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Development of microencapsulated rhizosphere growth promoting bacteria and evaluation of the growth promoting effects on corn
LIU Wenjia, LIU Tianyi, ZHANG Limin, XU Chang, MO Jixian
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  Objective  In order to improve the stability of PGPR in promoting crop growth, microcapsule microbial agents were developed using PGPR composite bacteria as raw materials, and its growth-promoting effects on corn were investigated.  Method  Isolation of PGPR strains from corn rhizosphere soil and preparation of composite liquid microbial agents. Sodium alginate (SA) and CaCl2 were used as embedding and crosslinking agents to embed the composite bacterial solution and prepare microcapsule microbial agents. Based on the difficulty level, strain embedding rate, and pre-proliferative and post-proliferative viable bacterial count in the preparation of composite microcapsule microbial agents, the optimal conditions for preparing microcapsule microbial agents were determined. The excellent characteristics of PGPR microcapsules were determined by analyzing the strains growth in PGPR microcapsules under different concentrations of PEG, temperature, pH, storage time, and salt concentration, as well as their effects on corn growth and root morphology.   Result  This study isolated two PGPR strains with various plant growth-promoting functions. The microcapsule microbial agents made with 1% SA-3% CaCl2 had good stability, with an embedding rate of 91.27%. After proliferation, the number of live bacteria in the microcapsule reached 8.73 × 109 cfu·g−1, with a proliferation rate of 7.79 times. Under different concentrations of PEG, temperature, pH, storage time, and salt concentration, the concentration of microencapsulated bacterial solution was higher than that of liquid bacterial solution. PGPR microcapsule microbial agents had significant promoting effects on corn growth. Applying microcapsule microbial agents increased corn plant height, seedling dry weight, and root dry weight by 91.83%, 81.82%, and 29.57%, respectively. At the same time, microencapsulated microbial agents significantly increased the total root length, root surface area, average root diameter, root volume, and root tip number of corn.   Conclusion  This study determined a preparation method for PGPR microcapsule microbial agents. The application of the PGPR microcapsule agents significantly promoted corn growth, improved corn root morphology, and had better growth-promoting effects on corn than liquid microbial agents.
Determination of enrofloxacin and ciprofloxacin residues in aquatic products by Surface-enhanced Raman Spectroscopy
LIU Wen-Jing, PAN Wei, LIN Hui-Zhen, CHEN Hong-Ju, YAN Shao-De
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:   Objective  A rapid method for detection of Enrofloxacin (ENR)and ciprofloxacin (CIP) residues in vegetables and fruits by Surface-enhanced Raman spectroscopy (SERS) was established.  Method  The best detection conditions were obtained by screening of enhancement reagent, optimizing extraction solvents and reaction time using fruit and vegetable products as the research objects.The silver nanoparticles with a particle size of 50±5 nm was used as the SERS enhancement reagent, 200 g·L−1 NaCl solution was used as the extraction solution, and the results were interpreted within 3 min. The linear regression equation of peak intensity and concentration was established, and the linear relationship and detection limit were established to evaluate the method.  Result  The results indicate that, the Raman spectrograms peaks of ENR and CIP are 532 cm−1、552 cm−1、651 cm−1、737 cm−1and 785 cm−1, of which 532 cm−1 is caused by C-N bending vibration, and 552 cm−1 is the out-of-plane bending vibration, 651 cm−1 is attributed to C-N, C-C-F out-of-plane bending vibration and C-C=O in-plane bending vibration .737 cm−1 is attributed to the stretching vibration of C-H on the benzene ring, C-C=O out-of-plane bending vibration, C-C-N bending vibration and C-H, C-N stretching vibration, and 785 cm−1 is attributed to C-H, C-N out-of-plane bending vibration. By quantifying the characteristic peak(737 cm−1), the Raman intensity and the concentration have a good linear relationship within 10 ng·mL−1~200 ng·mL−1, and R2 > 0.96, the recovery rate was 78.4%~106.7%, the RSD was 2.1%~6.7%, and the method detection limit was 10 μg·kg−1.   Conclusion  The method is accurate, rapid, stable, and can achieve ENR and CIP in aquatic products in the field for rapid screening and detection.
Preliminary Research on the Mechanism of SOCS3 Negatively Regulating the Expression of Interferons Induced by Influenza Virus
PENG Benqun, HU Jingyun, MAO Yanan, WANG Shulin, WANG Jiajun, CHEN Mengying, YOU Dongxue, WANG Song
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  Objective  The regulatory impact of SOCS3 on the interferon signaling pathway during influenza virus infection was studied.   Method  The A549 cell lines with SOCS3 overexpression and knockdown were constructed using lentivirus infection and siRNA technology, respectively. Subsequently, these cell lines, along with control cell lines, were infected with influenza virus. Samples were collected at different time points to assess the expression and activation of crucial molecules within the interferon signaling pathway using RT-PCR and Western blot analysis.   Result  The findings demonstrated a decrease in expression levels of type I interferon IFN-β and type III interferons IL-28 and IL-29 following the overexpression of SOCS3 in cells. Conversely, the expression levels of IFN-β, IL-28, and IL-29 were observed to increase upon the knockdown of SOCS3 expression. Subsequent investigations indicated that the overexpression of SOCS3 suppressed the expression of interferon regulatory factor IRF7, as well as pattern recognition receptors RIG-I, MDA5, and TLR3 responsible for detecting influenza virus RNA. Additionally, the phosphorylation level of STAT1 after virus infection was detected, and the results showed that overexpression of SOCS3 in cells inhibited the phosphorylation of STAT1 induced by influenza virus, while knocking down the expression of SOCS3 increased the phosphorylation level of STAT1.   Conclusion  Upon influenza virus infection, SOCS3 down-regulated the mRNA expression of pattern recognition receptors and interferon regulatory factor, inhibited the production of type I and III interferons, as well as the activation of STAT1. This ultimately led to the blocking of interferon signal transmission.
Study on the identification method of pollen viability of passionflower germplasm resources
XIANG Yanyan, HUANG Weijie, LAI Jiang Long, TANG Li Mei, LIN Tingxing, LI Liang, WEI Xiuqing, XU Jiahui
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:   objective  Clarifying the best in vitro medium for Passiflora plant pollens, the optimal observation time for in vitro culture, the optimal pollens staining method and pollens storage conditions will help to quickly and accurately identify the pollens viability of Passiflora germplasm resources. It provides scientific basis for the utilization of germplasm resources and the breeding of new varieties.  methods  The effects of sucrose, H3BO3, Ca(NO3)2 and polyethylene glycol-4000 (PEG-4000) on pollens germination rate in vitro were studied by orthogonal experiment with 6 passionflower pollens samples as experimental materials. The best pollens culture medium was used to screen the optimal observation time for pollens culture in vitro. I2-KI staining, TTC staining and Alexander staining were used to explore the most suitable passionflower powder staining method. The best storage conditions of pollen were selected by different storage temperature and time.  results  The optimum medium formula was selected by orthogonal test as follows: 100 g·L−1 sucrose+0.02 g·L−1 H3BO3+0.04 g·L−1 Ca(NO3)2+150 g·L−1 polyethylene glycol (PEG-4000)+200 g·L−1 MgSO4·7H2O+100 g·L−1 KNO3, pH 5.5. The optimum observation time for pollens in vitro culture was 1 h. TTC staining method had good staining effect and had no significant difference with in vitro germination rate, which could effectively detect the vitality of passionflower powder. Within 24 hours of pollens storage, the pollens viability at 25 ℃ and 4 ℃ was 30.48 % and 26.69 % respectively and the pollens viability remained 26 % ~ 29 % at 4 ℃ for 7 days.  conclusion  The optimum medium combination, in vitro culture time, pollens staining method and pollens storage conditions for passionflower pollens germination in vitro were selected. to provide a theoretical basis for rapid screening and variety improvement of passionflower pollens vigor in the future.
Construction and application of variety resources evaluation system for Crassulaceae plants
CHEN Yiquan, LUO Yuanhua, WU Jianshe, LIN Bing, YE Xiuxian
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:   Objective  To construct a comprehensive evaluation system for Crassulaceae variety resources, screen suitable varieties for cultivation in Fujian Province and provide theoretical basis for the scientific introduction and promotion of succulent plant cultivation.  Method  Based on the horticultural traits of 48 succulent plant varieties, the AHP hierarchical analysis method and K-Means clustering analysis method were used to construct a comprehensive evaluation system and classify the variety grades for Crassulaceae varieties from three aspects: ornamental quality, adaptability and potential application value.  Result  The AHP Analytic Hierarchy Process (AHP) determined 14 personality traits as evaluation indicators, with the weight of 5 factors including leaf color, degree of mesophyll texture, summer adaptability, shape and appearance, and flowering period accounting for 61.2% of the total ranking. They are the key factors for evaluating the comprehensive value.The K-Means clustering analysis method divided 48 varieties into four levels: excellent, good, medium, and average. Seven superior varieties, including Echeveri agavoides 'Ebony' , Graptopetalum 'Purple Delight', Echeveria ‘Alba Beauty’, Graptoveria 'Opalina', Echeveria nodulosa, x Sedeveria 'Blue Elf' and Echeveria 'Peach Pride' are excellent at level I, while 14 superior varieties including Crassula rupestris f. are good at level II, accounting for 43.75% of the total varieties. It can be focused on breeding and promoting cultivation.   Conclusion  Applying AHP and K-Means clustering analysis method to construct a comprehensive evaluation system for the Crassulaceae plants, which effectively meets the requirements of comprehensive evaluation for Crassulaceae varieties. The evaluation results are basically consistent with the field observation results of cultivation experiments. This evaluation system can provide reference for the planting and selection of succulent plants.
Identification and Expression Analysis of TIFY Gene Family in Isodon rubescens
LIAN Conglong, LAN Jinxu, YANG Jingfan, LI Jingjing, YANG Hao, CHEN Suiqing
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  Objective  TIFY proteins are key regulators of JA signaling pathway, which also play significant regulatory roles in plant growth and development, abiotic stress, and secondary metabolites accumulation. Study the TIFY genes of Isodon rubescens can provide a theoretical basis for the improvement breeding of stress resistance and the synthesis of secondary metabolites in I. rubescens.   Method  In this study, the TIFY gene family was identified and analyzed by bioinformatics methods based on the full-length transcriptome database of I. rubescens, and the expression characteristics of the TIFY gene family in different tissues were analyzed by RT-qPCR.   Result  (1) A total of 12 TIFY genes were identified in the full-length transcriptome data of I. rubescens. (2) Physicochemical properties analysis showed that the amino acid length of IrTIFYs ranged from 124 aa to 378 aa. The molecular weight ranges from 13924.89 to 39692.38 Da. The isoelectric points were rang from 5.05 to 9.69; All the proteins were belong to stable proteins except IrTIFY10. The subcellular location analysis showed that all IrTIFY proteins were located in the nucleus, and all TIFYs are belong to hydrophilic proteins without signal peptides. (3) The structural analysis showed that none of the IrTIFY proteins had a transmembrane structure, and random curling was the most abundant structure type in the secondary structure. All proteins contains multiple phosphorylation sites. (4) Codon preference analysis showed that the codon preference of IrTIFY gene family was weak, and the codon ending with A or U was slightly inclined to be used. (5) The cis-elements analysis of IrTIFYs promoter showed that there were many light responsive cis-elements, hormone responsive cis-elements, and stress responsive cis-elements in the TIFY family. However, there were differences in the numbers and types of cis-elements among different IrTIFY members.(6)The phylogenetic tree analysis showed that 12 IrTIFYs were divided into four subfamilies: PPD (IrTIFY2), ZML (IrTIFY3/8/10), TIFY (IrTIFY7/12) and JAZ (IrTIFY1/4/5/6/9/11). and IrTIFYs had the closest genetic relationship with Salvia miltiorrhiza, which also belong to the labiaceae family. (7) RT-qPCR analysis showed that the expression levels of 12 TIFYs exhibited same trend with leaf > stem > root of different tissues, and most of the members showed significant differences between different tissues.   Conclusion  Based on the above results, it is speculated that IrTIFYs play an important regulatory role in the growth and development of I. rubescens, and may be involved in regulating the biosynthesis of secondary metabolites of I. rubescens, which lays a foundation and provides ideas for further research on the function of TIFY gene family in I. rubescens.
Genetic Analysis and Comprehensive Evaluation of 47 Vegetable Spring Soybean Germplasms
LIN Wenlei, LV Meiqin, LI Mingsong, SHI Yingying, KANG Rongrong, ZENG Hongying
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:   Objective  In order to make better use of fresh spring soybean germplasms and screen out the varieties (lines) with good quality to apply to the breeding and production of fresh soybean in Fujian province.   Method  The descriptive statistics, variation analysis, correlation analysis, principal component analysis, cluster analysis and comprehensive evaluation on 7 phenotypic traits as well as 13 major agronomic characteristics of 47 fresh spring soybean germplasms were sutdy.   Result  The results showed that: there were differences in phenotypic traits except for plant type and podding habit. Abundant variations were observed among the 13 major agronomic traits, and the variation coefficient ranged from 7.49% to 38.07%. Among them, the coefficient of variation of bottom pod height was the largest, and the coefficient of variation of standard pod length was the smallest. The correlation analysis showed that: the plant height was significantly positively correlated with bottom pod height, number of effective branches, pod weight per plant and standard pod yield. The stem diameter was significantly positively correlated with pod weight per plant, standard pod yield and number of effective branches. The number of effective branches was significantly positively correlated with pod weight per plant, standard pod yield and fresh pod yield. The effective pod number per plant and standard pod number per plant were all significantly positively correlated with the pod weight per plant. The pod weight per plant was significantly positively correlated with standard pod yield and fresh pod yield. The results of CASAR normalized maximum variance rotational principal component analysis showed that the information content of 13 major agronomic traits was 73.677%, which could be described by 4 major factors: pod yield factor, pod number factor, plant type factor and pod shape factor. According to the principal factor characteristic vector and its corresponding characteristic root, the comprehensive principal component score of each variety was calculated. The euclidean distance was determined by the method of the sum of squared deviations and systematic cluster analysis based on comprehensive principal component scores and standardized data of 13 major agronomic traits. 47 germplasms were divided into 2 major categories and 2 sub-categories through the former method, and divided into 3 major categories and 2 sub-categories through the latter method. By comparison, the analysis results of the two clustering methods were generally consistent. The first 10 of the principal comprehensive principal scores were comprehensively analyzed with the standard pod yield and fresh pod yield, Nannong 1821, Mindou No.14, Xinghuadou No.8, Xinghua 618, Mindou No.10, Xinghuadou No.9, Xinghuadou No.4 and Xinghuadou No.3 were screened out.   Conclusion  47 vegetable spring soybean germplasms had high genetic diversity, and the 8 excellent vegetable spring soybean germplasms selected should provide excellent parents and intermediate materials for vegetable spring soybean breeding.
Isolation, identification, biological characteristics, and indoor drug screening of the pathogenic bacteria of Atractylodes macrocephala root rot disease
GU Qingyi, ZHANG Yaozhou, Wu Xiaoya, HUANG Yaqin, QIAO Xinrong, SHEN Jun
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:   Objective  In order to identify the pathogenic genera of Atractylodes macrocephala root rot disease in Xinyang, Henan Province, study their biological characteristics, and screen for Botanical fungicides that can be used for prevention and control.  Method  Morphological and multi gene analysis were used to identify the pathogens, and the biological characteristics of the pathogens were studied.   Result  The pathogen has white hyphae, oval shaped spores with slightly pointed ends, and large conidia with 3-5 septa and sizes of 7-10μm×3-4μm. Small conidia with 2 compartments or no compartments, size 3-5μm × 1-2μm. Multi gene joint analysis showed that the pathogen had high homology with Fusarium Fujikuroi. Combining morphological characteristics and molecular biology analysis, the pathogen of Atractylodes macrocephala root rot in Xinyang was identified as Fusarium Fujikuroi. This is the first report of Fusarium Fujikuroi causing Atractylodes macrocephala root rot. Research on biological characteristics shows that the optimal cultivation conditions for this pathogen are temperature 28 ℃, pH 7, sucrose as the optimal carbon source for growth, and potassium nitrate as the optimal nitrogen source for growth. Among the three botanical fungicides, the EC50 of 0.3% eugenol soluble agent is 6.906 mg/L, which is higher than other tested agents and has the strongest toxicity to pathogens..   Conclusion  The pathogen of Atractylodes macrocephala root rot in Xinyang, Henan Province is Fusarium fuciformis. The optimal cultivation conditions are temperature 28 ℃, pH 7, sucrose as the carbon source for growth, and potassium nitrate as the nitrogen source for growth. The research results provide a basis for the scientific prevention and control of root rot disease of Atractylodes macrocephala in Xinyang area.
Response Surface Optimization of baking parameters for high-quality tea charcoal
GAO Yusen, REN Jinbo, LIN Ting, WU Chuanyu
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  Objective  Using the charcoal baking process conditions of tea leaves as the research object, using the principle of fuzzy mathematical matrix analysis,To optimize the charcoal roasting process for tea leaves and obtain the best charcoal roasting technique,providing a reliable theoretical reference baking high-quality tea leaves and foundation for intelligent tea baking.  Methods  The fuzzy sensory evaluation score was taken as the response value,and the baking temperature (A),the spreading thickness (B) and the baking time (C) were taken as the independent variables to establish a combination of 3 factors and 3 levels,and a linear regression model was established to optimize the process of tea charcoal baking by response surface methodology.Conduct comparative analysis of actual and theoretical sensory evaluations through actual baking.The analysis of the impact of experimental techniques on the quality of tea further confirms the reliability of the quadratic regression model.  Results  Based on actual adjustments, the optimal combination parameters are as follows: baking temperature of 82 ℃, spreading thickness of 3 cm, baking time of 126 minutes. The main aroma components of tea include trans orange blossom tert alcohol, farnesene, plant alcohol, and indole, and their content is consistent with the aroma grade; The main biochemical components include tea polyphenols, soluble sugars, amino acids, and caffeine, and their content matches the color and taste level of the soup. The sensory evaluation results are consistent with the evaluation system.  Conclusion  The sensory evaluation standard of tea is reasonable and the sensory evaluation model is reliable,which can veritable reflect the best process of charcoal baking.
Analysis of Agronomic Characters and Agricultural Economic Benefits of Directly Replanting of the 3nd Year Ratoon Sugarcane
JIANG Zhuhui, ZHOU Huiwen, YAN Haifeng, LIANG Xueyou, SONG Shi, WEI Zhiying, QIN Lei, XIONG Faqian, QIU Lihang, YOU Jianhua
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Abstact:   Objective  In order to provide a basis for further promoting the new technology of extending the ratoon sugarcane seed stalk by direct replanting, the effects of extending the second year of ratoon on the traits and yield of sugarcane varieties were studied.   Method  GL2, GT44 and YT236, three major sugarcane varieties in Guangxi Province, were taken as the replanted population in the second year of ratoon. GF296 seed stalks were used for the direct replanting treatment on the ridge and furrow sugarcane beds with cultivated resistance and the non-replanted population was the control (CK), and the important agronomic traits and sucrose content of the replanted and non-replanted populations were determined at the harvest stage of the 3rd, 4th and 5th year of ratoon, respectively, and the economic benefits were measured.   Result  The number of effective stems of GL2, GT44 and YT236 after replanting was significantly higher than that of CK. The yield of CK decreased with the extension of permanent root years, and the population yield increased with the extension of permanent root years after replanting, but there was no significant difference in other agronomic traits. The effective stem number of GL2, GT44 and YT236 populations without replanting was negatively correlated with the ratoon years, which were −0.907, −0.931 and −0.756, respectively. The yield was also significantly negatively correlated with the years of ratoon roots, which were −0.915, −0.905 and −0.930, respectively. The results showed that the ratoon years of three varieties were not negatively correlated with yield and number of effective stems, indicating that the ratoon years of sugarcane could be extended after replanting. At the same time, the economic benefits of GL2, GT44 and YT236 groups also increased after replanting, and their total net income increased by 48120.80 yuan /hm2, 46765.80 yuan /hm2 and 42398.60 yuan /hm2 during the three-year ratoon period, respectively.  Conclusion  Directly replanting of GF296 could effectively extend the years of ratoon for 3 years, which was cost-effective in terms of economic benefit comparison and suitable for large-scale promotion. Therefore, this study provides a theoretical basis for constructing high and stable yield population, realizing the potential of sugarcane ratoon, and realizing stable and high yield.
Analysis of Nutritional Quality of a New Black Rice Variety Ziliangyou 7206
CHEN Linghua, CHENG Zuxin, XU Ming, HUANG Ronghua, LIN Lihui
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:   Objective  The nutritional quality of the newly bred two-line hybrid black rice Ziliangyou 7206 was analyzed to provide reference value for the breeding improved varieties of black rice and the development of functional foods.   Method  Using super rice Yiyou 673 as a control, the differences were analyzed in amino acid, fatty acid, anthocyanin, and total flavonoid content between Ziliangyou 7206 and Yiyou 673, as well as the differences in total antioxidant capacity and free radical scavenging ability.  Result  There were no significant differences in water, ash and fat content between Ziliangyou 7206 and Yiyou 673, and 17 amino acids and 8 fatty acids were detected. The protein content, total amino acid content, and essential amino acid content of Ziliangyou 7206 were 7.33%, 11.08%, and 9.71% higher than that of Yiyou 673, respectively. The essential amino acid content of Ziliangyou 7206 was not only higher than the essential amino acid pattern proposed by FAO, but also slightly higher than Yiyou 673. Ziliangyou 7206 had a higher TAV value of each flavor amino acid than Yiyou 673, enriching the taste quality of black rice. The content of saturated fatty acids (SFA) in Ziliangyou 7206 was lower than that in Yiyou 673, but its content of monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA) were slightly higher than that in Yiyou 673. The ratios of SFA, MUFA and PUFA of these two types of rice were basically close to the ratio of "<1:1:1" recommended by the Chinese Nutrition Association, and both contained essential fatty acids such as linoleic acid and linolenic acid. The content of anthocyanins, total flavonoids, total antioxidant capacity, and free radical scavenging ability of Ziliangyou 7206 were significantly superior to Yiyou 673.   Conclusion  The Ziliangyou 7206 obtained by cross breeding had richer nutritional value compared to Yiyou 673, which has a certain reference value for the cultivation of functional black rice and research and development of food.
Preparation and Application of Polyclonal Antibodies Recognizing the Capsid Protein of Chilli Veinal Mottle Virus
JIANG Jun, LI Xiquan, DAI Fei, ZHANG Fuqiang, LU Zhou, WANG Hancheng, SHI Caihua, FANG Shouguo, ZHANG Songbai, PAN Shouhui
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  Aim  Chilli veinal mottle virus (ChiVMV) is one of the most harmful viruses in the production of solanaceae palnts and a subject of quarantine inspections at ports. The aim of this study is to develop polyclonal antibodies against ChiVMV coat proteins with a high level of specificity, which can then be utilized for the field detection of infected plants.  Methods  The full-length (861 bp) and partial fragment (396 bp) of the coat protein gene from the ChiVMV GZ-tabacco isolate were amplified using RT-PCR, recombined into the prokaryotic expression vector pET28a, and subsequently transformed into E.coil BL21 for induced expression. After chromatographic isolation and dialysis purification of the expression products, they were used to immunize Dahl rabbits for the preparation of polyclonal antibodies. The antibody potency and specificity were assessed through enzyme-linked immunosorbent assay (ELISA) and Western blot..  Results  Two polyclonal antibodies against coat proteins, namely antiCP1-287aa and antiCP1-132aa, were successfully prepared with antibody titer of 1:6400 and 1:12800, respectively. Both antibodies, antiCP1-287aa and antiCP1-132aa, effectively detected the antigen by Western blot. In indirect ELISA tests of field strains, it was observed that the ChiVMV strain was specifically detectable by antiCP1-132aa, whereas the Potato virus Y (PVY) strain was not. However, antiCP1-287aa could not differentiate between these two strains.  Conclusion  The polyclonal antibody, antiCP1-132aa , exhibits high specificity, making it suitable for detecting ChiVMV-infected strains in the field. Additionally, it lays the foundation for the functional study of the coat protein of ChiVMV.
Gene Cloning and Functional Analysis of MebZIP2 in Cassava
CHEN Ganlu, YAN Yan, MENG Xianwei, FU Lili, QIU Xianjin, DING Zehong, HU Wei
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:   Objective  bZIP transcription factors play an important role in plant growth and development, starch synthesis, and abiotic stress. Cloning of cassava MebZIP2 and analysis of its subcellular localization, gene expression, and yeast one-hybrid can provide useful references to further study the function of MebZIP2.  Method  The coding sequence (CDS) of MebZIP2 was amplified from cassava SC205 and then used to perform bioinformatics analysis. A pNC-Green-SubN fusion expression vector was constructed and transferred into tobacco epidermal cells by Agrobacterium-mediated method, and fluorescence signals were observed to determine subcellular localization of the MebZIP2 protein. The expression profiles of MebZIP2 were analyzed in different tissues and at different development stages of cassava storage root. The interactions between MebZIP2 and the promoters of starch synthesis genes were studied by yeast one-hybrid.  Result  The CDS sequence of MebZIP2 was 465 bp in length and encoded 154 amino acids. The molecular weight was 17891.35 Da, the theoretical isoelectric point was 5.23, and MebZIP2 was an unstable hydrophilic protein. MebZIP2 contained a bZIP-conserved domain and showed the highest sequence similarity (74.22%) to a bZIP protein from the Hevea brasiliensis. The promoter region of MebZIP2 contained light-responsive elements, hormone (such as gibberellin, salicylic acid, and jasmonic acid) responsive elements, as well as endosperm expression and stress-responsive elements. MebZIP2 protein was localized both in the cell membrane and nucleus. MebZIP2 showed high expression levels in root apical meristems, fibrous roots, stems, and at early developmental stages of storage roots. MebZIP2 was co-expressed with several starch synthesis genes such as MeAPL5a, MeGBSS1, and MeISA1, and MebZIP2 could interact with their promoters.  Conclusion  MebZIP2 belonged to the bZIP gene family, and its expression showed tissue-specificity and root developmental stage-specificity. MebZIP2 protein could interact with the promoters of starch synthesis genes (MeAPL5a, MeGBSS1 and MeISA1), and it might be involved in storage root development and starch synthesis in cassava.
Identification and expression analysis of YUCCA gene family in Passiflora edulis
PAN Jiayi, PAN Ruoyun, JIANG Wenjie, REN Rui, FANG Ting
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  Objective  YUCCA gene encodes flavin monooxygenase, which is one of the main rate-limiting enzyme genes in the biosynthesis of Indole-3-acetic acid (IAA), and plays an important role in regulating plant growth and development. In this study, bioinformatics methods were used to identify members of the YUCCA gene family in Passiflora edulis Sims. In order to reveal the function of YUCCA family genes in hormone response, and provide references for bioinformatics studies of YUCCA family genes in other species.   Methods  The physicochemical properties and conserved domains of the encoded proteins, chromosome location, gene structure, phylogenetic tree and cis-acting elements of the genes were analyzed. qRT-PCR was used to explore the expression of some members treated with exogenous hormone IAA.   Results  A total of 29 YUCCA family members were identified in the passion fruit genome, unevenly distributed in 8 chromosomes. YUCCA gene structure analysis of passion fruit showed significant differences in the length of different genes, containing 1~8 introns and 8 conserved motifs. Through phylogenetic tree analysis, it was found that the YUCCA gene family of passion fruit can be divided into three distinct categories, and the YUCCA genes clustered in the same classification have high conservation. It was also found that the YUCCA gene of passion fruit was more closely related to alfalfa (Medicago sativa L.) and Arabidopsis thaliana, but more closely related to rice (Oryza sativa L.). Cis-acting element analysis showed that the promoter of YUCCA gene family of passion fruit was induced by various hormones and could respond to various stresses. Transcriptome data analysis showed that PeYUCCA6, PeYUCCA11 and PeYUCCA16 showed low or no expression in the leaves of Tainon and golden passion fruit, and the expression of PeYUCCA23 was the highest in Tainon and golden passion fruit, suggesting that this gene had a high effect on the development of passion fruit leaves. The results of qRT-PCR analysis showed that the expressions of PeYUCCA7, PeYUCCA13, PeYUCCA17, PeYUCCA24 and PeYUCCA26 genes were significantly increased after 100 μmol·L-1 IAA treatment.  Conclusion  The expression of YUCCA gene family members varies greatly under IAA treatment. YUCCA gene may play an important role in the growth and development of passion fruit and the resistance to adverse environment under growth regulator IAA.
Expression and Correlation Analysis of Citrate Synthase Gene (CS) in Pomelo and Citric Acid Accumulation
LI Sitong, LI Zhenqin, LIN Wantong, WANG Junning
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:   Objective  Rganic acids are one of the important flavor substances in fruits, Citrate synthase (CS) plays a pivotal role as a rate-limiting enzyme in the organic acid metabolism of fruits. The citric acid synthase gene AheCS was cloned, and the citric acid content of jackfruit postharvest under different treatments was determined. The biological function of AheCS gene and the correlation between citric acid content and relative expression of AheCS gene were analyzed, and the possible role of AheCS gene in the metabolism of organic acids in jackfruit was discussed.  Method  Haida 2 jackfruit were utilized, treated with 0.5 mg·L−1 1-MCP and 1000 mg·L−1 ETH (40%), and subjected to investigation into the dynamic changes in citric acid during jackfruit fruit ripening under room temperature (22±1°C) and 90% RH conditions. Three jackfruit CS genes (AheCS1, AheCS2, and AheCS3) were cloned, and their bioinformatics were analyzed. Its bioinformatics was analyzed, as well as AheCS1/2/3 under different post-harvest treatments (natural ripening, exogenous ethylene ripening, and 1-MCP delayed ripening).  Result  As jackfruit underwent natural ripening, citric acid content exhibited an initial increase followed by a decline. Exogenous ETH treatment accelerated the synthesis rate of citric acid, advancing the time of peak appearance. 1-MCP treatment inhibited the rate of citric acid decline in the early storage period, thus delaying the peak. Bioinformatics analysis revealed that the open reading frames (ORFs) of AheCS1/2/3 genes were 1422 bp to 1827 bp long. AheCS1/2/3 proteins contained the conserved 'WPNVDAHS' domain of citrate synthase, classifying them as members of the citrate synthase family. The amino acid sequences of AheCS1/2/3 showed the closest phylogenetic relationships with citrus CsCS (MH_048698.1), mulberry MnCs (XP010087965.1), and Anthurium AaCS (JAT55223.1), with similarities reaching 86.49%, 97%, and 86%, respectively. qRT-PCR results indicated that the expression of AheCS1/2/3 genes was low in the early stage of natural fruit ripening (CK) and increased in the later stage. Exogenous ETH treatment advanced the expression of AheCS1 and overall increased the expression of AheCS2/3. 1-MCP treatment delayed the increase in the expression of Peak occurrence time of AheCS1/2/3 but increased their expression in the later stages of maturity. Correlation analysis revealed a positive correlation between citric acid content and the expression of AheCS1/2/3 genes during jackfruit fruit ripening, with AheCS2 reaching a significant level.   Conclusion  The accumulation of citric acid during jackfruit maturation may be regulated by the AheCS2 genes, providing a foundation for further research on the functionality and genetic improvement of jackfruit AheCS genes.
Isolation and identification of a novel duck adenovirus B2 strain with insertion and deletion mutation characteristics
GUO Bolun, LI Yaru, ZHENG Xin, JIANG Dandan, ZENG Li, CHEN Shaoying, MA Yanmei, CHEN Shilong
[Abstract](0) [FullText HTML](0) [PDF 1825KB](0)
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:   Objective  In order to isolate duck adenovirus B2 (DAdV B2) strain from "Pale liver disease" and provide a reference for the DAdV B2 epidemiological study in Fujian Province.   Methods  The samples were detected by PCR, and the virus isolation was carried out and identified by the whole genome sequencing and ducklings challenge.  Result  A DAdV B2 strain, named DAdV B2/BG48, was successfully isolated from duckling pale liver disease. DAdV B2/BG48 infected LMH cells became large and round, and finally died and disintegrated. MDEF cells with with DAdV B2/BG48 inoculation became round and clustered together with intercellular space appeared. The results of next generation sequencing showed that BG48 genome had a 3 bp insertion in the pX gene region, a 33 bp insertion in the ORF19B gene domain, and 42 bp deletion at the junction of ORF64 and ORF67. There was no truncated mutation in the ORF67 gene. Other coding genes had no characteristic differences as compared to previously identified strains such as CH-GD-12-2014, BG27 and BG18. Duckling challenge experiments showed that the morbidity and mortality of BG48 to 2-day-old Muscovy ducks were 50% and 0%, respectively, and the clinical symptoms and pathological changes of diseased ducklings were similar to those of naturally infected ducks.   Conclusion  A novel DAdV B2 strain named BG48 was successfully isolated and identified from a case of "Pale liver disease" in Muscovy duck. BG48 has the characteristics of multi-site insertion and deletion, suggesting that DAdV B2 strain was prone to mutation with a complex clinical epidemic. These results provide a reference for the molecular epidemiological investigation and the genetic evolution of DAdV B2.
A Deep Learning-based Method for Detection of Precision Weeding Regions in Vegetables
LI Weili, JIN Xiaojun, YU Jialin, CHEN Yong
[Abstract](0) [FullText HTML](0) [PDF 5115KB](0)
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  Objective  Vegetables grow randomly and often infested with numerous weed species. Traditional algorithms are complex in weed detection, and typically fail to localize the weeding regions precisely. In the study, a deep learning-based method was proposed to identify weeds growing in vegetables in combination with weeding regions detection.   Method  Firstly, the original images were cropped into grid cells (sub-images). Afterwards, the deep learning models were utilized to classify the grid cells into vegetables, weeds and soil. The grid cells containing weeds were marked as weeding regions. ShuffleNet, DenseNet, and ResNet were employed to detect and classify the grid cells. Deep learning models were evaluated in terms of precision, recall value, F1 score, and overall and average accuracy.   Result  The experiment results show that the evaluated deep learning models can identify weeds and vegetables effectively. ShuffleNet model was proven to be the bestmodel, which achieves optimal balance between the precision and recall value, with the values of 95.5% and 97% respectively. Additionly, ShuffleNet achieved the highest detection speed, with the value of 68.37fps, making it suitable for real-time weed detection.   Conclusion  The results demonstrate that the proposed method is feasible and well-performed, which can be used for precision weeds control in vegetables.
Influences of organic manures application on rape seedling (Brassica napus L.) growth and photosynthetic fluorescence characteristics under aluminum stress in acid red soil
YU Juhua, WANG Limin, DING Hong, WANG Huangping, ZHENG Xiangzhou, ZHANG Yushu, ZHANG Yinlong
[Abstract](0) [FullText HTML](0) [PDF 869KB](0)
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  Objectives  Aluminum toxicity of acid soil is a major constraining factor for crop growth and development. The organic materials such as livestock and poultry manure are one of significant pathways to improve soil acidity and promote the national strategical plan of zero growth of chemical fertilizer. However, the effects of organic manures application on crop growth and photosynthetic fluorescence characteristics under aluminum stress and its mechanism are still unclear.   Methods  Using the pot experiment to explore the effects of pig and chicken manures application on biomass, photosynthetic parameter and chlorophyll fluorescence properties under aluminum stress in acid red soil.   Results  Under aluminum stress in acid red soil, the results showed aboveground biomass, photosynthetic pigment content, photosynthesis and chlorophyll fluorescence of rape (Brassica napus L.) were obviously improved under organic manures treatment (P<0.05), which on day 80 in treatment groups was significantly higher than those of in control group, suggesting that chelation with soil active aluminum and neutralization process of soil acidity by organic manures were capable of alleviating the toxic effect of aluminum stress on rape seedling growth in acid red soil. The same application rate of chicken manure was more beneficial to improve aboveground biomass and photosynthetic pigment content of rape seedling than those of pig manure, but not for photosynthetic parameters, Fv/Fm and qP. Moreover, the promoting effects of mixed application of chicken and pig manures than those of single usage (P<0.05). Above two direct evidences indicated chicken manure with higher pH was more helpful for rape growth, while pig manure with higher nutrient content was more helpful to improve photosynthetic fluorescence of rape seedling. The application rate of 50 g·kg−1 for pig manure was the most optimal content to alleviate the growth and photosynthetic fluorescence of rape seedling, whereas the optimal application rate for chicken manure was 30 g·kg−1.   Conclusions  In terms of growth and photosynthetic fluorescence of rape seedling, it is vital to alleviate the aluminum toxicity in acid red soil by scientific screening of manure species, optimal application rate in combination with soil acidity regulation.
Analysis of combining Ability and Correlation on Yield, Brix and Three-ear-leaves Characters of waxy maize
CUI Yang, LIU Junfeng, ZHANG Yao, SONG Qiaoheng, KONG Liangliang
[Abstract](0) [FullText HTML](0) [PDF 547KB](0)
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:   Objective  To study the genetic rule of waxy maize yield and sugar degree, and analyze the correlation between yield, sugar degree and three ear leaves traits, so as to provide reference for breeding of fresh waxy maize with high sugar degree and explore the mechanism of the influence of three ear leaves traits on quality traits.   Methods  In this study, 6 inbred lines were used as test species, and 15 waxy maize inbred lines were selected for incomplete diallel hybridization (NCII design). The yield of ear, soluble sugar content (sugar degree) of grain, leaf length, leaf width, leaf area, and other 10 ear and plant-related traits of the hybrid combinations were measured at the harvest period (21 days after pollination). Panicle weight and sugar content were the main yield and quality traits to evaluate the contribution of L. trefoil to yield and quality. The other characters were used as reference characters to evaluate the importance of three ear leave to yield and quality. According to the measured data, general combining ability (GCA) and special combining ability (SCA) of ear yield and sugar content were analyzed.   Results  The results showed that there was a strong correlation between leaf length and ear yield in three ear leave characters. There was a strong correlation between leaf width and sugar content. There was a significant negative correlation between ear yield and sugar content, so materials with moderate leaf length and wide leaf width should be preferred in breeding process to improve the selection efficiency of high-quality waxy maize.   Conclusion  The study indicated that the length and width of the leaves, especially the lower ear leaves and ear position leaves, could be used as the basis for selecting breeding materials of waxy maize.
Effects of electric field treatment on seed germination characteristics of Acacia cincinnata
ZHU Meng-tian, JIANG Rui-yi, ZHANG Ying, DENG Zhiwen, RONG Jun-dong, ZHENG Yu-shan, CHEN Li-guang
[Abstract](0) [FullText HTML](0) [PDF 1482KB](0)
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:   Objective  Physical technology is beneficial to the growth of plants and has a good application prospect. The effects of different electric fields, time and their interaction on seed germination characteristics were analyzed to provide theoretical reference for seed germination and sustainable management.   Method  This study used Acacia cincinnata seeds as the research object, using indoor simulation experiments, with different voltage intensities and treatment time as control factors, voltage intensities were 0~1.0 kV·cm−1, and time treatments were 0~60 min to study the effects of different voltage intensities and time interactions on seed germination characteristics.  Result  The results showed that with the increase of voltage intensity and treatment time, the germination rate, germination potential and germination index of A.cincinnata seeds showed varying degrees of change. Among them, the germination rate decreased first, then increased and then decreased with the increase of voltage intensity at 15 min, 30 min and 60 min, but the germination rate increased first and then decreased with the increase of voltage intensity at 45 min. The germination potential and germination index were the best under the voltage intensity of 0.6 ~ 0.8 kV·cm−1 and the treatment time of 45 min, and there was a significant difference compared with CK ( P<0.05 ). After nonlinear regression analysis, the optimal regression equation under different voltage intensity and time interaction was: Y=7.73+135.23X1+1.24X2−86.16X12−0.00911X22-0.75X1X2 (R2=0.81, P<0.05). Cluster analysis showed that the interaction of electric field and time had a very significant effect on germination index.   Conclusion  The interactive treatment of electric field and time had a certain effect on the germination characteristics of A.cincinnata seeds. The germination rate was the highest at 0.6 kV · cm−1 for 45 min, indicating that the appropriate voltage intensity and treatment time could improve the germination rate of A. cincinnata seeds.
Analysis of physicochemical properties of tomato vine biochar under different pyrolysis temperatures
REN Lihua, ZOU Xiufeng, HUANG Jiaqing, YE Jing, WANG Yixiang
[Abstract](0) [FullText HTML](0) [PDF 1803KB](0)
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:   Objective  Biochar utilization is a new direction of straw resource utilization and one of the ten modes of straw agricultural utilization. In order to explore the effect of pyrolysis temperature on tomato vine biochar.   Methods  Biochar was prepared by pyrolysis at 300 °C, 500 °C and 700 °C for 2 h. The surface structure, elements characteristics, surface functional groups of tomato vine biochar were comprehensively analyzed by means of electron microscope scanning, elemental analyzer and fourier transform infrared spectrometer.   Results  The results showed that the tomato vine biochar prepared by pyrolysis method was alkaline ( pH range 9.83-10.67 ). The yield of biochar decreased with the increase of pyrolysis temperature, while the ash content was the opposite. The total nitrogen content was the lowest at 500 °C, and the highest at 300 °C, but the fixed carbon content and C / N ratio were relatively high at 500 °C, which were 51.42 % and 36.63, respectively. The pore structure of tomato vine biochar was rich at low temperature pyrolysis. At high temperature pyrolysis, the pores were covered by ash and its molten structure, and the porosity decreased. With the increase of pyrolysis temperature, the degree of aromatization of biochar increased. Compared with 300 °C and 500 °C, the absorption peak of Fourier transform infrared spectroscopy at 700 °C decreased, especially in 500 ~ 800 cm−1.   Conclusion  Pyrolysis changed the physical and chemical properties and microstructure of tomato vine biochar. Considering all factors, tomato biochar prepared by pyrolysis at 300-500 °C for 2 h had better performance and higher benefits.
Research progress in effects of tillage methods on microbial residues in soil
LI Ruxin, GAO Qisong, YU Yingxin, LV Yi, HAN Huifang
[Abstract](0) [FullText HTML](0) [PDF 777KB](0)
Abstract:
The entombing effect of microbial residues contributes over 50% to soil organic carbon (SOC) in agricultural ecosystems. Reasonable tillage methods can promote the accumulation of microbial residues in soil by affecting microbial communities and aggregates. Therefore, studying the mechanism of action of microbial residues is crucial for analyzing the long-term and efficient farmland cultivation measures for SOC retention.The article reviews the research progress at home and abroad, including the contribution pathways and sizes of microbial residues to SOC, the impact of tillage methods on SOC and microbial residues, and the reasons for the impact of tillage methods on microbial residues. It deeply analyzes the mechanism of how tillage methods affect the regulation of SOC content through microbial residues, providing reference for carbon sequestration in farmland.
Cloning and expression analysis of RhMAX2A gene in Rosa hybrid
LI Shasha, DU Gaoqi, LI Xuejiao, GUAN Wenling, MENG Jing
[Abstract](0) [FullText HTML](0) [PDF 4149KB](0)
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:   Objective  To study the biological function and the transduction mechanism that regulates lateral branching of the RhMAX2A gene in Rosa, the cDNA sequence of RhMAX2A was cloned and was analysed for sequence characteristics and its expression in different tissues and after decapitation.  Methods  The cDNA sequence of RhMAX2A gene was cloned from Rosa hybrida ‘Dianhong’ by RT-PCR and the bioinformatics analysis was conducted. The subcellular location of RhMAX2A was analyzed by transient transformation of PC1300s-RhMAX2A-GFP in tobacco leaves. The expression levels of RhMAX2A in different tissues and the treatment of decapitation was detected by real-time quantitative PCR (qRT-PCR).   Results  The cDNA sequence of RhMAX2A gene (GenBank accession number: OP055810 ) was 1030 bp in length, encoding 246 amino acids. The calculated chemical molecular formula of RhMAX2A protein is C2910H4793N1029O1244S210, with 27.35 kD in molecular mass and the total atomic weight is 3909. The instability coefficient of this protein is 53.07, with fat coefficient-106.30 and GRAVY value 0.049, indicating that it is an unstable hydrophilic protein. The secondary structure of RhMAX2A protein is mainly composed of α-helix and random coil, and RhMAX2A is a presumed F-box domain, belonging to the α/β hydrolase family. Homologous sequence alignment and phylogenetic tree analysis showed that the amino acid sequence of RhMAX2A (OP055810) had the highest similarity with that of Rosa chinensis Old Blush (XP_024283944.1), followed by Fragaria vesca subsp. vesca (XP_004287076.1) of the same subfamily, which were closely related. The subcellular localization results showed that the encoded protein of RhMAX2A was located in the nucleus. The results of qRT-PCR indicated that RhMAX2A gene was expressed in roots, axillary buds and nodes, with the highest expression level in roots. The decapitation treatment significantly upregulated the expression of RhMAX2A in roots and axillary buds.   Conclusion  The RhMAX2A gene was successfully cloned from Rosa hybrida ‘Dianhong’, which encodes a protein that plays a role in the nucleus, mainly expressed in roots and axillary buds, and could be upregulated by decapitation.
Effect of Waterlogging on Quality of Maize Kernels at Filling Stage
GONG Xue, LU Shengqiao, CHEN Kun, QIN Hongyu, ZHOU Jinguo, WEI Zhengyi, ZHONG Changsong, YANG Yaojiong, JI Yuqi, LIU Yali, ZHANG Shukuan
[Abstract](204) [FullText HTML](49) [PDF 1123KB](5)
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  Objective  Effects of waterlogging on quality of maize kernels at filling stage were studied to decipher the mechanism and for breeding selection.  Method  Two backbone maize inbred lines in Guangxi, 88M-1-8 and Xian 21A, were subjected to normal water irrigation (CK) or artificial flooding treatments (W) after pollination. The experiment lasted 14d or 18d with a total of 4 variables, i.e., CK-14, CK-18, W-14, and W-18. Contents of soluble protein, soluble sugar, starch, sucrose, and abscisic acid (ABA) as well as activities of sucrose synthetase (SS-I in decomposition direction) and granule-bound starch synthase (GBSS) of the kernels were measured. Effect of waterlogging on maize quality at filling stage was evaluated using the principal component analysis and membership function method.   Result   Under W-14, the contents of protein, starch, and ABA and the activities of SS-I and GBSS in 88M-1-8 were significantly higher than those in Xian 21A, so were the soluble sugar, starch, ABA, and GBSS in 88M-1-8 under W-18. As the waterlogging prolonged, the soluble protein, starch, sucrose, SS-I, and GBSS in both inbred lines raised significantly. The protein, starch, ABA, and SS-I in 88M-1-8 increased more significantly, and the soluble sugar, sucrose and GBSS in Xian 21A more significantly in W-14 than CK-14. Furthermore, compared with CK-18, W-18 induced more significant increases on the soluble sugar and ABA in 88M-1-8, and the protein, starch, sucrose, SS-I, and GBSS in Xian 21A. Evaluated by the principal component analysis and membership function method on various kernel quality indicators, the tolerance of the two species to flooding were 88M-1-8 under W-14>88M-1-8 under W-18>Xian 21A under W-14>Xian 21A under W-18.  Conclusion   Artificially flooding the maize plants raised the soluble protein, soluble sugar, starch, sucrose, SS-I, and GBSS in kernels over regular irrigation. 88M-1-8 and Xian 21A differed in responses to the stress. Waterlogging-tolerant 88M-1-8 was significantly higher in the GBSS activity and ABA and starch contents than Xian 21A. Nonetheless, prolonged waterlogging reduced the stress tolerance of either cultivar.
Changes of Microbial Community in Agaricus bisporus Culture Substrate during Fermentation
YUAN Bin, KE Lina, LIAN Yanping, ZHAO Guanghui, FENG Weilin, CAI Weiming
 doi: 10.19303/j.issn.1008-0384.2022.009.018
[Abstract](252) [FullText HTML](192) [PDF 2371KB](31)
Abstract:
  Objective  Changes of the microbial community in fermentation of substrates for cultivating Agaricus bisporus were investigated.   Method  The microbial community characteristics of a compound A. bisporus culture substrates containing spent Flammulina velutiper and Pleurotus eryngii materials were monitored during the 7 stages, Ag1 to Ag7 from pile building to 1st and 2nd fermentation, using the high throughput 16S rDNA full-length sequencing of Pacbio platform.   Result  A total of 715 OTUs were obtained in the fermentation process (i.e., 328, 340, 294, 377, 364, 166, and 174 for each stage) with 161 OTUs commonly found in all stage. The microbes included 21 phyla, 299 genera, and 399 species. At phylum level, Fimicutes, Proteobacteria, Bacteroidetes, and Gemmatimonadetes had higher abundance in all 7 stages. The dominant genus in the substrate during the pile building and 1st fermentation stages was Ureibacillus, while Limnochordaceae, S0134_terrestrial_group, Thermobacillu, and Ruminiclostridium stood out in the 2nd fermentation stage. At species level, Ureibacillus thermophilus and Ureibacillus terrenus were dominant in the pile building and 1st fermentation stages. In the 2nd fermentation stage, the species in the genus of Limnochordaceae were the greatest in relative abundance. It appeared that the microbial abundance and diversity increased gradually before the onset of 2nd fermentation. The community structure of the substrate varied significantly between the 1st and the 2nd fermentation, but the deviation diminished significantly after the 2nd fermentation leaving mostly the dominant species that basically functioned to degrade the waste mushroom material enriching the fertilization effect.   Conclusion  The full-length sequencing technology clearly identified the dominant species unveiling many previously unclassified microorganisms. The results helped to better optimize the substrate fermentation process for an improved cultivation of A. bisporus.
Effects of coffee waste mulching on photosynthesis and water use efficiency of coffee seedlings
ZHANG Ang, DONG Yunping, LIN Xingjun, ZHAO Qingyun, SUN Yan, LONG Yuzhou, ZHONG Yiming, TAN Jun
[Abstract](261) [FullText HTML](79) [PDF 946KB](11)
Abstract:
  Objective  The effects of coffee peel and litter on the growth and photosynthesis of coffee seedlings were investigated to determine a suitable ecological cycle cultivation a suitable of coffee.   Method  One-year-old seedlings of seedlings were used as experimental materials. A randomized block design was used to study the effects of coffee peel and litter on their leaf photosynthesis and water use efficiency. Conventional mulching group was set up: C (control), L (litter mulching cultivation), P (peel mulching cultivation) and LP (litter and peel mulching cultivation).   Result  Coffee litter mulching significantly increased the specific leaf area of coffee by 45.46%, while coffee peel mulching significantly decreased the plant height by 12.11%. Coffee peel mulching significantly increased net photosynthesis, leaf respiration, total photosynthesis, net water use efficiency and total water use efficiency by 78.33%, 109.34%, 91.72%, 80.54% and 104.95%, but did not affect stomatal conductance, transpiration rate and carbon use efficiency. The comprehensive evaluation of coffee photosynthesis under coffee waste mulching treatment was P>LP>L>C.   Conclusion  Coffee peel mulching was better than other mulching treatments in promoting the growth and photosynthetic capacity of coffee seedlings. The suitable coffee ecological cycle cultivation with the peel amount of 239.05 g m-2 could to realize cost saving and efficiency increase in coffee planting process.
Carbon, Nitrogen, and Enzyme Activities in Saline-alkali Soil on Songnen Plain as Affected by Land Uses
LIU Qian, GUO Boya, WU Xiuyu, WANG Yue
[Abstract](280) [FullText HTML](156) [PDF 992KB](5)
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  Objective  Fertility and enzymatic activity of the saline-alkali soil in relation to land use were analyzed for ecological improvements and restoration.   Method  At Songnen Plain in western Jilin province, the effects on organic carbon, total nitrogen, invertase, urease, alkaline phosphatase, and catalase of the saline-alkali soils under different types of land use as paddy farming field (N1), dry farming field (N2), wetland (S), and grassland (C) were compared.   Result  The organic carbon contents in the soils ranged 9.70–16.27 g·kg1 under N1, 3.85–11.58 g·kg1 under N2, 2.14–2.97 g·kg1 under S, and 5.25–11.24 g·kg1 under C; and the total nitrogen, 1.83–2.32 g·kg1 under N1, 0.45–0.76 g·kg1 under N2, 0.34–1.28 g·kg1 under S, and 0.88–2.04 g·kg1 under C. The activities of various enzymes were urease>alkaline phosphatase>catalase>sucrase and decreased along the depth of the soil layers. The invertase significantly correlated with C/N at P<0.05, the urease with C/N at P<0.01, the alkaline phosphatase with the organic C at P<0.01 and with the total nitrogen at P<0.05, while the catalase with total nitrogen at P<0.01 and with C/N at P<0.05. The redundant analysis indicated that the activities of invertase and urease were mainly regulated by the pH and bulk density, while those of alkaline phosphatase and catalase largely affected by the moisture content and electric conductivity of the soil.   Conclusion  Land use exerted significant effects on the organic carbon, total nitrogen, and enzyme activity in the saline-alkali soils which gradually decreased from the surface to the deeper layers. Farming on the land fostered the nutrient accumulation and increased the enzymatic activities in soil. Thus, either paddy or dry field was more ecologically friendly than wetland or grassland for the regions of saline-alkali soil.
Role of pAPN and NEU3 in TGEV Infection on Pig
LI Zhaolong, FENG Zhihua, ZHANG Bingchen, FANG Zhou, LIANG Wangwang, CHEN Wenzhi
[Abstract](656) [FullText HTML](209) [PDF 1905KB](27)
Abstract:
  Objective  Role of aminopeptidase gene pAPN and sialic acid neuraminidase gene NEU3 in the transmissible gastroenteritis virus (TGEV) infection on pigs was investigated.   Methods  Being the main receptor of TGEV, pAPN was removed from pAPN and NEU3 in ST cells to verify its supposed key function on the disease. The CRISPR gene editing technique was applied to clip the target gene in ST cells prior to an artificial TGEV infection test. The resulting changes on the infection, virus copy number, cytopathic improvement, and fibronectin were monitored.   Results  Compared with control, the ST cells free of pAPN and NEU3 significantly attenuated TGEV infection-induced cytopathies and the virus copy number. In addition, at a same TGEV titer the mRNA immune responders induced by the knockdown ST cells were significantly lower than the wild-type counterparts.   Conclusion  It was confirmed that the removal of pAPN and NEU3 inhibited the TGEV infection in pigs with reduced viral induced cytopathies. Thus, an antiviral therapy and a guideline for breeding resistant pigs could be developed by targeting these two key genes in the ST cells.
Effects of different duration of waterlogging on physiological characteristics and yield of wheat at booting stage
DING Fugong, LU Yifei, JIA Baosen, ZHU Xudong, XIONG Zehao, WANG Chao, HOU Zehao, LIU Yike, ZHU Zhanwang, ZHANG Yingxin, WANG Shuping, FANG Zhengwu
[Abstract](480) [FullText HTML](241) [PDF 941KB](29)
Abstract:
  Objective  The stress of wheat waterlogging is the main abiotic stress factor in wheat production in the middle and lower reaches of the Yangtze River. To study the effects of different duration of waterlogging on physiological characteristics and yield of wheat at booting stage provides a theoretical basis for the research on the mechanism of wheat waterlogging resistance at booting stage and production.  Method  The effects of waterlogging duration on wheat growth and yield at booting stage were studied by pot pot water control method with wheat varieties Yangmai 16 and Zhongmai 895 as experimental materials.  Result  (1) Under the stress of waterlogging, the chlorophyll content of wheat leaves decreased significantly. The longer the waterlogging lasted, the greater the decline of SPAD value was. The SPAD value of the more heavily injured leaves decreased more, and the more severely injured the inverted two leaves than the flag leaves. (2) The activity of antioxidant enzymes such as CAT, SOD and POD in wheat showed a trend of type "∧" during the waterlogging period, while the content of reactive oxygen species (ROS) decreased or increased slowly in the early stage of waterlogging, while increased sharply in the late stage. (3) In the booting period, the effective number of ears, grain number of ears, 1000-grain weight and other yield factors increased slightly, which may be caused by the stress response of wheat. (4) The stress of waterlogging at the stage of heading had no significant effect on the height of wheat plant. Long-term waterlogging resulted in a significant decrease in wheat yield. The decrease of effective panicle number, grain number per panicle and 1000-grain weight was the main factor causing wheat yield reduction. After 15 d of waterlogging stress, the yield per plant of zhongmai 895 and yangmai 16 decreased by 51.47% and 43.99%, respectively, compared with CK.  Conclusion  Booting stage waterlogging stress significantly reduced the wheat leaf chlorophyll content, destroyed the plant active oxygen metabolism in the body and the balance between antioxidant enzyme system, excessive accumulation of reactive oxygen species causes cells to peroxide lipid membrane, causing cell structure and function is impaired, affect plant photosynthesis and nutrient transfer and accumulation, increase the biomass of wheat is reduced, resulting in lack of grain-filling, caused empty grain, grain and invalid number of flat significantly increased, resulting in wheat production. In addition, during the whole process of waterlogging stress, the resistance of the two wheat varieties tested was as follows: yangmai 16>zhongmai 895.
Effect of endogenous hormone content and gene expression related to hormone synthesis on tuber expansion of Bikeqi yam
AO Lan-ji-ya, JI Xiang, SHAO Ying, ZHAO Lingmin, ZHANG Yan-fang, HUO Xiu-wen
[Abstract](904) [FullText HTML](656) [PDF 693KB](36)
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  Objective  To investigate the changes of starch content, reducing sugar content and soluble total sugar content in tubers, the changes of endogenous hormone content in tubers, the morphological indexes, physiological indexes and the analysis of variance of endogenous hormones during the process of tuber expansion. And the correlation between these indicators and endogenous hormones, lay the foundation for further study of the physiological mechanism of yam growth and development and guiding the high yield and quality cultivation of yam.  Method  Six kinds of endogenous hormones such as ABA, GA3, IAA, JA, ZR and IPA were determined by enzyme-linked immunosorbent assay with different tuber lengths of Bikeqi yam. High performance liquid chromatography (HPLC) was used to determine the content of salicylic acid (SA), and to analyze the dynamic changes of source hormone content in different growth stages, and the correlation between morphological indicators of yam and endogenous hormones. The relationship between differentially expressed genes associated with endogenous hormones and endogenous hormones was explored for the effects of these factors on the expansion of yam tubers.  Result  The contents of IAA, ZR, ABA, JA and SA were positively correlated with the morphological indexes of yam tuber; the contents of GA3 and IPA were negatively correlated with the morphological indexes; the contents of IAA were positively correlated with the perimeter and diameter of yam tuber; the contents of GA3 were negatively correlated with the length of tuber; the genes correlated with IAA were negatively correlated with the contents of IAA  Conclusion  Endogenous hormones IAA, ZR, ABA, JA and SA promote the expansion of yam tuber; endogenous hormones GA3 and IPA inhibit the growth of yam tuber; endogenous hormones IAA promote the thickening of yam; endogenous hormones GA3 inhibit the elongation of yam; the down-regulation of IAA related genes can promote the synthesis of IAA, that is, regulate the content of IAA.
2024, 39(1): 1-2.  
[Abstract](0) [FullText HTML](0) [PDF 377KB](0)
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Animal Science
Role of Tail Fiber Protein in Attaching Riemerella anatipestifer to Phage
LIU Na, HUANG Yu, CHEN Hongmei, JIANG Nansong, LIU Rongchang, FU Guanghua, FU Qiuling, LIANG Qizhang, WAN Chunhe, LI Ang, CHENG Longfei
2024, 39(1): 1-6.   doi: 10.19303/j.issn.1008-0384.2024.01.001
[Abstract](39) [FullText HTML](36) [PDF 1812KB](4)
Abstract:
  Objective  Role of tail fiber protein (TFP) plays in adhering Riemerella anatipestifer by CRP2 phage was investigated.   Method  The putative TFP gene of ORF70 was cloned. The recombinant expression vector pET28a(+)-TFP was constructed and induced to express in E. coli BL21(DE3). The recombinant protein was purified by Ni-NTA affinity chromatography. ELISA was used to determine the binding effect of the recombinant protein with the outer membrane protein (OMP) of host bacteria. A competitive adsorption test was performed to determine the adhering effect of the CRP2 protein.   Result   The recombinant plasmids of pET28a(+)-TFP was successfully constructed and expressed in E. coli. The purified recombinant protein rTFP could bind to the OMP of or suppress the CRP2 adsorption to the host bacteria.   Conclusion  The TFP encoded by ORF70, one of the receptor binding proteins of CRP2 phage, was confirmed to adhere to the host bacteria OMP facilitating the subsequent bactericidal effect.
Interacting Proteins of Olfactroy Receptor OR1and OR2 with Co-IP Approach Followed by Mass Spectrometry Analysis in Apis cerana cerana
GUO Lina, SHEN Hongying, WANG Jue, YU Diandian, ZHANG Xufeng, GUO Yuan
2024, 39(1): 7-16.   doi: 10.19303/j.issn.1008-0384.2024.01.002
[Abstract](77) [FullText HTML](45) [PDF 1759KB](3)
Abstract:
  Objectives   Roles of the proteins interacting with the odorant receptors (ORs) in the highly sensitive olfactory system of Apis cerana cerana were investigated.   Methods   The eukaryotic expression vectors pFastBac of OR1 and OR2 in A. cerana cerana were constructed and transfected into Sf9 cells to extract the proteins. The cellular interacting proteins were identified by co-immunoprecipitation (Co-IP) and mass spectrometry, and GO function annotation, KEGG signal pathway analysis, and protein interaction network determination.  Results   The recombinant proteins of the IP and IgG groups were successfully expressed in the cells. There were 273 proteins interacting with OR1, and 204 with OR2. They were mainly tubulin, heat shock proteins, and ribosomal proteins involving in a variety of biological functions, such as RNA splicing and ribosome and energy transport. The KEGG analysis also showed their association with the regulation of several important intracellular pathways related to the ribosome, splicing body, RNA transport, pyruvate metabolism, thiamine metabolism, fatty acid biosynthesis, starch and sucrose metabolism, and FoxO and Hedgehog signals.   Conclusion  A unique honey bee species in China, A. cerana ceranahas evolved with a highly sensitive olfactory system capable of recognizing chemical signals in the bee population and distinguishing molecules emitted from food sources. OR1 and OR2 in A. cerana cerana regulated the olfactory transduction through direct or indirect interaction with a variety of proteins.
Crop Science
CRISPR/Cas9 Technology-generated High-amylose Rice Varieties
WU Min, HUANG Juan, SHI Taoxiong, ZHU Liwei, DENG Jiao, LIANG Chenggang, WANG Yan, LIU Fei, LI Rong, CAI Fang, CHEN Qingfu
2024, 39(1): 17-24.   doi: 10.19303/j.issn.1008-0384.2024.01.003
[Abstract](81) [FullText HTML](32) [PDF 2426KB](4)
Abstract:
  Objective  A series of new rice germplasms with high content of amylose derived from Zhonghua 11 using the CRISPR/Cas9 technology was generated.   Method   The gene of starch branching enzyme in rice, OsSBE3, was targeted for the genetic editing by CRISPR/Cas9. Homozygous T-DNA-free mutants were verified by PCR with starch content measured.   Result  Ten mutant lines were obtained from the T0 generation. From the T1 generation, 5 homozygous T-DNA-free lines were obtained that included 4 lines, i.e., sbe3-22-6, sbe3-25-3, Sbe3-25-4, and sbe3-25-6, showing significantly increased amylose content and amylose/amylopectin ratio over the wild type.   Conclusion  A series of new rice germplasms with high amylose content was created.
Breeding an Aromatic, High-quality, High-yield Hybrid Rice Yexiangyou 669
ZHU Yongsheng, WEI Yidong, LI Qixiang, ZHENG Feiyan, CAI Qiuhua, DONG Ruixia, CHEN Liping, JIANG Jiahuan, WU Fangxi, XIE Huaan, ZHANG Jianfu
2024, 39(1): 25-32.   doi: 10.19303/j.issn.1008-0384.2024.01.004
[Abstract](47) [FullText HTML](64) [PDF 697KB](5)
Abstract:
  Objective   An aromatic, high-quality, high-yield, disease-resistant rice variety was bred.   Method  Utilizing the genetic diversity and complementing agronomic traits between the male parent, Fuhui 669 with high yield and combining ability, and the female parent, the fragrant, high-quality, disease-resistant sterile line Yexiang A, a new breed of rice were generated. The hybrid was subjected to multiple ecological resistance identification, a quality analysis after high temperature maturation, and years of multi-point comparison test prior to final selection.   Result  The high-quality, high-yield, disease-resistant Yexiangyou 669 of a desirable plant type was obtained and submitted to the regional trial for mid-season rice in Fujian from 2019 to 2020. In the 2-year trial, it showed an average growth period of 139.8 d, which was 1.1 d shorter than the reference cultivar, II You 3301, and an average yield of 9 363 kg·hm−2, which was 2.24% higher than control but not statistically significant. It exhibited resistance to rice blast and up to the national Grade A quality. In 2021, it was certified by the Fujian Provincial Crop Approval Committee as the Min Approved Rice No. 20210014. Yexiangyou 669 had inherited 6 desirable genes from one parent and 16 common to both parents.  Conclusion  The new hybrid rice Yexiangyou 669 had high yield, strong disease resistance, and superb grain quality and was promoted as one of the main varieties in the province.
Horticultural Science
Cloning and Expression of BsDFR in Bougainvillea spectabilis
SUN Rong, LIU Tao, PAN Kaiyue, LIU Shan, DIAO Yi, ZENG Daoping
2024, 39(1): 33-39.   doi: 10.19303/j.issn.1008-0384.2024.01.005
[Abstract](95) [FullText HTML](46) [PDF 3486KB](12)
Abstract:
  Objective   The dihydroflavonol-4-reductase (DFR) gene in bracts of Bougainvillea spectabilis was cloned and characterized to study the role it plays in color formation.   Method  BsDFRwas cloned based on the transcriptome data on the ornamental plant to study the related bioinformatics. Molecular docking technology was employed to predict the substrate specificity, and qRT-PCR applied to examine the relative transcription levels of the genes in B. spectabilis of different colors.   Result   The full-length coding sequence of BsDFR (GenBank ID: ON417750) was 987 bp encoding 328 amino acids. The protein had a calculated molecular weight of 36.49 kDa and an isoelectric point of 6.33. It had the NADPH and substrate specific binding sites unique to DFR of Asn type without a transmembrane structure or signal peptide. The subcellular localization of the protein indicated it to be cytoplasmic. Alpha helices were the most abundant secondary structure of the protein, while the tertiary structure was a dimer. A substrate docking simulation, consistent with the structural analysis, predicted BsDFR to possess a catalytic activity on dihydrokaempferol, dihydroquercetin, and dihydromyricetin. The phylogenetic tree analysis grouped it along with caryophyllales plants. High expression of the gene was found in the orange B. spectabilis by qRT-PCR. It was speculated that the main substrate to be DHK, which was catalyzed by BsDFR into leucopelargonidin, a precursor of orange-colored anthocyanidin——pelargonidin.   Conclusion   BsDFR in B. spectabilis had typical molecular characteristics of the plant dihydroflavonol-4-reductase, which is associated with the pigment synthesis in the bracts of orange B. spectabilis.
Cloning and Expression of 4CLs in Impatiens uliginosa
CHEN Xinyi, WU Chengying, HE Haihao, HUANG Haiquan, QU Suping, HUANG Meijuan
2024, 39(1): 40-48.   doi: 10.19303/j.issn.1008-0384.2024.01.006
[Abstract](63) [FullText HTML](28) [PDF 6021KB](5)
Abstract:
  Objective  Genes of the enzyme in the phenylpropane metabolic pathway associated with anthocyanin synthesis of Impatiens uliginosa were studied for breeding purposes.   Method  RT-PCR was employed to isolate and clone Iu4CL1,Iu4CL2, Iu4CL3, and Iu4CL4 for bioinformatic analysis. Expressions of the genes in the flowers of different colors (i.e., white, pink, red, and deep red) at the budding (S1), initial flowering (S2), blooming (S3), and withering (S4) stages of I. uliginosa were determined using qRT-PCR.  Result  Iu4CL1, Iu4CL2, Iu4CL3, and Iu4CL4 had the cDNA lengths of 1620 bp, 1653 bp, 1698 bp, and 1638 bp encoding 539, 550, 565, and 545 amino acids, respectively. Iu4CL1 contained 2 introns and Iu4CL2, 4, but Iu4CL3 and Iu4CL4 had none. All 4 genes were signal peptide-free hydrophobic proteins, but only Iu4CL3 was unsterilized. Iu4CL2 had 3 transmembrane structures, but the other 3 had none. All of them belonged to the AMP-binding enzyme superfamily and the adenylate-forming domain class I superfamily with the highest homology with I. uliginosa. Iu4CL1 and Iu4CL2 were in one large branch, while Iu4CL3 and Iu4CL4 in another, suggesting a paraphyletic homology. The expressions of Iu4CL1 and Iu4CL3 peaked in S3 on the white flowers, Iu4CL2 on red flowers, and Iu4CL4 on dark red I. uliginosa.  Conclusion   It was evident that Iu4CLs played a crucial role in the anthocyanin biosynthesis of I. uliginosa.
Characteristics of SSRs in Zicaitai Mitochondrial Genome
XIAO Wanyu, ZHOU Xianyu, REN Hailong, HUANG Hongdi, ZHENG Yansong, ZHANG Jing, XU Donglin
2024, 39(1): 49-56.   doi: 10.19303/j.issn.1008-0384.2024.01.007
[Abstract](72) [FullText HTML](29) [PDF 691KB](6)
Abstract:
  Objective   The simple sequence repeats (SSRs) in the mitochondrial genome of zicaitai (Brassica rapa var. Purpuraria) were compared with those of other species in the genus.   Method  MISA software was used to search the mitochondrial genomes of zicaitai as well as 5 other major species and 3 mutants of Brassica rapa. The SSR sequence distributions of the specimens were compared and analyzed.   Result  In the mitochondrial genomes, 168, 170, 167, 168, 180, 280, 185, 165, and 179 complete SSRs were screened. The sequences showed relative densities of 764, 774, 760, 764, 775, 777, 721, 744, and 771 per Mb with total lengths of 1562, 1577, 1547, 1562, 1664, 2564, 1722, 1524, and 1646 bp that accounted for 0.71%, 0.72%, 0.70%, 0.71%, 0.72%, 0.71%, 0.67%, 0.69%, and 0.71% of the total sequence length of each specimen, respectively. Among the 1-6 different nucleotide repeat units in zicaitai and the other major Brassica species, most SSRs were mononucleotide, which was followed by dinucleotide, tetranucleotide, trinucleotide, and pentanucleotide, but no hexonucleotide. A/T, AG/CT, AT/AT, and C/G were the common repeat unit types in them.  Conclusion  The mitochondrial genome of zicaitai was 219779 bp with 168 SSRs at the relative density of 764 markers/Mb and 9 bp in length. Mono-, di-, and tetra-nucleotides were the most abundant repeat units, which could potentially be developed as highly polymorphic markers for species differentiation.
Volatiles and Amino Acids in Pericarp of Yellow Passion Fruit at Fruit Development Stages
WU Bin, HUANG Dongmei, XING Wenting, HUANG Lixiong, YANG Qijun, SONG Shun
2024, 39(1): 57-65.   doi: 10.19303/j.issn.1008-0384.2024.01.008
[Abstract](144) [FullText HTML](100) [PDF 1043KB](8)
Abstract:
  Objective  Studying the composition and dynamic changes of volatile components and free amino acids (FAAs) in the pericarp of Yellow Passion Fruit during different growth stages, and clarifying the key flavor amino acids and characteristic volatile components, can provide references for the aroma regulation, product development, and comprehensive utilization of Yellow Passion Fruit pericarp.  Method  Using amino acid analyzer and headspace solid-phase microextraction coupled with gas chromatography-mass spectrometry (HS-SPME-GC-MS), the FAAs and volatile compounds in the pericarp of different growth stages (T1–T7) were detected and analyzed for differences.  Result  A total of 15 types of FAAs were detected in the pericarp. Among the flavor amino acids, aromatic amino acids had the highest content, reaching a maximum of 98.48 mg·hg−1 at T7, accounting for 39.03%. OPLS-DA and TAV analyses indicated that cystine was the key flavor amino acid that affects aroma formation. A total of 90 volatile compounds were detected in the pericarp. Aldehydes were dominant during the T1–T6 stages, while terpenes were dominant at T7. OPLS-DA and ROAV analyses showed that there were 9 types of characteristic volatile compounds with differential expression. Among them, 1-octen-3-one was the dominant compound during the T1~T5 stages, followed by benzaldehyde (T1–T4 stages) and heptanal (T5 stage). During the T6 and T7 stages, (E)-β-damascone contributed the most.  Conclusion  Cystine was the crucial flavoring amino acid associated with the aroma formation of the pericarp. The volatile composition varied by fruit development stages, such as 1-octene 3-one was the dominant component when the pericarp was forming color in T1-T5, while (E)- β -damalenone when the fruit was maturing in T6 and T7.
Genetic Diversity and DNA Fingerprints of Tea Germplasms in Yongtai, Fujian
WANG Pan, YU Wentao, LV Shuiyuan, ZHU Yanyu, LAN Zhenliang, LI Qingying, LI Guolai, GUI Wenjing, YE Naixing
2024, 39(1): 66-74.   doi: 10.19303/j.issn.1008-0384.2024.01.009
[Abstract](98) [FullText HTML](30) [PDF 2318KB](15)
Abstract:
  Objective  Genetic diversity and population structure of tea germplasms in Yongtai County were analyzed for resource utilization.  Method  The EST-SNP molecular marker technology was applied to analyze the genetic relationships and diversity of 77 tea germplasms collected from Yongtai and surrounding areas.  Result   Forty-eight SNP loci for identifying the germplasms were selected. The average polymorphism information index of the loci was 0.407 with an average observed heterozygosity of 0.303 and an expected one of 0.271, as well as the average fixed index of −0.092 and the frequency of secondary alleles of 0.268. The principal coordinate, cluster, and structure population structure analyses revealed that the interactions within a same population of the tea plants were stronger than among different populations at Yongtai, and that the tea germplasms found in Wutong significantly differed from those in Fukou, Danyun, and other places with significant genetic distance but close to those from Youxi and Datian. Using the 48 selected high-quality SNP loci, a library of the DNA fingerprints on 54 germplasms from Yongtai was established to provide a means for accurate germplasm identification.   Conclusion   Yongtai County was rich in the resource of varieties of tea plants, which could be classified geographically into those from Wutong and those from Fukou, Danyun and other localities in the county. The cultivars from Wutong were more closely related to those found in Datian and Youxi, whereas those from Fukou, Danyun, etc. more to those existed in the north part of the county.
Formulation of Antrodia camphorata Culture Medium Incorporated with Pseudostellaria heterophylla By-product
ZHAO Shengyue, WU Xiaohuan, JIANG Chunling, ZHANG Yu, ZHENG Shizhong, LIU Shengrong, YE Zuyun
2024, 39(1): 75-82.   doi: 10.19303/j.issn.1008-0384.2024.01.010
[Abstract](51) [FullText HTML](24) [PDF 1060KB](1)
Abstract:
  Objective   To utilize a Pseudostellaria heterophylla by-product for increasing Antrodia camphorata yield and bio-effectiveness, formulation of a culture medium was optimized.   Methods   Powder of P. heterophylla fine roots was included in formulating a medium to maximize the mycelial growth of A. camphorata by a Plackett-Burman orthogonal experimental design.  Results   In addition to 6 g·L−1 of P. heterophylla powder, the medium was optimized to contain the ingredients critical to the mushroom growth, which included glucose 10 g·L−1, yeast extract 4 g·L−1, peptone 6 g·L−1, and KH2PO4 1.5 g·L−1. The A. camphorata culture on the formulated medium produced 4.73 g·L−1 biomass, which yielded 2.75% more intracellular triterpenoids than control and 24.85% more than PDB medium, and 161.11% more exopolysaccharides than control and 113.64% more than PDB medium. The fermentation broth also contained a high asexual spore at a density of 1.8×107 spores·mL−1.   Conclusion  P. heterophylla root powder could plausibly be added to the medium for A. camphorate culture as it promoted the mycelial growth, bioactive constituents yield, and asexual spore production of the mushroom.
Plant Protection
Lightweight Residual Networks for Diagnosis of Apple Leaf Diseases
ZHOU Hanmi, CHEN Jiageng, DAI Zhiguang, NIU Xiaoli, QIN Long, XIANG Youzhen, ZHAO Long
2024, 39(1): 83-92.   doi: 10.19303/j.issn.1008-0384.2024.01.011
[Abstract](0) [FullText HTML](0) [PDF 8364KB](0)
Abstract:
  Objective  An improved diagnostic model on apple leaf diseases was developed applying the lightweight residual networks.  Method  Data imbalance and overfitting of the original ResNet18 model was reduced by offline and online enhancements on its generalization ability. A scaling factor was introduced to minimize the number of network parameters and maximize the pooling layer in the constant mapping of down-sampled residual structure instead of 1×1 convolution. Redundant features in pictures were eliminated, and sensory field of the model increased. The first 7×7 convolutional layer of ResNet18 was replaced with a multi-scale feature extraction module to enrich fine lesions extraction. Finally, the DenseBlock module was inserted in the network to fully utilize valid shallow features.   Results  The improved ResNet18 model achieved an accuracy of 97.94%, which was a 0.88 percentage increase, with a significant 90.77% reduction on the program size of 3.97 MB. It performed superbly in comparison to other light-weight models, such as ShuffleNetv2, MobileNetv3, and EfficientNet, or the classical models, such as Inceptionv2, DenseNet, and ResNet.   Conclusion   The improved ResNet18 model could accurately identify the apple leaf diseases under complex circumstances. With fewer parameters than the original program, it could be more easily installed in a variety of devices for the diagnosis.
Resources and Environmental Science
Host Selection of Ectomycorrhizal Fungi at Pinus massoniana and Castanopsis carlesii Forests
ZHANG Xiaohui, XIE Rongzhang, ZHANG Jinyan, MAN Jiayin, SUN Daizhen, DENG Zongjie, WEI Lihua, LIAN Chunlan
2024, 39(1): 93-104.   doi: 10.19303/j.issn.1008-0384.2024.01.012
[Abstract](39) [FullText HTML](25) [PDF 1565KB](3)
Abstract:
  Objective   Host selection of ectomycorrhizal fungi (ECMF) in forest soil was studied.   Method  In the soils at forests of Pinus massoniana (Pm) and Castanopsis carlesii (Cc), various fungi were inoculated into Pm or Castanopsis uraiana (Cu) and designated as treatments of Pm-PmS, Pm-CcS, Cu-PmS, and Cu-CcS. After cultivating the inoculated seedlings for 6 months, mycorrhizal identification on the fungi was performed by ITS. The frequency, infection rate, relative abundance, relative frequency, richness, and diversity of ECMF in the roots of Pm and Cu plants were monitored or calculated. Seedling growth indexes and soil physiochemical properties were determined.   Result   The Cc forest soil (CcS) showed significantly higher pH and contents of total phosphorus, total carbon, and available phosphorus as well as the seedling shoot dry weight and root length than the Pm counterpart (PmS). The 19 OTUs of ECMF detected in these soil samples belonged to 7 families and 10 genera. Of which, Cenococum geophilum, Rhizopogon boninensis, and Tomentella sp. 2 were commonly found on the two soils. Out of the 13 ECMF identified in PmS, 8 infected Pm and 6, Cu; while among the 9 ECMF identified in CcS, 4 infected Pm and 7, Cu. Both C. geophilum and Sebacina sp. 2 were symbiotic with Pm and Cu. Hyaloscyphaceae sp., Lactarius inconspicuous, Rh. boninensis, Rh. flavidus, Tomentella sp. 1, Tomentella sp. 3 and Tomentellopsis submollis infected Pm, whereas Athelia sp., Amanita sp., L. atrofuscus, Russula minor, Russula sp., Sebacina sp.1, Thelephora sp. 1, Thelephora sp. 2 and Tomentella sp. 4 infected only Cu. The ECMF richness index (IV), Shannon index (H') and Simpson index (D) of PmS were higher than those of CcS. However, the Sorensen index on the PmS planted with host plants other than Pm was 0.14, which was lower than 0.36 on the CcS planted not with Cc. The infection rates of some ECMF were closely related to the physiological and ecological properties of the host.   Conclusion   The symbiosis between ECMF and trees has evolved in a long process. The ECMF in the soil at a Pm forest tended to infect Pm specifically, and so did those at a Cc forest to Cc, Cu or Fagaceae plants. However, the ECMF in PmS were more selective on their host plants than those in CcS. Even though soil physiochemical properties also affected the ECMF infection, the species of host plant largely determined the fungal colonization on the land.
Review
Research Progress on Biochar Application for Continuous Cropping of TCM Plants
GAO Hua, TU Haoze, ZHAO Yuyuan, KONG Wen, XIA Wenjian, WANG Feier, Muhammad Shaaban, LIN Shan
2024, 39(1): 105-114.   doi: 10.19303/j.issn.1008-0384.2024.01.013
[Abstract](0) [FullText HTML](0) [PDF 698KB](0)
Abstract:
As a major country in the world engaging in large-scale cultivation of traditional Chinese medicine (TCM) plants, China is facing a serious challenge in growth of the industry. Due to malpractices and poor management, undesirable consequences such as productivity decline and quality deterioration, relocation of authentic origins, insufficient resource protection, and uncoordinated material utilization have surfaced. The causes of the ill-effects may not be all clearly identified, nonetheless, rampant continuous cropping is deemed as one of the most urgent targets for correction. A generally alkali material, biochar is commonly applied to amend the soil in question to improve fertility, reduce acidification, and promote growth of the plants cultivated on it. It has been used by farmers in recent years, and the measure could be implemented to mitigate the damaging effects induced by the TCM continuous cropping as well. This article reviewed the benefits of biochar application on soil, such as adjusting the physiochemical properties, microbial community, and allelopathic environment. Some practical treatments for improvements are presented.
Research Progress on Glandular Trichome Secretions of Plants
QI Chaofan, LIU Yanhua, LIU Jing, DU Yongmei, LIU Xinmin, HAN Xiao, LEI Yunkang, ZHANG Hongbo, FU Qiujuan, OUYANG Yiming, LIU Guoxia
2024, 39(1): 115-124.   doi: 10.19303/j.issn.1008-0384.2024.01.014
[Abstract](121) [FullText HTML](61) [PDF 1503KB](14)
Abstract:
Plant glandular trichomes (GTs) are epidermal hairs on the surface of stems, leaves, floral organs, and fruits. Dubbed as the “biosynthesis factory,” they secret substances that can resist biotic and abiotic stresses to protect the plant. As secondary metabolites, they are potentially important in the development of and utilization for medicines, pesticides, and foods. This article reviewed the recent research progress on the types, extraction, isolation, and functionalities of various plant GTs and discusses the areas for future studies.
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