2019 Vol. 34, No. 5
Display Method:
2019, 34(5): 509-515.
doi: 10.19303/j.issn.1008-0384.2019.05.002
Abstract:
Objective Rice germplasms that are low in absorbing and accumulating Cd and Pb were selected for breeding resistant varieties. Method In a netted chamber, 60 genotypes of rice were planted in plastic pots in cement tanks and divided into 3 groups. Group Ⅰ was free of added heavy metals to be used as control; Group Ⅱ contained CdCl2·2.5H2O to simulate the 3rd grade quality of soil contaminated with Cd 1 mg·kg-1; and, Group Ⅲ of the same grade soil with Pb 500 mg·kg-1 by the addition of Pb(CH3COO)2·3H2O. Rice grains were harvested at maturity, dried, and stored for 3 months prior to Cd and Pb determinations in the unpolished rice samples using the inductively coupled plasma mass spectrometry (ICP-MS). Result (1) Significant variations in Cd absorption were found among the genotypes. Group Ⅰ had a Cd content ranging from 0.005 to 0.224 mg·kg-1, as compare to Group Ⅱ of 0.133-3.308 mg·kg-1, showing an average of 19.63-fold difference significant at P < 0.01. Among the varieties of rice, a significant correlation existed between the Cd contents of control and treatment at P < 0.01. The Cd-tolerant Taijing 8 only sustained a level below the acceptable threshold of Cd 0.2 mg·kg-1 for safe consumption. (2) The Pb absorption by the rice varied significantly as well. Group Ⅰ had Pb 0.010-0.288 mg·kg-1, while Group Ⅲ Pb 0.183-2.123 mg·kg-1, showing an average of 10.66-fold significant difference (P < 0.01). However, there was not an apparent correlation between the Pb contents of control and treatment. Jinzao 6 and Ningjing 216 exhibited in the trial a Pb tolerance with a sustained content below the 0.2 mg·kg-1 standard. (3) The accumulated Cd in Indica rice was significantly higher than that in Japonica, but not on Pb. (4) Based on color of the hull, the Cd and Pb absorption by red rice seemed to be the highest, followed by black, and the lowest white. Conclusion Basically, any soil below 3rd grade on the environmental quality is not recommended for rice planting. Since there were significant differences among rice genotypes on their tolerance to Cd and Pb contamination, a selection guideline was made available for breeding varieties that could be cultivated on lands moderately contaminated by Cd and Pb.
2019, 34(5): 516-524.
doi: 10.19303/j.issn.1008-0384.2019.05.003
Abstract:
Objective The core enzymes in anthocyanin anabolism were investigated, to provid a reference for further studies on the anthocyanin metabolism of G. bicolor and similar plants. Method Transcriptome of Gynura bicolor and G. divaricata were sequenced using the Illumina HiSeq 2 500 platform for the study. Result The Denovo mapping on 8 databases showed that 43 369 out of 80 798 unigenes annotated differences between the two wild vegetables. Among them, 104 unigenes were identified to relate to the anthocyanin anabolism based on two databases on the non-redundant and Swiss-Prot proteins. They included 10 chalcone synthases, 5 chalcone isomerases, 36 flavanone 3-hydroxylases, 19 dihydroflavonols, 9 anthocyanidin synthases, and 25 flavonoid 3-O-glucosyl transferases. According to the average FPKMs, 7 of the 15 differentially expressed genes (i.e., 1, 1, 3, 4, 2, and 4 genes from each of the 6 enzyme categories) between the 2 wild vegetables were found to be up-regulated, and 8 down-regulated. Conclusion There were 15 enzymes in the 104 unigenes appeared to closely associate with the anthocyanin synthesis in G. bicolor and G. divaricata.
2019, 34(5): 525-533.
doi: 10.19303/j.issn.1008-0384.2019.05.004
Abstract:
Objective This study was aimed to accurately reveal the genetic relationship among sun-cured tobacco germplasms, and improve the utilization and breeding efficiency of sun-cured tobacco resources. Method Genetic diversity of 36 sun-cured tobacco germplasms was analyzed using ISSR. Result From the samples, 113 DNA bands were amplified with 7 polymorphic primers selected from 100 ISSR primers. Among them, 91 bands (80.5%) were polymorphic. The average effective number of alleles (Ne), Nei's gene diversity index (H), and Shannon's information index (I) of the ISSR markers were 1.64, 0.37, and 0.55, respectively, indicating a high detection efficacy. The UPGMA analysis showed a low diversity among the germplasms with the genetic similarity coefficients (GS) ranging from 0.74 to 0.91. With the new method, the germplasms were clustered into 5 groups at the level of 0.78.The digital fingerprints effectively distinguish the germplasms. Conclusion The ISSR markers chosen for the DNA fingerprints of 36 sun-cured tobacco germplasms were appropriate for the otherwise difficult task of differentiating and identifying those genetically closely related varieties.
2019, 34(5): 534-543.
doi: 10.19303/j.issn.1008-0384.2019.05.005
Abstract:
Objective Salt tolerance of blueberry plants were studied for possible cultivation in saline-alkali lands. Method A pot experiment was conducted using varied salt additions [0(CK), 30, 60, 90, 120, 150, 180, and 210 mmol·L-1)] NaCl in a medium to artificially simulate the environmental stress. Two-year-old seedlings of semi-dwarf Vaccinium Northland blueberry were treated under the stress for 50d. Effects on the physiological and biochemical properties, as well as the photosynthesis, net photosynthetic rate (Pn), transpiration rate (Tr), intercellular CO2 concentration (Ci), and stomatal conductance (Gs), were determined. Result (1) A salt concentration at 30, 60 or 90 mmol·L-1 promoted the seedling growth, but the enhancing effect disappeared when it reached 120 mmol·L-1, where the result became similar to CK. Further increase to 150 or 180 mmol·L-1 inhibited the seedling growth. At 210 mmol·L-1, a threshold that caused plant death was observed. (2) When the salt concentration was less or equal to 90 mmol·L-1, the blueberry seedlings grew better than CK with increased plant height, crown width, dry and fresh weights of shoots, and soluble sugar content in leaves. The biochemical indices, such as Pro, superoxide dismutase (SOD) activity, relative permeability of cell membrane, and malondialdehyde (MDA) content, were elevated as well. (3) The daily variations on Pn, Tr and Gs of the treated plants were on an upward trend, but Ci downward. The responses of Pn, Tr and Ci to light exposure showed an upward trend, while that of Gs downward. These effects were most significant under 60 mmol·L-1 NaCl. Conclusion The semi-dwarf blueberry bushes exhibited a significant salt tolerance within 0-180 mmol·L-1. Northland blueberry was conceivably a variety that could be cultivated on soil that has a salt content below 90 mmol·L-1.
2019, 34(5): 544-550.
doi: 10.19303/j.issn.1008-0384.2019.05.006
Abstract:
Objective The ketoacyl-coenzyme A synthase (KCAS) gene relating to responses of Primulina spinulosa to drought and other stresses was studied based on the transcriptome sequencing. Method Total RNA of the gene was extracted by the improved Trizol method for sequencing with the established database. Physicochemical properties of the open reading frame (ORF), coding amino acids, and proteins of the gene were analyzed by DNAman, Finder, TMPRED, SWISS-MODEL, NetPhos 2.0, and SignaIP4.1. The phylogenetic tree was constructed accordingly. Results The full length of the KCAS gene was determined to be 1 935 bp (GenBank login number MH543314), ORF 1 599 bp, encoding 532 amino acids, molecular weight 59.82 kDa, and theoretical pI 9.16. It was a hydrophilic protein with 7 transmembrane regions, 30 possible phosphorylation sites, and one conservative domain similar to that of the chalcone synthase family. The homology between it and Dorcoceras hygrometricum (KZV788) was as high as 92%, and they were in a same phylogenetic tree. Conclusion The KCAS gene was postulated to relate to the leaf wax synthesis of P. spinosa and played an important role in drought resistance and biological response to stresses of the plant.
2019, 34(5): 551-559.
doi: 10.19303/j.issn.1008-0384.2019.05.007
Abstract:
Objective For introducing and breeding new varieties. Method Ornamental value of 28 Camellia cultivars was evaluated, key appealing factors of Camellia were selected using the correlation and principal component analysis to construct a comprehensive evaluation model. Result Significant correlations among the various traits were observed. For instance, the flower count and petal number correlated significantly with florescence, and flower color with new leaf color. Those 13 characteristic traits were grouped into 5 principal components:flower count, florescence, flower diameter, leaf color, and leaf shape with a total cumulative contribution of 85.48% for the differentiation. Flower count and florescence were critical with biological significance as well. Based on their ornamental values, the 28 varieties were classified into 3 grades. The 9 varieties judged to be excellent were Jinbihuihuang, Variegated Precious Pearl, Variegated Crane's Feather, Variegated Hibiscus, Kigirohi, Senritsu-Ko, Otome, Red Leaf Bella, and Masterpiece. The results were basically in agreement with field observation. Conclusion The ornamental value of Camellia was governed by the flower count, florescence, flower diameter, leaf color and leaf shape with the flower count and florescence as the determinant factors. Thus, varieties with large and abundant flowers, long florescence period, and attractive leaves should be the choice for cultivation and breeding.
2019, 34(5): 560-566.
doi: 10.19303/j.issn.1008-0384.2019.05.008
Abstract:
Objective The physiological and biochemical properties of Myrica rubra cv. Shuijingzhong fruits during development were studied. Method Indicators, including color, individual weight, dimensions, shape, TSS, and contents of soluble sugars and organic acids, of the fruits were determined in 18 consecutive days for the analysis. Results During the developmental stages, the fruit color changed slowly with increasing L* till reaching a constant and from a negative to a positive a*/b* ratio. The weight, length, and girth of the fruit all on an increasing trend as it grew. The contents of TSS, fructose, and glucose accumulated rapidly to peak at maturation. Malic acid increased initially followed by a decline and another increase, while citric acid and oxalic acid declined continuously. The L* and a*/b* of the fruits significantly correlated with TSS and sucrose contents, but inversely correlated with oxalic acid. The weight, length, and girth of a fruit significantly correlated with TSS, fructose, glucose, and sucrose contents, while significantly inversely correlated with citric acid and oxalic acid. Conclusion The color differences (L* and a*/b*) and quality indices (e.g., individual weight, length, and girth) closely related to the intrinsic properties of the fruits. Therefore, these external measurements reflected the fruit development and could be applied to determine the appropriate timing for harvest.
2019, 34(5): 567-574.
doi: 10.19303/j.issn.1008-0384.2019.05.009
Abstract:
Objective Enhancement on the development and application of exopolysaccharides produced economically from photosynthetic bacteria fermentation was investigated. Method A strain of polysaccharide-producing photosynthetic bacteria, HP1, was isolated from a shrimp aquaculture pond. Crude polysaccharides were prepared from fermentation broth and precipitated by ethanol. Using astragalus polysaccharides for comparison, effects of inclusion of the polysaccharides in feed on nutrients in the meat of Ophiocephalus argus fed for 30 d were examined. Result The physiological and biochemical characteristics as well as 16S rDNA sequencing of HP1 identified it to be Rhodopseudomonas faecalis. The polysaccharides produced from HP1 fermentation had no significant effect on muscle pH (P>0.05), but significantly decreased the drip loss and muscle fiber diameter of the fish after the feeding (P < 0.05). In the meat, the crude protein did not differ significantly by the diet change (P>0.05), but the amino acids did (P < 0.05). Compared with control without polysaccharide addition in the forage, the contents of total, essential, and delicious amino acids in the fish muscles increased by 8.7%, 11%, and 9%, respectively, crude fats by 10.7%, EPA+DHA by 12.8%, and the ratio of n-3/n-6. Conclusion It appeared that a diet included the exopolysaccharides not only improved the eating quality but also enhanced the nutritional value of O. argus with effects comparable to astragalus polysaccharides.
2019, 34(5): 575-580.
doi: 10.19303/j.issn.1008-0384.2019.05.010
Abstract:
Objective Activity of porcine antibacterial peptides (PAP) against avian Salmonella and Escherichia coli was determined for drug development. Method Inhibitive activity of PAP on avian pathogens was studied in vitro on an liquid medium and by an in vivo test. Results PAP at a concentration of 1.0 mg·mL-1 showed a higher bacteriostatic effect on the solution contaning 106 CFU·mL-1 of the pathogens than at 0.5, 0.2 or 0.1 mg·mL-1. Compared with 15 antibiotics, such as amoxicillin, erythromycin, tetracycline, etc., PAP induced the largest inhibition zone on the growth of avian Salmonella FJS13 and E. coli FJE61. In a challenging test on chicken, the PAP administration improved the survival rate of the subcutaneous injected birds. Conclusion PAP could be used as a bioagent to combat the infection on chickens by avian Salmonella and E. coli.
2019, 34(5): 581-586.
doi: 10.19303/j.issn.1008-0384.2019.05.011
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Objective Pathogenicity of various strains of Plasmodiophora brassicae on Brassica napus cultivars was investigated. Method Four different P. brassicaes strains were collected from the counties of Xixiang, Nanzheng, Mian, and Taibai in Shaanxi, and the disease-resistance of 15 varieties of B. napus toward them by inoculating in the roots determined in an indoor test. Result The pathogenicity of the various strains were significantly different.The P. brassicaes collected from Mian county was the strongest among all, while from Nanzheng county the weakest. The disease-resistance of the 15 rapeseed varieties differed significantly as well. The 5 significantly more resistant varieties, 62R, Huashuang 5R, 1628A×genkang, 171R, and 172R, were not native Shaanxi cultivars. The cluster analysis classified them as disease-resistant toward the clubroot disease and the 10 local germplasms disease-sensitive. Conclusion Physiological differences existed among the strains of P. brassicaes from the 4 regions in Shaanxi. The 5 identified clubroot disease-resistant rapeseed cultivars would be promising candidates for breeding new varieties for cultivation in the province.
2019, 34(5): 587-594.
doi: 10.19303/j.issn.1008-0384.2019.05.012
Abstract:
Objective To optimize the polyphenol extraction from leaves of jaboticaba and determine the α-glucosidase inhibition by the extract. Method The ethanol extraction with ultrasound application was optimized by an orthogonal experiment. Effects of the power and time of ultrasonic application as well as the ethanol volume fraction, material-solvent ratio, and processing temperature on the total polyphenols extracted from the leaves were analyzed. The inhibition on α-glucosidase activity by the extract was determined in vitro by an enzyme kinetic study and uv-vis absorption spectrometry. Result The optimal extraction conditions included the use of 120 W ultrasonic wave for 10 min, 40% ethanol volume fraction, 1:160(g:mL) material-solvent ratio, and 40℃ processing temperature to achieve a polyphenol extraction rate of 33.84%. The α-glucosidase activity was significantly inhibited by the extract with a low IC50 of 0.0031 mg·mL-1, as compared to IC50 of 8.110 8 mg·mL-1 on acarbose. The kinetic study by the Lineweaver-Burk plot showed that the polyphenol extract was an anti-competitive inhibitor of α-glucosidase. Conclusion The polyphenol extract from jaboticaba leaves significantly inhibited α-glucosidase and could be explored for use as a hypoglycemic agent.
2019, 34(5): 595-599.
doi: 10.19303/j.issn.1008-0384.2019.05.013
Abstract:
Objective To prepare monoclonal antibody of salidroside for the development of an indirect competitive enzyme linked immunosorbent assay (ic-ELISA) methodology to accurately and rapidly detect salidroside in medicinal materials. Method Balb/c mice of 6-8 weeks old were immunized with the prepared salidroside-BSA. Specificity of the anti-serum was determined by ELISA. Spleen cells from the mice with positive result were fused with SP2/0 myeloma cells. Monoclonal hybridoma cells were screened by indirect ELISA with a limited dilution. Ascitic antibodies were induced and prepared from the positive cell line. Specific antibody against salidroside was used to establish ic-ELISA detection method that was verified by its specificity, precision and recovery rate on a known standard. Result The sensitivity of ic-ELISA was 49.33 ng·mL-1 with a liner range of 4.07-598.45 ng·mL-1. The detection limit of the method was 1.77 ng·mL-1. The newly developed method detected salidroside in spiked samples with a recovery rate of 94.3% within the designed range. The relative standard deviation of the measurements was less than 4.5% on both intra- and inter-day assays. The determination by ic-ELISA agreed with that obtained by HPLC. Conclusion The newly established ic-ELISA method based on the salidroside monoclonal antibody was considered appropriate for the detection.
2019, 34(5): 600-605.
doi: 10.19303/j.issn.1008-0384.2019.05.014
Abstract:
Objective To construct a vector for expressing α-cyclodextrin glycosyltransferase (α-CGTase) gene in Bacillus subtilis. Method The α-CGTase gene from Gebacillius sp. CHB1 was amplified by PCR. The EcoR Ⅰ-digested pBES and Xho Ⅰ-digested α-CGT gene were connected and transformed into B. subtilis RIK1285. Subsequently, fermentation of the recombinant B. subtilis RIK1285/pBE-CGT was optimized. Result (1) The α-CGTase gene was expressed in a fermentation medium to show an enzymatic activity of 2.9 U·mL-1 by B. subtilis RIK1285/pBE-CGT. (2) Medium TB with the formula of 0.5% glycerol, 1.2% peptone, 2.4% yeast extract, 1.64% K2HPO4, and 0.23% KH2PO4 was found to be optimal for the fermentation. After fermentation in TB at 37℃ for 24h, the α-CGTase activity reached 5.3 U·mL-1, which was 8-fold of what the wild Gebacillius sp. CHB1 could generate. Conclusion The engineered B. subtilis RIK1285/pBE-CGT was successfully obtained and the fermentation process optimized.
2019, 34(5): 606-612.
doi: 10.19303/j.issn.1008-0384.2019.05.015
Abstract:
Objective To effectively manage nitrogen (N) fertilization for oil tea camellia (Camellia. Oleifera) plantations, various fertilizer types were evaluated. Method In addition to control (CK), the agricultural compounded fertilizer (NFF), specially compounded fertilizer (ZFF), and specially compounded organic fertilizer (ZYJ) were applied on plantation land to study the N utilization by the plants and the N pool in the soil. Result The fertilizers performed significantly different on the content of total N (TN), alkali-hydrolyzable N (AN), NH4+-N, NO3--N, dissolved organic N (DON), and microbial N (MBN) in soil, as well as on the N content in leaves and the yield of fresh fruits.Among the fertilizers, NFF was found to deliver the best result on NH4+-N in soil, while ZYJ on TN, leaf TN, and fruit yield as well as the soil N. Significant correlations among soil N, leaf TN, and fruit yield were observed. Conclusion Therefore, ZYJ seemed most desirable for the fertilization at C. oleifera plantations.
2019, 34(5): 613-620.
doi: 10.19303/j.issn.1008-0384.2019.05.016
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Objective Effect of adding soft rocks to aeolian sandy soil in varied ratios on the vertical distributions of carbon (C), nitrogen (N), and texture of the soil was studied for improving fertility of the wasteland. Method At Fuping, Shaanxi, soft rocks were blended to the local sandy soil in different ratios, i.e., 0:1 (CK), 1:5 (C1), 1:2 (C2), and 1:1 (C3) by volume to determine the C/N ratio and texture of soil in different depths. Result The organic C and total N contents decreased with depth upon the additions. The average C content ranged from 1.45 g·kg-1 to 2.70 g·kg-1, and it was significantly higher in the 0-10 cm layer than 20-30 cm layer (P < 0.05). The mixing ratio had a significant effect on C, as the soft rocks increased (i.e., C3) so was C content in the top layer. The total N ranged 0.28-1.31 g·kg-1, and that in the 0-10 cm layer significantly higher than in the deeper soils. In the top layer, C3 rendered significantly more N than other treatments. The average C/N ranged 1.72-5.92 with the lowest in the 0-10 cm layer and a decline upon increased addition of soft rocks. The varied mixing ratios did not affect the nitrate N and ammonium N contents, which ranged 33.56-197.00 mg·kg-1 and 5.51-70.02 mg·kg-1, respectively, with the top layer being the highest. Sand content in the soil gradually increased with depth, while silt and clay particles decreased. Along with the addition of soft rocks the soil texture changed from sand to loamy sand, and to sandy loam. There was a significant inversed correlation between C/N and nitrate N and ammonium N, and a correlation with the particle size distribution in soil. The correlations were most significant in the 10-20 cm soil layer. Conclusion The blending ratio of soft rocks and sand at 1:1 seemed to promote the C and N accumulation in the 0-10 cm layer of soil. It also enhanced the microbial degradation in the soil.
