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Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(1): 1-146.  
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Young Scientist Forum
Advances in animal models and treatment of dilated cardiomyopathy
Jiamin JIN,Qian GONG,Lenan ZHUANG
Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(1): 1-11.   https://doi.org/10.3785/j.issn.1008-9209.2023.05.081
Abstract( 69 )   HTML( 12 )   HTML (   PDF(940KB)( 67 )

Dilated cardiomyopathy (DCM) is a cardiovascular disease characterized by one or both ventricular dilation and systolic dysfunction. Its pathogenesis involves inherited genetic mutations and various secondary factors. Human DCM animal models have been developed using a variety of experimental animals such as mice, rats, zebrafish, and pigs, and are generally constructed by gene editing, drug induction, autoimmune deficiency induction, and viral infection. The previous studies have utilized DCM animal models to thoroughly investigate the pathogenic mechanisms and therapeutic targets of this disease. This paper briefly described the pathological features, clinical symptoms, and epidemiological characteristics of human DCM. Furthermore, it reviewed the types of DCM animal models and their construction methods used in recent years. This paper also presented new perspectives on optimizing modeling methods and promoting therapeutic research for DCM. Therapeutic studies based on DCM animal models can help us better understand the mechanisms of DCM and provide a basis for the development of new therapeutic approaches.

Advances in studies on sugar transporters in ornamental plants
Ziyang HUANG,Jie LIU,Jie KANG,Ziming REN,Qi CUI,Dongze LI,Yiping XIA,Si MA,Yun WU
Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(1): 12-24.   https://doi.org/10.3785/j.issn.1008-9209.2023.03.241
Abstract( 104 )   HTML( 11 )   HTML (   PDF(4869KB)( 69 )

Ornamental plants are widely used for beautifying the living environment because of their good ornamental value of flowers, leaves and fruits. Sugar transporters play critical roles in the growth and development, flowering and fruiting, and response to stress of ornamental plants. Therefore, it is of great significance to study the effects of sugar transporters on ornamental plants. Monosaccharide transporters, sucrose transporters/sucrose carriers and sugars will eventually be exported transporters are three major sugar transporters discovered in plants. The classification and basic characteristics of these sugar transporters in different ornamental plants were compared and discussed. We also summarized the function and regulation of these transporters in terms of recent research progress to provide a theoretical basis for the future use of genetic engineering technology to improve ornamental plants and enhance their ornamental and adaptive properties.

Research advances on regulating intestinal flora and their physiological functions by phytochemicals of goji berry
Ranran BI,Yuan ZHAO,Yujing SUN
Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(1): 25-34.   https://doi.org/10.3785/j.issn.1008-9209.2023.03.231
Abstract( 68 )   HTML( 4 )   HTML (   PDF(923KB)( 64 )

Goji berry contains several nutrients and active ingredients, with certain edible and medicinal value. The past studies have found that goji berry extracts have a variety of biological activities such as immune regulation and antitumor and anticancer activities. Recent research has shown that goji berry and its extracts can also regulate the intestinal flora. This paper reviewed the phytochemicals such as Lycium barbarum polysaccharides, anthocyanins, betaines, phenolic acids, flavonoids, and carotenoids in the fruits of L. barbarum and L. ruthenicum Murray, and outlined the effects of the whole fruit of goji berry and its extracts on the intestinal flora and their physiological functions. This paper provides prospects for future research.

Biological Sciences & Biotechnologies
Screening of small molecule inhibitors targeting DNA polymerase η
Jiajia CAO,Shumai YE,Ye ZHAO
Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(1): 35-41.   https://doi.org/10.3785/j.issn.1008-9209.2023.02.221
Abstract( 60 )   HTML( 6 )   HTML (   PDF(2598KB)( 67 )

The repair of DNA damage and maintenance of genomic stability are essential for the normal growth and adverse defense of plants and animals. In view of the genomic instability caused by the misincorporation of DNA polymerase, this study took DNA polymerase η as the research object and screened its possible small molecule inhibitors by computational simulated molecular docking and detected their enzyme kinetic parameters. The results showed that deoxyadenosine triphosphate (dATP) had an inhibitory effect on the activity of DNA polymerase η, resulting in a relative extension efficiency of 36% to 42%. Simulated molecular docking and in vitro experimental results showed that cyclic GMP-AMP (cGAMP) had a lower binding energy (with an affinity of -35.1 kJ/mol) than dATP (with an affinity of -26.7 kJ/mol) to DNA polymerase η. Enzyme kinetic experiments also showed that cGAMP had a stronger inhibitory ability than dATP and achieved the maximum effect at the concentration of 0.5 mmol/L (with a relative extension efficiency of 13%). Therefore, a potential small molecule inhibitor targeting DNA polymerase η was screened out in this study. At the same time, in view of the tolerance to antitumor drug (DNA damage agent) caused by high expression of this protein, these results provide a basis for the development of new drugs.

Horticultural Sciences
Study on influencing factors of bulblet enlargement in vitro of Lilium amoenum
Minhui ZHANG,Junji CHEN,Wei QUAN,Zhijia GU,Hongzhi WU
Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(1): 42-52.   https://doi.org/10.3785/j.issn.1008-9209.2023.03.161
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Lilium amoenum is a rare lily germplasm with an agreeable rose fragrance. This study aimed to explore the optimal culture conditions and medium formula for the enlargement of L. amoenum bulblets in vitro. Bulblets of L. amoenum with a diameter of 0.4-0.6 cm were inoculated on Murashige & Skoog (MS) medium supplemented with different concentrations of sucrose or sucrose combined with naphthalene acetic acid (NAA), N-(phenylmethyl)-9H-purin-6-amine (6-BA), indole butyric acid (IBA), or paclobutrazol (PP333). The culture conditions of 16 hours of light/8 hours of darkness and full darkness were set up. After 45 days of culture, the enlargement coefficient and diameter increase multiple of the bulblets were counted. The results showed that in the range of experimental concentrations, with the increase of sucrose and hormone concentrations, the enlargement coefficient and diameter increase multiples of L. amoenum bulblets overall first increased and then decreased. When the sucrose concentration was 50-120 g/L, the bulblet enlargement effect at 90 g/L sucrose was the greatest, and the bulblets were green and shiny, and had good cohesion. When 50-120 g/L sucrose was combined with 0.1-0.7 mg/L NAA/6-BA, 0.5-4.0 mg/L IBA or 5-40 mg/L PP333, the effect of bulblet enlargement was the greatest at 70 g/L sucrose and 1.0 mg/L IBA, and the enlargement coefficient and diameter increase multiple were 3.63 and 1.83, respectively. The synergistic effect of high concentrations of sucrose and PP333 significantly inhibited the enlargement of bulblets. Lilium amoenum bulblets cultured under 16 hours of light/8 hours of darkness grew better than those cultured under full darkness, but the rooting effect was greater under full darkness. This study provides a scientific basis for shortening bulb growth time in the field and for germplasm resource protection.

Plant Protection
Mitochondrial genome of Lipaphis pseudobrassicae (Davis, 1914) and phylogenetic analysis of Aphidinae
Xingyu LIN,Nan SONG
Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(1): 53-64.   https://doi.org/10.3785/j.issn.1008-9209.2023.02.161
Abstract( 49 )   HTML( 0 )   HTML (   PDF(1891KB)( 39 )

Lipaphis pseudobrassicae (Davis, 1914) is an important agricultural insect pest. In this study, we sequenced and analyzed the entire mitochondrial genome of L. pseudobrassicae, and reconstructed the phylogenetic relationships of Aphidinae based on the mitochondrial genome data. In phylogenetic analyses, the ingroup included 31 species of Aphidinae, while the outgroup selected three species of Eriosomatinae (Eriosoma lanigerum, Paracolopha morrisoni and Meitanaphis microgallis). The maximum likelihood method and Bayesian method were used to reconstruct the phylogenetic tree of Aphidinae, respectively. The results showed that the mitochondrial genome of L. pseudobrassicae was 16 743 bp in length (GenBank accession No. OP796483), which contained 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes and one non-coding control region. The phylogenetic tree constructed by the maximum likelihood method and Bayesian method both showed that the tribe Aphidini was a monophyletic group. The tribe Macrosiphini was recovered as a monophyletic group, and Lipaphis was a sister to Brevicoryne according to the maximum likelihood method. However, the tribe Macrosiphini was recovered as a non-monophyletic group, and Lipaphis was a sister to Brevicoryne+Diuraphis according to the Bayesian method. This study can enhance the understanding of the phylogenetic relationships among Macrosiphini, Lipaphis, and L. pseudobrassicae by utilizing mitochondrial genome data.

Preliminary evaluation of 110 peach/nectarine varieties on the gummosis disease resistance in Zhejiang Province
Yuqi WANG,Kexin GAN,Xiongwei LI,Jing JIN,Lan ZHAO,Zhengwen YE,Deli SUN,Li WANG,Huijuan JIA,Zhongshan GAO
Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(1): 65-74.   https://doi.org/10.3785/j.issn.1008-9209.2023.02.271
Abstract( 51 )   HTML( 0 )   HTML (   PDF(2954KB)( 30 )

This study aimed to screen highly resistant varieties by investigating field gummosis disease levels in 110 local peach/nectarine varieties and selected varieties of 4- to 7-year-old trees. Combined with pedigree analysis, the source of the gummosis disease resistant germplasms was traced. The results showed that there were significant differences in the levels of gummosis disease among different peach/nectarine varieties, and the resistance gradually decreased with the increase of tree age. Among the 80 varieties that showed relatively stable levels of gummosis disease, nine varieties showed high resistance, most of which were peach varieties, such as ‘Qiubaitao’ and ‘Nanshantiantao’; 40 varieties showed resistance, such as ‘Hakuri’ and ‘Yuanmeng’; 16 varieties showed moderate resistance, such as ‘Hujingmilu’ and ‘Jinxiu’; nine varieties were susceptible to the disease, such as ‘Feicheng Baili’ and ‘Kurakato Wase’; and six varieties showed high susceptibility, such as ‘Bailutao’ and ‘Huyou 278’. The peach varieties were generally more resistant than the nectarine varieties. According to the genetic pedigrees, some resistant and highly resistant varieties, which originated from the traditional Chinese local variety ‘Chinese Cling’, had a genetic relationship. This research provides a reference for the selection of resistant varieties in production, as well as for further genetic and breeding applications.

Food Sciences
Effects of volatile organic compounds on postharvest brown rot control and sensory quality of peach fruit
Zhihao LI,Siyin LIN,Ying GAO,Can YANG,Dan JIANG,Bo ZHANG
Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(1): 75-85.   https://doi.org/10.3785/j.issn.1008-9209.2023.03.011
Abstract( 55 )   HTML( 6 )   HTML (   PDF(5600KB)( 46 )

Peach fruit is susceptible to Monilinia fructicola during storage and logistics, leading to occurrence of brown rot and fruit quality deterioration. However, the regulatory effects of volatile organic compounds (VOCs) on brown rot remain unclear. This study analyzed the antifungal effects of major VOCs in ripe peach fruit on M. fructicola. The results showed that 12 VOCs involving carvacrol and (E)-2-hexenal were identified to have significant inhibitory effects on the growth of M. fructicola on media. Further research was conducted on the regulatory effects of 12 VOCs on postharvest brown rot of peach fruit. The results showed that the fumigation treatment with VOCs could effectively inhibit the growth of M. fructicola and delay the occurrence of brown rot. Among them, volatile aldehydes exhibited positive effects on maintaining fruit quality. Fumigation with 25 μL/L (E)-2-hexenal significantly inhibited the growth of M. fructicola and reduced brown rot symptoms in peach fruit. Meanwhile, the fruit appearance, ethylene release rate, hardness, total soluble solid content and sensory quality indicators were not affected, therefore maintaining the merchant ability of peach fruit during the postharvest storage. In summary, (E)-2-hexenal has the potential to be developed as a plant-based fungicide, offering quality assurance for the peach fruit supply chain.

Quality difference analysis of commercial dried hawthorn fruits from different producing areas
Xipan SHU,Ziqi CHAI,Xian LI,Chongde SUN,Jinhu TIAN,Xingqian YE
Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(1): 86-97.   https://doi.org/10.3785/j.issn.1008-9209.2023.04.041
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Taking ten kinds of commercial dried hawthorn fruits [seven kinds of Crataegus pinnatifida Bge. or C. pinnatifida Bge. var. major N.E.Br. (with the trade name “Beishanzha”), two kinds of Malus doumeri (Bois) Chev. (with the trade name “Guangshanzha”) and one kind of C. cuneata Sieb. et Zucc. (with the trade name “Nanshanzha”)] from different producing areas of China as research materials, the contents of main nutrients and biochemical indexes were determined, and the quality difference analysis and principal component analysis were carried out. The results showed that among the three types of commercial dried hawthorn fruits, Beishanzha had the highest titratable acid content, while Guangshanzha had the highest ratio of soluble sugar to titratable acid. The contents of soluble dietary fiber, α-tocopherol and β-carotene decreased in the order of Beishanzha, Nanshanzha and Guangshanzha. The results of principal component analysis indicated that ‘Yeshanzha’ from Huanggang of Hubei Province and ‘Tieshanzha’ from Jizhou of Tianjin City showed the best overall performance in the contents of main active components (total phenolics, total flavonoids, total procyanidins and total dietary fiber); Beishanzha, especially ‘Dajinxing’ from Linyi of Shandong Province and ‘Yubeihong’ from Xinxiang of Henan Province, had greater contents of titratable acid, soluble dietary fiber and soluble sugar compared with Guangshanzha and Nanshanzha. This study systematically sorted out the quality differences of commercial dried hawthorn fruits from different areas in China, and the results can provide data to support the diversified development and utilization of Chinese commercial hawthorn resources.

Resource Utilization & Environmental Protection
Composition and dynamic change characteristics of human pathogenic communities in dryland farmland with manure application
Minghui QI,Jianhua CHENG,Xiangyu TANG
Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(1): 98-108.   https://doi.org/10.3785/j.issn.1008-9209.2023.03.012
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To understand the community structure composition and dynamic change characteristics of human pathogenic bacteria (HPB) in soils after manure application, laboratory cultivation experiments were conducted on agricultural soils with long-term application of chicken manure, pig manure, or chemical fertilizer in five regions of Hangzhou, Jiaxing, Quzhou, Jinhua, and Longyou in Zhejiang Province, and the community compositions of the soil bacteria and HPB were analyzed by high-throughput sequencing and sequence alignment methods. The results showed that a total of 75 HPB were detected in 160 soil samples and two manure samples, and the dominant HPB were Bacillus_megaterium_QM_B1551 (24.2%) and Clostridium_beijerinckii_NCIMB_8052 (23.1%). The Shannon indexes of bacteria and HPB in the soils decreased after the application of pig manure, while the diversities of bacteria and HPB in the soils with the application of chicken manure or chemical fertilizer had no significant changes. The results of the principal coordinate analysis showed that there was a significant difference in the bacterial community composition of soils between the manure treatment and the unfertilized control, especially in the pig manure treatment (P<0.001); 22.7% of all HPB were shared among the soil samples; and the relative abundance of most HPB in the soils treated with manure was higher than that in the unfertilized control, and it decreased continuously with the extension of cultivation time. The results of the variance partitioning analysis showed that soil physicochemical properties, bacterial communities, and their interactions were important factors contributing to the variation of HPB in the soils. In summary, the HPB variation characteristics in soils treated with manure are influenced mainly by manure types, soil types, soil physicochemical properties, and inherent bacterial communities.

Effects of long-term non-flooding plastic film mulching and application of coated urea on rice yield, nitrogen use efficiency and soil nutrients
Tong QI,Sheng TANG,Jingjie ZHOU,Qingxu MA,Lianghuan WU
Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(1): 109-122.   https://doi.org/10.3785/j.issn.1008-9209.2023.02.131
Abstract( 56 )   HTML( 1 )   HTML (   PDF(1533KB)( 71 )

Non-flooding plastic film mulching cultivation (PM) for rice is a comprehensive and innovative technology that utilizes plastic film covering as the core to achieve water-saving rice production. However, after mulching with plastic film, nitrogen (N) fertilizer can only be applied once as a basal fertilizer before transplanting, which will lead to excessive vegetative growth at the early stage and potential N deficiency at the late growth stage, thereby limiting the high yield of rice. Polymer coated urea (CR) is a controlled release N fertilizer that has become one of the best management measures for improving crop yield and N use efficiency under a traditional flooding cultivation (TF) pattern, but it has not been evaluated in a long-term positioning test under the PM pattern. In this study, taking the high-yielding and medium-maturing indica hybrid rice cultivar ‘Liangyoupeijiu’ as a test material, the effects of applying CR and urea (UR) on rice yield, N use efficiency and soil nutrient contents were compared under the PM and TF patterns. The results showed that, compared with applying UR, applying CR under the TF and PM patterns improved the N use efficiency by 9.2% and 15.4%, respectively (P<0.05), and increased the rice yield by 8.6% and 15.0%, respectively (P<0.05). Compared with the TF pattern, the PM pattern accelerated the decomposition of soil organic matter and reduced the contents of total N and alkali-hydrolyzable N in the soil. Compared with applying UR, applying CR under the PM pattern alleviated the decrease of the total N and alkali-hydrolyzable N contents in the soil and increased the economic benefits by 16.8%. In summary, applying CR is an effective way to solve the problem of N deficiency at the late growth stage of rice under the PM pattern.

Animal Sciences & Veterinary Medicines
Characteristics of microbial communities and their association with immunity and metabolism at the early developmental stage of Protosalanx chinensis
Yifan ZHOU,Xizhao ZHANG,Yanfeng ZHOU,Dongpo XU,Chenhe WANG,Shiyue GUO,Shulun JIANG,Yang YOU
Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(1): 123-136.   https://doi.org/10.3785/j.issn.1008-9209.2023.03.281
Abstract( 58 )   HTML( 1 )   HTML (   PDF(3592KB)( 36 )

In order to understand the changes and characteristics of microbial communities during early development of Protosalanx chinensis, samples were collected at five developmental periods, which are the embryonic period (heartbeat stage, XT), the endogenous nutrition period (the first day after hatching, H1), the mixed nutrition period (the fourth day after hatching, H4), the open feeding period (the seventh day after hatching, H7), and the exogenous nutrition period (the tenth day after hatching, H10). Microbial community succession during early development was observed by 16S rRNA gene sequencing technology, especially the characteristics and key microbial genera before and after feeding. Combined with the transcriptome data of the same batch of samples, the microbial genera related to immunity and metabolism were analyzed based on the association network method. The results showed that there were significant differences in β diversity among the different periods at the early developmental stage (P<0.001). The dominant bacteria in the XT period were Flavobacterium and Chryseobacterium, and the dominant bacterium in the H1 period was Pseudomonas. The main bacteria in the H4 period were Flavobacterium and Pseudomonas. The main bacteria in H7 and H10 periods were Flectobacillus and Pseudomonas. The abundance of Pseudomonas was stable at all developmental periods. Node bacteria such as Flectobacillus were significantly associated with the expression of various immune and metabolic genes. This study has obtained the microbial succession information of the early developmental stage of P. chinensis for the first time, and screened out the dominant bacteria and node bacteria, which will provide references and ideas for the scaled cultivation of P. chinensis fry.

Preparation of polyclonal antibodies and subcellular localization analysis of chicken apolipoprotein A-Ⅰ
Shengwen WANG,Dan ZHANG,Yuqian WU,Jiyong ZHOU,Xiaojuan ZHENG
Journal of Zhejiang University (Agriculture and Life Sciences), 2024, 50(1): 137-146.   https://doi.org/10.3785/j.issn.1008-9209.2023.02.061
Abstract( 40 )   HTML( 1 )   HTML (   PDF(4557KB)( 30 )

Apolipoprotein A-Ⅰ (Apo A-Ⅰ) plays important roles in the regulation of atherosclerosis, viral infections, lipid metabolism and other aspects. However, there are few studies on chicken Apo A-Ⅰ (chApo A-Ⅰ), and its biological function is not well understood. Based on the bioinformatics analysis of chApo A-Ⅰ, this study further performed the expression and purification of the recombinant protein of chApo A-Ⅰ via the pET-28a prokaryotic expression system. Mouse polyclonal antiserum was prepared by immunizing mice with purified recombinant protein. Enzyme-linked immunosorbent assay (ELISA) was used to determine the titer of the polyclonal antiserum, and Western blot (WB) and indirect immunofluorescence assay (IFA) were used to determine its reactivity. Then, the polyclonal antiserum was further applied for subcellular localization analysis of chApo A-Ⅰ. Bioinformatic analysis revealed that the chApo A-Ⅰ protein contains signal peptide at 1-18 amino acids, which is composed of continuous alpha helix at the N-terminal. Homology analysis of amino acid sequences revealed that the chApo A-Ⅰ protein had the highest homology with turkey and the lowest homology with fish. The polyclonal antibody prepared using successfully expressed and purified recombinant protein His-chApo A-Ⅰ had an ELISA titer above 1×105 and specifically reacted with the eukaryotic expressed chApo A-Ⅰ protein in WB and IFA. Particularly, the antibody can recognize the Apo A-Ⅰ protein in chicken serum, but cannot cross-react with Apo A-Ⅰ proteins in the serums of mice, rabbits, cattle or pigs. This polyclonal antibody was further applied for subcellular localization analysis of full-length chApo A-Ⅰ (chApo A-Ⅰ-FL) and chApo A-Ⅰ without signal peptide (chApo A-Ⅰ-NS). Observed by confocal microscope, it was found that chApo A-Ⅰ-FL protein was mainly localized near the cell membrane, but chApo A-Ⅰ-NS protein was localized in the cytoplasm, and most of them were diffusely distributed. The specific polyclonal antibody and the results of subcellular localization of chApo A-Ⅰ provide a basis for further research on the biological function of Apo A-Ⅰ.

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