B-box（BBX）基因家族编码的蛋白是由包含锌指结构的转录因子组成，其N端有一个或两个高度保守的B-box基序。BBX蛋白在植物生长发育的各个方面起着至关重要的作用，包括幼苗的光形态发生、避荫、开花时间以及生物和非生物胁迫响应。目前,各国的研究者已经从几种植物中鉴定了BBX的家族成员，然而玉米中的BBX家族成员还知之甚少。通过对玉米BBX家族基因的全基因组鉴定、表达和互作的综合分析，可为了解其功能提供有用信息。本研究共鉴定出36个玉米BBX家族成员，进化分析显示其分布于三个主要分支。在每个主分支中ZmBBXs都具有相似的结构域、基序和基因组结构。基因重复分析表明，玉米BBX蛋白家族的扩张主要是通过片段重复来完成的。利用实时荧光定量PCR技术，本研究分析了ZmBBXs在不同器官组织和不同非生物胁迫条件下的表达。利用生物信息学工具，本研究建立了ZmBBXs蛋白的相互作用网络，并通过双分子荧光互补（BiFC）试验进行了验证。本研究的发现有助于理解ZmBBX家族的复杂性，并为揭示ZmBBX蛋白的生物学功能提供新的线索。

本研究提出一种统计数据空间化的方法构建多时像农作物种植格局空间数据集来解决数据缺失的问题。该方法采用两层嵌套结构实现土地利用层和农作物层模拟，其中第一层模拟的耕地数据用于控制第二层农作物种植格局空间模拟范围。第二层农作物层采用空间迭代的方法按分配规则进行农作物面积统计数据空间化，最终实现农作物空间格局动态模拟。该模型在中国黑龙江省地区进行2000-2019年农作物空间格局模拟，结果表明模型模拟精度较高，能够实现长时间序列的农作物种植面积统计数据空间化应用，未来该模型能广泛应用于农业土地系统各方面研究及生产应用。

本研究采用高通量测序法，对添加添加剂的农作物秸秆好氧堆肥过程（升温、高温、降温和腐熟四个阶段）中微生物多样性动态变化进行研究。此外，还对堆肥过程中理化参数的变化进行分析。添加尿素或尿素与微生物菌剂配合添加，可延长堆体高温发酵时间。C/N比和发芽指数变化表明，添加剂直接改变了堆肥的理化性质，影响了细菌和真菌的多样性和丰富度，有利于堆肥化。在高温阶段，秸秆中添加尿素+微生物制剂处理（SNW），其真菌和细菌丰度（OTU）、多样性指数（Shannon）和丰富度指数（Chao）较单独秸秆处理（S）显著增加。不同堆肥阶段，细菌和真菌门、属两级优势菌的相对丰度存在差异。在高温阶段，厚壁菌门和变形菌门的丰度依次为SNW>SN>S。葡萄球菌属、芽孢杆菌属和热裂菌属在中温阶段的丰度顺序相同。子囊菌占真菌总序列的92%以上。随着堆肥过程的进程，子囊菌的丰度逐渐降低。子囊菌在高温期的丰度顺序为S>SN>SNW。曲霉属的丰度占真菌总丰度的4-59%，并在前两个采样周期内丰度增加。曲霉丰度大小顺序为SNW>SN>S。此外，主成分分析（PCA）表明，稻草和稻草+尿素处理的群落组成相似，中温期（第1天）S、SN和SNW处理的细菌群落与其他3个阶段（分别在第5、11和19天）观察到的细菌群落不同，而真菌群落在堆肥过程只表现出轻微的变化。典型相关分析（CCA）和冗余分析（RDA）表明，总碳（TC）、NO₃-N（NN）、电导率（EC）和pH值与群落组成高度相关。因此，本研究表明添加剂有助于农作物秸秆堆肥腐熟，并且有利于堆肥品质的改善。

萝卜是一种重要的十字花科根菜类蔬菜作物，在其有色的根中有高水平的花青素累积。MYB转录因子(TFs)在植物发育和花青素代谢中起着重要作用，并且PAP1/2能促进花青素生物合成基因的表达。本研究在萝卜基因组中共鉴定出187个RsMYB基因，并将其分为32个亚家族；其中159个RsMYB基因被定位在9条染色体上。在4个不同颜色的萝卜品种肉质根发育阶段，14个RsMYB基因表现出差异的表达模式。一些RsMYB基因在成熟期有色根组织中高表达，这些基因包括RsMYB41，RsMYB117以及与PAP1/2同源的RsMYB132在‘NAU-YH’的红色根皮中高表达，RsMYB65和RsMYB159基因在‘NAU-YZH’的紫色根皮中高表达，表明这些RsMYB基因可能促进萝卜肉质根花青素积累。研究结果为进一步研究萝卜RsMYB基因功能特性提供有价值的信息，并有助于阐明萝卜花青素生物合成的分子机制。

Received  6 June, 2017    Accepted  13 October, 2017

Blast resistance and grain quality are major problems in hybrid rice production in China.  In this study, two resistance (R) genes, Pi46 and Pita, along with the gene Wx^b, which mainly affects rice endosperm amylose content (AC), were introgressed into an elite indica restoring line, R8166, which has little blast resistance and poor grain quality through marker-assisted selection (MAS).  Eight improved lines were found to have recurrent genome recovery ratios ranging from 88.68 to 96.23%.  Two improved lines, R163 and R167, were selected for subsequent studies.  R167, which has the highest recovery ratio (96.23%), showed no significant differences in multiple agronomic traits.  In contrast, R163 with the lowest recovery ratio (88.68%) exhibited significant differences in heading date and yield per plant compared with the recurrent parent.  At two developmental stages, R163 and R167 had greatly enhanced resistance to blast over the recurrent parent.  Similar trends were also observed for agronomic traits and blast resistance in R163- and R167-derived hybrids when compared with the counterparts from R8166.  In addition, R163, R167, and their derived hybrids significantly improved the grain quality traits, including amylose content (AC), gel consistency (GC), chalky grain rate (CGR), and degree of endosperm chalkiness (DEC).  It confirmed the success of efficiently developing elite restoring lines using MAS in this study.

    Homeobox 1 in Malus×domestica (MdHB-1) is a transcription factor that belongs to homeodomain-leucine zipper I (HD-Zip I) protein subfamily. According to previous reports, MdHB-1 could regulate ethylene synthesis by binding with the MdACO1 promoter, but other functions of MdHB-1 are still unknown. To reveal more clues concerning the characters of the MdHB-1 gene promoter and the functions of MdHB-1, the promoter region of MdHB-1 was cloned from the Royal Gala apple genome and recombined with the β-glucuronidase (GUS) gene in this study. This research was conducted in Nicotiana tabacum and supported by Agrobacterium-mediated transient transformation and bioinformatics analysis. Deletion analysis of the MdHB-1 promoter showed that the GUS gene could be activated by serially deleted promoters, and the activity promoted by 680 nucleotides (nt) was the lowest. The region, which is 266 nt upstream of the initiation code (ATG), was effective for GUS expression. Meanwhile, the activity of the MdHB-1 promoter (-1 057 nt), which was stronger than MdHB-1 promoter (-1 057 to -266 nt) and lack the 5´-untranslated region (5´-UTR), showed that 5´-UTR may have a positive effect on gene transcription. After the sequence analysis, the cis-acting elements that respond to hormones and environmental stresses were identified in the promoter region. The MdHB-1 promoter (1 057 nt) activity in Nicotiana tabacum was positively induced by ethrel and darkness, and it was suppressed by gibberellic acid (GA), whereas abscisic acid (ABA), salicylic acid (SA), wounding, and Pseudomonas syringae pv. tomato (DC3000) treatments revealed a slight auxo-action. These results reveal that the MdHB-1 promoter receive internal or external signals, and MdHB-1 may refer to many biological activities in apple, such as its stress response, development, and ripening.

    With increased cultivation of transgenic Bacillus thuringiensis (Bt) cotton in the saline alkaline soil of China, assessments of transgenic crop biosafety have focused on the effects of soil salinity on rhizosphere microbes and Bt protein residues. In 2013 and 2014, investigations were conducted on the rhizosphere microbial biomass, soil enzyme activities and Bt protein contents of the soil under transgenic Bt cotton (variety GK19) and its parental non-transgenic cotton (Simian 3) cultivated at various salinity levels (1.15, 6.00 and 11.46 dS m⁻¹). Under soil salinity stress, trace amounts of Bt proteins were observed in the Bt cotton GK19 rhizosphere soil, although the protein content increased with cotton growth and increased soil salinity levels. The populations of slight halophilic bacteria, phosphate solubilizing bacteria, ammonifying bacteria, nitrifying bacteria and denitrifying bacteria decreased with increased soil salinity in the Bt and non-Bt cotton rhizosphere soil, and the microbial biomass carbon, microbial respiration and soil catalase, urease and alkaline phosphatase activity also decreased. Correlation analyses showed that the increased Bt protein content in the Bt cotton rhizosphere soil may have been caused by the slower decomposition of soil microorganisms, which suggests that salinity was the main factor influencing the relevant activities of the soil microorganisms and indicates that Bt proteins had no clear adverse effects on the soil microorganisms. The results of this study may provide a theoretical basis for risk assessments of genetically modified cotton in saline alkaline soil.

    The structural characteristics of protein body accumulation in different endosperm regions of hard wheat cultivar (XM33) and soft wheat cultivar (NM13) under drought stress were investigated. Drought stress treatment was implemented from plant regreening to the caryopsis mature stage. Microscope images of endosperm cells were obtained using resin semi-thin slice technology to observe the distribution and relative area of protein body (PB). Compared with NM13, relative PB area of XM33 was significantly higher in sub-aleurone endosperm region. The amount of accumulation, including the size and relative area of PB, in two wheat cultivars was higher in sub-aleurone region than that in central region at 18 days post anthesis (DPA). Drought stress significantly enhanced the sizes and relative areas of PBs in the dorsal and abdominal endosperms in two wheat cultivars. Particularly for dorsal endosperm, drought stress enhanced the relative PB area at 18 DPA and NM13 (5.0% vs. 6.73%) showed less enhancement than XM33 (5.49% vs. 8.96%). However, NM13 (9.58% vs. 12.02%) showed greater enhancement than XM33 (10.25% vs. 11.7%) at 28 DPA. The protein content in the dorsal and abdominal endosperms of the two wheat cultivars decreased at 12 DPA and then increased until 38 DPA. Drought stress significantly increased the protein contents in the two main regions. From 12 to 38 DPA, the amount of PB accumulation and the protein content were higher in XM33 than those in NM13. The results revealed that PB distribution varied in different endosperm tissues and that the amount of PB accumulation was remarkably augmented by drought stress.

    Utilization of R (resistance) genes to develop resistant cultivars is an effective strategy to combat against rice blast disease. In this study, R genes Pi46 and Pita in a resistant accession H4 were introgressed into an elite restorer line Hang-Hui-179 (HH179) using the marker-assisted backcross breeding (MABB) procedure. As a result, three improved lines (e.g., R1791 carrying Pi46 alone, R1792 carrying Pita alone and R1793 carrying both Pi46 and Pita) were developed. The three improved lines had significant genetic similarities with the recurrent parent HH179. Thus, they and HH179 could be recognized as near isogenic lines (NILs). The resistance spectrum of the three improved lines, which was tested at seedling stage, reached 91.1, 64.7 and 97.1%, respectively. This was markedly broader than that of HH179 (23.5%). Interestingly, R1793 showed resistance to panicle blast but neither R1791 nor R1792 exhibited resistance at two natural blast nurseries. The results implied that the stacking of Pi46 and Pita resulted in enhanced resistance, which was unachievable by either R gene alone. Further comparison indicated that the three improved lines were similar to HH179 in multiple agronomic traits; including plant height, tillers per plant, panicle length, spikelet fertility, and 1 000-grain weight. Thus, the three improved lines with different R genes can be used as new sources of resistance for developing variety. There is a complementary effect between the two R genes Pi46 and Pita.

    The nuclear-encoded light-harvesting chlorophyll a/b-binding proteins (LHCPs) are specifically translocated from the stroma into the thylakoid membrane through the chloroplast signal recognition particle (cpSRP) pathway. The cpSRP is composed of a cpSRP43 protein and a cpSRP54 protein, and it forms a soluble transit complex with LHCP in the chloroplast stroma. Here, we identified the YGL9 gene that is predicted to encode the probable rice cpSRP43 protein from a rice yellow-green leaf mutant. A phylogenetic tree showed that an important conserved protein family, cpSRP43, is present in almost all green photosynthetic organisms such as higher plants and green algae. Sequence analysis showed that YGL9 comprises a chloroplast transit peptide, three chromodomains and four ankyrin repeats, and the chromodomains and ankyrin repeats are probably involved in protein-protein interactions. Subcellular localization showed that YGL9 is localized in the chloroplast. Expression pattern analysis indicated that YGL9 is mainly expressed in green leaf sheaths and leaves. Quantitative real-time PCR analysis showed that the expression levels of genes associated with pigment metabolism, chloroplast development and photosynthesis were distinctly affected in the ygl9 mutant. These results indicated that YGL9 is possibly involved in pigment metabolism, chloroplast development and photosynthesis in rice.

Myrosinase is a defense-related enzyme and is capable of hydrolyzing glucosinolates into a variety of compounds, some of which are toxic to pathogens and herbivores. Many studies revealed that a number of important vegetables or oil crops contain the myrosinase-glucosinolate system. However, the related promoter and genomic DNA sequences as well as expression profiles of myrosinase gene remain largely unexplored in radish (Raphanus sativus). In this study, the 2 798 bp genomic DNA sequence, designated as RsMyr2, was isolated and analyzed in radish. The RsMyr2 consisting of 12 exons and 11 introns reflected the common gene structure of myrosinases. Using the genomic DNA walking approach, the 5´-flanking region upstream of RsMyr2 with length of 1 711 bp was successfully isolated. PLACE and PlantCARE analyses revealed that this upstream region could be the promoter of RsMyr2, which contained several basic cis-regulatory elements including TATA-box, CAAT-box and regulatory motifs responsive to defense and stresses. Furthermore, recombinant pET-RsMyr2 protein separated by SDS-PAGE was identified as myrosinase with mass spectrometry. Real-time PCR analysis showed differential expression profiles of RsMyr2 in leaf, stem and root at different developmental stages (e.g., higher expression in leaf at cotyledon stage and lower in flesh root at mature stage). Additionally, the RsMyr2 gene exhibited up-regulated expression when treated with abscisic acid (ABA), methyl jasmonate (MeJA) and hydrogen peroxide (H2O2), whereas it was down-regulated by wounding (WO) treatment. The findings indicated that the expression of RsMyr2 gene was differentially regulated by these stress treatments. These results could provide new insight into elucidating the molecular characterization and biological function of myrosinase in radish.

Bone marrow mesenchymal stem cells (BMSCs) could differentiate into various cell types including adipocytes and myocytes, which had important scientific significance not only in the field of tissue regeneration, but also in the field of agricultural science. In an attempt to exhibit the characterization and differentiation into adipocytes and myocytes of porcine BMSCs, we isolated and purified porcine BMSCs by red blood cell lysis method and percoll gradient centrifugation. The purified cells presented a stretched fibroblast-like phenotype when adhered to the culture plate. The results of flow cytometry analysis and immunofluorescence staining demonstrated that the isolated cells were positive for mesenchymal surface markers CD29, CD44 and negative for hematopoietic markers CD45 and the adhesion molecules CD31. Cells were induced to differentiate into adipocytes with adipogenic medium containing insulin, dexamethasone, oleate and octanoate. Oil Red O staining demonstrated that the porcine BMSCs successfully differentiated to adipocytes. Moreover, the findings of real-time PCR and Western blotting indicated that the induced cells expressed adipogenic marker genes (PPAR-γ, C/EBP-α, perilipin, aP2) mRNA or proteins (PPAR-γ, perilipin, aP2). On the other hand, porcine BMSCs were induced into myoctyes with myogenic medium supplemented with 5-azacytidine, basic fibroblast growth factor, chick embryo extract and horse serum. Morphological observation by hochest 33342 staining showed that the induced cells presented as multi-nucleus muscular tube structure. And myogenic marker genes (Myf5, desmin) mRNA or proteins (Myf5, MyoD, myogenin, desmin) were found in the induced cells. In addition, the results of immunofluorescence staining revealed that myogenic marker (Myf5, MyoD, myogenin, desmin, S-MyHC) proteins was positive in the induced cells. Above all, these results suggested that the isolated porcine BMSCs were not only consistent with the characterization of mesenchymal stem cells, but also exhibited the multipotential capacity to form adipocytes and myocytes, which provided the basis to investigate the regulation mechanism involved in the selective differentiation of porcine BMSCs.

Sewage sludge amendment (SSA) is an alternative waste disposal technique and a potential way to increase fertility of mudflats for crop growth. The present study aimed to assess the suitability of SSA by assessing the nitrogen (N) and phosphorous (P) uptakes, heavy metal accumulation, growth, biomass, and yield response of ryegrass (Lolium perenne L.) at 0, 30, 75, 150, and 300 t ha-1 SSA rates at various growth stages. The results showed that the highest biomass of ryegrass at seedling and vegetative stages were at 300 and 150 t ha-1 SSA rate, respectively. The increments of ryegrass yield at reproductive stage at 30, 75, 150, and 300 t ha-1 SSA rates were 98.0, 122.6, 88.1, and 61.2%, compared to unamended soil. N and P concentrations in ryegrass increased with increasing SSA rates at all stages except N and P in roots dropped significantly at 300 t ha-1 rate at vegetative stage. The metal concentration for Mn, Cu, Zn, Ni, Cd, Cr, and Pb in shoot of ryegrass at 300 t ha-1 SSA rate increased by 0.63-, 2.34-, 15.02-, 0.97-, 10.00-, 0.01- and 1.13-fold, respectively, compared to unamended soil. However, heavy metal concentrations in shoot of ryegrass were lower than the standard for forage products in China. The study suggested that sewage sludge amendment in mudflat soils might be feasible. However, the impacts of sludge application on edible crop plants and soil environment need further investigations.

Regulator of G protein signaling proteins (RGS) accelerate the rate of GTP hydrolysis by Gα proteins, thus acting as negative regulators of G-protein signaling. Studies on Arabidopsis and soybean have proven that RGS proteins are physiologically important in plants and contribute to the signaling pathways regulated by different stimuli. Brassica napus is an important agriculturally relevant plant, the wildly planted oilseed rape in the world, which possesses an identified Gα, Gβ and Gγ subunits. In the present study, we identified and characterized a Brassica napus RGS gene, BnRGS1, which contained an open reading frame of 1 380 bp encoding a putative 52.6 kDa polypeptide of 459 amino acids, within seven putative transmembrane domains in the N-terminal and RGS box in the C-terminal. BnRGS1 is located on the membrane in onion epidermal cells and tobacco leaves, and interacts with BnGA1 in the mating-based split-ubiquitin system. The expression levels of BnRGS1 were quite different in different tissues and developmental stages, and induced by abscisic acid (ABA) and indole-3-acetic acid (IAA). The effects of gibberellin (GA3) and brassinolide (BR) on the expression of BnRGS1 were irregular under the concentrations tested. Moreover, the transcript level of BnRGS1 was also induced by polyethylene glycol (PEG), whereas remained little changed by 200 mmol L-1 NaCl. These results suggested that the BnRGS1 may be involved in B. napus response to plant hormone signaling and abiotic stresses.

Intramuscular fat (IMF) content is considered to be a key factor that affects the marbling, tenderness, juiciness and flavor of pork. To investigate the effects of myristic acid (MA) on the differentiation of porcine intramuscular adipocytes, cells were isolated from longissimus dorsi muscle (LDM) and treated with 0, 10, 50 or 100 μmol L-1 MA. The results showed that MA significantly promotes the differentiation of intramuscular adipocytes in a dose-dependent manner. MA also led to a parallel increase in the expression of peroxisome proliferator activated receptor-γ (PPARγ) and adipose-related genes, such as glucose transporter 1 (GLUT1), lipoprotein lipase (LPL), adipocyte fatty acid binding protein 4 (FABP4/aP2), fatty acid translocase (FAT), acetyl-CoA carboxylase α (ACCα), adipose triglyceride lipase (ATGL) and fatty acid synthase (FASN). However, no significant effects of MA were observed on the expression of CAAT enhancer binding protein-α (C/EBPα) or hormone sensitive lipase (HSL). The expression of pyruvate dehydrogenase kinase 4 (PDK4) was increased by MA during the early stages of differentiation (day 1-3). In addition, MA also increased the absolute content of C14 (P<0.001) and saturated fatty acids (SFA) (P<0.05) to varying degrees, but no effects were observed on other fatty acids. These results suggest that MA might be able to enhance the IMF content of pork and increase the accumulation of myristic and myristoleic acid in muscle, which might have beneficial implications for human health.

Different amino acids have been shown to affect feed intake when injected directly into the central nervous system of birds. In the present study, we investigated the effects of L-glutamine and L-alanine on feed intake and the mRNA expression levels of hypothalamic neuropeptides involved in feed intake regulation in broiler chicks. L-Glutamine or Lalanine was intra-cerebroventricularly (ICV) administered to 4-d-old broiler chicks and the feed intake were recorded at various time points. Quantitative PCR was performed to determine the hypothalamic mRNA expression levels of neuropeptide Y (NPY), agouti related protein (AgRP), pro-opiomelanocortin (POMC), melanocortin receptor 4 (MC4R) and corticotropin releasing factor (CRF). Our results showed that ICV administration of L-glutamine (0.55 or 5.5 μmol) significantly increased feed intake up to 2 h post-administration period and the hypothalamic NPY mRNA expression levels, while it markedly decreased hypothalamic POMC and CRF mRNA expression levels. In contrast, ICV administration of L-alanine (4 μmol) significantly decreased feed intake for the first 0.5 h post-administration period, and reduced the hypothalamic AgRP mRNA expression levels, while it remarkablely enhanced the mRNA expression levels of MC4R and CRF. These findings suggested that L-glutamine and L-alanine could act within the hypothalamus to influence feed intake in broiler chicks, and that both orexigenic and anorexigenic neuropeptide genes might contribute directly to these effects.