狗蔷薇PaWUS基因的超量表达诱导转基因烟草根尖形成不定芽

WUSCHEL是植物干细胞命运的决定基因,在植物发育过程中WUSCHEL基因的表达决定着干细胞的形成和器官的发生.从狗蔷薇的类原球茎中克隆出其同源基因；经过对其进行蛋白质序列比对和系统发育分析,将其命名为Ra WUS基因.构建在XVE系统下的化学调控的表达载体.在该系统下,Ra WUS基因的超量表达能诱导转基因烟草根尖形成不定芽；同时显微观察表明,根部的皮层中有圆球形的分生组织干细胞团的产生.结果表明:Ra WUS基因的过量表达能够促使根的皮层中薄壁细胞转变成为分生组织干细胞而形成不定芽在根尖异位发生,说明其能够在植物的根尖中诱导茎尖分生组织干细胞的活性和发育的可塑性.     

褪黑素的添加减轻猪精原干细胞的氧化损伤

旨在研究褪黑素（melatonin,MT）对体外培养猪精原干细胞（spermatogonial stem cells,SSCs）的作用机制。本研究采集3头7日龄健康大白公猪睾丸,利用差速贴壁法获得SSCs。后经形态学观察、碱性磷酸酶染色、标记基因检测及免疫荧光染色鉴定后以SSCs作为试验材料,设置MT浓度梯度（0、50、250、500、1 000 μmol·mL^-1）组处理SSCs,每组设3个重复（n=3）,空白对照组加入0.1% DMSO处理,分别检测添加MT后猪SSCs的细胞活力、活性氧（reactive oxygen species,ROS）水平、谷胱甘肽（glutathione,GSH）含量及凋亡基因表达变化。结果显示：1）分离的克隆团细胞具有SSCs的生长特性,可被碱性磷酸酶染色并表达干细胞标志基因OCT4、SOX2和SSCs标志基因NANOG、PLZF、UCHL1;2）50 μmol·mL^-1以上的MT在处理48 h后可显著提高SSCs的细胞活力（P<0.05）;3） MT可显著降低猪SSCs内ROS水平（P<0.05）,极显著增加细胞内GSH含量（P<0.01）;4） MT可显著抑制猪SSCs内凋亡蛋白Bax和Caspase3的表达（P<0.05）。MT具有清除猪SSCs中的ROS,提高总GSH含量,抑制凋亡基因表达进而提高细胞活力的作用,可为养殖过程中提高公猪繁殖性能提供参考。     

Production and Identification of Clone Sheep Using Bone Marrow Mesenchymal Stem Cells as Donor Cell

In order to obtain the cloned sheep using bone marrow mesenchymal stem cells（BMSCs）as the donor cells,the BMSCs of sheep were chosen and reconstructed embryos were built to transfer.10 published microsatellite markers were chosen,and the DNA samples from clone sheep,donor cells and surrogate ewes were amplified,and the relationship of father-son（RCP）was analyzed using the Quantity One for genotyping.The results showed that the reconstructed embryos were successfully built for electric fusion using sheep BMSCs as nuclear donor,and making nuclear transplantation into enucleated mature oocytes of which the fusion rate was 80.62%.20 surrogate ewe were chosen to be implanted with the reconstructed embryos at morula stage by implant surgery,and 5 lambs were born and only 3 were survived.The genotype of cloned sheep was in line with the dornor cell and the RCP were more than 99.999%.In conclusion,the first clone sheep were obtained successfully by using BMSCs as a nuclear donor in this experiment.     

一种快速判别梨果梗的方法

果梗完好与否是梨品质检测的指标之一,因而对果梗情况进行准确判别具有重要意义。为此,通过计算机视觉系统摄取梨的图像,利用图像处理技术提出了一种能快速判别梨果梗有无的算法。该算法的识别正确率达90%,识别速度大约在20～30ms,实现了对梨果梗进行高速检测的目标。     

Irrigation scheduling in a mature peach orchard using tensiometers and dendrometers

Three trickle irrigation schedules, two of which were scheduled according to soil water potential ( soil) (tensiometer method) and daily stem contraction (DSC) (dendrometer method) respectively and the other one was a schedule of restricted water supply, were applied to a mature peach orchard.The annual water application based on soil was greater than that based on DSC. However, tree growth, fruit size and leaf water potential (leaf) on the trees in the dendrometer scheduling plot did not differ from those in the tensiometer scheduling plot while the premature fruit drop and fruit bud initiation were greatly different. The restricted water supply treatment limited significantly both tree and fruit growth. In addition, the lower leaf was observed on the trees in this plot.Further study shows that use of the dendrometer method for scheduling irrigation satisfies the water needs of the plant and that the tensiometer method is less accurate.Abbreviations leaf leaf water potential - soil soil water potential - DSC daily stem contraction - LVDT linear variable displacement transducer - PET potential evapotranspiration     

Construction of mouse ESC line stably expressing Pdx1 by Tet-On system

AIM To construct the mouse embryonic stem cell （ESC） line with stable pancreatic and duodenal homeobox 1 （Pdx1） expression by Tet-On system, which may lay a foundation for further research on the differentiation of Pdx1⁺ definitive endoderm cells into pancreatic cells. METHODS The Pdx1-overexpressing lentiviral vector with green fluorescent protein marker and puromycin resistance was constructed by Tet-On system and was used to infect the mouse ESC. The cells were divided into 3 groups： blank control group （ESC group）, empty lentivirus control group （PDX1^- ESC group） and Pdx1 lentivirus transfection group （PDX1⁺ ESC group）. Flow cytometry was used to detect the transfected cells after screening by doxycycline （DOX）. The function of Tet-On system and the expression of Pdx1 gene were detected. The transfected cells in PDX1^- ESC group and PDX1⁺ ESC group were sorted by flow cytometry, and constructed ESC line with stable expression of Pdx1 and negative control ESC line were verified. RESULTS （1） The positive rates of transfected cells in PDX1^- ESC group and PDX1⁺ ESC group were 90.72% and 94.01% after screening by DOX, respectively. The positive rates of transfected cells in PDX1^- ESC group and PDX1⁺ ESC group was 97.84% and 98.13% after sorting by flow cytometry, respectively. （2） With DOX, green fluorescence was observed in PDX1^- ESC group and PDX1⁺ ESC group. The mRNA and protein expression of Pdx1 was significantly increased in PDX1⁺ ESC group （P<0.05）. Without DOX, no green fluorescence was observed in the cells of the 3 groups, and no significant difference in the mRNA and protein expression of Pdx1 was observed （P>0.05）. （3） After 3 months of cryopreservation, the cell lines still survived in resuscitation culture and were regulated by DOX. CONCLUSION Using Tet-On system, the mouse ESC line with inducible Pdx1 expression were successfully established and could be used as an effective cell model to research the differentiation of Pdx1⁺ definitive endoderm cells into pancreatic cells.     

“巨峰”葡萄热处理茎尖培养技术研究

以"巨峰"葡萄为试材,经热处理后取其茎尖进行离体培养,通过调整激素的种类和浓度,筛选出了适宜"巨峰"葡萄茎尖分化和生根的培养基;通过对不同移栽基质的研究,筛选出了移栽成活率较高的基质配方。结果表明:38℃热处理时葡萄苗发芽率最高,可达86.67%;以GS为基本培养基,附加BA 0.5mg/L与NAA 0.1mg/L,最适宜于"巨峰"葡萄茎尖分化成苗,分化率可达70.00%;生根培养基以GS+IBA 0.06mg/L的生根率最高。适宜"巨峰"葡萄茎尖组培苗移栽的基质为河沙1份+水苔1份。     

玉米茎结构参量与产量关联及最优化分析

高效高产农业是可持续生产的重点,为研究玉米茎结构参量综合利用效率及最优利用率产量提高幅度,本研究使用2015年北京市小汤山镇获取的玉米数据,将玉米茎结构参量作为输入指标,生物量、产量和经济系数为产出指标,运用数据包络分析探讨利用效率及最优利用效率。结果表明:‘京华8号’玉米茎结构参量利用效率都保持在0.7以上,仅67%达到最优利用效率水平;小喇叭口期和抽雄期茎结构参量利用效率中上部茎粗最差,下部茎粗最优但仍有提高潜力;玉米茎结构参量综合利用效率最优效率时资源充分利用产量有不同程度的提高,其中抽雄期提高产量最大为32%。玉米茎结构参量利用效率尚未达到最优,提高其利用效率尤其抽雄期可显著提高产量。     

不同生育期水稻干物质量与产量的关系研究

为明确不同生育期的干物重对粤晶丝苗2号、粤农丝苗、合美占3个水稻(Oryza sativa L.)品种的产量及产量构成因素的影响,设置了早造和晚造两次不同氮肥梯度随机区组栽培试验。结果表明,叶片干物质的分配比例在拔节孕穗期时最高,且其随着生育进程逐渐降低,在乳熟期达到最低值;茎干物质的分配比例在乳熟期降至最低;齐穗期的穗干物质分配比例不到20%,而乳熟期的穗干物质量将近占总干物质量的43%;早造和晚造水稻的结实率差异极显著,总干重差异显著,而穗数、每穗粒数、千粒重、产量差异不显著;齐穗期以后,结实率和千粒重均与干物重呈负相关,而叶干重与穗数呈显著正相关,且相关性较大,茎干重与各产量性状密切相关,总干重与产量呈正相关。水稻干物重是决定高产的重要因素。