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。

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。

    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.

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.

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.

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.