2021 Vol. 36, No. 1
Display Method:
2021, 36(1): 1-8.
doi: 10.19303/j.issn.1008-0384.2021.01.001
Abstract:
Objective The bPAG16 Methods Bioinformatics were applied to codon-optimize and synthesize the bPAG16 gene followed by connecting it to the proEM vector by T4 DNA ligase. The resulting proEM-bPAG16 recombinant vector was transfected into HEK293 cells. The expressed gene was purified by Ni2+ affinity chromatography and verified by SDS-PAGE and western blotting. Result With the optimization, the codon adaptation index (CAI) of the bPAG16 gene rose from 0.77 to 0.96 and the GC content from 48% to 58%. After enzymatic digestion, fragments of approximately 1 179 bp and 4 369 bp were obtained from the proEM-bPAG16 expression vector as expected. The nucleotide sequence of the inserted bPAG16 in the recombinant plasmid was same as that of the optimized gene and absent of any mutated amino acids. SDS-PAGE and western blotting showed that the 48kDa recombinant bPAG16 fusion protein was correctly expressed in the HEK293 cells with a purity greater than 90%. Conclusion The successfully executed codon optimization and synthesis of bPAG16 gene provided the base material for further study on early diagnosis of pregnancy in dairy cows.
2021, 36(1): 9-16.
doi: 10.19303/j.issn.1008-0384.2021.01.002
Abstract:
Objective Bainong Aikang-58 is one of the major wheat cultivars in China. To investigate the response mechanism of Bainon Aikang-58 seedling to Na2SO4 stress was discussed and to define its tolerance threshold to Na2SO4 stress was determined so as to provide basic data support for further research and scientific basis and and theoretical guidance for agricultural production practice. Method Effects of Na2SO4 stress at 0, 20, 40, 60, 80, and 100 mmol·L−1 on the growth and physiology of the wheat seedlings were studied in a hydroponic experimentation. Result As the Na2SO4 concentration in the medium increased, the plant height, fresh weight, and biomass of the seedlings decreased. The activity of each antioxidant enzyme in wheat seedlings would continuously increase with the increase of Na2SO4 concentration, and would decrease after exceeding a certain threshold. The membrane permeability and content of MDA in the synplasmic membrane increase with increasing of Na2SO4 concentration to maximize 3.2 and 4.94 times of the control. The chlorophyll content in the leaves were decreased with the increase of Na2SO4 concentration. The soluble sugar content in wheat increased first and then decreased with the increase of Na2SO4 concentration. The soluble sugar concentration was the highest at 80 mmol·L−1 Whereas, the proline content kept increasing to reach a maximum that was 8.51 times of control. Conclusion The tolerance threshold of Bainong Aikang-58 wheat seedling toward Na2SO4 stress was determined to be 80−100 mmol·L−1.
2021, 36(1): 17-23.
doi: 10.19303/j.issn.1008-0384.2021.01.003
Abstract:
Objective Functions of PYL gene in the post-harvest physiological deterioration (PPD) and under abiotic stresses of cassava (Manihot esculenta Crantz) were studied for further research on the key component of the plant abscisic acid (ABA) signaling pathway, PYR/PYL/RCARs family. Method The PYL gene, MePYL12 was cloned from cassava SC 124 by RT-PCR. Bioinformatics was used to analyze the physicochemical properties, conserved domain, genetic evolutionary relationship, protein structure prediction, promoter elements as well as the expressions of MePYL12 in PPD and under abiotic stresses. Result (1) The full-length cDNA of MePYL12 was 567 bp encoded a polypeptide of 188 amino acids with a predicted relative molecular mass of 20.8kD and an isoelectric point of 5.55. The predicted tertiary structure contained a helix. Its multiple protein sequence alignment showed high similarities with the PYL proteins in Hevea brasiliensis (94.68%) and Ricinus communis (86.77 %). MePYL12 consisted of the conserved motifs of the PYL family, such as the ABA binding region "Latch" and "Gate" suggesting it to be a genuine member from the family and highly conserved. (2) The expressions of MePYL12 in 10 different types of cassava tissue/organ were high in the root apical meristem, shoot apical meristem, and leaf. (3) The promoter element analysis showed that the gene contained the light-responsive, drought-induced MBS, and ABA responsive ABRE motifs. (4) The expression of MePYL12 was significantly upregulated by ABA treatment and drought stress as well as during PPD that peaked in 6h followed by downregulation. Conclusion MePYL12 might participate in the PPD process improving the ability of cassava plants in dealing with abiotic stresses. Further study on the functions of MePYL12 in cassava is in order.
2021, 36(1): 24-35.
doi: 10.19303/j.issn.1008-0384.2021.01.004
Abstract:
Objective Transcriptomes of differentially expressed genes related to flavonoids synthesis in tubers of two Huaiyushan cultivated varieties of Tetrastigma hemsleyanum Diels et Gilg were compared. Method Tubers from Huaiyu 1 (HY1) and Huaiyu 2 (HY2) were used for the transcriptome analysis. Result HY1 and HY2 had clean reads of 42 311 662 and 41 411 202, respectively, and no less than 95.75% of Q30 base. Their transcription factors basically belonged to the MYB-superfamily, bHLH, AP2/ERF, NAC, C2C2, WRKY, etc. The paired values of FPKM in HY1 and HY2 were between 0 and 2; the expression densities, between 0 and 0.7; the number of commonly expressed genes, 22 367; and, the number of uniquely expressed genes in HY1, 18 196, while 8 137 in HY2. The correlation between the expressions of the two had a coefficient of 0.913, and that between the samples was high. There were 12 199 differentially expressed genes between the two, with 3 551 upregulated and 8 648 downregulated in HY2 as compared to HY1. The GO enrichment analysis showed that the differential genes were mainly annotated into photosynthesis, light harvesting in photosystem I, photosynthesis, light harvesting, chlorophyll metabolic process, protein-chromophore linkage, generation of precursor metabolites and energy, chlorophyll biosynthetic process, response to oxidative stress, alpha-amino acid metabolic process, photosynthesis, plastoglobule, photosystem I, photosystem II, plastid nucleoid, photosystem, chlorophyll binding, monooxygenase activity, iron ion binding, heme binding, lyase activity, etc. Whereas, the KEGG enrichment analysis indicated the differential genes to be mainly annotated into photosynthesis-antenna proteins, ribosome, glyoxylate and dicarboxylate metabolism, phenylpropanoid biosynthesis, stilbenoid, diarylheptanoid and gingerol biosynthesis, flavonoid biosynthesis, photosynthesis, carbon fixation in photosynthetic organisms, glycine, serine and threonine metabolism, plant hormone signal transduction, glutathione metabolism, pyruvate metabolism, phenylalanine metabolism, circadian rhythm-plant, flavone and flavonol biosynthesis, cysteine and methionine metabolism, cyanoamino acid metabolism, carotenoid biosynthesis, alpha-linolenic acid metabolism, porphyrin and chlorophyll metabolism, and other metabolic pathways. Conclusion The differentially expressed genes related to flavonoids synthesis, such as stilbene synthase, leucoanthocyanidin dioxygenase, CHI protein, chalcone synthase 2, flavanone 3-hydroxylase and lleucoanthocyanidin reductase 1 and flavonoid 3’- hydroxylase gene were upregulated in HY2, while chalcone synthase, flavonol synthase and flavonoid 3’, 5’-methyltransferase downregulated. The variations apparently resulted in the differences shown on the total flavonoid content between the HY1 and HY2 tubers.
2021, 36(1): 36-40.
doi: 10.19303/j.issn.1008-0384.2021.01.005
Abstract:
Objective The rice cultivar Gufeng B confers strong, broad-spectrum, durable resistance against various rice blast isolates. The present study was aim to identify and map the blast resistance gene(s) in Gufeng B. Method The F1 and F2 population were obtained by crossing Gufeng B and Nipponbare, and the genetic model of blast resistance was analyzed after inoculating with 7 strains of Magnaporthe grisea. Subsequently, F2 population was used to construct a resistant pool and a sensitive pool respectively, and to map the associated loci via the method of bulked segregation analysis. Result Gufeng B exhibited high resistance to all of the tested strains, such as KJ201, RB22, CHNOS, RB6, 2Y838-1, 501-3 and IR16-1, suggesting that Gufeng B may carry the broad-spectrum and high resistance genes. The F1 progenies from the cross between Gufeng B and Nipponbare conferred resistance against the strains 501-3 and IR16-1, and the segregation ratio of resistance and susceptibility among F2 progenies does not fit 3:1, assuming that the resistance against the strains 501-3 and IR16-1 were controlled by multiple locus in Gufeng B. Whole genome re-sequencing of the two parental lines Gufeng B and Nipponbare identified 1,756,964 SNPs. Calculation results of △SNP-index showed that there were two candidate loci conferring resistance to rice blast disease, which were located at Chr.6: 10,082-11,397Kb, corresponding to the Pi2/9 locus, and Chr.11: 120-266Kb. 4006 SNPs and 623 InDels markers were searched within the interval of Chromosome 6, 752 SNPs and 195 InDels within the corresponding region of Chromosome 11, respectively. Conclusion The resistance of Gufeng B to 501-3 strain may be controlled by two resistance genes on chromosomes 6 and 11. Our results laid the foundation for finely mapping and cloning the resistance genes in Gufeng B, and provided marker resources for molecular marker-assisted selection.
2021, 36(1): 41-52.
doi: 10.19303/j.issn.1008-0384.2021.01.006
Abstract:
Objective Anthracnose-resistance of varieties of soybean in China was assessed and classified to facilitate the breeding of resistant cultivars. Method From 2011 to 2019, 590 varieties of domestic soybean cultivars were collected and challenged by an artificial inoculation of Colletotrichum truncatum to determine their degrees of resistance to anthracnose. Result The test showed 70 of the varieties belonged to the resistant (R), 143 to the medium resistant (MR), 219 to the medium susceptible (MS), 148 to the susceptible (S), and 10 to the highly susceptible (HS) categories, representing 11.86%, 24.24%, 37.12%, 25.08%, and 1.69%, respectively, of the entire sampled population. The overall disease resistant rate (RR) in a particular year varied from year to year, ranging from 13.64% to 68.00%, with the highest shown in 2013, followed by 52% in 2012 and the lowest in 2016. The RRs of the spring varieties in the tropical and subtropical areas (S1), the summer varieties in the tropical and subtropical areas (S2), the spring-sowing green varieties (S3), the summer-sowing green varieties (S4), the spring soybean from Yangtze river basin (S5), the new variety from Fujian (S6), and the new varieties from other regions (S7) ranked from the lowest of S3 at 5.89% to the highest of S2 at 69.23%, while S4 being the second highest at 57.69%. Conclusion Out of the 590 varieties, 36.10% were only resistant or moderately resistant to anthracnose with none exhibited high resistance. And, there was no spring-sowing green soybeans for fresh consumption being substantially resistant to the disease.
2021, 36(1): 53-58.
doi: 10.19303/j.issn.1008-0384.2021.01.007
Abstract:
Objective Possibility of applying Ailanthus altissima alkaloids for controlling 5 plant pathogenic fungi was studied. Method Crude ethanol extract of alkaloids from the bark of A. altissima was obtained. Extract was separated using the vacuum liquid chromatography. Antifungal activities of the crude alkaloids and the elution fractions on Fusarium oxysporum f. sp. cucumerinum, F. graminearum, F. oxysporum f. sp. benincasae, Botrytis paeoniae, and Gloeosporium sp. were determined according to the fungal mycelium growth rate. Result The crude ethanol-extracted alkaloids and canthin-6-one fraction exhibited inhibitory effects on the tested fungi, except B. paeoniae. The antimicrobial effects were particularly strong on Gloeosporium sp. and F. graminearum and especially toxic toward the latter with Conclusion The ethanol-extracted alkaloids from A. altissima barks could inhibit the growth and/or be lethal toward a variety of plant pathogenic fungi. Although the antifungal effect was not as potent as that of canthin-6-one, the crude extract perse appeared promising to be a new source for the development of a plant-based biocontrol agent.
2021, 36(1): 59-64.
doi: 10.19303/j.issn.1008-0384.2021.01.008
Abstract:
Objective Pathogens that cause serious diseases on Dioscorea batatas Decne grown on the mountain ridges in northern Hubei province were investigated to facilitate drafting effective disease prevention and control measures. Method Specimens of aboveground parts of the diseased plants were collected for pathogen isolation. The isolates were identified by colonial morphology, microstructure, and blasting sequence of ITS as well as the pathogenicity determination with tissue separation of phytopathology and ITS-PCR. Result The isolated pathogenic fungi included Alternaria alternate, Colletotrichum fructicola, C. gloeosporiaides, and Lasiodiplodia pseudotheobromae. Conclusion The pathogens causing the diseases, such as anthracnose, that can lead to withering or death of Chinese yams at the region were highly diversified and complex.
2021, 36(1): 65-70.
doi: 10.19303/j.issn.1008-0384.2021.01.009
Abstract:
Objective A newly modified analytical method for detecting pesticide residues in tea was developed to improve test efficiency and reduce cost. Method Fe3O4 nanoparticles were used to replace the conventional pigment-removing material prior to measurements on pesticide residues, including carbendazim, pyrimethanil, tricyclazol, imidachloprid, acetamiprid, methomyl, and thiamethoxam, in tea by liquid chromatography-tandem mass spectrometry. The methodology was evaluated according to its applicability and validated in a spiked residue test. Result The recovery rates on the 7 pesticides under the modified method using 300 mg Fe3O4 nanoparticles for pigment removal pretreatment were higher than graphitizing with carbon black (GCB). The pigment-removed tea extracts were separated with a Phenomenex Luna C8 (150 mm × 2.0 mm × 3.0 µm) by using 0.1 % formic acid and 5 mmol·L-1 ammonium acetate as Phase A and acetonitrile as Phase B for the mobile phase prior to the tandem mass spectrometric analysis under the multiple reaction monitoring mode with external standards. A linearity on the measurements by the new method was achieved within the pesticide concentrations of 0-50 µg·L-1 showing correlation coefficients (R2) greater than 0.995 for all 7 pesticides. On 5, 10, and 50 ng·g-1 spiked pesticide samples, average recovery rates of the new method ranged from 71.6 % to 107.7 % with relative standard deviations of 3.95%-13.62 %. The limits of detection (LOD) on the pesticides by the method were between 0.15 µg·kg-1 and 0.60 µg·kg-1, and the limits of quantitation (LOQ) between 0.5 µg·kg-1 and 2.0 µg·kg-1. Conclusion The newly developed pesticide detection method was stable, repeatable, and accurate for the designed purpose.
2021, 36(1): 71-77.
doi: 10.19303/j.issn.1008-0384.2021.01.010
Abstract:
Objective Effects of peanut shell replacing peat soil as the cultivation substrate on the substrate properties and the growth quality of Anoectochilus roxburghii were studied to obtain the optimal substrate ratio for Anoectochilus roxburghii cultivation. Method Five ratios of peanut shell to peat soil, including CK (100% peat soil), N75 (75% peat soil + 25% peanut shell), N67 (67% peat soil + 33% peanut shell), N50 (50% peat soil + 50% peanut shell) and N33 (33% peat soil + 67% peanut shell), were setted to study the physical and chemical properties of the soil and the growth and quality of Anoectochilus roxburghii by pot experiment. Result (1) The replacement of peat soil by peanut shell improved the aeration, water holding capacity and fertility of cultivation substrate. Compared with control, the bulk density of 33% peat soil + 67% peanut shell treatment reduced by 53.84%, maximum water holding capacity and non-capillary porosity increased by 97.14% and 148.35%, and total nitrogen, available phosphorus, and vailable potassium content increased by 7.85%, 67.76%, 1830.53%, resectively, with significant difference. (2) The growth and quality of Anoectochilus roxburghiithe was the best under 33% peat soil + 67% peanut shell treatmentm, the plant height, root length, leaf length, and leaf width of which were significantly better than those of the control group; and the contents of total flavonoids, polysaccharides, free amino acids, and total phenols were significantly increased by 41.57%, 30.59%, 45.08%, and 53.16%, respectively, with significant differences. Conclusion 33% peat soil + 67% peanut shell was the optimal ratio as substract for Anoectochilus roxburghiithe by increasing the soil fertility and improving the growth quality.
2021, 36(1): 78-90.
doi: 10.19303/j.issn.1008-0384.2021.01.011
Abstract:
Objective To ensure a low-carbon agriculture for Anhui province, temporal and spatial variations on the carbon sources/sinks at the farmlands were analyzed based on the carbon footprints at selected localities. Method Statistical data on the related agricultural inputs and crop biomass in the province from 2008 to 2016 were applied to calculate the carbon sources/sinks and footprints using the emission coefficient method. ArcGIS and Geoda software helped visualize and auto-correlate the spatial and temporal characteristics of the selected 16 cities in 9 years. Results The emission intensity at the two major contributing municipalities, Huangshan and Suzhou, showed a trend of "high-in-the-north/south, low-in-the-central, and balanced-on-east/west", whereas, the absorption a mode of "heavy-on-both-ends and light-in-the-center" at Fuyang and Bengbu, the major contributing cities. The carbon emission of the ecosystem was declining during the study period, but the absorption irregular. The varied land uses over the years might contribute to the results. Conclusion The carbon emission intensity and absorption in the province varied with characteristic patterns. In general, the carbon footprints in the area were below the ecological tolerance capacity indicating rooms for spatial autocorrelation. Such surplus, thus, would allow the surrounding prefectures and cities a better situation in dealing with the environmental carbonization. Based on the finding, it appeared that the Action Plan for Implementing Green Development could be adequately carried out in Anhui with a scientifically planned low-carbon emission operation taking local conditions into consideration. The desirable land use and agricultural practices could be well accomplished without jeopardizing the target ecological improvement for the province.
2021, 36(1): 91-103.
doi: 10.19303/j.issn.1008-0384.2021.01.012
Abstract:
Objective Bacteriostatic property of Lactobacillus plantarum LV02 bacteriocin produced on an optimized fermentation medium was determined. Method Based on a single-factor design, the PB and the steepest ascent experiments were conducted to locate the center point of the step length and direction of the influencing factors for a CCD test. Subsequently, effects of different media on the bacteriostatic property and OD600 of the cultured L. plantarum LV02 were evaluated for formulation optimization. The Oxford cup method was employed to determine the bacteriostatic capacity and stability under application conditions of the resulting LV02 bacteriocin produced on the optimized medium. Result The optimized formula for the LV02 fermentation medium in 1 L of water constituted 34.07 g of glucose, 18.12 g of yeast extract, 2 g of dipotassium hydrogen phosphate, 0.16 g of manganese sulfate, 5 g of sodium acetate, 0.20 g of magnesium sulfate, 1 g of ammonium citrate, 1 mL of Tween 80, and 50 mL of carrot juice. On the optimized medium, LV02 grew to yield a 12% increase on OD600 and bacteriocin with a 26% increase on the diameter of inhibition zone against Escherichia coli YS. For the crude LV02 bacteriocin extraction, 80% saturation concentration of ammonium sulfate was used. The Conclusion L. plantarum LV02 cultured on the optimized fermentation medium was bacteriostatic against E. coli YS. The fermentation produced bacteriocin with desirable stabilities to heat, acid, and slight alkaline condition and was considered promising for the development of an antibacterial agent.
2021, 36(1): 104-114.
doi: 10.19303/j.issn.1008-0384.2021.01.013
Abstract:
Objective TPreparation of an spray-dried encapsulated anthocyanins from Hibiscus sabdariffa was optimized. Method Single factor experiment and response surface method were employed for the process optimization. Structure of the microcapsules was characterized with scanning electron microscopy (SEM). Product stability under exposure of light, heat, and metal ions was determined. Results The optimum encapsulation used 7% total solids that consisted of 70% maltodextrin, 3% gum Arabic, and 7% H. sabdariffa anthocyanin, and the dehydration applied a spray-drying inlet-air temperature at 143 ℃. The resulting encapsulation rate reached 97.11%. Observed under SEM, the spherical microcapsules were smooth on the surface with no cracks or voids. Exposed to temperature or light, the microcapsules were significantly more stable than the purified anthocyanins. And, the product stability in the presence of Cu2+ or Zn2+ was also significantly improved by the process. A maximum shelf life of the product could be achieved by storing it at 4 ℃ in darkness and avoiding direct contact with Cu2+ or Zn2+. Conclusion H. sabdariffa anthocyanin microcapsules with a significantly improved shelf life were successfully prepared by the optimized microencapsulation and spray-drying.
2021, 36(1): 115-123.
doi: 10.19303/j.issn.1008-0384.2021.01.014
Abstract:
Variants of anthocyanins content in grain hull gives brown rice the appearance of varied colors such as black, purple, red, green, yellow, etc. These varieties of rice are, therefore, categorically termed as “colored rice”. The beneficial and functional components in colored rice include amino acids, functional lipids, dietary fiber, vitamins, minerals, anthocyanins, phenolic compounds and γ-oryzanol. Such substances are either nutritionally or medicinally advantageous to the human well-being and valued by the consumers making colored rice highly popular on the market. For instance, in comparison to research activities on the regular varieties, programs specifically targeting colored rice are relatively few and commercial products emphasizing the functionality sparse. This article reviews the status on the germplasm resource, evaluation, utilization, functional ingredients, characteristics, genetics, breeding and application of colored rice. Obstacles hinder the industrial development are analyzed. Hence, in the attempt to expand the utilization and meet the market demand of colored rice, systematic and effective studies on the germplasm genetics for advancements in breeding and the rice chemistry for new product development would be indispensable.
Variants of anthocyanins content in grain hull gives brown rice the appearance of varied colors such as black, purple, red, green, yellow, etc. These varieties of rice are, therefore, categorically termed as “colored rice”. The beneficial and functional components in colored rice include amino acids, functional lipids, dietary fiber, vitamins, minerals, anthocyanins, phenolic compounds and γ-oryzanol. Such substances are either nutritionally or medicinally advantageous to the human well-being and valued by the consumers making colored rice highly popular on the market. For instance, in comparison to research activities on the regular varieties, programs specifically targeting colored rice are relatively few and commercial products emphasizing the functionality sparse. This article reviews the status on the germplasm resource, evaluation, utilization, functional ingredients, characteristics, genetics, breeding and application of colored rice. Obstacles hinder the industrial development are analyzed. Hence, in the attempt to expand the utilization and meet the market demand of colored rice, systematic and effective studies on the germplasm genetics for advancements in breeding and the rice chemistry for new product development would be indispensable.
2021, 36(1): 124-134.
doi: 10.19303/j.issn.1008-0384.2021.01.015
Abstract:
Adverse external conditions commonly affect plant growth and development which directly or indirectly cause decline on crop yield and even death of the plants. The homeodomain-leucine zipper (HD-ZIP) transcription factors have been known to involve in stress responses of plants. Belonging to the homeobox (HB) protein family, the factors are unique to plants and tightly connected by the highly conserved HD and ZIP. The formation of protein dimers mediated by the LZ domain allows HD to bind to the target DNA and regulate the expression of the target gene. The HD-ZIP transcription factors not only play an important role in regulating plant growth and development but also in responding to external stresses. This article focuses on the published reports of recent studies concerning the roles of the 4 subfamilies Ⅰ-Ⅳ of the HD-ZIP transcription factors in response to pathogenic attacks and/or abiotic stresses such as drought, salt, extreme temperature, wounding, low R/FR light, and heavy metals. Through the internal molecular response mechanisms initiated by HD-ZIP, a plant could ward off the imposed adversities. With an in-depth understanding of the functions, means to improve the growth and stress resistance of plants could be realized.
Adverse external conditions commonly affect plant growth and development which directly or indirectly cause decline on crop yield and even death of the plants. The homeodomain-leucine zipper (HD-ZIP) transcription factors have been known to involve in stress responses of plants. Belonging to the homeobox (HB) protein family, the factors are unique to plants and tightly connected by the highly conserved HD and ZIP. The formation of protein dimers mediated by the LZ domain allows HD to bind to the target DNA and regulate the expression of the target gene. The HD-ZIP transcription factors not only play an important role in regulating plant growth and development but also in responding to external stresses. This article focuses on the published reports of recent studies concerning the roles of the 4 subfamilies Ⅰ-Ⅳ of the HD-ZIP transcription factors in response to pathogenic attacks and/or abiotic stresses such as drought, salt, extreme temperature, wounding, low R/FR light, and heavy metals. Through the internal molecular response mechanisms initiated by HD-ZIP, a plant could ward off the imposed adversities. With an in-depth understanding of the functions, means to improve the growth and stress resistance of plants could be realized.
