Supervisor(s): Ministry of Agriculture Sponsor(s): Chinese Academy of Agricultural Sciences；Chinese Association of Agricultural Science Societies CN：11-1328/S
Scientia Agricultura Sinica, the 1st in Comprehensive Agricultural Science, is supervised by Ministry of Agriculture of PRC, and sponsored by Chinese Academy of Agricultural Sciences; Chinese Association of Agricultural Science Societies. Scientia Agricultura Sinica, launched in 1960, is a leading peer-reviewed and mufti-disciplinary journal and published semi-monthly in Chinese with English title, abstract, figures, tables and references. It aims to publish those papers that are influential and will significantly advance scientific understanding in agriculture fields worldwide. The scope covers Crop Genetics, Breeding, Germplasm Resources; Physiology, Biochemistry, Cultivation, Tillage Plant Protection; Soil & Fertilization, Agro-Ecology & Environment, Bio-energy; Animal Science, Veterinary Science, Agricultural Information Science; Food Science; Agricultural Economics and Management; Agricultural Sustainability.
The journal is included in JST, CA and CSCD.
Editor-in-Chief Wan Jianmin Associate Editor-in-Chief Zou Ruicang Tang HuaJun Wu Kongming Guo YuYuan Geng Xu Sun Tan Executive Editor Lu Wenru
[Objective] Octopaminergic signaling system plays a crucial role in the reregulation of behavioral and physiological processes in insects. The red flour beetle (
Tribolium castaneum) is a model insect which has been widely used in the study of insect growth, development and physiology. The objective of this study is to utilize
T. castaneum as the research insect and elucidate the vital functions of octopamine receptors involved in the physiology and behavior. [Method] Based on the sequence information in GenBank (XP_008198078), the cDNA of an octopamine receptor (
TcOctβR3) was cloned by RT-PCR. The open reading frame (ORF), deduced amino acid sequence and the transmembrane domains were predicted by using online service, and the phylogenetic tree associated with OctβR3 from other insects was constructed by using neighbor-joining method to clarity its phylogenetic relationship. In addition, the RNA was extracted from different developmental stages (egg, larva, pupa, and adult), different tissues (central nervous system, fat body, midgut, hindgut, malpighian tubule, testis and ovary), as well as the larvae under stress of starvation, respectively. Ribosomal protein S3 gene (
TcRPS3) was used as an internal reference. Real-time quantitative PCR (qRT-PCR) was employed to determine its expression patterns at different developmental stages and in different tissues as well as the induced expression profiles under the stress of starvation.
TcOctβR3 was transiently expressed in human embryonic renal cell (HEK293) by using heterologous expression system, and cAMP measuring method was performed to determine the activity of its ability to combine with ligands. Finally, the double-stranded RNA of TcOctβR3 was synthesized
in vitro, and the physiological functions were verified by track ball behavior analysis as well as RNA interference (RNAi) technology. [Result] A complete sequence of
TcOctβR3 was cloned with open reading frame (ORF) of 1 305 bp, encoding 434 amino acids, with a signature of seven transmembrane domains which belonged to the superfamily of G-protein coupled receptors. This gene exhibited a close relationship with
AtOctβR3 from
Aethina tumida based on neighbor-joining phylogenetic tree. The qRT-PCR results indicated that
TcOctβR3 was expressed at all tested developmental stages, particularly high at the early larval stage. However, the expression of
TcOctβR3 was not significantly different among other developmental stages. Besides, the expression of
TcOctβR3 was remarkably higher in the central nervous system than that in other tissues. In the process of starvation for 24 h, the expression of
TcOctβR3 significantly fluctuated and then returned to the normal level. The lowest expression of TcOctβR3 was 0.47 folds at 6 h and the highest expression was 1.80 folds at 16 h compared with control, respectively. Based on cyclic AMP response assay, it was found that TcOctβR3 could be activated by octopamine in a dose-dependent manner with a median effective concentration (EC
50) of 8.68 × 10
−7 mol·L
−1 after heterogenous expression in HEK293 cells. Naphazoline strongly activated this receptor with EC
50 of 8.56 × 10
−8 mol·L
−1 compared with other ligands. Generally, the rank order for the potency of four tested ligands was naphazoline > tyramine > octopamine > dopamine. The RNAi results indicated that the transcript level of TcOctβR3 was significantly knocked down by 61.5%. However, no effect on the walking speed and fecundity of
T. castaneum adults was observed. [Conclusion] TcOctβR3 plays an important role in the central nervous system, which modulates the response of the beetle larvae upon starvation. The molecular characterization was performed based on a cAMP assay, which will provide a solid basis for the future screening of its high efficient agonists and inhibitors.
[Objective] The objective of this study is to isolate an apple auxin response factor gene
MdARF5, to analyze its expression of exposing to auxin, to identify its role in regulating anthocyanin biosynthesis, then to reveal its biological functions and to provide a theoretical basis for auxin-mediated anthocyanin accumulation. [Method] The apple auxin response factor gene
MdARF5 was cloned by PCR technology from apple (
Malus ×
domestica ‘Royal Gala’). The phylogenetic tree was constructed by MEGA 5.0 software. The transgenic apple calli were generated via
Agrobacterium-mediated transformation. The difference in the anthocyanin accumulation was compared between wild-type and transgenic apple calli. The transient expression assays in tobacco leaves were carried out to test the transcriptional regulation of
MdMYB1 gene by
MdARF5. [Result]
MdARF5 gene (MDP0000143749) was obtained. The open reading frame (ORF) of
MdARF5 contained 2 691 bp, encoding a protein of 896 amino acid residues. The phylogenetic tree analysis showed that the homology of MdARF
5 was close to the PbARF5. The transcriptional analysis results indicated that
MdARF5 was induced by auxin treatment. On the contrary, the expression of anthocyanin biosynthesis genes was repressed. The
MdARF5-overexpressing apple calli exhibited decreased anthocyanin content, suggesting that
MdARF5 gene might play an important role in regulating anthocyanin accumulation. The sequence of
MdMYB1 promoter region was analyzed and a putative ARF binding motif was found. Meanwhile, the transient expression assays were performed in
Nicotiana benthamiana leaves and the results showed that MdARF5 could repress the expression of
MdMYB1. [Conclusion] It is speculated that MdARF5 down-regulates anthocyanin accumulation by directly repressing the transcript of
MdMYB1.
[Objective] The purpose of this study was to clarify the mechanism of photosynthetic morphology of soybean at seedling stage in maize/soybean intercropping from the perspective of transformation of related sugars and cellulose synthesis. [Method] Under soybean monoculture and maize/soybean intercropping systems, the strong shade-tolerant soybean (Nandou12) and weak shade-tolerant soybean (Nan 032-4) were used as the experimental materials. The photosynthetic rate of leaf and the total carbon, cellulose, soluble sugar, sucrose, and β-1,3-glucan of stem were measured and analyzed. [Result] Compared with soybean monoculture, the leaf photosynthetic rate of intercropping soybean was significantly decreased due to maize shading, but the degree of response was different for different soybean cultivars. In intercropping system, the decreasing degree of strong shade-tolerant soybean Nandou 12 was relatively small, and Nandou 12 showed stronger photosynthetic ability than Nan 032-4. The total carbon content in the leaf and stem of shaded soybean decreased, but the reduction degree of Nandou12 was significantly lower than that of Nan 032-4. The correlation analysis showed that the photosynthetic rate of leaf was in highly significantly positive correlation with the total carbon content in the leaf and stem, and the cellulose content in the stem (
r = 0.952, 0.935, 0.825, respectively,
P < 0.01). The result explained that shade of intercropping affected the photosynthetic rate, reduced the accumulation and distribution of photosynthates in the stem. Finally, the cellulose content in soybean stem was decreased. However, the strong shade-tolerant soybean Nandou 12, with the higher photosynthetic rate and accumulation of photosynthate, was more suitable for planting in intercropping. At seedling stage, the soluble sugar content of intercropping soybean stem was significantly lower than that of monoculture soybean. The contents of β-1,3-glucan and sucrose in intercropping soybean stem from 30–51 days after germinating were significantly higher than those in monoculture, and in intercropping, the carbohydrate transformation rates of two soybean were significantly different. Under the same planting pattern, the contents and transformation rates of soluble sugar, sucrose and β-1,3-glucan of soybean stem with strong shade-tolerant soybean Nandou 12 were significantly higher than those in Nan 032-4. The analysis of cellulose depositionion modes showed that, for the same soybean material, the accumulation time and accumulation rate in monoculture system were higher than those in intercropping system. In the same planting pattern, the cellulose accumulation time in the stem of Nandou 12 was shorter than that of Nan 032-4, but the difference was small, and the accumulation rate was higher than that in Nan 032-4. It resulted in that the cellulose content in the stem of Nandou12 was significantly higher than that in Nan 032-4. [Conclusion] Shading from maize in intercropping system decreases the photosynthetic capacity of soybean leaves, slowes the transportation of photosynthate from leaf to stem, and reduces the content of stuffing in stem. Shading changes the accumulation modes of cellulose in soybean stem, and makes the cellulose content decrease. In intercropping system, the strong shade-tolerant soybean Nandou12 can maintain higher photosynthetic capacity and stronger cellulose synthesis ability in the stem, so its lodging resistance is strong.
[Objective] Starch grain density affects maize kernel test weight. In this study, we used a mutant
Mrd of maize to identify and fine map the gene controlling starch grain density, which is helpful for the cloning and functional verification of the related kernel test weight gene. [Method]
Mrd is a starch grain density mutant which was identified during maize breeding practice. Scanning electron microscope and near-infrared spectrophotometer (NIRS) were used to observe changes of chemical composition in the
Mrd kernels. The segregation populations F
2 and BC
1 were derived from the cross of
Mrd and B73, which were planted in Zhengzhou and Yuanyang, Henan Province, and Sanya, Hainan Province, and used for genetic analysis. Bulked segregation analysis (BSA) was used to identify linkage markers of the target gene with the 1 000 pairs of SSR primers from maize GDB (http://www.maizegdb.org). The BC
1 segregation population of 38 000 individuals were used to fine map the target gene
tw1. The candidate genes were sequenced and functionally predicted by bioinformatics. Allelism test was performed for
tw1 and
su2. [Result] Compared with the normal kernels, the
Mrd had smaller kernel size, no change in kernel length, and increased specific gravity. The crude protein content decreased and the crude starch content did not change significantly in the mutant, but the
Mrd has irregular shape, smaller starch grains, increased density, and increased kernel test weight. Observation of the starch grain structure inside the kernels on different days after pollination showed that the starch grain density in the mutant kernel increased with the progress of development and was always higher than that in the normal kernel. The genetic analysis of F
2 and BC
1 populations showed that the mutation of kernel test weight was controlled by a single recessive gene
tw1 which was firstly located between SSR markers umc1105 and bnlg1154 on chromosome 6. After screening and analyzing the recombinant from the BC
1 segregation population, it was found that the gene was located between B3 and A47 (physical distance 0.2 Mb). There were three protein-coding genes in the 0.2 Mb candidate physical interval. Allelism test excluded
su2 gene and sequence analysis of the other two candidate genes verified that GRMZM2G042607 should be the primary candidate gene of
tw1, which encodes the protein with a carbohydrate recognition domain and deposits the deposition of carbohydrates in seeds. [Conclusion] The
tw1 gene was fine mapped and the candidate gene was found to be GRMZM2G042607 that encodes the β-1,3-galactosyltransferase.
[Objective] The ZBED6 gene is a transcription factor that regulates muscle growth and development. In order to investigate the regulatory mechanism of ZBED6 in myocardium growth and development, this study was designed to compare the transcriptome of cardiac tissue obtained from Bama miniature pig between the ZBED6 knockout group (ZBED6-KO) and the wild type group (ZBED6-WT) and to find out the effect of ZBED6 gene knockout on the development and gene expression profile of pig heart tissue. [Method] The phenotypic characteristics of ZBED6-KO and ZBED6-WT pig tissues were analyzed by
t-test and the expression level of the target gene IGF2 of the ZBED6 gene was quantified by real-time quantitative PCR (q-PCR). The differences in histological level of tissue structure were compared by paraffin section method. The total RNA was isolated from ZBED6-KO group and ZBED6-WT group. RNA-Seq analysis was performed on Illumina Hiseq 2000 platform. The differentially expressed genes between ZBED6-KO and ZBED6-WT heart tissues were screened by the method of RNA-Seq Pig Sus_scrofa10.2 was used as the reference sequence. Differentially expressed genes were enriched and analyzed with IPA software. Nine differentially expressed genes were randomly selected to verify the reproducibility of RNA-Seq results by q-PCR. [Result] The heart of ZBED6-KO pig had a larger measured data in weight than ZBED6-WT, which also had a higher expression profile of IGF2 than the latter. Compared with ZBED6-WT pig, ZBED6-KO pig had wide muscle fiber width and less connective tissue. The results suggested that the knockout of ZBED6 gene could promote the growth and development of porcine heart. The sequencing results showed that at least 10 G data were obtained and the clean ratio, Q30 data were more than 90%, of which 62.8%–80.1% of the reads mapped to the pig genome. All of these indicated sequencing was well saturation and the sequencing data was reliable. By analyzing sequencing data, 184 differential genes were indicated which contained 114 up-regulated genes and 70 down-regulated genes. Among the 184 genes, there were 141 intact function annotation genes and 43 non-annotated genes. Differential gene hierarchical clustering analysis showed that, the similar expression pattern was detected in ZBED6-KO group (x1, x3, x6), which was also happen to ZBED6-WT group (x2, x4, x5). GO and IPA enrichment analysis obtained 13 significant GO terms, 33 pathways. The differential gene was mainly enriched in the immune response, muscle development, RhoA signal and other related channels. The expression pattern of 9 differentially expressed genes was consistent with the results of RNA-Seq analysis. [Conclusion] In this study, ZBED6-KO Bama miniature pig was for the first time used as the model. We used RNA-Seq technique to explore the effects of ZBED6 gene knockout on cardiac development, which enriched the research of ZBED6 on myocardial development and function.
[Objective] The soil will be saturated by H
+and Al
3+ under the process of electrodialysis. Theoretically, the acidification characteristic of soil can be studied with the method of electrodialysis. [Method] Purple soil, yellow soil, and latosol were selected to undergo the electrodialysis at a potential gradient of 15 V·cm
−1. The electrodialysis was performed 30 times and the duration of each time was 8 h. The acidification characteristics of the three kinds of soil were analyzed based on soil exchangeable acidity, base cations before and after the electrodialysis, and the release regulation of base cations in the process of electrodialysis. [Result] The results showed that the tested soils could be acidified quickly with electrodialysis in short time. The pH value of each soil was below 4.5, which meant the soils were strongly acidified. After electrodialysis, the exchangeable acidity, exchangeable H
+, and exchangeable Al
3+ increased significantly. The exchangeable acidities of purple soil, yellow soil, and latosol increased from 3.35, 0.23, and 0.76 cmol(+)·kg
−1 to 18.9, 7.0, and 5.8 cmol(+)·kg
−1, respectively. On the contrary, the contents of water soluble and exchangeable base cations, especially Ca
2+ and Mg
2+, base exchange capacity, and base saturation decreased. For example, the base saturation of purple soil, yellow soil and latosol decreased from 96.8%, 82.6%, and 47.3% to 20.5%, 11.8%, and 12.2%, respectively. The acidified purple soil had higher soil exchangeable acidity and base saturation. Because the contents of Ca
2+ and Mg
2+ were higher than those of K
+ and Na
+ and the electrostatic force between Ca
2+/Mg
2+ and negatively charged soil surface was stronger, the monovalent base cations K
+ and Na
+ were quickly released from the soil in the process of electrodialysis, and the divalent base cations Ca
2+ and Mg
2+ were released from the soil slowly and wavily. The exchangeable acidity of purple soil was significantly higher than those of yellow soil and latosol because of the high cation exchange capacity (CEC), which increased the risk of Al
3+ toxicity of plant in the acidified purple soil. Meanwhile, the content of base cations and base saturation of purple soil were higher than those of yellow soil and latosol, because the cations in purple soil could be supplied from the decomposition of soil minerals. The high content of base cations in the purple soil ensured the demand of base cation for plant growth and alleviated the further acidification of soil. [Conclusion] Therefore, the electrodialysis could be as a useful tool to research the acidification characteristics of soil. The acidification characteristic of soil was affected by the content of negative charge on the surface of soil and the soil renewed ability of base cations. Compared with yellow soil and latosol, the purple soil had a dual effect on the process of acidification.
