2023 Vol. 38, No. 3
Display Method:
2023, 38(3): 253-261.
doi: 10.19303/j.issn.1008-0384.2023.03.001
Abstract:
Objective Correlation between the variation sites on the multiple epidermal growth factor-like domains protein 9 gene (EGFL9) and the growth of Ictalures punctatus was investigated. Method The targeted gene sequencing of EGFL9 was performed to identify the variation sites by alignment with the reference genome. Correlation between the sites and the growth traits of the fish was statistically analyzed. Result There were 27 sites with polymorphic variations on EGFL9 with 22 effective mutations obtained after filtering. Among which, 4 sites, i.e., g.142, g.573, g.3079, and g.7409, were significantly associated with the growth traits of channel catfish. The SNP sites, g.142, g.3079, and g.7409, were in introns, while the InDel site, g.573, in exon 2. The mean body length of the fish with A/A type at g.142 was significantly longer than that of fish with A/G type (P＜0.05), while that of the fish with A/G type at g.7409 significantly longer than that of fish with G/G type (P＜0.05). Both mean body mass and length of the fish with C/C type at g.3079 were significantly greater than those of the fish with A/A type (P＜0.05). The InDel site, g.573, reduced one serine residue in the protein encoded by EGFL9 significantly altereed the tertiary structure. Consequently, the mean body mass of the fish with TACC/T type at this site was significantly heavier than that of the fish with TACC/TACC type (P＜0.05), and the mean body length significantly longer than that of the fish with TACC/TACC or T/T type (P＜0.05). Conclusion The 4 growth-related markers, g.142, g.573, g.3079, and g.7409, revealed by this study showed significant associations with the growth traits of I. punctatus. The information would possibly lead to the development of breeding chips and molecular marker-assisted breeding on channel catfish in the future.
2023, 38(3): 262-270.
doi: 10.19303/j.issn.1008-0384.2023.03.002
Abstract:
Objective Effects of substituting refined meal in forage with beer brewing residues on the digestion and microbial diversity of rumen of Hu sheep were studied to explore the feasibility of utilization of the waste material. Methods Twenty-four healthy Hu sheep with similar body weight of (15.90±0.36) kg were randomly divided into 4 groups. The animals were fed with a basic forage that consisted of corn and soybean meal at 36% as control (Group I) or one that used beer lees to replace 1/3 (Group II), 2/3 (Group III), or total (Group IV) of the meal. At end of the feeding period, 3 sheep were randomly selected from each group to determine the digestion indicators and microbial diversity in the rumen fluid. Results (1) None of the substitutions showed significant differences on volatile fatty acids, pH, ammonia nitrogen, or acetic acid in the rumen (P＞0.05), but significantly higher on isobutyric acid in Group II than others (P＜0.05). (2) At phylum level, no significant differences on richness of diversity were introduced by the substitutions on Firmicutes, Bacteroidetes, Proteobacteria, Synergistetes, Fibrobacteres, Actinobacteria, Epsilonbacteraeota, Spirochaetes, and Tenericutes (P＞0.05). However, the diversity of Verrucomicrobia in Group I was significantly lower (P＜0.05). At genus level, the beer lees replacements in the forage did not significantly alter the diversity on Prevotella, Succiniclasticum, Intestinimonas, Tyzzerella, Ruminococcus, Faecalicatena, Pseudobutyrivibrio, and Hungateiclostridiumhad (P＞0.05). Conclusion The replacement of the refined meal in forage with beer lees reduced the relative richness of Verrucobacteria but did not significantly changed the microbial community structure and diversity of the rumen fluid.
2023, 38(3): 271-280.
doi: 10.19303/j.issn.1008-0384.2023.03.003
Abstract:
Objective Contents of active ingredients and volatiles in Anoectochilus roxburghii of different strains at different growth stages were determined. Method SPME-GC-MS analysis was conducted to determine the chemical compositions in Fujian small-leaf Tuhaojin and large-leaf Dayuanbao A. roxburghii. Result The highest total polyphenols of 265.07 mg·kg−1 was found in Dayuanbao 18 months after planting, so were the total flavonoids of 32.68 mg·g−1 and soluble sugars of 27.71 mg·kg−1 in 12-month-old and soluble protein of 600.62 mg·g−1 in 18-month-old Dayunbao plants. There were 6 alcohols, 8 aldehydes, and 4 ketones and 2-pentyl-furan identified with the greatest total amount in Dayunabao grown for 24 months. The volatile content of the 6-month-old tissue culture in Tuhaojin was merely 22.9% of the peak value in the respective cultivated seedlings, and in Dayuanbao, 16.7%. The odor activity value (OAV) and principal component analysis (PCA) concluded that (E)-2-hexenal, octanal, and nonanal were the major aromatic compounds in Tuhaojin, while 1-octen-3-ol, 3,7-dimethyl-1,6-octadien-3-ol, and β-ionone in Dayuanbao. The PCA and PLS-DA indicated that these volatiles could adequately distinguish A. roxburghii of different strains harvested at different times. Conclusion Active ingredients and volatiles in A. roxburghii significantly varied by the strains and growth stages. It was plausible that a chemical analysis could suffice for quality control and authentication purpose on the products in commerce.
2023, 38(3): 281-293.
doi: 10.19303/j.issn.1008-0384.2023.03.004
Abstract:
Objective Characteristics and biotic stress response of WRKY transcription factors (TFs) in tomato plants were investigated. Method The latest available bioinformatics and genomics methods were employed to identify the tomato WRKY TFs. RNA-seq of disease-resistant and susceptible tomato inbred lines before and after artificial Ralstonia solanacearum infection were obtained to identify the TFs associated with the pathogenic resistance of the plants. Result Eighty-five tomato WRKY TFs were identified and divided into I, IIa+b, IIc, IId+e, and III categories. The IIe group had the highest number of the TFs. The conserved motif of 9 TFs had one single amino acid variation, and WRKYGKK was the dominant mutant. The TFs, especially those in the IIe group, were mainly found on chromosome 5, at the ends, and in clusters. In them, 45.88% showed collinearity and 58.82% (mainly in I and IIc groups) formed 73 pairs of orthologs with those in Arabidopsis and chili pepper at a Ka/Ks ratio below 1. Sixteen of them, mainly belonging to IIa+b and IIc, responded significantly to the biotic stress with expressions largely in the roots. There were 12 differentially expressed WRKY TFs identified mainly in III and IIb. Of which, the interaction between Solyc03g095770.3 (III) and Solyc09g014990.4 (I) played a significant role in the response of the tomato plant to bacterial wilt. Conclusion The WRKY TFs were identified in tomato plants. Twelve genes responded to the bacterial wilt were isolated.
2023, 38(3): 294-301.
doi: 10.19303/j.issn.1008-0384.2023.03.005
Abstract:
Objective F-BOX genes responsible for the resistance of Brassica rapa to sclerotinia were investigated. Method Suspected members of F-BOX family were identified at the transcriptome level to identify their subcellular localization, chromosomal localization, and conserved domains as well as expressions by q-PCR under artificial induction of sclerotinia. Result There were 32 BraF-BOX genes classified into 4 subgroups with the relative molecular masses varying from 34 751.13 Da to 105 942.22 Da. The predicted subcellular localizations of 26 F-BOX were in the nucleus and cytoplasm, and 6 in the chloroplast. The genetic structures of all 32 genes contained Motif 1 and exon in the DNA sequences with expressions differed in different parts of Chinese cabbage. The relative expressions by q-PCR of 6 genes agreed with the transcriptome data at 36 h. The expressions of Bra037120, Bra011427, and Bra009835 were upregulated and increased with prolonged induction. Conclusion The bioinformatics and transcriptome data analysis on the BraF-BOX genes suggested that Bra037120, Bra011427, and Bra009835 were potentially associated with the sclerotinia resistance of Chinese cabbage.
2023, 38(3): 302-311.
doi: 10.19303/j.issn.1008-0384.2023.03.006
Abstract:
Objective Universality of simple sequence repeat (SSR) markers of members of Malvaceae family and roselle herbal plants was examined to extend the genetic information bank. Method A total of 62 pairs of SSR primers were selected from Hibiscus cannabinus, Gossypium spp, Hibiscus esculentus L, and Hibiscus syriacus L. of Malvaceae family. They included 17 pairs from rosemallow and 15 pairs from each of the other crops to compare for polymorphism with those of 8 accessions of Hibiscus sabdariffa L. germplasms. Result Of the 62 SSR primers, 58, or 93.5%, were found common and 48, or 82.8%, polymorphic between the two groups. And out of 246 bands amplified, 239, or 97.2%, were polymorphic. Among the Malvaceae family members, cotton had 14 primer pairs polymorphic at the highest rate of 86.7%, while the rate with rosemallow primers 82.4%, with okra primers 73.3%, and with kenaf primers 66.7%. The PIC of rosemallow primers was the highest at 0.685 among all. A cluster analysis classified the 8 roselle accessions into 3 groups at a similarity coefficient of 0.49. The strain, 19FZ-76, was clustered by itself in the genetic relationship significantly far from other accessions that was reflected by the phenotypic traits as well. The greatest genetic similarity coefficient was observed in the 19FZ-74 cultivars from Myanmar and Zhangzhou, indicating a close genetic relationship of the two accessions and the likely origin of the roselle in Fujian. Conclusion The SSR primers of Malvaceae plants including roselle showed a certain universality. The selected markers could provide a reliable venue for genetic studies, such as variety identification, relationship analysis, and map construction on the herbal plant.
2023, 38(3): 312-321.
doi: 10.19303/j.issn.1008-0384.2023.03.007
Abstract:
Objective Genetic differences and mycelial morphology of imported and locally cultivated Lyophyllum decastes (Lyd) were compared. Methods Sequence homology, multiple sequence alignment, and phylogenetic tree were examined and constructed based on the sequences of the fungal 18S rRNA (V4) and ITS (ITS1-ITS4) (18S rRNA-ITS) of the imported and locally cultivated Lyd strains. Mutation analysis and genetic distance determination were performed to identify the phylogenetic relationship, while mycelium growth and scanning electron microscopic images observed to differentiate the morphology of the strains. Results The 18S rRNA-ITS sequence alignment between the two strains showed the imported Lyd-LR1, Lyd-LR6, Lyd-LR10, Lyd-LR15, and Lyd-LR17 to be higher on the ratio of base deletion and base substitution mutations. The ITS sequence homology between them decreased to 80.09%–89.72% with Lyd-LR1 and Lyd-LR6 being higher than Lyd-LR10, Lyd-LR15, Lyd-LR17 as well as the local Lyd-LRX and Lyd-LRY on the proportions of base mutation. The phylogenetic tree analysis indicated a distant genetic relationship between Lyd-LR1 and Lyd-LR6 and Lyd-LR10, Lyd-LR15, Lyd-LR17, Lyd-LRX, Lyd-LRY, and some Lyd or Lyophyllum (Ly) registered by NCBI. The loose and radial fusiform mycelia of Lyd-LR1 grew slowly and irregularly, whereas the pitted mycelia of Lyd-LR6 tiled with high margin thickness proliferated rapidly and polymerized considerably. The growth rates and morphology of Lyd-LR1 and Lyd-LR6 mycelia differed from those of Lyd-LR10, Lyd-LR15, Lyd-LR17, Lyd-LRX, and Lyd-LRY, which had full cylindrical mycelia and thick and fast-growing protuberant mycelia. Compared to Lyd-LR1, Lyd-LR6 exhibited more apparent changes in gene sequence base mutation location, base mutation proportion, and mycelial morphology. Conclusion The gene sequence base mutation location (base substitution and base deletion), 18S rRNA-ITS sequence homology, and phylogenetic relationship of Lyd-LR6 significantly differed from those of the other imported and the locally cultivated Lyd strains. A high proportion of gene sequence base mutations induced significant changes in the growth and morphology of Lyd-LR6 mycelia indicating a potential use of it in breeding new varieties of Lyd mushrooms.
2023, 38(3): 322-328.
doi: 10.19303/j.issn.1008-0384.2023.03.008
Abstract:
Objective The polyclonal antibody against MiPDCD6 protein in the esophageal glands of Meloidogyne incognita was prepared to study the pathogenic mechanism of the root-knot disease on plants transmitted by the nematode. Method The functional fragment of MiPDCD6 was amplified by PCR to construct recombinant plasmid pET-32a-MiPDCD6 and transform it into Escherichia coli BL21 cells for MiPDCD6 fusion protein induction and expression. Polyclonal antibodies were prepared by immunizing male New Zealand white rabbits with purified MiPDCD6 expression protein. Titer and purification of the obtained antibody were verified using ELISA and SDS-PAGE techniques. Result Under IPTG concentration of 1.0 mmol·L−1 at 37℃with constant rotation of 150 r·min−1 for 5 h, MiPDCD6 transformed in the E. coli BL21 was clearly expressed. The secured polyclonal antibody was highly specific with a high titer of approximately 1∶50000 as shown by ELISA and SDS-PAGE. Conclusion The prokaryotic expression conditions of MiPDCD6 were determined. The polyclonal antibody obtained had a high titer and specification against MiPDCD6 and was considered adequate for studying the pathogenesis of the root knot disease on plants infected through M. incognita.
2023, 38(3): 329-338.
doi: 10.19303/j.issn.1008-0384.2023.03.009
Abstract:
Objective Safety and efficacy of applying vomitoxin to control soilborne diseases on potato plants were investigated. Method In a laboratory experiment, vomitoxin (aka deoxynivalenol, DON) was applied on the Atlantic variety potato to determine the disease resistance to and physiological alternations induced by some major soilborne pathogens on the plant for treatment optimization. Result Within a range of concentrations, DON caused no significant ill-effects on the potato plants. A treatment of it at 5 ng·mL−1 for 4 h effectively inhibited the spread of bacterial wilt caused by Ralstonia solanacearum, soft rots by Erwinia chrysanthemi, or scabies by Streptomyces scabies on the leaves, and at a dosage of 3 ng·mL−1, the growth of dry rot pathogen Fusarium sambucinum was retarded. The strongest DON-induced resistance against the bacterial wilt or dry and soft rots of the plant was found at 25 ℃ and pH 7, and against scabies at pH 9. With an exposure of 12 000 lx light, the DON-induction on the plants reached the greatest efficacy against the dry rots. Whereas, under 15 000 lx, that could be achieved on the bacterial wilt, soft rots, and scabies. A DON treatment significantly raised the SOD activity and lignin content but lowered MDA in the leaves. Conclusion Application of DON as an elicitor in low concentrations exerted no adverse effect but significantly inhibit some major soilborne diseases on potato plants. A potential venue for preventing and treating the diseases seemed merit further investigation.
2023, 38(3): 339-345.
doi: 10.19303/j.issn.1008-0384.2023.03.010
Abstract:
Objective An efficient and rapid detection method was developed to monitor the spread of a newly discovered bacterial canker disease infected by Xanthomonas perforans that seriously impacted the production of water spinach (Ipomoea aquatica), a recently introduced and increasingly important cash crop in southern China. Method Two pairs of primers were designed to detect X. perforans based on the specific sequences in the genomes including TC2-1_002562 and TC2-1_002580, which encode a phage terminator large subunit family protein and a phage family protein, respectively. The applicability of the proposed methodology with a single PCR reaction was verified. Result The newly developed PCR method exhibited a high specificity for detecting X. perforans. In the first sampling batch of water spinach plants, planting soils, and water collected from regions in Guangdong Province, the pathogen was only detected in 5 specimens of water spinach leaves from Dongguan. However, during the subsequent second sampling 10 days later, the pathogen was found in numerous specimens from all sources. Conclusion The established PCR method could rapidly detect X. perforans for early disease diagnosis on water spinach. As the field monitoring indicated, the endemic was prevalent in Dongguan and Shenzhen, possibly some other localities, but spreading fast. The origin of the infection might come from the stored water spinach seedlings and/or seeds. The area water and soil could also link to the spread of the disease. Urgent study to timely develop effective prevention and control measures is in order.
2023, 38(3): 346-351.
doi: 10.19303/j.issn.1008-0384.2023.03.011
Abstract:
Objective Appropriate methods for efficient tissue disinfection, explant induction, and culture medium formulation were established for a high-performance program to propagate pyrethrum. Method Unopened flower buds of white flower pyrethrum were sterilized and used as explants on an MS solid medium for the experiment. Effects of various plant growth regulators on bud induction, proliferation, and rooting of the seedlings were monitored. Result The optimum conditions for disinfecting the explants were found to be a treatment of 75% alcohol for 30 s followed by one of 0.10% mercuric chloride solution for 10 min and another of 15% hypochlorite for 15 min. For bud induction, the choice medium was formulated with MS + 2.0 mg·L−1 6-BA + 0.5 mg·L−1 TDZ+ 0.2 mg·L−1 IBA; for bud proliferation, MS + 1.0 mg·L−1 6-BA + 0.1 mg·L−1TDZ + 0.1 mg·L−1 IBA; for rooting, MS + 0.1 mg·L−1 IAA + 0.1 mg·L−1 IBA; and for transplanting seedlings, peat:perlite at 6:1. A survival rate greater than 90% as well as adequate transplanting was achieved. Conclusion The newly developed in vitro regeneration system materially lessened the pressure of the recently encountered white flower pyrethrum germplasm degradation. In addition, a supply of high-quality seedlings could be assured with the proposed propagation program.
2023, 38(3): 352-359.
doi: 10.19303/j.issn.1008-0384.2023.03.012
Abstract:
Objective Organic matter contents in the latosolic red soils under various long-term fertilization practices were measured to analyze the effects and to establish a prediction model for efficient management. Method Two long-term experiments were conducted on separate uplands in Fujian of peanut-sweet potato rotating cultivation fields with latosolic red soil. The designated lots were under either a continuous application of different chemical fertilizers for 16 years or of chemical/organic fertilizations for 14 years. Content of soil organic matters (SOM) was monitored, and a grey prediction model constructed based on the collected data. Result The fertilizations boosted SOM content in the soils in comparison to the lot without fertilizer application. The use of the Recommended Fertilizer (RF) increased the average SOM to (19.83±0.77) g·kg−1, which was significantly higher than the applications of other chemical fertilizers. The content further increased to (22.53±1.69) g·kg−1, i.e., 2.8 times of RF treatment on an annual basis, when the chemical/organic manure combination (RF+OM), especially the decomposed pig manure (RF+PM), was applied. The grey prediction model on SOM yielded fitting errors ranging from 1.226% to 3.307% for all fertilizations. While the predicted result of the non-fertilization was on a continuously downward trend, and the RF treatment increased to (20.220±0.002) g·kg−1, which was superior to all other fertilizations using chemicals. More important, the long-term SOM would be on a increasing trend under chemical/organic fertilization, especially RF+PM that ranked the top among all treatments reaching the significantly higher level than RF at (23.777±0.017) g·kg−1. Conclusion Based on the past records and the predicted trend on SOM, RF undoubtedly improved the fertility of the latosolic red soil. However, RF+OM, especially RF+PM, would bring even more impressive results, and thus deserved serious consideration for the agricultural practice in the area.
2023, 38(3): 360-366.
doi: 10.19303/j.issn.1008-0384.2023.03.013
Abstract:
Objective Effect of encapsulation to protect Bacillus mucilaginosus from thermal shock was evaluated for formulation optimization. Method Rates of survival and activation of B. mucilaginosus encapsulated with trehalose, skimmed milk powder, and/or modified H-MS medical stone after spray-drying were determined. Optimal formulation for the encapsulation was obtained using the response surface method. Result The survival rate of the naked B. mucilaginosus was (34.15±1.07)%. In contrast, the encapsulation with trehalose significantly improved the rate to (52.17±1.08)% (P＜0.01), while with H-MS, (51.64±0.77)% (P＜0.01) and with skimmed milk powder, (43.67±1.62)% (P＜0.05). Furthermore, a combined use of trehalose at 4.33%, skimmed milk powder at 2.90%, and H-MS at 7.57% for the encapsulation, the survival reached (73.32±0.76)% with a microbial count of 1.12×109 CFU·g−1. Conclusion The three encapsulation materials, especially, when they were applied in combination, significantly raised the B. mucilaginosus survival rate under a heat treatment. The scanning electron microscopic image revealed that trehalose and skimmed milk powder immobilized the bacteria cells in H-MS pores producing an effective thermal insulation that sheltered the encapsulated microbes.
2023, 38(3): 367-375.
doi: 10.19303/j.issn.1008-0384.2023.03.014
Abstract:
Objective Composition and diversity of the ectomycorrhizal (ECM) fungi communities at natural and cultivated Castanea henryi forests in Taining, Fujian were studied. Methods Samples of C. henryi root and rhizosphere soil at the sites were collected for physiochemical analysis, mycorrhizal morphology observation, and molecular identification. A correlation between the ECM fungi in the roots and environmental conditions was analyzed. Results The physicochemical properties of the rhizosphere soil significantly differed between the natural forests and the plantations. The contents of organic matters, total nitrogen, total potassium, and water in the soil of a natural forest were significantly higher than those of a plantation. Five types of ECM fungi belonging to 2 phyla, 3 families, and 5 genera were identified in the collected samples. Among the fungi, the highest infection rate of Cenococcum geophilum at 33.62% was found on the C. henryi roots at the natural forests and that of Scleroderma citrinum at 65.61% at the plantations; while the other dominants included Lactarius kesiyae, Xerocomus sp. and Russula sp. . The contents of total phosphorus, total potassium, and water as well as pH in the soil significantly correlated with the fungal infection on the C. henryi roots. Conclusion Significant differences existed in the ECM fungi communities at the natural C. henryi forests and plantations in Taining. The fungal diversity in rhizosphere soil was higher at the natural forests than the plantations. The information gathered from this study provided a guideline for improved C. henryi cultivation management and plantation soil restoration.
2023, 38(3): 376-386.
doi: 10.19303/j.issn.1008-0384.2023.03.015
Abstract:
Infectious bovine rhinotracheitis is an important infectious disease of cattle. The clinical symptoms of the disease were principally respiratory ones that may be accompanied by conjunctivitis, mastitis, abortion, etc. The pathogenic virus is also known as bovine herpesvirus type 1. It encodes 30 to 40 structural proteins with 11 envelope glycoproteins, which play an important role in the process of virus adsorption and host cell invasion. Glycoprotein gB is essential for the virus to invade, spread, and replicate on host cells. Glycoprotein gD is critical in viral replication, transmission, and infection with strong immunogenicity that induces neutralizing antibodies. Studying gB and gD not only helps decipher the infection mechanism at the molecular level but also leads to new clinical diagnosis and prevention method developments on rhinotracheitis. This article summarizes recent research results on glycoprotein gB and gD concerning the biological functions and applications of these proteins in producing vaccines and generating advanced diagnosis methodologies for the infectious disease in cattle.
Infectious bovine rhinotracheitis is an important infectious disease of cattle. The clinical symptoms of the disease were principally respiratory ones that may be accompanied by conjunctivitis, mastitis, abortion, etc. The pathogenic virus is also known as bovine herpesvirus type 1. It encodes 30 to 40 structural proteins with 11 envelope glycoproteins, which play an important role in the process of virus adsorption and host cell invasion. Glycoprotein gB is essential for the virus to invade, spread, and replicate on host cells. Glycoprotein gD is critical in viral replication, transmission, and infection with strong immunogenicity that induces neutralizing antibodies. Studying gB and gD not only helps decipher the infection mechanism at the molecular level but also leads to new clinical diagnosis and prevention method developments on rhinotracheitis. This article summarizes recent research results on glycoprotein gB and gD concerning the biological functions and applications of these proteins in producing vaccines and generating advanced diagnosis methodologies for the infectious disease in cattle.
