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2024, 39(6): 623-632.
doi: 10.19303/j.issn.1008-0384.2024.06.001
Abstract:
Objective Chronic ammonia nitrogen stress on juvenile crucians were studied with observations on liver and kidney histology and non-specific immunity of the fish for an improved aquaculture water quality control. Method Juvenile Carassius auratus with a body mass of (3.10±0.15) g were raised in an environmentally controlled pond. By an acute toxicity test on the fish, the semi-lethal (LC50) and safe concentrations (SC) of ammonia nitrogen over 96 h were determined. Accordingly, a control at 0 mg·L−1 and three treatments groups (i.e., low concentration L at 6 mg·L−1, medium concentration M at 15 mg·L−1, and high concentration H at 24 mg·L−1) of ammonia nitrogen in water were applied to the pond with the juvenile fish for 60 d. Histopathological changes induced by the treatments in the liver and kidney as well as the alternations occurred to the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), catalase (CAT), and superoxide dismutase (SOD) of the fish were monitored continuously on the 15th, 30th, 45th, and 60th days. Result The acute ammonia nitrogen toxicity test on the fish showed the LC50 of 289.29 mg·L−1 and SC of 28.9 mg·L−1. The microscopic examination revealed the increasing ammonia nitrogen concentration and stress duration caused symptoms in the fish included blurred and disorganized hepatocytes with some vacuolated and cell nuclei dissolved in hepatic tissue, tumefacient renal tubular epithelial cells, narrowed renal tubular lumen, and atrophied glomerulus in nephridial tissue. The ALT and AST activities in the liver and the kidney rose with increasing ammonia nitrogen in water. The ALT in the livers of fish in L and M groups did not significantly differ from control on the 15th day (P＞0.05). On the other hand, the ALT and AST in livers and kidneys of all 3 treatment groups were significantly different from control at all testing times (P＜0.05). The specific activity of CAT and SOD in liver of the fish increased initially and decreased after a period of time of the treatments; that of CAT in treatment groups significantly differed from control on the 45th day; that of SOD significantly differed on the 15th day, and that of CAT in renal tissue significantly higher than that in control group on the same day; while that of SOD in all treatment groups significantly higher than that in control at all time periods. Conclusion The stress of prolonged high concentrations of ammonia nitrogen in aquacultural water caused severe damage to the liver and kidney tissues in juvenile crucians. The histopathological injuries could result in malfunctioned metabolism, compromised detoxification capacity, and reduced non-specific immunity of the fish. Consequently, for a healthy and productive aquaculture, it was deemed imperative the ammonia nitrogen content in the water be continuously monitored and strictly controlled.
2024, 39(6): 633-642.
doi: 10.19303/j.issn.1008-0384.2024.06.002
Abstract:
Objective To study the effects of modified Yupingfeng decoction on antioxidant, anti-inflammatory and immune function and anti-coccidial effect of Dehua black chickens infected with Eimeria tenella. Methods The treated 120 45-day-old Dehua black chickens were randomly divided into 6 groups, namely, blank control group (K group), infected control group (G group), low-dose group of Radix dichroa and Artemisia annua (CQL group), low-dose group of modified Yupingfeng decoction (YL group), high-dose group of Radix dichroa and Artemisia annua (CQH group), and high-dose group of modified Yupingfeng decoction (YH group). Except for the K group, all other groups were infected with Eimeria tenella. Treatment with the respective decoctions began 24 hours after infection and continued for 7 days. On the 8th day, various indices including anti-coccidial index, immune function, and antioxidant capacity were measured. Results Compared with the infection control group (G group), the symptoms of bloody stool and cecal lesions were alleviated in the CQH, YH, CQL, and YL groups in order. Except for the CQH group, the cecal lesion scores and submucosal lymphocyte density in the cecum were significantly or extremely significantly reduced (P＜0.05 or P＜0.01); oocyst per gram (OPG) was deduced extremely in the other three traditional Chinese medicine groups (P＜0.01), while OPG in CQH group was reduced significantly (P＜0.05); the cecuml epithelial cell shedding score was decreased extremely in YL group and YH group(P＜0.01). Compared with the CQL and CQH groups, the YL and YH groups showed extremely significant reductions in MDA, cecal lesion scores, epithelial cell shedding scores, and OPG levels (P＜0.01). Compared with G group, the contents of IL-6, IFN-γ, TNF-α and malondialdehyde (MDA) in the four Chinese medicine groups were significantly decreased (P＜0.01). In contrast, IL-2 content, SIgA level, and the activity of glutathione peroxides (GSH-Px), superoxide dismutase (SOD), Catalase (CAT) and total antioxidant capacity (T-AOC) were significantly increased (P＜0.01). IL-2 content, SIgA level, and the activity of GSH-Px, SOD, and CAT were significantly increased in the YL group (P＜0.01), and GSH-Px and SOD activities were significantly increased in the YH group (P＜0.01). Compared with K group, the relative weight gain rate of the G group, CQL group, YL group, CQH group, and YH group were 79.88%, 103.49%, 107.27%, 95.05%, and 96.06%, and the ACI were 96, 165.5, 177, 133.5, and 162.5, respectively. Conclusion The modified Yupingfeng decoction has better immunomodulation effect of improving anti-inflammatory, antioxidant and immune function on Dehua black chickens infected with Eimeria tenella, and correspondingly has better anti-coccidia effect, while the Yupingfeng modified low-dose group (YL) has the best anticoccidia effect.
2024, 39(6): 643-651.
doi: 10.19303/j.issn.1008-0384.2024.06.003
Abstract:
Objective Members of the Na+/H+ antiporter (NHX) family in Brassica napus L. were identified, and genes related to salt stress screened. Methods The genome of B. napus cv. Zhongshuang 11 was used to obtain the candidate genes, and NHXs sequences by homologous alignment. Physicochemical properties, evolutionary relationship, and expressions under salt stress of the genes were analyzed. Results Twenty-one BnNHXs were identified with 71 to 1265 amino acids, an isoelectric (pI) ranging from 5.54 to 7.68, and 0 to 24 introns. Among the BnNHXs, BnNHX1 was in the plasma membrane, BnNHX5 and BnNHX12 in the cell and vacuole membranes, and the remainders in the vacuole only. Phylogenetically, NHXs could be divided into 3 subfamilies scattered on 11 chromosomes containing a variety of abiotic stress response elements. The qRT-PCR expressions on most BnNHXs were upregulated under salt stress, with overall higher expression in the leaves than in the roots. Notably, BnNHX2, BnNHX6, BnNHX8, BnNHX11, and BnNHX19 had relatively more pronounced changes in the expression. Conclusion Five NHXs, namely, BnNHX2, BnNHX6, BnNHX8, BnNHX11, and BnNHX19, could be the candidate genes for breeding salt-tolerant new varieties as well as for studying molecular mechanisms of rapeseed.
2024, 39(6): 652-661.
doi: 10.19303/j.issn.1008-0384.2024.06.004
Abstract:
Objective Genetic variations in the southern high-protein soybeans were revealed by re-sequencing the whole genome. Method High-throughput sequencing was conducted on the whole genome of the southern high-protein soybean Fudou 234 to detect genetic variations. Result In the 64757037 clean reads, the sequencing depth was 17× with genome coverage of 98.08% (1×) and 96.25% (5×). There were 1478393 single nucleotide polymorphisms (SNPs) and 356739 small insertion-deletions (Small InDels) identified. Among them, 14323 non-synonymous SNP mutations and 4186 Small InDel mutated genes were found in the coding sequence (CDS). Analysis by COG (Clusters of orthologous groups of proteins) revealed that signal transduction mechanisms, transcription, carbohydrate translocation and metabolism and KEGG(Kyoto encyclopedia of genes and genomes) analyses revealed that the pathways of carbon metabolism, starch and sucrose metabolism, amino acid biosynthesis, phytohormone signalling, and endoplasmic reticulum protein processing were associated with genetic variation in Fudou 234. In addition, by studying the candidate genes in two major segments of soybean kernel protein quantitative trait locus (QTL), 10 SNP and 7 Small InDel type variations were discovered in 65 genes. Conclusion The genetic variations in the southern high-protein soybeans deviated from the regular varieties were unveiled to provide new venue for breeding and developing molecular markers in studying soybeans.
2024, 39(6): 662-670.
doi: 10.19303/j.issn.1008-0384.2024.06.005
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Objective Physiobiochemical changes and differentially expressed genes in Polygonatumcyrtonema Hua seeds after a post-ripening treatment were studied. Method Contents of superoxide dismutase, α-amylase, β-amylase, soluble protein, crude fat, and soluble sugar at 6 seed development stages were monitored. Differentially expressed genes were analyzed with high-throughput sequencing followed by a qRT-PCR verification. Result (1) The post-ripening treatment raised the soluble sugar content in the seeds gradually in the initial 4 stages and slightly declined in the last two stages, while induced a declining trend on the crude fat and protein. Meanwhile, the activities of α-amylase and β-amylase increased. (2) Significantly enriched differentially expressed genes were observed in the sucrose and starch metabolic pathways as well as the hormone signal transduction pathway during the process. Conclusion The post-ripening treatment on the P. cyrtonema seeds brought about the conversion and metabolism of starch and other nutrients as well as endogenous hormones interactions promoting the physiological changes and dormancy release.
2024, 39(6): 671-679.
doi: 10.19303/j.issn.1008-0384.2024.06.006
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Objective Indicators for distinguishing differentiate physiological traits of naturally occurring variants of Iris hollandica were determined. Methods According to the DUS testing guidelines on bulbous irises, botanical and agronomic traits of 4 wild-type variants of Chunzhimeng , namely MHS5-3, XCQ5-1, MZB7-2, and CZL7-3, were examined. Major ornamental characteristics of the plants were analyzed for the indicator selection. Results The morphological, flowering, and growth characteristics of these variants were not significantly deviate from those of the wild-type plants. On the other hand, among the 40 investigated traits, 7 pseudo-qualitative characteristics, including colors of bud, flag pedal, filigree, style, feathercrown, and upper and lower pendant petals , groud color of upper side of blade in outer tepal, color of Inner tepal, filament color, color of upper side in bridge, color of upper side in crest, and groud color of upper side of claw in outer tepal, significantly differed between the wild-type the variants. These traits showed a potential of being used as criteria for differentiating between the two, while the remaining 33 traits were phenotypically similar. Conclusion Being an apparent property of an ornamental flowering plant, red and blue, in particular, appeared superior to purple and white I. hollandica flower color for separating variants from the wild-type counterparts.
2024, 39(6): 680-688.
doi: 10.19303/j.issn.1008-0384.2024.06.007
Abstract:
Objective PropagatingStropharia rugosoannulata in liquid culture medium was studied. Methods S. rugosoannulata No. 8 was cultivated in experimental media to determine the mycelial biomass increase in a single factor and orthogonal test L9(34) for formulation and culture conditions optimization. Mycelial biomass, reducing sugars, and amino nitrogen content as well as the extracellular enzyme activities of carboxymethyl cellulase, amylase, acid protease, and laccase of the culture were monitored to determine cultivation end point. Average full-bag filling time was used as indicator for the inoculation amount, substrate particle size, and carbon-to-nitrogen ratio in a maximized reproduction and yield. Results The optimized liquid culture medium was formulated with 20 g·L−1 glucose, 30 g·L−1 wheat flour, 0.75 g·L−1 yeast powder, 1.00 g·L−1 potassium dihydrogen phosphate, and 0.50 g·L−1 magnesium sulfate at an initial pH of 5. On the 8th day of culture, the mycelial biomass reached a maximum at 1.66 g·h mL−1. As the mycelia grew, the reducing sugar in the medium decreased from 12.23 mg·mL−1 to 1.38 mg·mL−1 and the amino nitrogen from 0.09 mg·L−1 to 0.06 mg·L−1 during the culture process. The activities of carboxymethyl cellulase and amylase were the highest on the 4th day with the activities of 6.49 U and 5.16 U, respectively. The activity of acid protease peaked at 1.80 U on the 2nd day; and that of laccase, at 1.63 U on the 6th day. The inoculum production was best carried out with an inoculation of 15 mL, a coarse-to-fine ratio of 7∶3 on the substrate particle size, and a carbon-to-nitrogen ratio of 50∶1. Conclusion Since the viability of the mushroom propagation correlated with some of the physiological and biochemical indicators of the liquid medium, 7 d was determined to be the peak for the cultivation. For filling a hyphal bag, 23.7d of culture was required, which was 2.7d shorter than what needed without the optimization. The results provided the basis for the development of a scale-up industrial operation.
2024, 39(6): 689-699.
doi: 10.19303/j.issn.1008-0384.2024.06.008
Abstract:
Objective Role of VfNHX1 of faba bean in response to salt stress was studied. Methods A Na+/H+ reverse transporter protein-encoding gene, VfNHX1, was cloned from Vicia faba L. by 3' and 5' RACE for a bioinformatics analysis, subcellular localization determination, expression under salt stress, and a preliminary verification on the function. Results (1) VfNHX1 sequence was 2255 bp with a CDS coding region of 1629 bp encoding 542 amino acids. (2) As a stable transmembrane protein, VfNHX1 had 10 transmembrane regions without signal peptide and contained a typical Na-H exchanger conserved functional domain of NHX family. It was in the vacuolar membrane. (3) In the leaves, the expression of VfNHX1 under NaCl stress determined by qRT-PCR showed a trend of decreasing, then increasing, followed by decreasing with a peak reached in 12 h. In the roots, the expression declined initially and then rose significantly at 48 h (P＜0.01). (4) The presence of VfNHX1 in a yeast growth experiment indeed heightened the NaCl tolerance of the salt-sensitive yeast mutant AXT4K. Conclusion VfNHX1 in V. faba L. was able to respond to salt stress and could be a functional gene to boost the salt resistance of the plant.
2024, 39(6): 700-710.
doi: 10.19303/j.issn.1008-0384.2024.06.009
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Objective NACs in petunia responsible for the growth, floral senescence, and stress response were identified and analyzed. Method Based on the Petunia axillaris genome, transcriptomes on the flower in senescence as well as some other organs under the stress of inoculated tobacco rattle virus (TRV), low phosphorus, low temperature, NaCl, copper ion, or drought were obtained. Expression under stress, cis-acting elements, and transcription factor binding sites in promoters of differentially expressed PaNACs were analyzed. The expression in flower senescence was determined using qRT-PCR, and putative target genes of proteins encoded by them predicted. Result Of the 131 PaNACs, 59 (i.e., 45.04% of all) were identified as differentially expressed genes (DEGs) during flower senescence and in response to stresses. PaNAC72, PaNAC22, PaNAC29, PaNAC40, PaNAC2, PaNAC90, PaNAC83, PaNAC56, PaNAC36, and PaNAC35 exhibited significant differential expressions in response to at least 3 stress treatments. Among them, PaNAC29, an orthologue of the Arabidopsis key senescence-related gene AtNAP, was highly upregulated during flower senescence and in response to low temperature, low phosphorus, or copper ion treatment. PaNAC72 was significantly affected by all except copper ion treatment. PaNAC22 was upregulated during flower senescence and in responses to low temperature and low phosphorus treatments but downregulated in the presence of copper ion or under drought condition. Multiple stress responsive elements presented in the promoters of the 10 PaNACs, and many senescence and stress responsive DEGs containing NAC binding sites in their promoters. Conclusion NAC (NAM, ATAF and CUC) involved widely in the growth, development, and stress responses of plants. PaNACs in petunia, such as PaNAC29 appeared to be a key positive regulator of floral senescence, and PaNAC72 responsive to a wide variety of stresses.
2024, 39(6): 711-719.
doi: 10.19303/j.issn.1008-0384.2024.06.010
Abstract:
Objective Liupao Tea (Camellia sinensis var. sinensis cv. Liupao) was studied for the genes associated with anthocyanin synthesis for target breeding. Methods Anthocyanin were extracted with ethanol hydrochloride from Liupao Tea plants bearing purple or green buds. Transcriptome sequencing was performed using Illumina Hiseq 2500 high-throughput platform to identify differentially expressed genes, determine expression of the genes related to anthocyanin synthesis, and conduct GO and KEGG enrichment analyses on the extracts. The sequencing results were subsequently verified by fluorescence quantification PCR. Results The young leaves on the tea plants borne with purple buds contained sevenfold higher anthocyanin than those on the plants with green buds. The transcriptome of the genes of the purple buds had 165570 unigenes with an average length of 1450 bp. Of them, 243 related to the anthocyanin biosynthesis pathway, and 43 significantly differentially expressed between the two types of plants. The 43 differentiated genes encoded 14 key enzymes, i.e., phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), chalcone isomerase (CHI), cinnamate acid 4-hydroxylase (C4H), anthocyanidin reductase (ANR), 4-coumarate-CoA ligase (4CL), acetyl-CoA carboxylase (ACC), flavonol synthase (FLS), flavonoid-3′,5′-hydroxylase (F3′5′H), flavonoid 3′-hydroxylase (F3′H), flavanone-3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), anthocyanidin synthase (ANS), and leucoanthocyanidin reductase (LAR). Conclusion Thirty-four genes significantly upregulated in Liupao Tea plants with purple buds were found to be associated with 14 key enzymes encoding anthocyanin biosynthesis. They were speculated to play an important role in the bud color differentiation between the two varieties.
2024, 39(6): 720-729.
doi: 10.19303/j.issn.1008-0384.2024.06.011
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Objective Effects of pine wood nematode (PWN) invasion on Pinus massoniana were quantified based on multiomics. Method Among 4-year-old Masson pine plants, a treatment, PWN(+), by inoculating 5 000 Bursaphelenchus xylophilus and a control, PWN(−), with the injection of sterile distilled water into the tree trunks were subjected to a 14 d post-inoculation followed by a quantified study including phenomics, transcriptome, microbiome, and metabolome. Result The pine trees in the PWN(+) group significantly increased over control on the contents of the reactive oxygen species (ROS) by 3.2-fold and hydrogen peroxide (H2O2) by 1.7-fold (P＜0.05), on the expressions of c60547.graph_c0 and c82953.graph_c0 in the unigenes encoding stress responsive pathways (P＜0.05), as well as on the expressions of c64867.graph_c0 and c68789.graph_c0 in the terpenoid biosynthesis pathways; and significantly lower on the c81022.graph_c0 in syncytium formation (P＜0.05) and the microbial diversity (P＜0.05). Hypocreales was the dominant order of microorganisms in the PWN(+) group. The functional annotations and abundance information from the KEGG database were mainly related to the replication and repair pathways, DNA replication pathway (PATH:ko03030), and DNA replication protein pathway (BR:ko03032). There were 365 upregulated and 351 downregulated differentially regulated metabolites in the metabolome. Metabolites like phloretin, ursodiol, and carbenpenicillin were notably enriched in the ABC transporters pathway, arachidonic acid metabolism, flavonoid biosynthesis, and glycerophospholipid metabolism in the PWN-infected pine wood to resist the invasion. Conclusion After PWN was inoculated in Masson pine, the host responded with a series of complex defensive reactions initiated by various mechanisms. Through multiomics, some such interactions between the species were unveiled providing clues for combating forestry declination by the infestation.
2024, 39(6): 730-737.
doi: 10.19303/j.issn.1008-0384.2024.06.012
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Objective Being crucial to the development of rhizobium inoculants and the understanding on the associated mechanisms of soil nitrogen fixation, community structure of the endophytic microorganisms in root nodules of Astragalus sinicus L. was investigated. Method Endophytic microbial community in the root nodules of 7 cultivars of A. sinicus planted in Fujian was analyzed with the Illumina Miseq PE300 sequencing platform using 16S rRNA as the target gene. Result The microbial composition of endophytic community in the nodules of the different cultivars varied. For instance, the predominant genus in ZZ5 root nodules was Pseudomonas, but it was Mesorhizobium in other cultivars. Furthermore, high Shannon indices were found in the root nodules of MZ8, XY, and ZZ5 with diverse non-rhizobial endophytes like Pseudomonas, Stenotrophomonas, Duganella, Erwinia, etc. In contrast, high relative abundances of pathogens, such as Erwinia rhapontici and Pantoea ananatis PA13 existed in the root nodules of MZ8, XY, and ZZ5. Conclusions Significant microbial variations in the endophytic communities were evident among the different varieties of A. sinicus planted in Fujian. The predominant genus in the root nodules of ZZ5 was non-rhizobial, growth-promoting Pesudomonas, while it was rhizobia for the other cultivars.
2024, 39(6): 738-751.
doi: 10.19303/j.issn.1008-0384.2024.06.013
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Objective Explore the spatial distribution and influencing factors of soil organic carbon (SOC) in cultivated land in Fujian Province. Method Based on the data generated from over 30,000 survey sites on farmland in Fujian, Pearson correlation coefficient and random forest model were employed to derive key factors affecting the SOC. The geodetector model was used to analyze the spatial SOC distribution in the province. Result The data on SOC of the province in 2008 ranged between 0.12 and 67.28 g·kg−1 with a spatial pattern of being low in the southeast coastal areas and high in the west and central regions. The geodetector model was shown to render the most comprehensive and objective analysis among the three models tested. It concluded the climate-related conditions to be the major factors affecting the spatial differentiation of SOC on the farmland with top 6 rankings of: annual precipitation (0.168 5)＞annual average temperature (0.167 7)＞altitude (0.144 9)＞climate type (0.135 9)＞soil type (0.082 4)＞landform type (0.073 1). The interactive detectors further revealed the interaction between the annual precipitation and annual average temperature to exert the greatest influence on the SOC spatial differentiation (0.194 1), while the annual precipitation and soil type (0.192 3) and the annual precipitation and cultivated land use type (0.1918) followed. Conclusion Multiple factors affected the SOC on the farmland in Fujian in the past. For improving the spatial utilization efficiency and bettering the agriculture production layout on the land, it seemed imperative that all various factors highlighted in this study be taken into serious considerations.
