Ubi1 intron-mediated enhancement of the expression of Bt cry1Ah gene in transgenic maize (Zea mays L.)

The cry1Ah gene was one of novel insecticidal genes cloned from Bacillus thuringiensis isolate BT8. Two plant expression vectors containing cry1Ah gene were constructed. The first intron of maize ubiqutin1 gene was inserted between the maize Ubiquitin promoter and cry1Ah gene in one of the plant expressing vectors (pUUOAH). The two vectors were introduced into maize immature embryonic calli by microprojectile bombardment, and the reproductively plants were acquired. PCR and Southern blot analysis showed that foreign genes had been integrated into maize genome and inherited to the next generation stably. The ELISA assay to T1 and T2 generation plants showed that the expression of Cry1Ah protein in the construct containing the ubi1 intron (pUUOAH) was 20% higher than that of the intronless construct (pUOAH). Bioassay results showed that the transgenic maize harboring cry1Ah gene had high resistance to the Asian corn borers and the insecticidal activity of the transgenic maize containing the ubi1 intron was higher than that of the intronless construct. These results indicated that the maize ubi1 intron can enhance the expression of the Bt cry1Ah gene in transgenic maize efficiently     

玉米自交系幼胚离体培养及植株再生研究

采用4种不同类型的玉米自交系为供试材料，用MS和AG培养基进行诱导培养。获得胚性愈伤组织，结果表明，基因型和外植体的发育时期对愈伤组织的诱导率有很大影响；不同的2，4-D浓度及山梨醇浓度影响愈伤组织的保持和质量改善；6-BA浓度为2mg／L时，有利于再分化培养，在再分化培养中，获得了再生植株。     

Gus基因在佛手叶片中的瞬间表达

采用基因枪法对佛手(Citrus medica L.var.sarcodactylis(Noot)Swingle)嫩叶和老叶叶盘进行了Gus基因转化和瞬时表达研究,分析了轰击距离、叶片组织特性对转化效果的影响.结果显示,在7.584 5×106Pa轰击压力下,采用6 cm轰击靶距转化嫩叶可获得比较高的Gus基因瞬间表达频率;老叶的转化效果比嫩叶差;增加轰击次数并不一定能提高瞬间表达频率.     

用基因芯片技术分析干旱胁迫下生殖生长期大麦的基因表达

耐旱性是干旱地区稳定和增加大麦产量的一个关键因素。鉴定出与耐旱性相关的功能基因,一方面可了解大麦的耐旱机理,同时还可以促进利用生物技术来改良大麦的耐旱性。在研究中,2个在耐旱性上具有明显差异的大麦品种Tadmor(耐旱)和WI2291(干旱敏感)被选作材料,采用22000个ESTs(基因表达序列标签)的Affymetrix大麦基因芯片Barley1来分析生殖生长期干旱胁迫下2个大麦材料的差异表达基因。研究结果表明,干旱胁迫下2个大麦材料中有77个共调节基因,其中部分基因已被报道过可能与抗旱性相关。这些基因中已有功能注释的基因按其生物学功能被分为14组,猜测它们是干旱胁迫的响应基因,在抗旱性上可能不起重要作用,或者是必需的但单独不足以提高大麦的抗旱性。进一步比较2个材料差异表达的基因,发现二材料之间有372个受干旱调节基因的差异。这些基因中有功能注释基因的生物学功能中可分为15组,其中一些已被认为与抗旱性相关;而对那些未知功能的基因,推测可能亦在大麦的抗旱性上扮演一定的角色。研究所得结果可为阐述生殖生长期大麦的耐旱性机理提供新的认识。     

ICP-MS法测定银合欢种和种皮中27种金属元素含量

采用电感耦合等离子体质谱（ICP-MS）法测定了云南省建水县产银合欢种和种皮中27种金属元素的含量．其中13中金属元素V、Cr、Mn、Fe、Cu、Zn、As、Se、Mo、Ag、Cd、Tl、Pb等采用普通模式检测,14种金属元素Li、Be、B、Mg、A1、Co、Ni、Ga、Rb、Sr、Te、Ba、Bi、U等采用碰撞／反应池技术（CCT）模式测定,以消除样品溶液中潜在的干扰．实验条件下,Co、Ni、Cr、Zn、Se、Mo、Ag、Pb等8种金属元素未检出．方法检出限Fe为9．789ng／mL、Cr为2．691ng／mL,Zn为1．803ng／mL,B为2．076ng／mL,Mg为1．977ng／mL,A1为3．024ng／mL,Ni为1．824ng／mL,其它元素为0．003-0．921ng／mL,银合欢种精密度的RSD为0．141％-11．86％,银合欢种皮精密度的RSD为0．0449／6-31．14％,加标回收介于90．8％-107．1％之间．