禽偏肺病毒（Avian metapneumovirus，aMPV）为副黏病毒科肺病毒亚科偏肺病毒属家族的成员，其主要引起火鸡鼻气管炎（Turkey rhinotracheitis，TRT）和肉鸡肿头综合征（Swollen head syndrome，SHS）。目前，B亚型aMPV是我国鸡群中主要的优势流行毒株，由于缺乏aMPV反向遗传操作技术，有关该病毒的致病与致弱机制及是否可作为病毒载体的研究相对较少。为此，本研究将B亚型aMPV弱毒株LN16-A株全长分为5个cDNA片段进行扩增，并在基因组的3′端和5′端分别添加了T7启动子和丁型肝炎核酶序列，构建了全长cDNA感染性克隆质粒pOKLN16-A。将pOKLN16-A与4个辅助质粒pCAGGS-N、pCAGGS-P、pCAGGS-M21和pCAGGS-L共转染至表达T7 RNA聚合酶的BSR-T7/5细胞中，拯救出了病毒，成功建立了基于T7 RNA聚合酶的aMPV反向遗传操作系统。为进一步探究aMPV作为疫苗载体的潜力，利用反向遗传操作技术将增强型绿色荧光蛋白基因（Enhanced green fluorescent protein，EGFP）插入aMPV基因组的不同位点，并比较了其表达水平，结果显示，EGFP在B亚型aMPV的G和L基因之间的表达水平显著高于另外两个插入位点（前导基因和N基因之间及替换SH基因），因此确定外源基因表达的最佳插入位点为G和L基因之间。为进一步验证该插入位点的可用性，以鸡传染性法氏囊病病毒超强毒株（Very virulent infection bursal disease virus，vvIBDV）为模式病毒，利用反向遗传操作技术在该位点插入了其保护性抗原VP2基因，成功拯救了稳定表达vvIBDV VP2蛋白的重组B亚型aMPV，命名为rLN16A-vvVP2株。将rLN16A-vvVP2株以5000 TCID₅₀/只的剂量免疫SPF鸡，免疫3周后使用B亚型aMPV LN16-F4强毒株及vvIBDV HLJ0504强毒株进行攻毒。结果显示，单次免疫rLN16A-vvVP2株可同时诱导机体产生针对B亚型aMPV及vvIBDV两种病毒的中和抗体，免疫3周后的中和抗体效价分别为8.7及8.2 log2。此外，单次免疫rLN16A-vvVP2株对B亚型aMPV强毒及vvIBDV强毒的攻毒保护率均为100%，并能有效预防vvIBDV攻击后引起的法氏囊损伤。本研究成功建立了B亚型aMPV的反向遗传操作系统并鉴定了外源基因表达的最佳插入位点，首次评价了B亚型aMPV作为疫苗载体的潜力，研究结果为进一步研究aMPV的致病机制和安全有效的新型载体疫苗提供了技术支撑。

次生代谢物质与植物的营养品质和保健功能密切相关。本研究首次收集了国内23个野生石榴和27个栽培石榴的成熟果实，比较分析了不同种质石榴果皮和果汁中黄酮和鞣质的含量差异，并采用液相色谱-电喷雾串联质谱法（LC-ESI-MS/MS）法测定了果汁高黄酮种质‘则拉4’果皮（ZLP）和果汁（ZLZ）中的次生代谢物质。方差分析（P<0.05）表明，不同石榴种质间的黄酮和鞣质含量存在显著差异。Pearson相关分析表明，影响石榴黄酮和鞣质含量的主要环境因子是纬度和海拔。本研究在‘则拉4’果皮和果汁中共鉴定出279种次生代谢物质，其中227种次生代谢物首次在石榴中发现。通过正交偏最小二乘判别分析法，在ZLP和ZLZ中筛选出了90种差异代谢物。本研究还筛选了8种特异性种质资源（果皮高黄酮，‘军拥3’；果皮低黄酮，‘胭脂红’；果汁高黄酮，‘则拉4’；果汁低黄酮，‘豫大籽’，果皮高鞣质，‘军拥4’，果皮低鞣质，‘安巴1’，果汁高鞣质，‘叶巴1’，果汁低鞣质，‘白花玉石籽’。本研究结果可为我国野生石榴资源的开发利用和石榴育种提供参考。

CCCH（C3H）锌指转录因子(Zinc finger, Znf)是一种新型的Znf基因，其通过绑定于基因的mRNA上调控基因的表达，并在植物生长发育和抗非生物胁迫中发挥重要作用。龙眼是一种具有重要的经济价值热带、亚热带果树。然而，龙眼C3H的基因组信息及功能仍不清楚。本研究对龙眼C3H (DlC3H)基因家族进行了全基因组鉴定及表达分析。从龙眼基因组数据库中共鉴定出分布于3个进化枝中的49个DlC3H基因，并对其基因结构、motif组成、系统发育树和潜在功能等方面进行了基因特征分析。可变剪接事件(alternative splicing, AS)分析表明，DlC3H基因AS事件可能参与龙眼非胚性培养物向胚性培养物的转换。启动子分析显示，大多数DlC3H基因包含与激素和胁迫响应相关的顺式作用元件。实时荧光定量PCR（qRT-PCR）分析显示，26个具有MeJA和ABA响应顺式作用元件的DlC3Hs，在ABA、MeJA及其内源性抑制剂的作用下表现出不同的表达模式，提示DlC3Hs可能参与了ABA和MeJA信号通路。同时，在龙眼非胚性愈伤组织和3个胚胎培养阶段的表达谱显示，17个DlC3Hs成员中只有5个DlC3Hs的表达模式与转录组数据FPKM相同；DlC3H07/14/16/36/49在胚性愈伤组织中表达较高，而DlC3H04/38在球形胚中表达较高，说明它们在胚胎发育中具有不同的作用。通过改良RLM-RACE验证了DlC3H01/03/05/11/19/39被sRNAs调控。本研究首次对龙眼的C3H基因进行了系统分析，特别是C3H基因可能参与激素、胁迫反应以及体细胞胚胎形态建成有关。本试验的初步结果以期为进一步研究龙眼C3H基因家族的特征和功能提供线索。

本研究为了量化不同“C饥饿”水平下玉米叶片C固定及其分配，并分析其与植株适应性生长之间的关系，利用室内液体培养玉米植株，在6叶展期进行连续三天的延长黑暗（ED）处理。结果表明，ED处理显著降低了植株生长和叶片叶绿素水平，但单位叶片CO₂气体交换速率没有明显变化。由于延长黑暗缩短了光合时长，降低了日光合同化产物积累，成熟叶片中的淀粉和可溶性碳水化合物（TSC）日累积量也呈下降趋势。然而，ED处理下叶片淀粉和TSC积累量占日同化C总量的比例却有所增加。这些“暂储性”C大部分以TSC形式存在，并且主要为增加的夜间呼吸消耗所用而非转运至库器官。另一方面，随着时间的推移，不同处理叶片中的“暂储性”C累积量及其占日同化C总量的比例均呈下降趋势，这主要是由于叶片淀粉合成下降所致。叶片中的腺苷二磷酸葡萄糖焦磷酸化酶和可溶性淀粉合酶活性随时间推移显著下降。因此，我们认为淀粉和TSC都参与了C突然短缺时植株生长和C供应之间的协调，但可能存在不同的作用方式。在突然的“C饥饿”情况下，植株将更高比例的同化产物留存在叶片中，以维持叶片功能。同时，成熟叶片中的“暂储性”C量及其占日同化C总量的比例随时间推移不断下降，以满足库器官的持续性生长需求。

Nsf1(Nutrient and stress factor 1)是典型的C2H2型锌指蛋白，酿酒酵母中Nsf1在非发酵碳源或者盐胁迫的条件下才会表达。进化树分析发现该基因在不同物种间比较保守，然而，Nsf1的功能在丝状真菌中研究得并不是很透彻。为了探索FgNsf1在小麦赤霉病的病原菌禾谷镰孢菌中的功能，我们构建了FgNSF1基因敲除体（ΔFgNsf1）和包含GFP标记的回复体（ΔFgNsf1-C），亚细胞定位表明FgNsf1蛋白集中于细胞核。进一步研究发现，与野生菌株PH-1和回复体ΔFgNsf1-C 相比，敲除体ΔFgNsf1的菌丝体生长速率明显减慢，分生孢子产量及萌发率显著下降，且有畸形孢子产生，子囊壳产量也显著降低。但是红色镰刀菌素和黄色镰刀菌素的产量明显增加，为了验证这一结果，我们利用实时荧光定量PCR技术检测了相关基因(AurJ, AurF, AurO, AurR2)的表达量，研究结果发现，相关基因的表达量都显著上调。此外，使用不同浓度的NaCl处理时，野生菌株PH-1中FgNSF1基因的表达量均上调，而在使用不可发酵碳源乙醇、甘油或醋酸盐作为唯一碳源时，FgNSF1基因的表达量都显著下调。另外我们发现敲除体对渗透，细胞壁，氧化和部分金属离子等胁迫因子的敏感性显著下降，只对0.2M镁离子胁迫的敏感性显著提高。药敏性实验发现，敲除体对咯菌腈和抑菌脲的抗药性明显增强，而对戊唑醇和多菌灵的敏感性显著提高。随后，我们在小麦胚芽鞘和麦穗上进行了致病力实验，结果发现ΔFgNsf1的致病力显著减弱，在产毒培养基中DON（脱氧雪腐镰刀烯醇）产量也显著下降，以及DON毒素合成相关基因TRI5和TRI6的基因表达量也显著下调。结论：FgNSF1在禾谷镰孢菌的生长发育，有性和无性生殖，应对外界胁迫，产毒和致病的过程中扮演着重要的角色。创新性：我们首次系统地报道了FgNSF1在禾谷镰孢菌中的功能。

 

细胞自噬通过维持细胞内物质与能量的动态平衡，进而调控很多发育过程。已知延伸复合物蛋白Elp3具有多种功能并参与调控自噬，但其在稻瘟病菌中的功能仍不清楚。为此，构建了稻瘟病菌Elp3编码基因PoELP3（MGG_05481）的敲除突变体，对其功能进行研究。表型分析结果显示，PoELP3基因的敲除导致稻瘟病菌菌丝生长受抑制，产孢量下降，对细胞壁胁迫剂和盐胁迫剂的敏感性增强，附着胞膨压和致病性显著下降。这些结果表明稻瘟病菌PoElp3在生长发育、胁迫响应和致病过程中均具有重要作用。亚细胞定位结果表明GFP-PoElp3融合蛋白定位于细胞核和细胞质中。为检测稻瘟病菌Elp3在细胞自噬中的作用，将自噬标记GFP-PoAtg8分别导入野生型和突变体中，并通过计算总蛋白中游离GFP占GFP-PoAtg8和游离GFP总量的比例，评估野生型和突变体中自噬的水平。结果表明，无论是在营养充分还是营养不足的条件下，△Poelp3突变体均呈现较高的自噬水平。这可能是导致菌丝生长缓慢的原因。此外，△Poelp3突变体营养菌丝和侵染菌丝的生长均对雷帕霉素更加敏感，但是PoELP3基因的缺失并不影响雷帕霉素对TOR-信号途径下游基因的转录抑制，暗示其并不参与TOR-信号的传导。综上所述，稻瘟病菌Elp3可以通过调控自噬影响无性发育和致病性。但是PoElp3调控自噬的机制仍有待进一步研究。

Blumeria graminis f. sp. tritici, the pathogen that causes wheat powdery mildew, is one of the most important diseases affecting wheat production in China, and the oversummering is the key stage of wheat powdery mildew epidemic.  The more oversummering regionalization of wheat powdery mildew has played an important role in disease prediction, prevention and control.  In this study, we analyzed the correlation between oversummering data of wheat powdery mildew and the meteorological factors over the past years, and determined that temperature was the key meteorological factor influencing oversummering of wheat powdery mildew.  The average temperature at which wheat powdery mildew growth was terminated (26.2°C) was used as the threshold temperature to regionalize the oversummering range of wheat powdery mildew.  This regionalization was done using the GIS ordinary kriging method combined with the Digital Elevation model (DEM) of China.  The results showed that annual probability of oversummering region based on Model 26.2 were consistent with the actual survey of the more summer wheat powdery mildew.  Wheat powdery mildew oversummering regions in China mainly cover mountainous or high-altitude areas, and these regions form a narrow north-south oversummering zone.  Oversummering regions of wheat powdery mildew is mainly concentrated in the high-altitude wheat growing areas, including northern and southern Yunnan, northwestern Guizhou, northern and southern Sichuan, northern and southern Chongqing, eastern and southern Gansu, southeastern Ningxia, northern and southern Shaanxi, central Shanxi, western Hubei, western Henan, northern and western Hebei, western Liaoning, eastern Tibet, eastern Qinghai, western Xinjiang and other regions of China.

Genomic selection has been demonstrated as a powerful technology to revolutionize animal breeding.  However, marker density and minor allele frequency can affect the predictive ability of genomic estimated breeding values (GEBVs).  To investigate the impact of marker density and minor allele frequency on predictive ability, we estimated GEBVs by constructing the different subsets of single nucleotide polymorphisms (SNPs) based on varying markers densities and minor allele frequency (MAF) for average daily gain (ADG), live weight (LW) and carcass weight (CW) in 1 059 Chinese Simmental beef cattle.  Two strategies were proposed for SNP selection to construct different marker densities: 1) select evenly-spaced SNPs (Strategy 1), and 2) select SNPs with large effects estimated from BayesB (Strategy 2).  Furthermore, predictive ability was assessed in terms of the correlation between predicted genomic values and corrected phenotypes from 10-fold cross-validation.  Predictive ability for ADG, LW and CW using autosomal SNPs were 0.13±0.002, 0.21±0.003 and 0.25±0.003, respectively.  In our study, the predictive ability increased dramatically as more SNPs were included in analysis until 200K for Strategy 1.  Under Strategy 2, we found the predictive ability slightly increased when marker densities increased from 5K to 20K, which indicated the predictive ability of 20K (3% of 770K) SNPs with large effects was equal to the predictive ability of using all SNPs.  For different MAF bins, we obtained the highest predictive ability for three traits with MAF bin 0.01–0.1.  Our result suggested that designing a low-density chip by selecting low frequency markers with large SNP effects sizes should be helpful for commercial application in Chinese Simmental cattle.

The aim of this study was to evaluate the growth of rape (Brassica napus L.) seedlings under different light intensities to select appropriate conditions for cultivation in an indoor system.  Seedlings were grown under different light intensities of red and blue light provided by light-emitting diodes (LEDs) and their self-adjustment ability and changes in leaf microstructure were evaluated.  Light was supplied by red LEDs with peak wavelengths of 630 (R₁) and 660 nm (R₂) and by blue LEDs (B) with a peak wavelength of 445 nm (the light intensity ratio of R₁:R₂:B was 3:3:2), at intensities of 400 (R₁R₂B400), 300 (R₁R₂B300), and 200 μmol m^–2 s^–1 (R₁R₂B200).  Natural solar light served as the control (C).  Plant height, stem diameter, root length, leaf area, and dry weight of rape seedlings gradually increased with increasing light intensity.  The seedlings in the R₁R₂B400 treatment grew more vigorously, while those in the R₁R₂B200 treatment were weaker.  The photosynthetic pigment contents did not differ significantly between the R₁R₂B400 treatment and C, but were significantly lower in the R₁R₂B300 and R₁R₂B200 treatments.  The highest intercellular CO₂ concentration, stomatal conductance, and transpiration rate were in the R₁R₂B300 treatment.  The highest photosynthetic rate was in the R₁R₂B400 treatment, and was related to more compact leaves, thicker and tidier palisade and spongy tissues, and well-developed chloroplasts.  In contrast, the seedlings in the R₁R₂B200 treatment had disordered mesophyll cells, round chloroplasts, and fractured and fuzzy grana lamellae, all of which inhibited plant growth.  In conclusion, the seedlings in the R₁R₂B400 treatment had well-developed leaves, which favored photosynthesis.  Compared with the light intensities below 300 μmol m^–2 s^–1, the light intensity of 400 μmol m^–2 s^–1 provided by a combination of red and blue LEDs was beneficial for cultivating strong and healthy rape seedlings in an artificial system.  

    The antioxidant of reduced glutathione (GSH) is the most abundant thiol in cells for the maintenance of the intracellular redox balance. The study aimed to assay the effect of sperm treatment with GSH before incubation with oocytes on the development potential of embryos obtained by in vitro fertilization (IVF). Also the mitochondrial membrane potential (ΔΨm), plasma membrane integrity (viability), DNA fragmentation, reactive oxygen species (ROS) content, superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activities, methane dicarboxylic aldehyde (MDA) level as indices of lipid peroxidation in sex-sorted and unsorted sperm from three bulls were investigated using flow cytometry and enzyme-labeled instrument individually. The results showed that 2 mmol L^–1 GSH increased significantly the cleavage rate (86.68% vs. 82.78%), 4- to 8-cell rate (82.30% vs. 73.43%) and blastocyst rate (43.15% vs. 35.24%) of IVF embryos compared with untreated group. Furthermore, addition of GSH increased significantly the ΔΨm and viability, decreased the ratio of DNA fragmentation in sex-sorted or unsorted semen (P<0.05), except the sex-sorted semen from bull 019. Similarly, activities of SOD, CAT and GPx were increased significantly. However, the contents of MDA were decreased significantly both in sex-sorted and unsorted semen treated with GSH (P<0.05). These results suggest that sperm pretreatment with GSH during IVF can maintain better the viability and fertility of sperm through reducing apoptosis and increasing the antioxidant capacity, which improves the IVF embryos development.

  Omega-3 polyunsaturated fatty acids (ω-3 PUFAs) are essential components required for normal cellular function and have been shown to have important therapeutic and nutritional benefits in humans. But humans or mammals cannot naturally produce ω-3 PUFAs, due to the lack of the ω-3 fatty acid desaturase gene (fat-1 gene). Previously, fat-1 gene has been cloned from Caenorhabditis elegans and transferred into mice, pigs and sheep, but not yet into beef cattle. We attempt to transfer it into beef cattle. The object of this paper is to edit the fat-1 gene from C. elegans to express more efficiently in beef cattle and verify its biological function in mice model. As a result, the fat-1 gene from C. elegans was modified by synonymous codon usage and named it Bfat-1. We have demonstrated that degree of codon bias of Bfat-1 gene was increased in beef cattle. Moreover, Bfat-1 gene could be transiently expressed in mouse liver and muscle, the ω-6/ω-3 PUFAs ratio of 18 and 20 carbon was decreased significantly in liver (P<0.05), and the ratio of 20 carbon decreased significantly in muscle 24 and 72 h after injection (P<0.05). This confirms that the Bfat-1 gene modification was successful, and the protein encoded was able to catalyze the conversion of ω-6 PUFAs to ω-3 PUFAs.

The objective of this study was to establish an efficient system of producing early monozygotic twin bovine embryos in vitro using the blastomere separation and coculture technique. In this study, early eight-cell embryos were chosen to optimize the separation method, and multi-coculture tactics were applied to improve the efficiency of this production system. Bovine embryo blastomeres (groups of at least 30 at the eight-cell stage) were separated into eight segments (to regard an eight-cell embryo as a tangerine, a blastomere as one segment) and one, two and four segments (blastomeres) were cultured respectively in microwells on the bottom of the four-well dish (Nunc, Denmark) with 400 μL of culture medium under paraffin oil. Four different types of coculture tactics (cocultured with nothing, intact embryos, bovine cumulus cells (bCCs), intact embryos & bCCs) were applied to the group of four segments (blastomeres). Finally, diameter and inner cell mass (ICM):trophectoderm (TE) cell ratio was measured as a criterion to assess the quality of the twin embryos which were derived from bovine separated blastomeres. Our results showed that rate of blastocyst formation of the four segments group was significantly greater than one or two group (P<0.05). In addition, rate of blastocyst formation was significantly increased when the four segments were cocultured with intact embryo & bCCs (P<0.05). Although the ICM, TE and total cells of blastocysts derived from separated blastomeres was less than the control group from intact embryo (P<0.05), more important quality indicator of the blastocyst diameter and ICM:TE cell ratio was similar between our experimental group and the control group (P>0.05). Thus, these results suggest that combined with intact embryos & bCCs coculture system, culturing four isolated segments (blastomeres) per microwell is an efficient system of producing early monozygotic twin bovine embryos. Furthermore, our results also indicate that the quality of blastocysts derived from separated blastomere may be similar to those derived from intact eight-cell embryos.

In order to clarify the impact posed by wheat powdery mildew (Blumeria graminis f. sp. tritici) on the yield and yield components in different epidemic seasons, field trials were conducted in three growing seasons, 2009-2010, 2010-2011 and 2011-2012, in Langfang City, Hebei Province, China. The relationships between 1000-kernel weight, crude protein content of grain and yield and disease index (DI), as well as area under disease progress curve (AUDPC) were studied. The models of the percentage of loss of 1000-kernel weight, crude protein content and yield were constructed using DI at critical point (CP) of growth stages (GS) and AUDPC in the three growing seasons, respectively. The CPs for estimating 1 000-kernel weight, crude protein content of grain and yield of wheat caused by powdery mildew were GS 11.1, GS 10.5.3 and GS 10.5.3, respectively. Models based on DI at CP to estimate the percentage of loss of 1 000-kernel weight, crude protein content of grain and yield were better than models based on AUDPC. And models of the percentage of loss of 1 000-kernel weight, crude protein content and yield for 2011-2012 season were significant different from these for 2009-2010 and 2010-2011 seasons. These results indicated that besides powdery mildew, weather conditions also had influence on 1 000-kernel weight, crude protein content of grain and yield loss of wheat when powdery mildew occurred.

Wheat (Triticum aestivum L.) often experiences photoinhibition due to strong light during the grain filling stage. As such, increasing the tolerance of wheat to photoinhibition is very desirable in breeding efforts focused on increasing grain yields. Previous reports have suggested that PROTON GRADIENT REGULATION 5 (PGR5) plays a central role in the generation of a proton gradient across the thylakoid membrane (DpH) and in acclimation to high light intensity conditions. Three PGR5 homoeologues were isolated from wheat, and mapped onto chromosomes 7A, 7B and 7D, respectively. The TaPGR5s shared highly similar genomic sequences and gene structures. The transcripts of TaPGR5s were found to be abundantly expressed in the flag leaves, and were transiently up-regulated by treatment with high light. High light treatment inhibited the net photosynthetic rate (Pn) and the maximal quantum yield of photosystem II (Fv/Fm). Further, these inhibitions were more evident in the leaves with reduced expression of TaPGR5s achieved using virus-induced gene silencing methods. Moreover, reducing TaPGR5 expression impaired the induction of non-photochemical quenching (NPQ), which caused more severe cell membrane damage and lipid peroxidation in high light. Additionally, we observed that TaPGR5s transcripts were more abundantly expressed in the wheat genotypes with higher ms-delayed light emission (ms-DLE), a value reflecting transthylakoid DpH. These results suggested that TaPGR5s play important roles in the tolerance of wheat to photoinhibition.

Phosphorus (P) is an essential element for plant growth and yield. Improving phosphorus use efficiency of crops could potentially reduce the application of chemical fertilizer and alleviate environmental damage. Soybean (Glycine max (L.) Merr.) is sensitive to phosphorus (P) in the whole life history. Soybean cultivars with different P efficiencies were used to study P uptake and dry matter accumulation under different P levels. Under low P conditions, the P contents of leaf in high P efficiency cultivars were greater than those in low P efficiency cultivars at the branching stage. The P accumulation in stems of high P efficiency cultivars and in leaves of low P efficiency cultivars increased with increasing P concentration at the branching stage. At the late podding stage, the P accumulation of seeds in high and low P efficiency cultivars were 22.5 and 26.0%, respectively; and at the mature stage were 69.8 and 74.2%, respectively. In average, the P accumulation in whole plants and each organ was improved by 24.4% in high P efficiency cultivars compared to low P efficiency cultivars. The biomass between high and low P efficiency cultivars were the same under extended P condition, while a significant difference was observed at late pod filling stage. At the pod setting stage, the biomass of high P efficiency cultivars were significant greater (17.4%) than those of low P efficiency cultivars under high P condition. Meanwhile, under optimum growth conditions, there was little difference of biomass between the two types of cultivars, however, the P agronomic efficiency and P harvest index were significant higher in high P efficiency cultivars than those in low P efficiency cultivars.

Super high-yielding soybean cultivar Liaodou 14, soybean cultivars from Ohio in the United States, and the common soybean cultivars from Liaoning Province, China, with similar geographic latitudes and identical pod-bearing habits were used as the study materials for a comparison of morphological traits and production characteristics to provide a theoretical basis for the breeding of improved super high-yielding soybean cultivars. Using a randomized block design, different soybean cultivars from the same latitude were compared under conventional and unconventional treatments for their production characteristics, including morphological traits, leaf area index (LAI), net photosynthesis rate, and dry matter accumulation. The specific characteristics of the super high-yielding soybean cultivar Liaodou 14 were analyzed. The results showed that the plant height of Liaodou 14 was significantly lower than that of the common cultivars from Liaoning, whereas the number of its main-stem nodes was higher than that of the cultivars from Ohio or Liaoning. A high pod density was observed in Liaodou 14 under conventional treatments. Under both conventional and unconventional treatments, the branch number of Liaodou 14 was markedly higher than that of the common cultivars from Liaoning, and its branch length and leaf inclination angle were significantly higher than those of common cultivars from Liaoning or Ohio. Only small changes in the leaf inclination angle were observed in Liaodou 14 treated with conventional or unconventional methods. Under each treatment, Liaodou 14 exhibited the lowest amplitude of reduction in SPAD values and net photosynthesis rates from the grain-filling to ripening stages; the cultivars from Ohio and the common cultivars from Liaoning exhibited more significant reductions. Liaodou 14 reached its peak LAI later than the other cultivars but maintained its LAI at a higher level for a longer duration. Under both conventional and unconventional treatments, Liaodou 14 produced a higher yield than the other two cultivars, with significant differences from the Ohio cultivars. In summary, super high-yielding soybean cultivars have several main features: suitable plant height, high pod density, good leaf structure with strong functionality, and slow leaf senescence at the late reproductive stage, which is conducive to the accumulation of dry matter and improved yield.

Wheat dwarf bunt, caused by Tilletia controversa Kühn (TCK), is an important quarantine wheat disease throughout the world. Based on published research results of the biology and the epidemiology of the disease, the main factors including temperature, humidity, snow cover, and their parameters relating to teliospore germination, infection and epidemics of TCK were determined. The geophytopathological models for the risk analysis of wheat dwarf bunt establishment were modified. Fifty-year meteorologic data from about 500 weather stations in China were used to calculate the probabilities of TCK establishment in different geographic sites with the models. The map that displays the establishment risk of TCK in winter wheat growing regions in China was generated by using geographical information system (GIS). The zones showing high, moderate, low, and very low, including no risk, of TCK establishment accounted for 27.33, 27.69, 38.12, and 6.86% of total winter wheat growing areas in China, respectively. These results will provide useful information to formulate quarantine regulations and wheat importation policy in China.

The ultra-structure of mother and outer daughter scales of Lilium Oriental hybrid Sorbonne were studied using transmission electron microscope to examine the sub-cellular localization of starch and lipid droplets during growth and development from shoot emergence to senescence. The contents of starch granules and lipid droplets in the cell of the mother scales decreased significantly from shoot emergence to anthesis, indicating that these scales served as a source for growth and development. After flowering, the number of starch granules and lipid droplets increased dramatically, and finally the cells were filled with the above molecules indicating that the bulb becomes a major sink during bulb enlargement. Ultrastructure observation also showed that symplastic pathway is the main pathway in cells in the exchange and transportation of material during bulb development. The activity of β-amylase, one of the key enzymes catalyzing starch breakdown, showed a similar trend. The enzyme sub-cellular localization via immune-gold electron-microscopy showed that β- amylase was predominantly located together with starch granules, while the gold particles were scarcely found in other sub-cellular compartments. The result suggested that this enzyme is compartmented together with its functional substrate supporting its function in catalyzing starch breakdown in living plant cells.

EST-derived SSR marker has been developed in many species, but few methods of high efficiency have been reported for the exploitation of EST-SSR markers. Thus, a high efficiency method for mining millions of redundant EST data is needed. A modified method for the EST-SSR development with high efficiency was established based on the redundant EST data of soybean in this study. The method achieved its function through classifying ESTs according to the same SSR motif and detected candidate loci with redundant sequences. In this study, a total of 80 polymorphic EST-SSR markers of soybean were developed, 50 of them were exploited by this modified method which proved the higher speed and efficiency of this method. All the 80 polymorphic EST-SSRs were mapped on soybean physical map through in silico mapping and 15 markers were integrated on a genetic map constructed in previous study. A software named hpSSR (high polymorphic SSR) was programmed based on the concept of the up-built method for EST-SSR development. This method is not only pragmatic for EST-SSR exploitation in soybean, but also effective for the development of the marker in other species if the redundancy EST data is available.

To our knowledge, no single study has systemically compared cryopreservation efficiencies of bovine blastocysts derived from in vitro fertilization (IVF), intracytoplasmic sperm injection (ICSI) and somatic cell nuclear transfer (SCNT) by controlled freezing and vitrification. This experiment, therefore, was designed to compare the cryopreservation of these blastocysts with controlled freezing and OPS vitrification. Adenosine-5´-triphosphate (ATP) content and reactive oxygen species (ROS) level in blastocysts were also analyzed. Firstly, for each type of blastocyst (IVF, ICSI or SCNT), significant differences were observed between the survival rates of the controlled freezing ((81.56±2.33), (68.18±4.72) or (47.89±5.83)%) and OPS vitrification groups ((92.24±4.54), (82.40±3.76) or (78.71±5.91)%; P<0.05). Secondly, for each type of blastocyst (IVF, ICSI or SCNT), ATP content was significantly decreased after controlled freezing or vitrification, and the ATP content in the controlled freezing group (0.43±0.06), (0.35±0.05) or (0.21±0.02) pmol) was significantly lower than that found in the OPS vitrification group (0.62±0.04), (0.46±0.03) or (0.30±0.01) pmol; P<0.05). Thirdly, ROS level in fresh IVF ((47.33±3.56) c.p.s (counted photons per second), ICSI ((36.51±2.58) c.p.s) or SCNT blastocysts ((26.44±1.49) c.p.s) was significantly lower than that found in the OPS vitrification group ((72.14±4.31), (58.89±3.89) or (40.11±5.73) c.p.s; P<0.05), but higher than that of the controlled freezing group (34.41±3.32), (23.13±1.26) or (15.46±2.45) c.p.s; P<0.05). The present study indicated that vitrification is more efficient in the cryopreservation of bovine blastocysts derived from IVF, ICSI or SCNT than controlled freezing. Furthermore, both vitrification and controlled freezing significantly altered the ATP content and ROS level in those blastocysts.

The aim of this study is to reveal the regulation mechanism of the effect of Semen vaccariae and Taraxacu mogono on the cell-cell adhersion molecule, E-cadherin and b-catenin on the proliferation role and secretion function of bovine mammary epithelial cells cultured in vitro. Firstly, the epithelial character of bovine mammary epithelial cells was authenticated using immunofluorescence, then the cell grow curve was observed and investigated after S. vaccariae and T. mogono treatment. On the effect of S. vaccariae and T. mogono, cell adhesion molecules E-cadherin, b-catenin and CycinD1 mRNA and protein were detected by qRT-PCR and Western blotting, respectively. The results showed that the cellular keratin 18 expressed positively and proliferated vigorously after S. vaccariae and T. mogono treament. The mRNA and protein levels of E-cadherin and CycinD1 were remarkably higher (P<0.05) in 36 h after S. vaccariae and T. mogono treatment. The cell proliferation at 36 h was increased significantly (P<0.05). In conclusion, S. vaccariae and T. mogono have a positive impact on the cell proliferation and an effect on the adhesion molecules E-cadherin, b-catenin and CycinD1 in the Wnt signaling pathway.

Mitosis of mammary epithelial cell is foundation of mammal lactation. We developed a strategy of combined application of generation of longer cDNA fragments from the serial analysis of gene expression (SAGE) tags for gene identification (GLGI) to screen and identify genes influencing lactating ability of mammary epithelial cell in dairy goat. GLGI as a new tag identification technique was brought about with SAGE. Bzw2 was found as a candidate gene related to lactation by screening Long-SAGE library of mammary gland in dairy goat. Bzw2 cDNA was synthesized by switching mechanism at 5´-end of RNA transcript (SMART) technology. The mRNA level of Bzw2 was relatively higher in early lactation than in other development stages of mammary gland. The proliferation of mammary epithelial cell was inhibited by transfecting specific shRNA of Bzw2. The mRNA levels of Stat5, Csn2 and Prlr were also down-regulated, suggesting the lactating ability of mammary epithelial cell was attenuated after Bzw2 RNAi. The reduction of mammary epithelial cell growth and lactation by Bzw2 RNAi was rescued through over-expression of Bzw2. These results revealed that Bzw2 might play an important role in lactation though the molecular mechanism was still unclear.

In previous experiment, we isolated a compound dibutyl phthalate (DBP) from Vaccaria segetalis which had galactopoieticfunction on mammary gland epithelial cells of dairy cow (DCMECs). In this experiment, we ascertained the metabolicregulation function of DBP on DCMECs. Many genes related to lactation including Stat5, AMPK, â-casein, Glut1, SREBP-1,PEPCK, and ACC were detected by real-time PCR. Furthermore, Stat5 and AMPK were detected by Western blot andimmunofluorescence co-localization, respectively. The results showed that DBP stimulates the expression of Stat5 andp-Stat5, thus activates Stat5 cell signal transduction pathway and stimulates â-casein synthesis. DBP also raises theactivities of Glut1 and AMPK to stimulate glucose uptake and glycometabolism and activates the expression of AMPKdownstream target genes PEPCK and ACC and expression of SREBP-1 to stimulate milk fat synthesis. In addition, theactivities of HK, G-6-PDH, ICDH, ATPase, and energy charges were stimulated by DBP to increase the energy metabolismlevel of DCMECs. The results showed DBP stimulates energy metabolism related to galactopoietic function in DCMECs.