents of alcohol and lycopene in the fermentation broth. Conclusion During the fermentation, gradually in the broth, Saccharomyces cerevisiae dominated the fungal community, while soluble solids broke down and pH lowered by the organic acids produced by the microbial metabolism, and alcohol and lycopene contents fluctuated. The revealed regulation roles of fungal flora played on the natural fermentation provided a guideline for future development of technology for an industrial utilization of tomatoes.
