重组K88黏附素亚单位蛋白在大肠杆菌中的诱导表达

采用响应面分析方法,对肠毒素大肠杆菌(EnterotoxigenicEscherichiacoli)K88黏附素亚单位重组蛋白FaeG诱导表达的主要影响因素进行了优化研究。响应面分析结果表明:不同的诱导时机、IPTG浓度及诱导时间对目的蛋白表达量均有显著影响(P<0.01),三因素之间的交互作用对目的蛋白表达量也有极显著的影响(P<0.01)。通过进一步的模拟优化,得到了最佳诱导表达条件为培养BL21(K88ac)重组菌株2.5h后,加入IPTG至终浓度0.8mmol/L,37℃诱导培养5h。采用以上参数进行验证实验,得到目的蛋白在全菌蛋白中的表达含量为35.4%;与单次单因子法得到的最佳表达条件下的表达量(27.5%)相比,提高了28.7%。采用响应面分析法可以确定IPTG的诱导时机、诱导浓度及诱导时间等多个因素对重组蛋白在大肠杆菌中表达的最佳作用条件,有利于提高目的蛋白的表达量。     

The effect of colchicine on triticale anther-derived plants: Microspore pre-treatment and haploid-plant treatment using a hydroponic recovery system

In cereals, chromosome doubling of microspore-derived haploid plants is a critical step in producing doubled haploid plants. This investigation was undertaken to study the effect of incorporation of colchicine in the induction medium for anther culture, and the effect of colchicine on anther culture-derived plants of triticale grown under controlled greenhouse conditions. In the latter case, chromosome doubling of adult sterile plants derived from anther culture of fourteen triticale populations was attempted, where androgenetic plants with non-dehiscent anthers were cloned and subjected to the colchicine treatment, and then grown with the aid of hydroponics. The hydroponic system provided optimal conditions for recovery of the affected haploids from the toxic effects of colchicine treatment and all colchicine-treated plants survived. A topcross-F₁ (TC₁F₁) population with timopheevii cytoplasm produced the highest percentage of plants with seed-set either due to chromosome doubling by colchicine (98%) or spontaneous doubling of chromosome number (15%). Colchicine-treated anthers performed inferior than control in both induction and regeneration phases. One of the key observation of this study was the reversal from reproductive stage back to the vegetative stage which in turn enabled further cloning of haploid plants under hydroponic conditions once they were identified as sterile. The one hundred percent survival rate of in vitro-derived plants, 100% survival rate of colchicine treated haploid plants and the high chromosome doubling success rate (X = 82.3) observed in this study imply that a temperature-controlled greenhouse with an hydroponic system provides an efficient environment for inducing chromosome doubling of haploid plants in cereals. This revised version was published online in August 2006 with corrections to the Cover Date.     

水稻剑叶性状的遗传分析和基因定位

本研究以水稻籼粳杂交(窄叶青8号×京系17)F1经花药培养, 产生的双单倍体(DH)群体和应用该群体已构建的分子图谱为基础。 采用QTL区间作图法对5个剑叶性状进行定位分析, 结果表明, 在DH群体中, 剑叶长、 宽、 长宽比和叶面积呈连续分布, 受微效多基因控制, 并且各性状均存在一定数量的超亲遗传类型。 4个性状共检测出13     

基于电磁感应的典型干旱区土壤盐分时空变异快速诊断

选取南疆典型干旱区研究其土壤盐分时空变异,在研究区域选取校正点测定表观电导率(ECa),并同时采集土壤样品,研究表明土壤盐分含量与大地表观电导率具有良好相关性(R =0.935),可由此建立土壤盐分电磁感应解译模型.构建移动式磁感调查系统对研究区域进行调查,获取足量表观电导率信息,结合GIS和地统计等相关知识研究土壤盐...     

基于最小平方误差的单倍体和二倍体分类方法研究

基于玉米油分的花粉直感效应,采用核磁共振鉴别单倍体,快速、准确、低成本地确定单倍体与二倍体之间的含油率阈值,是实现大批量自动鉴别的前提和关键。提出一种基于最小平方误差的单倍体和二倍体分类方法,通过少量籽粒确定含油率阈值的研究,对单倍体和二倍体进行识别。以10个玉米组合为研究对象,利用核磁共振分析仪测定各组合一定数量单倍体和二倍体籽粒的含油率,依据最小平方误差方法确定各组合油分阈值,并通过逐级递增训练集样本数量的方法,确定油分阈值所需训练集规模,获得含油率阈值,能够满足单倍体正确识别率及对二倍体正确拒识率的均值达到90%以上的要求,为核磁共振全自动单倍体鉴别系统的后期大规模分类提供参数指导,提高分选效率。同时因单倍体生产效率低,运用最小平方误差的方法,通过少量的单倍体及二倍体籽粒来构建油分鉴别单倍体模型,以较小的投入获得更优的分类效果在玉米单倍体自动化分选中具有非常重要的现实意义,将该分类方法应用到全自动核磁分选单倍体系统中,可以达到较好的效果,为加快单倍体工程化育种提供保障。     

辐射诱变选育特优63糯的研究

用60 Coγ射线 35 0Gy分别照射杂交稻保持系 (B系 )和恢复系 (R系 )干种子 ,M2 获得相应的糯性基因wx突变体 ,并育成糯不育系wxA和糯恢复系wxR ,组配出杂交糯稻新组合。特优 63糯与特优 63具有相似的生育期、农艺性状、产量水平和抗性。     

籼粳交加倍单倍体后代性状遗传的研究

对一个典型的籼/粳组合、窄叶青8号/京系17的F1花药进行培养,所获二倍体花粉植株除约20%自然结实率为0外,其余植株的结实率在1%至正常结实之间,65%株系在75%以上,并稳定地传至自交后代。对130多个加倍单倍体(DH)多种性状的遗传进行了研究与分析,结果表明,花药培养可以克服籼/粳后代包括育性在内的各种性状长期分离不易     

Potential,constraints and applications of in vitro methods in improving grain legumes

Grain legumes, the important constituents of sustainability‐based cropping systems and energy‐limited vegetarian diets have long been the subject of scientific research. Tremendous technological strides were made in the so‐called orphan crops, in terms of both varietal improvement and generation of basic information. Despite recalcitrancy and high genotype dependency, in vitro culture techniques such as organogenesis, in vitro mutagenesis, embryo rescue and in vitro gene transfer have been deployed for improvement of several grain legumes and these played an important role in introgression of desirable genes from related and distant species and creation of additional genetic variability. Stable and reproducible regeneration protocols resulted in the development of genetically modified chickpea, pigeon pea, cowpea, mungbean, etc., while embryo rescue was deployed successfully for recovery of interspecific recombinants, a few of them exploited for the development of commercial cultivars. Nevertheless, doubled haploidy witnessed limited success and protoplast regeneration and in vitro mutagenesis remained of academic interest. The present review focuses on the progress, achievements, constraints and perspectives of using in vitro technology in grain legume improvement.     

水稻花培一次成苗的研究──Ⅱ．影响水稻花培一次成苗各主要因素的探讨

影响水稻花培一次成苗的主要因素为：（１）基本培养基、激素配比和基因型对一次成苗有重要作用．基本培养基以Ｎ′６的为最好；激素配比以２，４—Ｄ０．０２＋ＮＡＡ２．５０＋ＫＴ４－６ｍｇ／Ｌ为佳；３种基因型一次培养的绿苗率大小顺序为粳型品种Ｃ５７＞籼粳交（早香１７×Ｃ５７）杂种Ｆ１＞籼型品种早香１７．（２）培养基保持ｐＨ值５．８、蔗糖浓度８．５％，并添加丝氨酸、脯氨酸、精氨酸、天冬氨酸、丙氨酸各４０ｍｇ／Ｌ和谷氨酰胺４００ｍｇ／Ｌ，所获一次培养的绿苗率最高；添加０．１％活性炭的有抑制作用．（３）低温（９－１０℃）预处理对一次成苗有重要作用．早香１７以预处理５－１０ｄ的为宜，Ｃ５７以１０—１６ｄ的较好．恒温处理优于变温处理的．高温（３０℃）预培养的作用不大．接种后进行暗培养对一次成苗有利．