用原生质体融合进行冬菇育种的研究

用灭活的〔Flammulinasubnudus (Pr .)Sing〕双核菌株原生质体与〔Flammulinavelutipes (Fr.)Sing〕双核菌株原生质体融合 ,在 2 2℃的条件下筛选得融合子。融合频率为 0× 10 -5～ 4 1× 10 -5。融合子与双亲有明显的拮抗性。融合子的菌丝体形态 ,氨基酸含量 ,子实体的形态 ,以及酸性磷酸酶的测定都与双亲不同。     

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.     

不同寄主来源的葡萄霜霉病菌致病力测定及孢子囊大小比较

对采自我国不同地区、不同主栽葡萄品种‘红地球’、‘巨峰’及‘赤霞珠’上的12份葡萄霜霉病菌的致病力及孢子囊大小进行了比较和分析,结果表明:采自不同葡萄品种的病原菌对感病葡萄品种‘奥迪亚无核’叶片的致病力具有明显差异,其中‘巨峰’及‘红地球’来源的病原菌致病性较强,明显大于‘赤霞珠’来源的病原菌致病力;采自不同葡萄品种的霜霉病菌孢子囊大小存在一定的差异,‘赤霞珠’来源的病原菌孢子囊明显大于‘巨峰’及‘红地球’来源的霜霉病菌孢子囊。上述结果表明,采自不同寄主的葡萄霜霉病菌的致病力及孢子囊大小均存在一定程度的分化。     

早期限饲对肉鸡肌肉生长及生产性能的长期影响

本研究旨在比较不同生长阶段隔日限饲(早期限饲:1~14日龄限饲,后期限饲:50~63日龄限饲)对肉鸡肌肉生长及相关激素水平的影响,并探讨早期限饲对肉鸡肌肉生长的程序化作用。试验结果:1)早期限饲组整个试验阶段体重均极显著低于对照组,后期限饲组限饲一周后体重显著低于对照组并于63日龄实现完全补偿生长;2)早期限饲组14日龄和63日龄胸肌、腓肠肌重均显著低于对照组;后期限饲组63日龄胸肌、腓肠肌重均显著低于对照组,且二者与早期限饲组均差异不显著;3)早期限饲显著降低了14日龄肉鸡肌肉中DNA、RNA和蛋白含量,63日龄恢复至正常水平,且核酸、蛋白水平在胸肌和腓肠肌内的趋势表现不一致;4)早期限饲组14日龄血清中T3、T4水平均显著低于对照组,63日龄血清T3水平仍显著低于对照组;后期限饲组63日龄血清T3水平显著低于对照组,T4水平与对照组差异不显著。结果表明:1)限饲可显著阻碍肉鸡肌肉的生长;2)不同部位的肌肉对限饲的敏感度不同;3)早期营养因子对肉鸡肌肉的生长具有"程序化"作用,甲状腺激素可能参与这些过程的调节。     

LED白光处理对番茄贮藏品质的影响

以番茄为试材,研究了LED白光照射处理对番茄的保鲜效果的影响,为改善采后品质提供参考。结果表明:LED白光处理可有效维持番茄的感官品质,延缓其水分损失,并且不影响番茄的色泽。除此之外,还能够较好地抑制番茄可溶性固形物、维生素C含量等营养物质的降解,提高番茄的过氧化物酶(POD)和多酚氧化酶(PPO)活性,增强番茄的抗氧化能力。     

橘皮叶黄素提取及性质研究

以橘皮为试材,采用超声波辅助提取工艺对橘皮中叶黄素进行了研究。结果表明:橘皮叶黄素在可见光范围内的最大吸收峰为448nm,L9(34)正交实验确定了影响叶黄素提取效果的最主要因素是提取温度,并得出橘皮叶黄素提取的最佳工艺条件是丙酮浓度60%、料液比1∶15kg·L~(-1)、提取时间60min、提取温度60℃。同时研究光照强度、pH、金属离子、温度、氧化剂和还原剂对叶黄素稳定性的影响。强光、较低pH、不同金属离子、高温、维生素C均对橘皮叶黄素有一定的降解作用。     

重组H5N3亚型禽流感细胞毒灭活疫苗免疫佐剂效果的评价

为评价佐剂对疫苗免疫效果的影响,本试验以反向遗传学技术拯救的重组H5N3亚型禽流感病毒为种毒,对国产矿物油以及赛比克公司提供的MontanideTM ISA 70M VG(简称7OM)和MontanideTM ISA 206 VG(简称206)3种佐剂制备的灭活疫苗进行了SPF鸡免疫攻毒试验.结果表明国产矿物油佐剂组与70M佐剂组都能对SPF鸡100%保护,206佐剂组仅能保护40%,攻毒的SPF鸡发病,但不死亡;从抗体水平上比较发现70M佐剂组稍好于国产矿物油佐剂组,但两组之间差异不显著,而这两组与206佐剂组差异极显著.这3种佐剂组免疫鸭均能够诱导产生较高的抗体水平,70M佐剂组优于国产矿物油佐剂组和206佐剂组,但3者差异不显著.用70M系的8种佐剂MontanideTM ISA 70 essai Mi(i-1-8)(简称70Mi)佐剂乳化的疫苗免疫SPF鸡,结果表明70M佐剂组效果最好.攻毒后免疫鸡不排毒,而且无临床症状;统计学分析显示70M佐剂组与所有70Mi佐剂组差异均显著.这些结果表明70M佐剂可以作为新型疫苗佐剂用于禽流感疫苗的制备.     

高寒草地凋落物覆盖对狼毒生长微环境及种苗定居的影响

种子萌发和幼苗定居是植物生活史中最为脆弱的两个阶段.为了探明高寒草地狼毒(Stellera chamaejasme)种苗定居的生境选择,本研究在其原生境下通过人工控制试验,研究凋落物覆盖对土壤微环境(温、湿度)的影响以及狼毒种子在不同生境条件下的出苗和定居表现.结果表明:凋落物覆盖总体上能够降低土壤温度,提高土壤含水量...