vvi-miR160s介导VvARF18应答赤霉素调控葡萄种子的发育

【目的】研究vvi-miR160家族(vvi-miR160s)及其靶基因在‘魏可’葡萄种子发育过程中的作用,探究其应答赤霉素(GA)调控葡萄果实无核的潜在机理。【方法】采用miR-RACE、生物信息学分析、RT-qPCR、RLM-RACE等方法,鉴定vvi-miR160家族成员及其靶基因,分析vvi-miR160s及其靶基因应答GA的时空表达模式及其潜在功能。【结果】花前GA处理强烈抑制‘魏可’葡萄胚珠及种子发育,高效诱导其无核,且无核率达99.8%。克隆鉴定了VvMIR160s前体基因序列(501 bp)及成熟体序列,且它们在不同物种间具有较高的保守性;基于vvi-miR160s序列,预测到靶基因VvARF18,利用RLM-RACE技术检测到vvi-miR160s对VvARF18的裂解位点在第10和第11位之间,裂解频度9/17,证明VvARF18是vvi-miR160s的真实靶基因。该靶基因编码683个氨基酸,在398—411位存在核定位信号,且该蛋白亚细胞定位于细胞核上。VvARF18与其他物种间序列的同源保守性较高,基因结构相似,其中VvARF18蛋白与茶、烟草、梅花等物种亲缘...     

Effect of quinoa flour on gluten-free bread batter rheology and bread quality

A new gluten-free bread formulations composed of quinoa, buckwheat, rice flour and potato starch were developed in the present study. Rheological characteristics of the bread batter with increasing amount of quinoa were determined; storage (G′) and loss modulus (G″) values were also measured for investigation of viscoelastic properties. To evaluate the quality of breads; technological and physical (bake loss %, specific volume, texture, microstructure, color), chemical (protein, moisture, ash) and sensory properties were determined. All batter formulations independent of the quinoa amount exhibited pseudoplastic behavior, and G′ values were found to be higher than G″ values in expressing the solid like characteristics of the batter. Amount of quinoa flour addition did not present significant difference on bake loss%, specific volume and protein content (p>0.05); however, 25% quinoa flour bread displayed better results with its higher sensory scores and softer texture. Quinoa and buckwheat flour mixture therefore will be a good alternative for conventional gluten-free bread formulations.     

教育美育化刍议

传统教育由于存在种种弊端已远不适应现代化发展的要求,时代需要的是德、智、体。美全面发展的人,是富于想象力和创新精神的人,美育的特质恰恰与这一需要相吻合。因此,教育改革意味着实现教育现代化,必须伴随教育美育化。     

Synthetic tetramer of a Phytophthora sojae-inducible fragment from soybean GmaSKTI36 promoter improves its pathogen induction activities

Using pathogen-induced promoters to control expression of the functional genes in transgenic plants may greatly increase the chances of boosting disease resistance. However, the number of the inducible promoters is limited. Here, we found that soybean GmaSKTI36 gene is strongly induced upon Phytophthora sojae infection. Functional analysis showed that its promoter could mediate rapid and strong induction of GUS expression upon pathogen infection in both Nicotiana benthamiana leaves and soybean hairy roots. Then, a 122 bp fragment that was critical to the activity was successfully identified by a progressive 5′ deletion analysis. Importantly, we found that a synthetic promoter by tetramerizing this fragment could confer strong P. sojae induction activities. Overall, the results suggested that the GmaSKTI36 promoter, the 122 bp fragment, and the synthetic promoter are potentially useful pathogen-inducible promoters.     

哈克尼西棉细胞质陆地棉雄性不育系orf160的克隆及遗传转化

以哈克尼西棉细胞质陆地棉雄性不育系及其保持系、恢复系为材料,克隆并测序线粒体功能基因的侧翼序列。通过生物信息学分析,在atp4下游发现一个哈克尼西棉细胞质不育系特有的orf160。orf160全长480 bp,N端与atp6序列部分同源,C端序列与核序列部分同源;编码的159个氨基酸序列,与膜蛋白、细胞周期蛋白具有部分同源性。以含有线粒体导肽的GFP瞬时表达载体转化后,在洋葱表皮的细胞膜上观察到绿色荧光,表明线粒体定向载体正常表达。构建酵母和植物表达载体,分别转化酵母、烟草和拟南芥;同对照相比,转基因酵母菌斑畸形且生长缓慢;转基因拟南芥和烟草种子大部分畸形,T₁和T₂代植株的结实率和种子发芽率降低,花粉着色比野生型浅。结果表明,orf160基因的表达影响受体正常发育。     

茉莉酸对苹果气孔运动及抗旱性的影响

50、75、100、150 mg／L JA处理苹果叶片表皮，使开放气孔的气孔开度分别减小27．4％、37．5％、56．9％和63．8％，使关闭后的气孔在光下开放时气孔开度分别减小15％、18％、28．8％、52．5％、61．3％；处理植株后干旱胁迫，叶片相对电导率分别降低15％、18％、31％、58％，脯氨酸含量分别提高0．09％、0．40％、0．42％、0．54 ng／gFW，可溶性糖含量分别提高1．89％、2．53％、3．80％、5．06％。25 mg／L JA处理与对照均无显著差别。JA的作用表现出明显的浓度效应。     

哲罗鱼胰岛素样生长因子-II的原核表达与活性分析

在鱼类胚胎发育过程中,胰岛素样生长因子-II(insulin-like growth factors-II,IGF-II)起着关键的作用。本研究根据Gen Bank收录的鲑鳟鱼IGF-II基因序列设计引物,以哲罗鱼(Hucho taimen)肝RNA提取物为模板,利用RT-PCR方法扩增出哲罗鱼IGF-II基因开放阅读框。将目的基因IGF-II与原核表达载体p SUMO连接构建出重组表达载体p SUMO-IGF。将该重组质粒转化到大肠杆菌Rosetta中进行目的蛋白的诱导表达。SDS-PAGE电泳分析显示,约在40 k D处含有清晰的条带,与预期结果相符;目的蛋白以包涵体形式存在。对包涵体进行变性/复性后获得较纯的目的蛋白,利用ELISA和MTT方法对目的蛋白进行免疫学活性及生物学活性分析。ELISA结果显示该目的蛋白能够与商品化的抗鲑鳟鱼IGF-II的抗体发生特异性反应,并且呈现抗原浓度依赖性,该结果说明本研究获得了具有良好免疫原性的IGF-II蛋白;MTT方法测定IGF-II蛋白对鲤(Cyprinus carpio)上皮细胞(epitheliaoma papulosum cyprini,EPC)和虹鳟(Oncorhynchus mykiss)性腺细胞(rainbow trout gonad,RTG-2)的增殖效果来鉴定IGF-II蛋白的生物学活性,结果显示所表达的哲罗鱼IGF-II蛋白能够有效的刺激EPC细胞和RTG-2细胞增殖。该结果表明利用原核表达系统获得的哲罗鱼IGF-II蛋白具有良好的生物学活性。本研究为哲罗鱼的生长模式和生长繁殖的研究奠定了基础。     

常温泡沫箱加冰运输条件下荔枝的温度、品质、呼吸和乙烯释放变化

常温泡沫箱加冰密封技术已经广泛应用于荔枝的国内运输。以淮枝为材料，探讨加冰与预冷处理对果实温度、品质以及某些生理指标的影响。结果表明：29～31℃室温下72　h后，对照与5℃预冷处理的果实温度相近，达33℃，高于室温。加冰处理使果实温度先下降，至最低点后回升。加冰量达50％时，果实最低温度降至6℃，72　h后回升至27℃，低于室温。对不同加冰比例的荔枝可溶性固形物含量、好果率、果实失重、酶活性以及箱内乙烯、二氧化碳等气体成分动态进行了比较研究。