Loading...

Table of Content

    01 September 2016, Volume 49 Issue 17
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Genetic Diversity in Broomcorn Millet (Panicum miliaceum L.) from China and Abroad by Using SSR Markers
    LIAN Shuai, LU Ping, QIAO Zhi-jun, ZHANG Qi, ZHANG Qian, LIU Min-xuan, WANG Rui-yun
    Scientia Agricultura Sinica. 2016, 49(17):  3264-3275.  doi:10.3864/j.issn.0578-1752.2016.17.002
    Abstract ( 630 )   HTML ( 10 )   PDF (3084KB) ( 867 )   Save
    References | Related Articles | Metrics
    【Objective】The objective of this study is to assess the genetic diversity of broomcorn millet accessions which collected from China and abroad.【Method】Five hundred pairs of SSR primers developed in the authors’ laboratory by high-throughput sequencing were used to identify polymorphisms in six representatives randomly selected from the total of accessions. A total of 63 primer pairs produced clear and reproducible polymorphic fragments among the six accessions and then were used to analyze the genetic diversity and relationship of 192 broomcorn millet landraces and wild accessions.【Result】A total of 161 alleles were detected with an average of 2.56 alleles per locus, and the mean Shannon-Weaver index (I), mean Nei and mean PIC were 0.6275, 0.3874 and 0.4855, respectively. The results indicated that there is a significant difference among the 10 populations of broomcorn millet resources in genetic diversity from diverse geographic origins. The variance range of effective alleles number is 1.2407 (South region) - 1.8846 (Inner Mongolia). In domestic populations, the rank of Shannon-Weaver index is Inner Mongolia Plateau>Tohoku>Loess Plateau>Northwest>southern regions, and the rank of foreign populations is the former Soviet Union>Europe>Mongolia>India>United States. The results of Nei’s genetic heterozygosity analysis showed that the minimum and maximum of observed (Ho) and expected heterozygosity (He) is 0.2372 from India and 0.3966 from Inner Mongolia as well as 0.3114 from the Unite State and 0.4622 from Inner Mongolia Plateau, respectively. The effective number of alleles (1.9285±0.5101), Shannon-Weaver index (0.6948±0.2852) and Nei gene diversity index (0.4373±0.1773) of the wild germplasm are much higher which in domestic and foreign accessions. For domestic population and alien population, the effective number of alleles (1.8145±0.4519) of domestic resources, Shannon-Weaver index (0.6657±0.2413), and Nei gene diversity index (0.412± 0.1574) of domestic accessions were higher than that in foreign resources (1.6862±0.4527, 0.5897±0.2469, 0.3652±0.1655). UPGMA cluster analysis showed that the 10 geographic populations could be clustered into three categories, the accessions from the Inner Mongolia Plateau, the Loess Plateau, northeast, northwest, Mongolia area were clustered as one group, the former Soviet Union, the United States, India, Europe together as one group, and southern region of China clustered as one independent group. The wild millet (34) which is from Qiqihaer is separated from others at 0.37, the wild millet from Gansu (19) was divided into an independent individual at 0.34, indicating that there are significant genetic variances between the two wild accessions and others. In general, genetic division of population is not significant for 192 domestic and foreign accessions, and there have material interpenetration among different groups.【Conclusion】The Inner Mongolian Plateau, northeast area and the Loess Plateau with the most abundant genetic diversity is the most complex area of genetic relationship, which further confirms that China is the origin center of Panicum miliaceum.
    Response of Millet Transcription Factor Gene SibZIP42 to High Salt and ABA Treatment in Transgenic Arabidopsis
    QIN Yu-hai, ZHANG Xiao-hong, FENG Lu, LI Wei-wei, XU Zhao-shi, LI Lian-cheng, ZHOU Yong-bin, MA You-zhi, DIAO Xian-min, JIA Guan-qing, CHEN Ming, MIN Dong-hong
    Scientia Agricultura Sinica. 2016, 49(17):  3276-3286.  doi:10.3864/j.issn.0578-1752.2016.17.003
    Abstract ( 459 )   HTML ( 8 )   PDF (3752KB) ( 650 )   Save
    References | Related Articles | Metrics
    【Objective】 This study was conducted to analyze the molecular characteristics and biological function of the transcription factor SibZIP42 in foxtail millet and to discuss the regulation pathway of SibZIP42 in improving salt tolerance. At the same time, the study would provide a potential gene for improving abiotic stress resistance in crop molecular breeding. 【Method】 Bioinformatics methods were used to analyze the molecular characteristics of SibZIP42. DNAMAN and MEGA 5.05 softwares were used to do multiple sequences alignment and construct the phylogenetic tree of SibZIP42. The 2 000 bp sequence upstream of SibZIP42 was downloaded as the promoter sequence from databases Phytozome, and the cis-acting elements of SibZIP42 promoter were analyzed in PLACE database. NetPhos 2.0 Server database was used to predict phosphorylation sites of SibZIP42 protein. The real-time PCR was used to analyze the expression patterns of SibZIP42 under various stress treatments. SibZIP42 was fused with GFP to detect its subcellular localization in protoplast cells. SibZIP42 was overexpressed in Arabidopsis to analyze its function under high salt and ABA stress conditions. 【Result】 The full length of SibZIP42 was 546 bp with one exons and encode a hydrophilic protein with 181 amino acid residues, and the protein molecular weight was about 20.3 kD. Phylogenetic tree showed that the gene is located in the S subgroup of bZIP family. It is the highest sequence homology with AtbZIP42. Promoter cis-acting element analysis showed that there are many stress-related response elements including ABRE, MYB and MYC in promoter sequence of SibZIP42. Phosphorylation site analysis showed that there are 14 serine, 4 tyrosine and 1 threonine phosphorylation sites in SibZIP42 protein sequence. The expression pattern analysis showed that SibZIP42 is involved in responses to various abiotic stresses and exogenous hormones. The gene expression profile results indicated that the expression level of SibZIP42 in root was higher than stem and leaf, and it was induced by drought, salt and ABA treatments in millet. The protein subcellular localization analysis revealed that SibZIP42 protein was localized in nucleus. Functional analysis of SibZIP42 showed that there is no significant difference of germination rate between transgenic lines and wild-type Arabidopsis WT on the normal MS medium, whereas on the MS culture medium with NaCl concentration of 90, 120 and 150 mmol·L-1, germination rate of SibZIP42 transgenic Arabidopsis was significantly higher than WT. Under 90 mmol·L-1 NaCl treatment condition, cotyledon green rate of SibZIP42 transgenic Arabidopsis was significantly higher than WT, whereas cotyledon green rate of SibZIP42 transgenic Arabidopsis was significantly lower than WT when the concentration of ABA was 0.5, 1 and 2 μmol·L-1 in MS culture medium. Some ABA-related response genes including HIS1-3, RD29B and RAB18 and drought-related response gene including PIP2A were up-regulated in SibZIP42 transgenic Arabidopsis, which suggested that the overexpression of SibZIP42 improved salt stress tolerance in transgenic Arabidopsis by ABA signaling pathway. 【Conclusion】 The salt tolerance of SibZIP42 transgenic Arabidopsis was significantly stronger than WT during the period of seed germination, while SibZIP42 transgenic Arabidopsis seeds were more sensitive to ABA treatment than WT which indicated that SibZIP42 positively regulated salt tolerance in transgenic Arabidopsis by ABA signaling pathway.
    Evaluation of Nitrogen Efficient Cultivars of Foxtail Millet and Analysis of the Related Characters at Seedling Stage
    CHEN Er-ying, YANG Yan-bing, QIN Ling, ZHANG Hua-wen, LIU Bin, WANG Hai-lian, CHEN Gui-ling, YU Shu-ting, GUAN Yan-an
    Scientia Agricultura Sinica. 2016, 49(17):  3287-3297.  doi:10.3864/j.issn.0578-1752.2016.17.004
    Abstract ( 500 )   HTML ( 4 )   PDF (778KB) ( 516 )   Save
    References | Related Articles | Metrics
    【Objective】The objective of this experiment was to probe genetic variation of nitrogen uptake efficiency (NupE) and nitrogen utilization efficiency (NutE) of foxtail millet cultivars at seedling stage and screen N efficient cultivars, which could provide a theoretical basis for the development of new N efficient varieties and N efficient mechanism study. 【Method】In this study, a sand culture pot experiment was conducted with 79 foxtail millet cultivars from three typically ecological types, shoot biomass, nitrogen concentration, nitrogen content, NutE and NupE were assessed under low nitrogen (0.2 mmol·L-1) and high nitrogen (6 mmol·L-1) supply, nitrogen use efficiency (NUE) types were also classified. 【Result】Large genetic variation was observed in shoot biomass, shoot N concentration, shoot N content, NupE, NutE and NUE at seedling stage. Among the three ecological types, northwest spring foxtail millet cultivars had the highest genetic variation of NupE, followed by north summer and northeast spring foxtail millet cultivars, while the genotypic variation of NutE in north summer type was more than that in northwest and northeast spring foxtail millet cultivars. Shoot biomass was significantly and positively correlated with NupE and NutE (P<0.01), and correlation coefficients were R2N0.2=0.1827** and R2N6=0.1027**, R2N0.2=0.8985**and R2N6=0.9442**, respectively. NUE was significantly and positively correlated with nitrogen content and negatively correlated with nitrogen concentration, and correlation coefficients were R2N0.2=0.8985**and R2N6=0.9442**, R2N0.2=0.1962** and R2N6=0.0998**, respectively. NupE was negatively correlated with nitrogen concentration, and the correlation coefficients were R2N0.2=0.9924** and R2N6=0.9910**. There was no significant correlation between NupE and nitrogen concentration, and between NupE and NutE. According to means of shoot biomass and NUE of 79 foxtail millet cultivars, foxtail millet cultivars from three ecological regions were classified into four types, respectively, both higher than the average under low nitrogen and high nitrogen level (HLHH), both lower than the average under low nitrogen and high nitrogen level (LLLH), higher than the average under low nitrogen and lower than the average under high nitrogen level (HLLH), and lower than the average under low nitrogen and higher than the average under high nitrogen level (LLHH). HLHH and LLHH types were dominant cultivar types in northeast spring foxtail millet, and the percentage of LLLH types was the lowest in northeast spring foxtail millet (PNW17.6%<PNS 32.4%<PNE36.0%). However, HLLH types were the main cultivar types in northwest spring foxtail millet (PNW 24.0%>PNS 18.9%>PNE 5.9%). 【Conclusion】There was a significant NUE genetic variation of seedlings in different foxtail millet cultivars. Genetic variation of NupE was the highest in northwest spring foxtail millet cultivars, and north summer foxtail millet cultivars had the highest genetic variation of NutE. There was no significant correlation between NupE and NutE, indicating that the evaluation and improvement of N uptake and utilization should be undertaken independently.
    Response of Leaf Defensive Enzymes and Antioxidant to Smut Fungus Stress in Broomcorn Millet
    ZHOU Yu, LIU Jia-jia, ZHANG Pan-pan, QU Yang, ZHANG Ji-ru-fei, ZHU Ming-qi, FENG Bai-li
    Scientia Agricultura Sinica. 2016, 49(17):  3298-3307.  doi:10.3864/j.issn.0578-1752.2016.17.005
    Abstract ( 347 )   HTML ( 3 )   PDF (406KB) ( 535 )   Save
    References | Related Articles | Metrics
    【Objective】 Head smut is an important disease that threats the yield of broomcorn millet seriously and the best way to control the disease is to plant resistant varieties. In order to screen the physiological and biochemical indexes of broomcorn millet for resistance to head smut, the activity of defensive enzymes and content of antioxidant were measured under the smut fungus stress. Furthermore, this will also provide theoretical supports for the breeding of broomcorn millet smut resistant varieties. 【Method】Artificial inoculation of seed saturated inoculation was adopted to infect broomcorn millet and inoculated broomcorn millet plants were planted in field. A field experiment of resistance levels identification to screen the varieties with different resistance was conducted in 2012-2013. In 2014, the broomcorn millet varieties with different resistance levels were used to measure the activity of defensive enzymes and content of antioxidant at seedling stage (SS), elongation stage (ES), heading stage (HS) and filling stage (FS) under the smut fungus stress. The defensive enzymes include the phenylalanine ammonia lyase (PAL), ascorbate peroxide (APX), glutathione reductase (GR) and the antioxidant contained the ascorbate (AsA) and glutathione (GSH). 【Result】 After 2 consecutive years of resistance identification, the average incidence rate of Heigezao (R1), Lvtuochuan (R2) and Xiaomaimizi (R3) were 0, 0 and 0.73%, respectively, indicating that they were disease-resistant varieties; the average incidence rate of Huangyingshu (S1), Ning04-262 (S2) and Yym0965 (S3) was 19.71%, 19.86% and 32.28%, respectively, therefore, they belong to disease-susceptible varieties. With the stress of smut fungus, the PAL activity of susceptible varieties changed greater than that in resistant varieties, since the PAL activity of susceptible varieties was significantly higher at elongation stage (3 610.8 U·g-1 FW), but significantly lower at filling stage (2 425.0 U·g-1 FW) compared to that (2 520.7 and 2 946.0 U·g-1 FW, respectively) of resistant varieties. The APX activity showed the same trend in both kinds of varieties, which was decreased first and then increased; the minimum value appeared at the elongation stage. At heading stage and filling stage, the APX activity of susceptible varieties was 461.1 U·g-1 FW and 516.7 U·g-1 FW, which was significantly higher than that (361.5 U·g-1 FW and 428.2 U·g-1 FW) in resistant varieties. The GR activity was increased first and then decreased in all the varieties. At heading stage, the value of GR activity was significantly higher than that of other 3 stages. The difference between the 2 kinds of varieties was that the GR activity of susceptible varieties was obviously higher than that of resistant varieties at heading stage, the value of which was 271.9 U·g-1 FW and 167.4 U·g-1 FW, respectively. After inoculation with smut fungus, the content of AsA ranged from 147.7 μg·g-1 FW to 344.8 μg·g-1 FW without obvious regularity in all the varieties. Also there was no significant difference between resistant and susceptible varieties. The GSH content of resistant varieties was decreased significantly from seedling stage to heading stage, and then increased significantly at filling stage. The GSH content of susceptible varieties was also decreased significantly from seedling stage to heading stage, but there was no significant difference compared to filling stage. Therefore, the GSH content was obviously higher in resistant varieties (984.7 μg·g-1 FW) than that in susceptible varieties (676.0 μg·g-1 FW) at filling stage. 【Conclusion】 Different broomcorn millet varieties have different resistance levels to head smut disease. Smut fungus stress can cause the changes of defensive enzymes activity and antioxidant content in leaves of broomcorn millet. The PAL activity of broomcorn millet leaves at elongation stage and filling stage, APX activity at heading stage and filling stage, GR activity at heading stage, GSH content at filling stage are obviously different between resistant and susceptible varieties, which can be used as the physiological and biochemical indexes to identify head smut resistant germplasms in broomcorn millet.
    Determination of Standard Varieties for Identifying Physiological Races of Foxtail Millet Blast Fungus
    LI Zhi-jiang, JIA Guan-qing, LI Xiang-yu, LI Yi-chu, MA Jin-feng, ZHI Hui, TANG Sha, ZHANG Shuo, CHAI Yang, LI Yan-dong, DIAO Xian-min
    Scientia Agricultura Sinica. 2016, 49(17):  3308-3318.  doi:10.3864/j.issn.0578-1752.2016.17.006
    Abstract ( 455 )   HTML ( 3 )   PDF (841KB) ( 485 )   Save
    References | Related Articles | Metrics
    【Objective】Foxtail millet blast (Pyricularia setariae Nisikado) is an important disease, which could cause reduced production and quality of foxtail millet, and the identification of different millet blast physiological races are very important for evaluating the resistant resources, breeding of resistant varieties and reasonable allocation of different resistant varieties for cultivation. The goal of this study is to determine standard varieties for the identification of physiological races of millet blast fungus, which would establish a basis for the evaluation of resistance of foxtail millet to different blast races. 【Method】 Sixty foxtail millet varieties collected from different geographical regions in China were inoculated using ten foxtail millet blast isolates sampled from different eco-regions. Statistical results of the sensitive reaction of 60 foxtail millet varieties to 10 blast isolates were recorded into excel format according to 0-9 grade standards of millet blast. The different resistant results were recorded as high resistance, resistance, middle resistance, sensitivity and high sensitivity. The high resistance, resistance, middle resistance were denoted with 0, and the sensitivity and high sensitivity were denoted with 1. The results were changed to 0/1 to construct the phylogenetic tree by NTSYS. Based on the resistant and infected results of sixty varieties to ten blast isolates, the standard identification system of foxtail millet blast were constructed. 【Result】Clustering analysis based on the pathogenicity of ten blast isolates and resistance of sixty varieties was conducted, respectively. The foxtail millet blast isolates were grouped into three clusters by similarity coefficient 0.70 as boundary, and foxtail millet varieties were divided into five categories by similarity coefficient 0.65 as boundary. Seven varieties, including Danpinian, Jinpingguzi, Jingu5, Damaomaogu, Longzhuagu, Jiajinmiao and Niutougu, were applied as standard identification varieties. The resistant varieties, Huangbangtou, Qitouhuang, Danpinian, Baigu, Weizinagu, and Zhenggu4 with broad spectrum resistance were screened, which could be used as the resources of resistant gene in foxtail millet blast disease breeding. 【Conclusion】This study screened a set of standard varieties for foxtail millet blast fungus identification, and identified several genetic stocks with broad-spectrum resistance to foxtail millet breeding.
    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    Impact of Ozone Stress on Growth and Yield Formation of Rice Genotypes with Different Ozone Sensitivity
    SHAO Zai-sheng, SHEN Shi-bo, JIA Yi-lei, MU Hai-rong, WANG Yun-xia, YANG Lian-xin, WANG Yu-long
    Scientia Agricultura Sinica. 2016, 49(17):  3319-3331.  doi:10.3864/j.issn.0578-1752.2016.17.007
    Abstract ( 356 )   HTML ( 3 )   PDF (503KB) ( 607 )   Save
    References | Related Articles | Metrics
    【Objective】 In order to cope with the increasing tropospheric ozone concentration, the responses of the growth and yield formation of rice with different ozone sensitivities were investigated. 【Method】 By using glasshouse-type fumigation chambers, 23 rice cultivars or lines were grown in 2013 at two ozone levels: Low ozone concentration as control (C-O3, 10 nL·L-1) and high ozone concentration as elevated O3 treatment (E-O3, 100 nL·L-1) from around a week after transplanting until maturity. Based on the decrease in the above-ground biomass under high ozone concentration, these rice genotypes were clustered into three types by the MinSSw (dynamic clustering method-minimum sum of squares within groups) method, namely A (low sensitivity type rice), B (moderate sensitivity type rice), and C (high sensitivity type rice) in the order of ozone sensitivity from low to high, respectively. The effects of ozone stress on plant height, tillering dynamics, grain yield, and yield components were determined, and relationships between their responses to ozone stress and the ozone-induced changes in the above-ground biomass at maturity stage. 【Result】 Compared to the control, ozone stress decreased the above-ground biomass of three rice types A, B, and C at grain maturity by 19%, 38%, and 52%, respectively, and significant treatment effects were detected in B and C. Ozone stress decreased plant height at all growth stages except the earliest stage, and the reduction in plant height was increased gradually with the plant development, but the magnitudes of decline among different rice types were comparable. On the contrast, the influence of ozone stress on tiller formation varied among different rice types. Averaged over the whole growth period, ozone stress had no significant effect on tiller number of rice type A, but significantly reduced those of B (-17%) and C (-23%). Ozone stress significantly reduced grain yield and yield components, and the different rice types exhibited different reduction in panicle number per unit area (A had no response, but B and C decreased by 16% and 26%, respectively), spikelets per panicle (A, B, and C decreased by 16%, 19%, and 27%, respectively) and spikelets per unit area (A, B, and C decreased by 11%, 31%, and 46%, respectively). However, the ozone-induced reduction in filled grain weight, filled grain percentage, and final grain yield showed small differences among three rice types. The ozone-induced reduction in grain yield was mainly caused by the sharp decline of filled grain percentage and total spikelets, and secondly associated with the decline of panicle number and filled grain weight. There were significant positive correlations between the ozone-induced response in above-ground biomass and the tiller number at middle and late growth stages, final panicle numbers, spikelets per panicle, as well as spikelets per unit area, but no close correlation with plant height, filled grain percentage, filled grain weight, or the final grain yield at each growth stage. 【Conclusion】Ozone fumigation of 100 nL·L-1 during plant growth shortened rice plant height, inhibited tiller production, spikelet formation, and especially grain filling, which led to a dramatic decline in grain yield. Rice ozone sensitivity was closely associated with the ozone-induced changes in tiller number at middle and late growth stage, final panicle numbers, spikelets per panicle, and spikelets per unit area, which can be used as screening parameters in ozone-tolerant rice breeding.
    Identification of the Chilling Resistance from Germination Stage to Seedling Stage in Upland Cotton
    WANG Jun-juan, WANG De-long, YIN Zu-jun, WANG Shuai, FAN Wei-li, LU Xu-ke, MU Min, GUO Li-xue, YE Wu-wei, YU Shu-xun
    Scientia Agricultura Sinica. 2016, 49(17):  3332-3346.  doi:10.3864/j.issn.0578-1752.2016.17.008
    Abstract ( 448 )   HTML ( 7 )   PDF (1415KB) ( 453 )   Save
    References | Related Articles | Metrics
    【Objective】 Low-temperature is one of the main factors affecting the normal growth and development of cotton plants. Identification of chilling resistance of cotton is the prior condition for the mechanism of cold resistance, which also can provide certain theoretical references for cold resistance breeding. 【Method】 The temperature conditions for cotton cold resistance identification were screened at germination stage following 7 d recovery at normal conditions (28℃, day/night, 14 h/10 h) after 7 d treatment at different low temperatures of 0℃, 4℃, 10℃, 15℃ with 4 upland cotton varieties (lines) with different cold resistance levels. Screening of days of cold resistance identification for 0℃ low-temperature treatment at germination stage was performed by 7 d recovery after the respective treatments of 1, 2, 3, 4, 5, 6, and 7 d under 0℃ low temperature. Two upland cotton varieties (lines) with different cold resistance levels was treated for 1, 2, 3, 4, 5, 6, and 7 d under 4℃ low temperature and recovered 7 d in order to evaluate cold resistance of cotton at budding stage. Evaluation of cold resistance of cotton at cotyledon stage was carried out at 4℃for 24 h and normal growth was recovered for 7 d with 47 cotton varieties. The biochemical indexes of bud and cotyledon under low temperature were measured at budding and cotyledon stages with the reagent kit developed by Nanjing Jiancheng Bioengineering Institute. 【Result】 The study established the standards of identification of cold-resistance by screening the stress conditions at different growth periods, including germination stage, budding stage and cotyledon stage. The relative cotyledons unfolding rate under normal condition growth for 7 d after 0℃ treatment for 4 d can be served as a cold resistance identification index of germination stage. The relative cotyledons unfolding rate under normal condition growth for 7 d after 4℃ treatment for 5 d can be used as chilling resistance identification index of the budding stage. Chilling resistance index under normal condition growth for 7 d after 4℃ treatment for 24 h can be used for evaluating the chilling resistance of cotton at cotyledon stage. The activities of SOD and POD in the bud of cold-resistant variety Yu2067 were increased at the early stage of low-temperature treatment and then decreased to a steady, while the activities of two enzymes in the bud of chilling-sensitive cotton line Hengmian No.3 decreased at first and then increased to steady. The activity of CAT in the bud of two varieties all showed a uptrend at first and then downtrend, but the activity of CAT in Yu2067 was always higher than that in Hengmian No.3 at later stage of low-temperature treatment. Besides, the soluble protein contents in the bud of two varieties were similar at first, but the content in Yu2067 started to be higher than that in Hengmian No.3 at later stage of low-temperature treatment. After the treatment of 4℃ for 24 h at cotyledon stage, the activities of SOD, POD and CAT were all showed a down-trend in Yu2067 while they all went up in Hengmian3. But the soluble protein content in Yu2067 went up while declined in Hengmian No.3. 【Conclusion】 Specific identification ways are needed at different growth stages correspondingly. The standards of identification of cold-resistance levels at different stages are different. Different materials have different cold resistance at different growth stages. Based on the variation trends of enzyme activity of three enzymes, it was speculated that the cold-resistance of cotton at budding stage had relation with antioxidant enzymes activity, while the cold-resistance of cotton at cotyledon stage had little relation with three antioxidant enzyme activities, but may be closely with the soluble protein content.
    PLANT PROTECTION
    Analysis of RNA-Seq-Based Expression Profiles of Δznf1 Mutants in Magnaporthe oryzae
    YUE Xiao-feng, QUE Ya-wei, WANG Zheng-yi
    Scientia Agricultura Sinica. 2016, 49(17):  3347-3358.  doi:10.3864/j.issn.0578-1752.2016.17.009
    Abstract ( 428 )   HTML ( 1 )   PDF (2651KB) ( 822 )   Save
    References | Related Articles | Metrics
    【Objective】Magnaporthe oryzae is the causal agent of rice blast, which is one of the most important disease threatening the production of cultivated rice worldwide. Appressorium-mediated penetration is a key step in the disease cycle of the fungus. Previously, it was reported that a C2H2 zinc finger transcription factor encoded by ZNF1 is essential for appressorium development, penetration and pathogenicity in the rice blast fungus. The objective of this study is to understand the regulatory mechanism of Znf1 and reveal the genes transcriptionally regulated by Znf1, thus providing new clues for further investigating molecular mechanism of pathogenicity in this fungus. 【Method】The transcriptome profiles of vegetative mycelia of the wild-type strain Guy11 and a Δznf1 mutant were assayed with the RNA-Seq technique. The gene expression levels were calculated using the FPKM method. The criteria of false discovery rate (FDR)≤0.001 and absolute value of log2 ratio≥1 were used to identify differentially expressed genes (DEGs). The sequences of the DEGs were subjected to BLAST queries against the gene ontology (GO) database and KEGG pathway database to predict their biological function and pathways. In order to define in more detail about the sub-set of genes regulated by Znf1, transcriptome profiles of a mutant lacking the PMK1 MAP kinase-encoding gene was also analyzed based on the RNA-Seq technique. To identify the genes regulated by both Znf1 and Pmk1, the DEGs between Δznf1 and Δpmk1 were compared. In addition, the genes highly expressed during appressorium formation but down-regulated in either Δznf1 or Δpmk1 were obtained by comparison with the previous transcriptional profile data. 【Result】 Totally, 709 DEGs in the Δznf1 mutant, including 299 up-regulated and 410 down-regulated genes, were identified by comparison with the wild-type strain Guy11. Gene ontology enrichment analysis showed that 118, 299 and 308 DEGs were classified into cellular component, molecular function and biological process, respectively. KEGG pathway enrichment analysis revealed that the DEGs were mainly involved in metabolic pathways, biosynthesis of secondary metabolites and glycerophospholipid metabolism. Several known pathogenicity-related genes, including LPP3, HOX7, PBS2 and MPG1, were found down-regulated in Δznf1. The comparison of DEGs showed that about 56% DEGs in Δznf1 shared identical to those in Δpmk1. Three isotrichodermin C-15 hydroxylase encoding genes, MGG_03825, MGG_02329 and MGG_08498, were significantly down-regulated in both Δznf1 and Δpmk1. In addition, 48 genes up-regulated during appressorium formation were down-regulated in the two mutants, indicating that these putative appressorium-associated genes were regulated directly or indirectly by Znf1 and Pmk1. To confirm the reliability of the RNA-Seq data, 10 DEGs were randomly selected for qRT-PCR. The results showed that the expression patterns in qRT-PCR were consistent with those in RNA-Seq.【Conclusion】 The expression profiling data and predicted molecular function of Znf1-dependent DEGs were obtained by RNA-Seq technique. Several pathogenicity-associated genes were regulated by Znf1. Additionally, several genes highly expressed during appressoria formation were also regulated by Znf1 as well as Pmk1. This study provided valuable information for further research on Znf1 downstream gene regulatory network.
    Cloning and Expression Analysis of Fusarium Wilt Resistance- Related Gene ClMYB Transcription Factor from Citrullus lanatus
    HAN Jin-huan, WANG Li-xia, GAO Hong-bo, Lü Gui-yun
    Scientia Agricultura Sinica. 2016, 49(17):  3359-3369.  doi:10.3864/j.issn.0578-1752.2016.17.010
    Abstract ( 474 )   HTML ( 3 )   PDF (5070KB) ( 600 )   Save
    References | Related Articles | Metrics
    【Objective】The objective of this study is to clone ClMYB, which is a gene encoding MYB transcription factor, analyze its sequence features and expression characteristics, and to provide new insights into the understanding of the molecular mechanism in response of watermelon plants to Fusarium oxysporum f. sp. niveum infection.【Method】In a recent study, a novel EST of MYB transcription factors was obtained from data analysis of microarray and suppression subtractive hybridization (SSH) of incompatible interaction between watermelon and F. oxysporum f. sp. niveum. cDNAsequenceof the MYB was isolated and cloned using cDNA from watermelon roots with F. oxysporum f. sp. niveum infection by RT-PCR method, designated as ClMYB. The conserved domains and sequence features of ClMYB protein were analyzed by bioinformatics methods. multiple sequence alignments and a phylogenetic tree of homologous species were done between ClMYB protein and its homologous ones from other species using MEGA5.0. The subcellular localization of ClMYBwas analyzedusing fusion expression vector PYBA1332-GFP. Then ORF of the cloned gene was inserted into the expression vector pCzn1 by Nde I and Xba I digestion. The recombinant plasmid was transformed into E. coli Arctic Express system and induced expression by 0.5 mmol·L-1 IPTG for 4 h. The expression of the fusion protein was detected by SDS-PAGE. The expression pattern of target genes in watermelon with F. oxysporum f. sp. niveum were detected by real-time fluorescent quantitative PCR (qrt-PCR).【Result】Using RT-PCR method, cDNA sequence of ClMYB (GenBank: KT751229) was cloned from watermelon (PI296341-FR) root. Sequence alignment and bioinformatics analysis revealed that the deduced amino acids of ClMYB had common characteristics of MYB transcription factors with two MYB domains of R2 and R3 at the N-terminal and highly variation at the C-terminal. Phylogenetic tree analysis suggested that the encoded protein had the closest genetic relationship with Cucumis melo MYB (GenBank: XM_008440304) and Cucumis sativus MYB (GenBank: XM_011652633). Subcellular localization results showed thatClMYB protein was located in the nucleus which belongs to a typical transcription factor. The expression of fusion protein was obtained by inducing with IPTG in E. coli and its relative molecular weight was 36 kD. qrt-PCR showed that the expression of ClMYB was induced by F. oxysporum f. sp. niveum, and appeared peak earlier and higher in PI296341-FR than Black diamond. MeJA at 50 μmol·L-1 could significantly improve the resistance of Black diamond to F. oxysporum f. sp. niveum and induce the expression of ClMYB. In PI296341-FR, the expression of ClMYB was also induced by MeJA, while its expression level was lower than Black diamond. 【Conclusion】ClMYB is a typical R2R3-MYB transcription factor and is located in the nucleus. The expression of fusion protein was obtained by prokaryotic expression and its relative molecular weight is 36 kD. The expression of ClMYB was induced by F. oxysporum f. sp. niveum and JA. It is speculated that ClMYB may be involved in JA-mediated resistance signal transduction network of F. oxysporum f. sp. niveum. It might play an important role in disease resistance of Citrullus lanatus.
    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    The Effect of Urea-Formaldehyde Fertilizer Under Different Components by Summer Maize
    NI Lu, BAI You-lu, YANG Li-ping, LU Yan-li, WANG Lei, ZHOU Li-ping
    Scientia Agricultura Sinica. 2016, 49(17):  3370-3379.  doi:10.3864/j.issn.0578-1752.2016.17.011
    Abstract ( 414 )   HTML ( 3 )   PDF (414KB) ( 577 )   Save
    References | Related Articles | Metrics
    【Objective】This experiment was carried out to study the effect of urea-formaldehyde fertilizer with different components on summer maize and to provide a theoretical basis for one time fertilization in summer maize, thus simplifying fertilization steps, saving fertilization cost, increasing fertilizer efficiency, reducing loss of nutrients and achieving environmental protection.【Method】Maize variety "zhengdan958" was used as materials in a field experiment. Under field conditions in two consecutive years, three urea-formaldehyde fertilizer treatments (UF1, UF2 and UF3), conventional fertilization (CF) and no fertilization (CK) were designed. Research mainly focused on the influence of different urea-formaldehyde treatments on yield of summer maize, nitrogen use efficiency, accumulation amount of above-ground portion nitrogen and content of inorganic nitrogen in soil.【Result】Results of the experiment showed that urea-formaldehyde of different components had different fertilizer efficiency. All slow release fertilizers of UF1, UF2 and UF3 could increase yield of summer maize and nitrogen use efficiency, among which UF2 was the best. Compared with conventional fertilization, UF2 had increased yield of summer maize by 7.63% and nitrogen use efficiency by 16.43 percentage points. UF3 increased yield by 4.98% and nitrogen use efficiency by 0.82 percentage points. In terms of effect, UF1 was only next to it. By using it, the average yield increase of summer maize was 6.53% and nitrogen use efficiency was improved by 12.30 percentage points. In addition, urea-formaldehyde with different components showed different releasing rates. Compared with conventional fertilization, use of urea-formaldehyde fertilizer could keep relatively high content of inorganic nitrogen in 0-20cm soil layer. And for UF2, the content was the highest, which was up to 80.09 mg·kg-1. UF1 took the second place which was up to 66.47 mg·kg-1 and UF3 had the lowest content of 51.18 mg·kg-1. The inorganic nitrogen content of conventional fertilization increased because of topdressing urea at bell-mouthing stage, which was up to 27.46 mg·kg-1 in filling period. In 20-40 cm soil layer, the inorganic nitrogen contents of UF1 and UF2 were stable between bell-mouthing stage and filling stage and decreased after filling stage. The inorganic nitrogen content of UF1 was as low as 13.40 mg·kg-1and lower than conventional fertilization by 1.05 mg·kg-1 in harvesting period. The inorganic nitrogen content of UF2 was 14.13 mg·kg-1, which was lower than conventional fertilization by 0.32 mg·kg-1 in harvesting period. But the inorganic nitrogen content of UF3 was slightly higher than other treatments and higher than conventional fertilization by 1.51 mg·kg-1 during the harvest period. Therefore the one-time urea-formaldehyde fertilization met the demand for nutrients during the whole maize growth period.【Conclusion】Relatively higher output and nitrogen use efficiency were obtained from UF2 treatment among different urea-formaldehyde fertilizers. It was conclude that this fertilizer application method can be served as the special slow release fertilizer in the north of North China Plain where low and medium productive soil dominates the land.
    Effects of Aerated Irrigation on CO2 Emissions from Soils of Tomato Fields
    CHEN Hui, HOU Hui-jing, CAI Huan-jie, ZHU Yan, WANG Chao
    Scientia Agricultura Sinica. 2016, 49(17):  3380-3390.  doi:10.3864/j.issn.0578-1752.2016.17.012
    Abstract ( 342 )   HTML ( 2 )   PDF (750KB) ( 795 )   Save
    References | Related Articles | Metrics
    【Objective】Carbon dioxide (CO2) is a significant greenhouse gas that also contributes to the global warming. Lots of studies have reported that aerated irrigation would increase crop production, quality and water use efficiency by changing the aeration of root zone, while few studies have focused on the environmental effects of aerated irrigation, especially the greenhouse gas emissions from soils of greenhouse vegetable fields. Hence, it is of significance to analyze the effects of aerated irrigation on CO2 emissions from soils of greenhouse tomato fields for assessing farmland ecological effects of aerated irrigation. 【Method】Venturi meter (Mazzei 287, America) was used as the aeration equipment to irrigate the experimental plots. The method of static chamber/gas chromatography was used to study the effects of aerated irrigation on CO2 emissions. Two factors (irrigation and aeration) were designed in the experiment to reveal the effects of aerated irrigation on soil CO2 emissions. Four treatments were designed in the experiment: aerated deficit irrigation (AI1), unaerated deficit irrigation (CK1), aerated full irrigation (AI2) and unaerated full irrigation (CK2). 【Result】Throughout the whole growth period of tomato, soil CO2 emissions followed the same pattern in different treatments, and peaks occurred at the blooming and fruit setting stages. Treatments with aeration and full irrigation increased average value of CO2 fluxes and cumulative emissions of soil CO2 throughout the whole growing period compared to relative non-aeration and deficit irrigation, but the difference was not significant (P>0.05). The average value of CO2 fluxes was 229.31, 193.66, 259.10 and 224.76 mg·m-2·h-1, respectively. The maximum cumulative emission of CO2 was 6 383.43 kg·ha-1 in AI2 treatment, which was 1.12, 1.32 and 1.13 times as that in AI1, CK1 and CK2 treatments, respectively. Soil water filled pore space (WFPS) during the whole growing season showed a decreasing trend in different treatments, soil temperature (T) showed a increasing trend and the difference between treatments was relatively small at the same time, while soil organic carbon (SOC) showed a fluctuant pattern. In addition, aerated irrigation decreased T and WFPS, but increased SOC compared to unaerated irrigation, and the treatment effects was not significant. Full irrigation increased WFPS and SOC indistinctively compared to deficit irrigation, while the effects of full irrigation and deficit irrigation on T was different. A negative correlation between soil CO2 emissions and WFPS was observed, while a positive correlation between soil CO2 emissions and T and SOC was observed, but the difference was not significant (P>0.05). 【Conclusion】Compared to non-aeration, aeration increased soil CO2 emissions, but the difference was not significant (P>0.05). This study provided some scientific basis and a useful reference for assessing farmland ecological effects and mitigating greenhouse gas emissions from greenhouse soils of aerated irrigation.
    HORTICULTURE
    Metabolic Profiles of Sugar Metabolism and Respiratory Metabolism of Korla Pear (Pyrus sinkiangensis Yu) Throughout Fruit Development and Ripening
    PAN Yan, MENG Xin-tao, CHE Feng-bin, XUE Su-lin, ZHANG Ting, ZHAO Shi-rong, LIAO Kang
    Scientia Agricultura Sinica. 2016, 49(17):  3391-3412.  doi:10.3864/j.issn.0578-1752.2016.17.013
    Abstract ( 353 )   HTML ( 4 )   PDF (3008KB) ( 473 )   Save
    References | Related Articles | Metrics
    【Objective】The responding relationship of sugar metabolism and respiratory metabolism throughout the fruit development and ripening of Korla pear (Pyrus sinkiangensis Yu) was analyzed with an attempt to assess the effect of metabolic interaction on fruit sugar accumulation.【Method】Korla pears were sampled at 10-day intervals from 60-150 DPA days post anthesis and were sliced into pulps, cores and peels, then the 12 enzyme activities of sugar metabolism and 9 enzyme activities of respiratory metabolism as well as the quality, vertical and horizontal diameter, contents of starch and 4 soluble sugars, and respiration rates of different respiratory pathways were measured. The relationship between quality and sugar components, respiratory main pathway and the effect of clustered metabolic enzymes interaction on the sugars composition at different stages of fruit development and ripening were determined from the results of regression analysis, hierarchical clustering and principal component analysis for discussion and profile of the metabolic pathways and features of different fruit parts in Korla pear.【Result】EMP (Embden-Meyerhof-Parnas) is the respiratory main path of core. EMP and TCAC (Tricarboxyficacid cycle) alternately compose the respiratory main path of pulp. EMP is integrated with TCAC in the respiratory main path of fruit peel. Either core or pulp, the metabolic interaction between sugar metabolism and respiratory metabolism differs in 2 patterns at 90 DPA before and after, and differs at 120 DPA of peel, respectively. The enzymes of core involved in metabolism of starch, sucrose and sorbitol were clustered with the enzymes involved in metabolism of EMP, PPP (phosphopentose pathway) and CP (Cytochrome pathway) at maturation, as well as the hierarchical cluster of pulp grouped the enzymes involved in metabolism of starch, sucrose, EMP, TCAC, CP and AP (alternative pathway), and the cluster of peel grouped the enzymes involved in metabolism of sucrose, sorbitl, EMP, TCAC, CP and AP. The clustered enzyme activities increased dramatically or peaked that was relating significantly to the fructose and glucose accumulations of core and pulp, and relating to the increase of fructose and glucose with sorbitol conversion of fruit peel also.【Conclusion】Korla pear forms typical internal quality by sugar accumulation composed of principal components with fructose and glucose, which has a heterostructure presence of sugar composition and sweetness, that is the result of different metabolic profiles between sugar metabolism and respiratory metabolism respond progressively in different structures at different stages throughout the fruit development and ripening. GPI (glucose phosphate isomerase) and COX (Cytochrome oxidase) are key enzymes of common crosslink in sugar metabolism and respiratory metabolism at fruit maturation in three parts of Korla pear.
    Effect of Carbonized Apple Branches on Bacterial and Fungal Diversities in Apple Root-Zone Soil
    CAO Hui, LI Yan-ge, ZHOU Chun-ran, NING Liu-fang, YANG Hong-qiang
    Scientia Agricultura Sinica. 2016, 49(17):  3413-3424.  doi:10.3864/j.issn.0578-1752.2016.17.014
    Abstract ( 396 )   HTML ( 3 )   PDF (2584KB) ( 796 )   Save
    References | Related Articles | Metrics
    ObjectiveThe microorganisms in soil of root-zone are important factors affecting root environment, carbonized apple branches are the low oxygen pyrolysis products of the abandoned fruit trees. The purpose of this study was to understand the structure of soil bacteria and fungi in apple root-zone and the response of their diversities to carbonized apple branches, and to provide a theoretical basis for the reasonable application of carbonized apple branches and the improvement of soil biological characters in orchard. 【Method】 In the spring, the 2-year-old ‘Fuji’ apple trees (rootstock for Malus hupehensis Rehd) in similar growth were transplanted to the potting soil, the soil was mixed with different mass ratios (0-4%) of carbonized apple branches beforehand. Soil samples were collected after 120 days of transplanting, genomic DNA was extracted, and PCR amplification was made to establish libraries. In this study, the 16S rRNA genes V3+V4 regions of soil bacteria and fungal ITS1 regions were sequenced by Illumina high-throughput sequencing technology on Miseq platform, and related biological analysis was conducted to explore the changes of soil bacterial and fungal abundances, diversities and structures.ResultA total of 16 656 bacterial operational taxonomic units (OTUs) and 435 fungal OTUs were obtained from 15 apple root-zone soil samples, among them, Proteobacteria, Bacteroidetes and Acidobacteria were the dominant bacteria which the total relative abundance was 70.68%-72.80%, and Basidiomycota, Ascomycota and Zygomycota were dominant fungi which the total relative abundance was 68.00%-75.14%. The richness indices of Chao and Ace showed that 1% (w/w) carbonized apple branches increased the abundance of bacteria by 15.42% and 3.89% compared with the control, respectively. 0.5% (w/w) carbonized apple branches increased the richness of fungi by 2.80% and 3.61%,respectively. Simpson and Shannon diversity index analysis showed that 0.5%-4% (w/w) carbonized apple branches reduced the diversity of soil bacteria, increased the diversity of soil fungi; bacteria Shannon diversity index was the lowest at 1% carbonized apple branches and fungi Shannon diversity index was the highest at 0.5% carbonized apple branches. Application of 1%, 2% and 4% carbonized apple branches to the soil, the relative abundance of Proteobacteria, Acidobacteria and Basidiomycota all decreased and the relative abundance of Bacteroidetes and Zygomycota all increased in apple root-zone soil at different degrees. The relative abundance of Hymenochaetaceae (31.99%-46.74%) was the highest in Basidiomycota. 1%, 2% and 4% carbonized apple branches all made it decrease. Carbonized apple branches at 0.5%-4% dosage could increase the number of bacteria unique OTUs significantly and affect the bacteria groups, wherein the number of bacterial unique OTU was one to three times of the common OTU numbers, but they had no significant effect on the fungal groups.ConclusionThe 0.5%-4% (w/w) carbonized apple branch applied to the soil of apple root-zone, the abundance and diversity of soil bacteria and fungi had obviously changed, and the specific bacterial species under each dosage increased too. The soil bacterial richness increased significantly in root-zone applied 1% carbonized apple branches.
    STORAGE·FRESH-KEEPING·PROCESSING
    Analysis and Comparison of Calpain in Mutton with Different Levels of Tenderness
    DU Man-ting, LI Xin, LI Zheng, GAO Xing, ZHANG Cai-xia, ZHANG De-quan
    Scientia Agricultura Sinica. 2016, 49(17):  3425-3432.  doi:10.3864/j.issn.0578-1752.2016.17.015
    Abstract ( 297 )   HTML ( 2 )   PDF (599KB) ( 334 )   Save
    References | Related Articles | Metrics
    【Objective】Calpain plays an important role in postmortem tenderization through the proteolysis ability. The activity of calpains, especially μ-calpain, is closely related to the tenderness of muscles. The objective of this study was to investigate the activation process and the autolysis rate of μ-calpain in mutton during postmortem aging by analyzing and comparing the differences of calpain in mutton with different levels of tenderness, thereby provide a theoretical basis for regulating tenderness through regulating calpain activity. 【Method】 The longissimus dori of WuZhuMuQin Fat Tailed sheep were divided into tender group and tough group based on myofibrillar fragmentation index, then the pH value, the existent state of μ-calpain large subunit and the content of native μ-/m-calpain during postmortem were analyzed. 【Result】The variation trend of pH at postmortem in tender/tough group was identical, all the pH values in two groups at the same postmortem time showed no difference. The 80 kDa subunit of μ-calpain degraded gradually after slaughter. The content of μ-calpain 80 kDa subunit of tender group was significantly higher than tough group at 30 min postmortem. The intact 80 kDa subunit of μ-calpain could not be detected after 48 h in tender group, while it could be detected until 5 d postmortem in tough group. The content of native μ-calpain was decreased gradually, and this content of tender group was significantly higher than tough group at 30 min, 2 h postmortem, however, at 12 h, 5 d, and 7 d postmortem the tough group was significantly higher than the tender group. The autolysis and degradation of μ-calpain in tender group was apparent at 6 h postmortem.【Conclusion】Compared with tough group, tender group had a higher initial μ-calpain content, but the autolysis and degradation rate of μ-calpain in tender group was much faster, leading higher autolysis and degradation degree in tender group. The results indicated that meat tenderness during the postmortem was directly affected by μ-calpain expression quantity and autolysis. In addition, changes of μ-calpain during 24 h postmortem, especially 12 h, had the most impact on mutton tenderness.
    ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT
    Influences of Pax6 PAI Subdomain on MITF, TYR, TYRP1 and TYRP2 in Melanocytes
    NIE Rui-qiang, YANG Yu-jing, XIE Jian-shan, FAN Rui-wen, XU Dong-mei, YU Xiu-ju, DUAN Zhi-cheng, DONG Chang-sheng
    Scientia Agricultura Sinica. 2016, 49(17):  3433-3442.  doi:10.3864/j.issn.0578-1752.2016.17.016
    Abstract ( 263 )   HTML ( 1 )   PDF (2496KB) ( 327 )   Save
    References | Related Articles | Metrics
    【Objective】 Highly conserved PAX family has important effects on the differentiation of melanocytes and production of melanin. It has mainly binding sites with target gene in PD domain that is included by all of PAX family, on the other hand, the PAI subdomain, which is located in amino terminal of PD domain, has most important effects on PD domain which bound with target gene. Many reports show that Pax6 has the most important effects on the differentiation of retinal pigment epithelial cells. This experiment studies the function of PD domain and PAI subdomain of PAX family by analysing the function of Pax6 PAI subdomain.【Method】The structure of Pax6 PD domain was analyzed by Psipred. The target gene binding sites of Pax6 PD domain was analyzed by NCBI. The binding sites of Pax6 PD domain to the promoter of MITF, TYR, TYRP1, and TYRP2 were analyzed by Jasper. The coding sequences of Pax6 PAI subdomain was amplified by PCR and the Pax6 PAI subdomain was cloned into the T-Vector, meanwhile, confirmed by sequencing. The fragment was then subcloned into a mammalian expression vector, resulting in a construction that contained a promoter driving the expression of green fluorescent protein (GFP). The plasmid vector was confirmed by sequencing. Then, the mouse melanocytes were transfected with the vector using Liposome 2000. Three methods were used in the result test, they were quantitative real-time PCR, western blot and melanin content measurement. 【Result】The target gene binding sites of Pax6 PD domain was mainly distributed in PAI subdomain which is located in amino terminal of PD domain. There was a binding site of Pax6 PD domain at -695 base of MITF promoter; Two binding sites of Pax6 PD domain at -873 base and -1133 base of TYR promoter; One binding site of Pax6 PD domain at -629 base of TYRP1 promoter; And one binding site of Pax6 PD domain at -655 base of TYRP2 promoter. The RT-PCR and western blot results showed that the four target genes and melanin content were significantly increased. MITF mRNA was significantly increased by 2.05 times (P<0.01), TYR mRNA was increased by 2.09 times, TYRP1 mRNA was increased by 2.93 times(P<0.05), TYRP2 mRNA was increased by 3.62 times (P<0.01). Compared with the control group, MITF protein was significantly increased by 1.7 times (P<0.01), TYR protein was increased to 2 times (P<0.05), TYRP1 protein was increased by 1.9 times(P<0.01), TYRP2 protein was increased to 1.37 times. Meanwhile, the melanin content was significantly increased by 1.33 times (P<0.001).【Conclusion】Results of the study demonstrated that the Pax6 PAI subdomain still promoted the expression of MITF, TYR, TYRP1, and TYRP2, while increased the production of melanin of melanocytes.
    Effect of Free Stall Barn-Feeding and Castration on Fattening Performance of Holstein Bulls
    DU Liu-liu, LI Qiu-feng, LI Yan, CAO Yu-feng, YU Chun-qi, LI Xiao-meng, WANG Xiao-ling
    Scientia Agricultura Sinica. 2016, 49(17):  3443-3452.  doi:10.3864/j.issn.0578-1752.2016.17.017
    Abstract ( 409 )   HTML ( 3 )   PDF (781KB) ( 405 )   Save
    References | Related Articles | Metrics
    【Objective】This study was designed to investigate the effects of free stall barn-feeding and castration on growth and serum biochemical parameters of Holstein bulls to provide a scientific basis for fattening and management of Holstein bulls. 【Method】 Twenty-six healthy Holstein bulls were randomly selected and castrated (steers). Twenty days later, another 26 healthy Holstein bulls were randomly selected as controls (bulls). The bulls and steers were randomly divided into 2 groups with 13 heads in each group. The body weight of cows from each group were similar (about 270 kg). Four groups were conducted with this trial: group I (bulls with free stall barn-feeding, BF), group II (bulls with tie stall barn-feeding, BT), group III (steers with free stall barn-feeding, SF), group IV (steers with tie stall barn-feeding, ST). The experiment was lasted for 206 days. 【Result】The results showed as follows: (1) Compared with tie stall groups, the average daily gain (ADG) of free stall groups increased by 13.59% (P<0.01). Compared with the steers groups, the ADG of bulls groups increased by 13.59% (P<0.01). The interaction of free or tie-stall (FOT) × non-castration or castration (NOC) showed significant effects on the ADG (P<0.05). Compared with group II, III and IV, the ADG of group I increased by 5.26% (P>0.05), 5.26% (P>0.05) and 31.87% (P<0.01) respectively. (2) The apparent digestibility of the neutral detergent fiber (NDF) (P<0.01), calcium (Ca) (P<0.01), crude fat (EE) (P<0.05), phosphate (P) (P<0.05) of tie stall groups were lower than that of free stall groups. The apparent digestibility of Ca of steers groups was lower than that of bulls groups (P<0.01). (3) In comparison with tie stall group, the free stall groups showed higher serum concentrations of total protein (TP) (P<0.01), albumin (ALB) (P<0.01), triglyceride (TG) (P<0.01), total cholesterol (TC) (P<0.01), triiodothyronine (T3) (P<0.01), lysozyme (LYS) (P<0.05), NO (P<0.05) as well as the activity of serum of glutamic-pyruvic transaminase (AST) (P<0.01) and glutamic-oxalacetic transaminase (ALT) (P<0.05). While the serum concentrations of urea nitrogen (BUN) (P<0.01) of the free stall groups were lower than that of the tie stall groups. In comparison with bulls groups, the serum concentrations of TP (P<0.01), AST (P<0.01), growth hormone (GH) (P<0.01), T3 (P<0.01), testosterone (T) (P<0.01), and immunoglobulin G (IgG) (P<0.01) were lower, but the serum concentrations of TG (P<0.01), TC (P<0.01), leptin (LP) (P<0.01), and BUN (P<0.05) were higher in steers groups. The serum concentrations of TP, TG, TC, T3 significantly affected by interaction between FOT×NOC (P<0.01). The serum concentrations of TP and T3 in group I were the highest, while TG and TC in group III were the highest. 【Conclusion】 In conclusion, bulls with free stall barn-feeding could improve the fattening performance and immune function, and steers with free stall barn-feeding could increase the accumulation of adipose tissue. Therefore, the results suggested that for fair quality beef production, bulls with free stall barn feeding could be applied to the Holstein bulls at 270-500 kg live weight, but for top grade beef production, steers with free stall barn-feeding could be used. 
    Digital Gene Expression Analysis of Silkworm Infected by Bombyx mori Bidensovirus Zhenjiang Strain
    GAO Kun, SHANG Meng-ke, QIAN He-ying, QIN Guang-xing, GUO Xi-jie
    Scientia Agricultura Sinica. 2016, 49(17):  3453-3464.  doi:10.3864/j.issn.0578-1752.2016.17.018
    Abstract ( 335 )   HTML ( 3 )   PDF (892KB) ( 486 )   Save
    References | Related Articles | Metrics
    【Objective】 The objective of this study is to screen differentially expressed genes in the Bombyx mori larvae infected with BmBDV-ZJ (B. mori bidensovirus Zhenjiang strain) and identify regulatory genes related to the virus infection and the host response so as to provide important clues for better understanding of the mechanism of B. mori resistance against BmBDV-ZJ infection. 【Method】 The differential gene expression profiles in JS B. mori larvae after oral infection with BmBDV-ZJ were constructed using Illumina Genome Analyzer platform. In order to exclude the effects of individual differences, 10 larvae were dissected and pooled as one sample for digital gene expression (DGE) analysis, respectively. The differential expression detection of genes across samples was performed using a rigorous algorithm method. False discovery rate (FDR) was used to determine the P value threshold in multiple tests and analyses. The significance of the gene expression difference was obtained through a FDR≤0.001 and the absolute value of log2 ratio≥1. The gene ontology (GO) classification system was used to determine the possible functions of all differentially expressed genes. P value was calculated by GO (http://www.geneontology. org/) and corrected by Bonferoni. A corrected P value≤ 0.05 was selected as a threshold for significant enrichment of the gene sets. WEGO (web gene ontology annotation plot) software was used for visualizing, comparing and plotting GO annotation results. Pathway enrichment analysis was conducted to further identify the significantly enriched metabolic pathways or signal transduction pathways by using the KEGG database. Pathways with a Q value≤0.05 were designated as significantly enriched pathways in DGEs. Then some of the differentially expressed genes were verified by quantitative real-time PCR (qRT-PCR). 【Result】 Totally, 4 850 663 and 4 875 307 raw tags were generated in the control and BmBDV-ZJ infected DGE (digital gene expression) libraries, respectively. There were 4 757 934 and 4 788 406 clean tags corresponding to 62 436 and 63 680 distinct clean tags were filtered from the raw tags. The distribution of the total and distinct tags over the different tag abundance categories showed highly similar patterns in each DGE library. The sequencing depths reached approximately 3.5 and 3.7 million in the two DGE libraries, respectively, which satisfied the requirement for the experiment. So the two DGE libraries were reliable. The tag sequences of the two DGE libraries were mapped to the reference database of B. mori. In the control and BmBDV-ZJ-infected DGE library, 36.39% and 45.30% of the clean tags were mapped to a gene in the reference database, 50.02% and 43.34% of the clean tags could be mapped to genome of B. mori, while 13.59% and 12.35% of the clean tags were unknown tags. A total of 447 differentially expressed genes were detected, of which 306 were upregulated and 141 were downregulated. There were 218, 147, 179 differentially expressed genes have GO categories according to molecular function, cellular component and biological process, respectively. KEGG (http://www.genome.jp/kegg) ontology assignments were used to classify the functional annotations of the identified genes. Among the differentially expressed genes, 330 were mapped to 151 pathways in the KEGG database. Nineteen terms was significantly enriched (Q value≤0.05) and the cytosolic DNA-sensing pathway was significantly enriched. Moreover, 24 differentially expressed genes were verified using qRT-PCR, showing that 20 genes were concordant in the expression with DGE. Among the 9 differentially expressed genes related to cytosolic DNA-sensing pathway, BGIBMGA009408-TA, BGIBMGA004913- TA, BGIBMGA011753-TA, which were the DNA-directed RNA polymerase III genes in B. mori, were all up-regulated in the BmBDV-ZJ infected B. mori with 4.3, 2.3, 3.4-fold change, respectively. 【Conclusion】 The results of this study may serve as a basis for future research not only on the molecular mechanism of BmBDV-ZJ invasion but also on the mechanism of B. mori resistance against BmBDV-ZJ infection.