2024 Vol. 39, No. 1
Display Method:
2024, 39(1): 1-6.
doi: 10.19303/j.issn.1008-0384.2024.01.001
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.
2024, 39(1): 7-16.
doi: 10.19303/j.issn.1008-0384.2024.01.002
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.
2024, 39(1): 17-24.
doi: 10.19303/j.issn.1008-0384.2024.01.003
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.
2024, 39(1): 25-32.
doi: 10.19303/j.issn.1008-0384.2024.01.004
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.
2024, 39(1): 33-39.
doi: 10.19303/j.issn.1008-0384.2024.01.005
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.
2024, 39(1): 40-48.
doi: 10.19303/j.issn.1008-0384.2024.01.006
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.
2024, 39(1): 49-56.
doi: 10.19303/j.issn.1008-0384.2024.01.007
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.
2024, 39(1): 57-65.
doi: 10.19303/j.issn.1008-0384.2024.01.008
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.
2024, 39(1): 66-74.
doi: 10.19303/j.issn.1008-0384.2024.01.009
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.
2024, 39(1): 75-82.
doi: 10.19303/j.issn.1008-0384.2024.01.010
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.
2024, 39(1): 83-92.
doi: 10.19303/j.issn.1008-0384.2024.01.011
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.
2024, 39(1): 93-104.
doi: 10.19303/j.issn.1008-0384.2024.01.012
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.
2024, 39(1): 105-114.
doi: 10.19303/j.issn.1008-0384.2024.01.013
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.
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.
2024, 39(1): 115-124.
doi: 10.19303/j.issn.1008-0384.2024.01.014
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.
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.
