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Monthly, Started in1993
ISSN 1674-7968  CN 11-3342/S
Administrated by: Ministry of Education of People's Republic of China
Sponsored by: China Agricultural University and Chinese Society of Agricultural
WU Wei-Hua
Associate editor-in-chief:CHEN Hua-Lan   LI Kui   LI Yi   LIN Min   PENG Yu-Fa
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2024 Vol. 32, No. 2  Published: 01 February 2024
Articles and Letters
Study on the Function of TaGRF1-A in the Interaction Between Wheat (Triticum aestivum) and Puccinia triticina
GU Jia, NIU Ze-Lin, WANG Qian, WANG Dong-Mei
2024 Vol. 32 (2): 247-258   DOI:10.3969/j.issn.1674-7968.2024.02.001 |  Full text (HTML) (1 KB)  |    PDF  (6604 KB)  ( 22 )
The cloning of genes related to wheat (Triticum aestivum) resistance leaf rust and the in-depth study of their expression characteristics are of great significance for elucidating the molecular mechanisms of wheat resistance Puccinia triticina. By analyzing the RNA-seq transcriptome database obtained in the laboratory in the early stage, a gene belong to 14-3-3 family closely related to resistance to P. triticina infection was discovered, with significant differences in expression between compatible and incompatible combinations. In order to deeper understanding of the mechanism of this gene in wheat resistance to P. triticina infection, this study cloned the gene from wheat near isogenic lines TcLr26, the result of sequence alignment showed that the gene was TaGRF1-A in wheat. Preparation of polyclonal antibodies against TaGRF1-A protein, Western blot analysis showed that the antibody was able to bind specifically to TaGRF1-A protein in wheat, and showed an upregulation and then decrease expression trend induced by Puccinia triticina in the incompatible combinations. The virus induced gene silencing (VIGS) technology was used to reduce the expression of TaGRF1-A in incompatible combinations, inoculate with leaf rust showed a significant increase in the area of hypersensitive reaction (HR) and the number of haustorium mother cells (HMCs) at the site of P. triticina infection. After 15 d of inoculation with leaf rust, a spore of P. triticina appeared on the surface of the leaves, and the expression of pathogenesis-related (PR) genes were reduced. The results indicated that TaGRF1-A was positive regulating the wheat resistance to leaf rust infection. This study provides reference for further exploring the mechanism of TaGRF1-A in the interaction between wheat and leaf rust.
Genome-wide Association Study and Candidate Gene Prediction for Plant Height in Wheat (Triticum aestivum)
CHE Zhuo, WANG Peng, TIAN Tian, ZHANG Pei-Pei, CHEN Tao, LIU Yuan, YANG De-Long
2024 Vol. 32 (2): 259-272   DOI:10.3969/j.issn.1674-7968.2024.02.002 |  Full text (HTML) (1 KB)  |    PDF  (4118 KB)  ( 28 )
Plant height (PH) is one of the important agronomic traits in wheat (Triticum aestivum) and is highly correlated with wheat yield. In order to further illustrate the genetic basis of wheat PH, 132 wheat cultivars (lines) were phenotyped under 4 environmental conditions in this study, and genome-wide association analysis (GWAS) of PH was performed based on the mixed liner model (MLM) using a wheat 35K single nucleotide polymorphism (SNP) assay. The results showed that the phenotypic variation of wheat PH was significantly affected by environment and was significantly correlated with the environment, with a broad-sense heritability (h2B) of 0.60. A total of 22 SNPs significantly associated with PH (P≤0.001) were identified in this study, among which 7 SNPs were identified in 2 or 3 environments, which were stably associated SNPs and distributed on chromosomes 1A, 1B, 1D, 2D, and 4A. One to 3 stably associated SNPs identified on each chromosome, and a single stably associated SNP could explain 9.15%~15.07% of the phenotypic variation. 4 stably associated SNPs overlapped with reported QTLs for PH, and the remaining 3 stably associated SNPs were new loci probably. 265 genes in the 1 Mb interval upstream and downstream of the stably associated loci were analyzed for functional homology in rice (Oryza sativa), and 7 candidate genes were screened for possible effects on PH development. This study may provide new markers and genetic resources for wheat PH improvement.
Effects of Phosphorus Transporter StPHO1.3 on Heat Resistance of Potato (Solanum tuberosum) and Screening of Its Interacting Proteins
LI Wan, LI Cheng, CHENG Min, WU Fang
2024 Vol. 32 (2): 273-282   DOI:10.3969/j.issn.1674-7968.2024.02.003 |  Full text (HTML) (1 KB)  |    PDF  (9081 KB)  ( 10 )
Plants absorb and transport phosphorus through phosphorus transporters (PHO), and phosphorus is one of the essential nutrients for plant growth and development, which has an important effect on crop yield, quality and stress resistance. High temperature is an important environmental factor affecting potato yield and quality. In order to explore the influence of phosphorus transporter on potato (Solanum tuberosum) heat resistance, this study constructed a transgenic potato strain with overexpression of phosphorus transporter gene StPHO1.3 mediated by Agrobacterium tumefaciens. By comparing the growth and development of transgenic strain and wild-type strain at high temperature (35 ℃), it was found that overexpressing StPHO1.3 could enhance the heat resistance and promote the growth and development of potato. In addition, the results of subcellular localization showed that StPHO1.3 was expressed on the cell membrane. Therefore, in this study, the interacting proteins of StPHO1.3 were screened in the membrane system library plasmid by yeast two-hybridization and further verified by bimolecular fluorescence complementation (BiFC). The results showed that, StPHO1.3 interacted with both the photosystem Ⅱ protein subunit and calcium transport-associated proteins. In conclusion, overexpression of StPHO1.3 could enhance the resistance to high temperature stress and promote the growth and development of potato by affecting photosynthesis and signal transduction. These results provide theoretical basis and reference for further understanding of the function of phosphorus transporters and promote the breeding of new potato varieties.
Identification of Gossypium hirsutum PLC Gene Family and Its Expression Analysis During Fiber Developmental Stages
DOU Ling-Ling, NIU Huan-Huan, ZHENG Kai
2024 Vol. 32 (2): 283-298   DOI:10.3969/j.issn.1674-7968.2024.02.004 |  Full text (HTML) (1 KB)  |    PDF  (8629 KB)  ( 23 )
Phospholipase C (PLC) is an enzyme involved in the hydrolysis of phosphatidylinositol 4,5-diphosphate, and its downstream products are involved in the signal transduction of Ca2+ and protein kinase C. In this study, 28 members of the GhPLC gene family were identified from the genome of Gossypium hirsutum 'Nongdamian No. 8' (NDM8) by using the Hidden Markov Model (PF09279 and PF04185). According to the phylogenetic analysis of protein sequences, GhPLC family members were divided into 2 subgroups: PIPLC (16) and nPLC (12). Gene duplication analysis showed that 50% of GhPIPLC and 92% of GhnPLC genes originated from segmental duplication and subjected to purification selection. The cis-acting elements analysis of the upstream 2 000 bp sequence of GhPLC family genes as promoters found that all GhPLC gene promoters contained plant hormone or abiotic stress responsive cis-acting elements, indicating that GhPLCs could be induced by plant hormone or abiotic stresses. qPCR analysis showed that 15 GhPLC gene promoters contained gibberellin responsive-element, and 14 of them could be induced by gibberellin to up-regulate the expression. 15 GhPLC gene promoters contained auxin-responsive elements, among which 14 GhPLC could be induced by auxin analogues to up-regulate the expression. 25 of the 28 GhPLC gene family members were up-regulated in the initiation and elongation stages of cotton fiber developmental stages, accounting for 89% of the GhPLC gene family members. This study provides a reference for further study on the function of GhPLC gene in cotton fiber initiation and elongation stages.
Analysis of Transcriptomics and Hormone Levels of Jiaobai 'zhenong 7' in Different Growth Stages
ZHANG Jue-Feng, LI Fang, ZHONG Hai-Ying, CHEN Jian-Ming
2024 Vol. 32 (2): 299-310   DOI:10.3969/j.issn.1674-7968.2024.02.005 |  Full text (HTML) (1 KB)  |    PDF  (4193 KB)  ( 8 )
Jiaobai is formed when Ustilago esculenta infects and stimulates the Zizania latifolia plant, causing endogenous hormones changes that induce the host pregnancy and stimulate the stem of the plant to swell. So determination of dynamic changes of hormones and transcriptome is important to clarify the regulation mechanism of key genes in different stages of growth and development of Jiaobai. Illumina platform sequencing technology was used to sequence the transcripts of 4 growth stages (pre tillering stage, tillering stage, pre pregnancy stage and pregnancy stage) of Jiaobai varieties 'zhenong 7' (ZN7) in this study. The results showed that total of 194 differentially expressed genes (DEGs) were screened in 4 growth stages, Gene Ontology (GO) function analysis showed that the DEGs in 4 growth stages of ZN7 were mainly involved in cell response to high light intensity and cell response to ultraviolet-A (UV-A) biological process. DEGs were mainly involved in photosynthesis, photosynthesis-antennal protein, carbon fixation in photosynthetic organisms etc. in KEGG enrichment analysis. Compared the contents of endogenous hormones in the 4 growth stages of ZN7, it was found that the trend of content changes of gibberellin (GA3), cytokinin (CTK), auxin (IAA) and abscisic acid (ABA) in each growth stage of Jiaobai were not consistent. The content of GA3 in pregnant period was significantly higher than that in tillering stage (P<0.05), contents of IAA and ABA in each period were basically the same, which was late tillering stage>early tillering stage>pregnant stage, the CTK content reached the peak in the early tillering stage, which was significantly higher than that in the late tillering stage and pregnant stage (P<0.05), but there was no significant difference among the other 3 stages. The results provide scientific basis for key genes screening and regulatory mechanisms illustrating at different growth stages of Jiaobai.
Identification and Expression Analysis of 4CL Gene Family in Rubus chingii
ZHENG Fei-Xiong, CHEN Jun-Yu, JIANG Lin-Qi, ZHAO Jia-Ying, LYU Jia-Yi, YU Zhen-Ming
2024 Vol. 32 (2): 311-321   DOI:10.3969/j.issn.1674-7968.2024.02.006 |  Full text (HTML) (1 KB)  |    PDF  (4790 KB)  ( 19 )
4-coumarate coenzyme A ligase (4CL) is the last key enzyme of the phenylpropane pathway, which can regulate the biosynthesis of lignin, anthocyanin, and flavonoids, and has an important effect on stress resistance. In order to reveal the biological role of 4CL gene family in Rubus chingii, the present study identified the 4CL gene family through an approach of bioinformatics, and investigated the physicochemical properties, subcellular localization, chromosomal localization, systematic evolution, gene structure, conserved motifs, protein secondary structure and cis-acting elements of each member. Besides, their expression profiles in different tissues and at different growth stages were detected. The results showed that 6 4CL gene family members were presented in R. chingii, with similar physicochemical properties and conserved gene structure, and were divided into 3 subfamilies. Six 4CL genes were unevenly distributed on chromosomes 2, 3, 4, 5 and 7. The promoter regions of 6 4CL genes harbored auxin, gibberellin, jasmonic acid, abscisic acid, light, anaerobic, and drought responsive cis-acting elements. The 4CL genes differentially expressed in different R. chingii tissues, among them, Rc4CL1, Rc4CL2, Rc4CL3 and Rc4CL4 were highly expressed in roots, while Rc4CL5 and Rc4CL6 were highly expressed in fruits. Simultaneously, they also differentially expressed at different fruit growth stages, among them, Rc4CL1, Rc4CL2 and Rc4CL3 showed a downward trend at the late stage of fruit development. However, Rc4CL2, Rc4CL5 and Rc4CL6 were highly expressed at the late stage of fruit development, indicating that they might play a crucial role in fruit ripening. The results provide a reference for in-depth study of the 4CL gene and its biological role in R. chingii.
Effects of Forest Canopy Density and Litter Manipulation on Corydalis yanhusuo Growth and Soil Enzyme Stoichiometry
ZHANG Wen-Zhuo, ZHANG Qian-Qian, YU Ye-Fei, CHI Xiao-Li, HE An-Guo, WANG Yu-Qi, LYU Qiang-Feng, LI Yong-Chun
2024 Vol. 32 (2): 322-333   DOI:10.3969/j.issn.1674-7968.2024.02.007 |  Full text (HTML) (1 KB)  |    PDF  (3281 KB)  ( 9 )
Canopy density is the main factor affecting herbaceous plant growth because it leads to insufficient light, thick litter layer, and gives allelopathy effect. The main objective of this study was to explore the canopy density (low, medium, and high) and litter effects on the growth of C. yanhusuo with the restriction of soil enzyme stoichiometry. Results revealed that low canopy density of Cunninghamia lanceolata significantly increased height (24% to 27%), SPAD (soil and plant analyzer development) value (11% to 33%), net photosynthetic rate (28% to 39%), stomatal conductance (28% to 35%) of C. yanhusuo. Also, low canopy density increased the intercellular carbon dioxide concentration (10% to 24%) in C. yanhusuo plants. Although, litter manipulation had no significant impacts on growth of C. yanhusuo. With low canopy density, litter retention significantly increased the content of soil-soluble organic nitrogen (N) (11% to 21%), and enzyme activity of β-1,4-glucosidase (BG), β-1,4-n-acetylglucosaminidase (NAG) and leucine aminopeptidase (LAP). Canopy density, litter manipulation and their interaction showed significant effects on values of SPAD, soil C, N, phosphorus (P) content and related enzymes. The interaction between canopy density and litter had significant effects on plant height and SPAD value (P<0.01), which had a significant effect on the soil C, N and P content, and had a significant effect on the soil BG, NAG, LAP, and acid phosphatase (AP) activity (P<0.01). Microbial P limitation was demonstrated among all treatments via BG/(LAP+NAG) and (LAP+NAG)/AP, which indicated that the soil lacked P for microorganisms availability. In litter retention treatment, the soil BG/AP ratio decreased with increasing canopy density, and suggested that reducing canopy density and retaining litter could alleviate the restriction of soil phosphorus. It was evident that there was significant positive correlation between soil BG/AP and plants growth parameters. The low canopy density with litter retention significantly improved the soil enzyme activities and phosphorus supply that increased the growth of C. yanhusuo. This study could possibly provide the theoretical basis to explore the habitat suitability of C. yanhusuo and other herbaceous wild plants endangered.
Temporal and Spatial Expression Characteristics of Leptin and Its Receptor in Yak (Bos grunniens) Epididymis
WANG Xiao-Yan, CUI Yan, YU Si-Jiu, MA Rui, HE Jun-Feng, ZHANG Qian, LONG-Min
2024 Vol. 32 (2): 334-343   DOI:10.3969/j.issn.1674-7968.2024.02.008 |  Full text (HTML) (1 KB)  |    PDF  (18166 KB)  ( 10 )
Leptin and leptin receptor (LEPR) play important roles in the regulation of male reproduction. This study was conducted to investigate the distribution characteristics and possible roles of Leptin and LEPR in the caput, corpus, and cauda of yak (Bos grunniens) epididymis. Epididymal tissues of yaks of different ages were collected to detect exact distribution and expression levels of Leptin and LEPR in the caput, corpus, and cauda of epididymis using qPCR, Western blot and immunohistochemistry techniques. Immunohistochemical results showed Leptin and LEPR mainly expressed in the epididymal epithelial cells, basal cells, and smooth muscle cells. qPCR results showed that the gene expression levels of Leptin and LEPR in the caput, corpus and cauda of the epididymis of juvenile yak were significantly higher than those of adult and eged yak (P<0.05), and the expression levels of Leptin and LEPR in the cauda of the juvenile yak were significantly higher than those in the caput and corpus of the epididymis of juvenile yak (P<0.05) and there was no significant difference in the caput and corpus of the epididymis of juvenile yak (P>0.05); Leptin and LEPR were not significantly different in the caput, corpus and cauda of the epididymis of adult and aged yaks (P>0.05). Western blot results showed that Leptin and LEPR were expressed in the epididymis of yaks at different ages, and the expression levels of Leptin and LEPR were the highest in juvenile yaks, and decreased successively from juvenile, adult and aged yaks. Western blot results showed that Leptin and LEPR proteins were attached in yaks at different ages. The expression level was the highest in the young group, and showed a decreasing trend from juvenile, adult and old age, with significant differences among different ages (P<0.05); Leptin and LEPR proteins were found to have the highest expression levels in the cauda of juvenile epididymis. There was no significant difference between the caput of the epididymis and the corpus of the epididymis (P>0.05), but there was significant difference between the cauda of the epididymis and the two parts of the epididymis (P<0.05). At the same time, the expression levels of Leptin and LEPR proteins were the highest in the cauda of juvenile epididymis, and there was no significant difference between the two proteins in the caput of juvenile epididymis and the corpus of juvenile epididymis (P>0.05). The expression levels of Leptin and LEPR proteins in adult and aged epididymis were cauda>caput>corpus, and there were significant differences among different parts (P<0.05). The expression levels of Leptin and LEPR were also different in different parts of the epididymis, and the caudal epididymis was relatively the highest, indicating that the caudal environment had a high concentration of Leptin. The temporal and spatial expression differences of Leptin and LEPR in yak epididymis suggest that Leptin and LEPR play important roles in the maturation, storage and energy conservation of yak sperm. This study provides certain research data for the reproductive regulation mechanism of male yaks in plateau environment.
Clone of STAG3 Gene and Its Expression and Localization in Yak (Bos grunniens) Testis with Different Ages
SHI Jun, CHEN Wen-Li, BAI Xu, LI Jian-Fu, YUAN Bao, ZHAO Xing-Xu, ZHANG Quan-Wei
2024 Vol. 32 (2): 344-354   DOI:10.3969/j.issn.1674-7968.2024.02.009 |  Full text (HTML) (1 KB)  |    PDF  (10341 KB)  ( 12 )
Stromal antigen 3 (STAG3), one component of adhesin complexes, plays important role in regulation of follicular development, premature ovarian failure and carcinogenesis in animals. However, its function and regulatory mechanisms are incompletely understood in yak (Bos grunniens). In the present study, the yak testicular tissues with different ages (2, 4, 6 and 8 years old) were collected. The coding sequence (CDS) of STAG3 gene was cloned using reverse transcription-PCR (RT-PCR). The interacted proteins and biological functions of STAG3 were predicted using STRING (functional protein association networks) and Gene Ontology (GO) procedures. Histomorphological structure of yak tissues was observed using hematoxylin-eosin (HE) staining. The localization and expression pattern analysis of STAG3 were evaluated using immunohistochemical (IHC) staining and qRT-PCR. The results showed that the CDS of yak STAG3 was 3 679 bp with a 1 659 bp open reading frame, and encoding 552 amino acids without transmembrane structure region. STAG3 might involve in the meiosis Ⅰ phase, synaptic re-combination and chromosome concentration. The STAG3 protein was widely expressed in testicular tissues, and were located mainly in primary spermatocyte and secondary spermatocyte. The highest expression level of STAG3 was presented in the yak testis of 2 years old, and was gradually decreased with ages. In conclusion, STAG3 was conservative in animal evolution, and widely expressed in the yak testises, which might play an important role in yak reproductive processes. This study provides basis for uncovering the function and mechanism of STAG3 in yak reproduction.
Study of Apoptosis Mode of Porcine Kidney Cells Induced by Low Concentration Suilysin
DAI Shi-Xin, ZHAO Qin, XU Kui, MOU Xin-Ran, DU Sen-Yan, WU Rui, WEN Yi-Ping, HUANG Xiao-Bo, YAN Qi-Gui, CAO San-Jie
2024 Vol. 32 (2): 355-366   DOI:10.3969/j.issn.1674-7968.2024.02.010 |  Full text (HTML) (1 KB)  |    PDF  (11045 KB)  ( 4 )
Streptococcus suis is a serious zoonotic pathogen. Suilysin (Sly) plays a key role in the invasion and pathogenesis of S. suis. It has toxic effects on various types of cells, but there are few studies on the cytotoxic mechanism of Sly. In order to explore the death modes of the porcine kidney cells 15 (PK-15) induced by low concentration of Sly (0.3~0.6 μg/mL), in this study, recombinant suilysin (rSly) was obtained by prokaryotic expression and purification, the toxicity of rSly on PK-15 cells was detected by lactate dehydrogenase (LDH) release assay. Then, for 4 common programmed cell death modes, qPCR, immunofluorescence, Western blot, and flow cytometry assay were used to verify the ways of rSly-induced cell death in PK-15 cells for 12 and 24 h. The results showed that rSly with high hemolytic activity (214 hemolytic units) was successfully expressed, and its toxic effect on PK-15 cells was confirmed to be concentration-dependent. The transcription levels of apoptosis-related genes-cysteine aspartic acid specific protease 3 (Caspase 3) and Caspase 9 were significantly increased after rSly treatment for 12 and 24 h (P<0.05). The apoptosis rate was positively correlated with the concentration of rSly, and the apoptosis rate of PK-15 cells increased from 9.35% to 49.9% after 0.6 μg/mL rSly treatment for 24 h. There was no significant change in the transcription level of microtubule associated protein 1 light chain 3β (LC3β), a key autophagy gene, and no autophagosome formation. The transcription level of gasdermin D (GSDMD), a pyroptosis-related gene had no significant change, and no obviously cleavage of GSDMD protein. Necroptosis-related genes were down-regulated, and the expression level of phosphorylated mixed lineage kinase domain-like protein (p-MLKL) was inhibited. The results suggested that low concentration of rSly induced PK-15 cell death mainly by apoptosis. This study enriches the cellular model of Sly toxicity and helps to reveal the molecular mechanisms underlying the development of S.suis infection.
Effects of Transport Stress on Apoptosis and Expression of Apoptosis-related Genes Bcl-2 and Bax in the Gastric Wall Tissues of Goat (Capra hircus)
HU Wei, LIU Ben, ZHENG Wen-Ya, FANG Man-Xin, HU Ying-Dong, ZHANG Zhi-Tao, ZHANG Wen-Yuan, YE Tian
2024 Vol. 32 (2): 367-378   DOI:10.3969/j.issn.1674-7968.2024.02.011 |  Full text (HTML) (1 KB)  |    PDF  (15033 KB)  ( 5 )
With the development of goat (Capra hircus) breeding, transportation-induced health problems in goats cannot be ignored. The aim of this study was to investigate the effects of transport stress on cell apoptosis and the expression of apoptosis-related genes, such as B-cell lymphoma-2 (Bcl-2) and Bcl-2 associated X protein (Bax) in gastric wall tissue of Ganxi goat. Twelve healthy one-year-old Ganxi male goats were randomly divided into control group (n=4), 2 h transport group (n=4) and 6 h transport group (n=4). During the transportation process, food and water restrictions were carried out. After the transportation, tissues from stomach wall of the goats were collected and analyzed for relevant histology and quantification. Terminal dexynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) results showed that the cell apoptotic rate of each stomach wall tissue in the 2 h transport group and the 6 h transport group was significantly increased (P<0.05) compared to the control group. Additionally, the cell apoptotic rate of other gastric wall tissues, except for omasum, was obviously higher in the 6 h transport group than in the 2 h transport group (P<0.05). Immunohistochemical results showed that Bax and Bcl-2 were mainly expressed in the mucosa epithelium of the anterior stomach (rumen, reticulum and omasum) and in the lamina propria of the abomasum. Bax expression in the 2 transport groups was clearly higher in the aforementioned areas of the stomach wall, with the decrease of Bcl-2 expression, but which was not obvious down-regulated in omasum. qPCR analysis revealed that the mRNA expression level of Bax was significantly up-regulated in other gastric wall tissues, except for omasum, in the 2 transport groups (P<0.05), whereas the Bcl-2 mRNA expression was significantly down-regulated (P<0.05). More importantly, Bax/Bcl-2 ratio in mRNA levels was significantly increased after transportation (P<0.05). Western blot results also showed that there was a similar pattern in the Bax and Bcl-2 protein expression to that of mRNA. These findings indicated that transport stress could lead to cell apoptosis in goat stomach wall tissue through regulating the expression of Bax and Bcl-2. This work may be able to serve as a theoretical foundation for investigating strategies to reduce the damage of goat organs and tissues caused by transport stress.
Comparative Transcriptome Analysis of Three Types of Gonads in Northern Snakehead (Channa argus)
HUANG Su-Jing, WU Yu-Xia, XIA Wei-Wei, ZHANG Xiao-Tian, ZHAO Jian, LI Kai-Bin, LUO Qing, LIU Hai-Yang, FEI Shu-Zhan, CHEN Kun-Ci, OU Mi
2024 Vol. 32 (2): 379-395   DOI:10.3969/j.issn.1674-7968.2024.02.012 |  Full text (HTML) (1 KB)  |    PDF  (7690 KB)  ( 9 )
Northern snakehead (Channa argus) is an important freshwater fish in China with significantly sexual dimorphism. In order to illuminate the molecular mechanism of sex differentiation and gonadal development of C. argus, and explore the functional genes and signaling pathways that differentially express in testis and ovary, RNA-seq was performed on gonads of 6-month-old normal XX females (XX-F), normal XY males (XY-M), and YY super-males (YY-M) by Illumina HiSeq 2000. Correlation analysis of pairwise samples of gonadal tissues indicated that similar expression patterns were found in XY-M and YY-M testis, which were different from that in XX-F ovary, and the result was consistent with the histological observation of gonads. A tremendous amounts of differentially expressed genes (DEGs) were found in the pairwise comparison among XX-F, XY-M, and YY-M gonads. 12 317 DEGs were obtained in XY-M testis compared with XX-F ovary, in which 5 427 were up-regulated and 6 890 were down-regulated. 212 DEGs were obtained in XY-M testis compared with YY-M testis, in which 59 were up-regulated and 153 were down-regulated. 11 282 DEGs were obtained in YY-M testis compared with XX-F ovary, in which 5 038 were up-regulated and 6 244 were down-regulated. qRT-PCR was used to verify DEGs that were related to sex differentiation and gonadal development with significant differences among testis and ovary. It displayed that 10 genes had relatively high expression in testis of XY-M and YY-M, such as anti-müllerian hormone receptor type 2 (Amhr2), anti-müllerian hormone (Amh), doublesex-mab3-related transcription factor1 (Dmrt1) and gonadal somatic cell-derived factor (Gsdf), 9 genes had relatively high expression in ovary of XX-F, including bone morphogenetic protein 15 (Bmp15), cytochrome P450 family 19 subfamily A member 1a (Cyp19a1a) and forkhead transcription factor gene 2 (Foxl2). The results of qRT-PCR validation were consistent with the results of the RNA-seq, illustrating that Amhr2, Amh, Dmrt1 and Gsdf genes were related to the regulation of testicular development, and Bmp15, Cyp19a1a and Foxl2 genes were related to ovarian development regulation in C. argus. KEGG pathway enrichment analysis showed that most DEGs were enriched in the pathways related to sex differentiation and metabolism, such as mitogen-activated protein kinase (MAPK), wingless-type MMTV integration site family (Wnt) and transforming growth factor-β (TGF-β) signaling pathway. In this study, a large number of candidate genes and signaling pathways related to sex differentiation and gonadal development were screened out, which would provide preliminary basis and data support for further studies on the molecular mechanism of sex differentiation and gonadal development and the gene function of the sex-determining genes in C. argus.
Screening of SNP of MYF6 Gene in Nile Tilapia (Oreochromis niloticus) and Its Association Analysis with Growth Traits
GAO Feng-Ying, TONG Yan-Nan, LIU Zhi-Gang, CAO Jian-Meng, WANG Miao, YI Meng-Meng, KE Xiao-Li, LU Mai-Xin, ZHU Hai
2024 Vol. 32 (2): 396-416   DOI:10.3969/j.issn.1674-7968.2024.02.013 |  Full text (HTML) (1 KB)  |    PDF  (2929 KB)  ( 13 )
The myogenic factor 6 (MYF6) is integral to the initiation and development of skeletal muscle and the phenotype maintenance. Thus, it is candidate gene for growth-related traits. The objectives of this study were to investigate SNPs in MYF6 gene and to evaluate whether these polymorphisms were associated with growth-related traits in Nile tilapia (Oreochromis niloticus). Initially, the SNPs of MYF5 gene were screeded by PCR and Sanger sequencing, and they were confirmed by Sanger sequencing and SNaPshot method. Sixty one SNPs were identified in MYF6 gene, and 36 loci with relative high polymorphism (>3%) were selected from 61 loci for genotyping analysis in Gaoyao parent population using Sanger sequencing and SNaPshot method. The general linear model (GLM) and one way analysis of variance (ANOVA) of SPSS 19 software were used to conduct the association analysis between SNP loci and growth traits. The result showed that 2 of the 36 loci were related to body weight, 4 to full length, 7 to head length, and 2 to body height. These loci, which related to growth-related traits in Gaoyao parent population, were genotyped in Gaoyao offspring and Panyu population. Association analysis between SNP loci and growth-related traits in Gaoyao offspring population showed that S7 (5620-GAGACGGAGA5629) locus was correlated with body weight, body length and body height; S24 (C-7388G) was related to body weight, body height and body thickness; S56 (A-11815C) was correlated with body weight, total length, body length, head length, body height and body thickness. In Panyu population, 2 SNP loci were related to body weight, 1 to body length and full length, 1 to head length, and 2 to body thickness. The above loci, which were related to growth-related traits of Gaoyao offspring and Panyu population, were further verified in Hainan population, and 1 locus associated with body weight was obtained in male population. The correlation analysis between the diploid and the growth-related traits of each population showed that 1 diploid related to body weight was obtained from each of the Gaoyao parent population, the Gaoyao offspring population and the Panyu population, respectively. This study provides effective molecular markers for improving growth traits and breeding of tilapia.
Effects of Intermittent Fasting on Microbial Community Structure, Water Quality and Growth of Litopenaeus vannamei in a Biofloc System
QU Yin, ZHANG Xiao-Dong, REN Gang, SHEN Wen-Ying
2024 Vol. 32 (2): 417-426   DOI:10.3969/j.issn.1674-7968.2024.02.014 |  Full text (HTML) (1 KB)  |    PDF  (2174 KB)  ( 18 )
Diet restriction has been used as a strategy to reduce the feed and labor cost in the aquaculture. The compensatory growth is widely present in mammals, fish and crustaceans. Based on full compensatory growth ability of Litopenaeus vannamei, the biofloc technology combined with diet restriction were applied to L. vannamei breeding. The effects of intermittent fasting on the microflora structure, water quality and growth parameters of L. vannamei in a biofloc system were investigated. The L. vannamei were randomly assigned into 4 groups (each with 3 replicates): Group 1 was the control group, normally fed with basal L. vannamei feed. Group 2 was the biological floc group, fed with L. vannamei feed and brown sugar (70% of the feed), and the C/N ratio was 12 in water. Group 3 was the probiotic and biological floc group, besides feed and brown sugar, Bacillus licheniformis (concentration was 1.2×105 CFU/mL in water) was also added; Group 4 was the intermittent starved group, starved for 2 d and re-feeding for 5 d. The results indicated the biological floc sedimentation in water showed an increasing trend. At the 4th week, the formation of biological floc in group 2, 3 and 4 were significantly higher than that in group 1 (P<0.05), respectively. The operational taxonomic units (OTU) of microflora in group 1, group 2, group 3 and group 4 were 3 501, 6 386, 5 387 and 6 577, with high throughput sequencing, respectively. The unique OTU numbers were 165, 463, 362 and 592, respectively, and 173 OTUs were shared by these 4 groups. The α diversity order of the bacterial community from high to low was: Group 4>group 2>group 3>group 1. The abundance analysis at genus level showed that the most dominant flora of group 1 was Georgenia (20.2%), followed by Hydrogenophaga (13.5%) and Pseudomonas (13.2%). In group 2, the highest abundance was Pseudomonas (39%), followed by Georgenia (9.1%) and Brevundimonas (7.0%). In group 3,the highest abundance was Brevundimonas (38.4%), followed by Microbacterium (5.7%) and Methyloversatilis (5.7%). In group 4, the highest abundance was Alishewanella (21.0%), followed by Pseudomonas (15.5%) and Exiguobacterium (6.8%). The concentration of ammonia nitrogen and nitrite nitrogen significantly decreased in groups 2, 3 and 4 (P<0.05). The feed conversion rate (FCR) of group 4 was significantly higher than 3 other groups (P<0.05). The results showed the biofloc technology based on compensatory growth could effectively increase the richness and diversity of microflora, optimize the composition of the microbial community, reduce the concentration of ammonia nitrogen and nitrite nitrogen in aquaculture water of L. vannamei, thereby to optimize aquaculture water quality, and effectively increase the feed conversion rate.The results were conducive to elucidating the mechanism of biofloc optimizing water quality, and provide experiental data for the application of the biofloc technology based on compensatory growth in L. vannamei culture.
Effects of Paenibacillus terrae NK3-4 on Gene Expression in Fusarium fujikuroi
LIU Wen-Zhi, WU Xin-Yu, DU Jia-Peng, PENG Zhi-Hong, GONG Sha-Sha, YU Wen-Qing, LI Peng
2024 Vol. 32 (2): 427-443   DOI:10.3969/j.issn.1674-7968.2024.02.015 |  Full text (HTML) (1 KB)  |    PDF  (12072 KB)  ( 6 )
Paenibacillus terrae NK3-4 strain is found to be antagonistic to plant pathogens. However, the response mechanism of NK3-4 interaction with the pathogen remains unclear. In this study, the inhibitory effect of NK3-4 on rice (Oryza sativa) bakanae disease pathogenic Fusarium fujikuroi was measured, and a transcriptome sequencing analysis for the NK3-4 inhibited F. fujikuroi was taken. The results showed that NK3-4 inhibited the growth, pigment synthesis and conidia formation of F. fujikuroi, and damaged the mycelia. Among the 2 291 differential expressed genes in F. fujikuri induced by NK3-4, the number of down-regulated genes was 2.44 times of up-regulated genes. And the down-regulated genes were mainly enriched in nucleus (GO:0005634), protein-containing complex (GO:0032991), intracellular non-membrane-bound organelles (GO:0043232) and other functional groups. Among the differential expressed genes, the number of down-regulated expressed genes related to pathogenesis (GO:0009405), toxin biosynthetic process (GO:0009403), aflatoxin biosynthetic process (GO:0045122), reproduction (GO:0000003 ) and other relative functions were more than that of the up-regulated genes. Meanwhile, of the differential expressed genes which involved in cell wall chitin biosynthetic process (GO:00060382), asexual reproduction (GO:0019954), conidia formation (GO:0048315), conidia development (GO:0061794), pigment biosynthetic process (GO:0046148) and other relative functions were all down-regulated expressed. The result of reverse transcription fluorescence quantitative PCR detection was consistent with transcriptome sequencing result (Pearson correlation index 0.90), indicating NK3-4 down-regulated the expression levels of genes in F. fujikuroi. Synthetic analysis showed that NK3-4 may inhibit F. fujikuroi by down-regulating the expression of genes, which related to growth and reproduction, pigment synthesis and spore production, and may reduce its virulence by inhibiting the biosynthesis of toxin in F. fujikuroi. The results provide a basis for further elucidating the molecular mechanism of NK3-4 interacting with F. fujikuroi.
Reviews and Progress
Advances of Light-harvesting Chlorophyll a/b Binding Protein (CAB) in Higher Plants
ZHONG Si-Tong, ZHANG Ya-Zhen, KONG Xiang-Rui, JIN Shan, CHEN Zhi-Hui, CHEN Chang-Song
2024 Vol. 32 (2): 444-457   DOI:10.3969/j.issn.1674-7968.2024.02.016 |  Full text (HTML) (1 KB)  |    PDF  (1030 KB)  ( 16 )
Light-harvesting chlorophyll a/b binding protein (CAB) is a kind of membrane protein encoded by the CAB gene family. It plays an important role in capturing and transmitting light energy, photoprotection and excess energy dissipation, regulating the distribution of light energy between the two photosystems, and maintaining the structure of the thylakoid membrane. CAB genes widely participate in the whole biological processes in higher plants, including the growth and development of leaf, flower and fruit, and in response to various stresses. In this review, the research progresses of CAB family in their classification, protein structure, physiological functions, and stress expression characteristics were summarized. The future research directions of CAB were also prospected. This paper provides references for further studies on the CAB gene family.
Research Progresses on the Regulation of Sucrose on Plant Branching
ZHOU Hui-Wen, LUO Han-Min, XIONG Fa-Qian, GAO Yi-Jing, LIU Jing, YANG Tai-Yi, YAN Hai-Feng, QIU Li-Hang
2024 Vol. 32 (2): 458-470   DOI:10.3969/j.issn.1674-7968.2024.02.017 |  Full text (HTML) (1 KB)  |    PDF  (1657 KB)  ( 20 )
Shoot branching is an important agronomic trait of plant architecture because it substantially affects the yield and value of the plant. Branching characteristics are affected by many factors, and sucrose is the most popular research topics affecting the branching characteristics of plants. A certain number of studies have shown that sucrose not only serves as a carbon source to provide energy for the meristematic growth of plants, but also is an important signaling molecule. In this review, the main pathways through which sucrose promotes branching are summarized. Among them, strigolactone, auxin, and cytokinin are the most important hormones in the process of sucrose promoting branching, and transcription factors such as teosinte branched 1 (TB1) and branched 1 (BRC1) also play an important role. This paper provides a basis for the theory of plant branching and constructing a reasonable plant population.
Research Progress on Application of Genome Sequencing in the Meat Quality Improvement in Domestic Animals
YU He, LI Rui, YIN Xu-Dong, PANG Wei-Jun
2024 Vol. 32 (2): 471-483   DOI:10.3969/j.issn.1674-7968.2024.02.018 |  Full text (HTML) (1 KB)  |    PDF  (1872 KB)  ( 248 )
The focus of China's breeding work previously focused on improving important economic trait indicators such as meat production, daily weight gain, and litter size of livestock. With the improvement of consumption level, breeding livestock and poultry breeds with excellent production performance to produce meat products with good flavor and delicate taste is the key to the high-quality development of China's livestock industry in the future. Skeletal muscle fibers and intramuscular fat are closely related to livestock meat quality and directly affect the juiciness, color, and flavor of meat. In recent years, research methods based on genome sequencing technologies have been evolved, including a selection signaling method that can mine important candidate genes, genome-wide association analysis, and a genome-wide selection method that uses genomic information for variety improvement, which have incomparable advantages over traditional meat quality improvement methods. Based on the summary of livestock meat quality evaluation indexes and important influencing factors, in this paper the application of mainstream genome sequencing technologies in livestock meat quality improvement are reviewed, and the advantages and disadvantages of different methods are analyzed. In addition, the latest technical tools and theoretical studies that can be combined with genome sequencing technology and may play a role in livestock meat quality improvement are introduced. This review provides scientific guidance for the development of genome-wide selection technology and theory for livestock meat quality trait improvement.
Resources and Updated Technology
Construction and Antibacterial Effect Evaluation of Nano TiO2 Loaded Moxifloxacin Hydrochloride System
HUANG Han-Ye, GUO Xue-Ping, XU Jia-Wei, LI Qian-Qian, CAI Xin-Kai, GUAN Xiong, PAN Xiao-Hong
2024 Vol. 32 (2): 484-492   DOI:10.3969/j.issn.1674-7968.2024.02.019 |  Full text (HTML) (1 KB)  |    PDF  (7077 KB)  ( 7 )
Moxifloxacin hydrochloride (MOX) is a broad-spectrum quinolone drug and is widely used in clinic as a antibacterial agent. In order to explore the effect of nanocarriers on its antibacterial effect, MOX was loaded onto nano titanium dioxide (TiO2) by surface adsorption in this study, and then the chemical characterization, antibacterial activity against Escherichia coli and pathological observation of the nano TiO2-MOX were investigated. Nano TiO2 combined with MOX in the form of physical embedding. When the content of nano TiO2 increased from 0.5 mg to 1.0 mg, the drug loading and the encapsulation rate increased by about 20% and 40% respectively. Nano TiO2 in nano TiO2-MOX could help release the effective components of MOX. The results of the in vitro activity against E. coli showed that the synthesized nano TiO2 had certain antibacterial activity (22.8%), and the antibacterial activity of MOX increased from 28.0% to 73.4% with the support of nano TiO2. Pathological observation also indicated that the damage of the cell membrane of E. coli was not obvious by MOX, but nano TiO2 and nano TiO2-MOX could cause obvious damage to the cell membrane and then cause cell death. In this study, the synthesized nano TiO2-MOX showed good drug loading and releasing effect, and significantly improved the antibacterial activity of MOX, which provides a scientific basis for the application of nano TiO2 as a new drug carrier in further clinical research.
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